Research Trends in

Food Technology
and
Nutrition
Volume - 4

Chief Editor
Dr. Deepa Verma
Assistant Professor: Home Science (F&N),
G.P.S. Government Mahila P.G. College, Ambari, Azamgarh,
Uttar Pradesh, India

AkiNik Publications
New Delhi
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 Contents

Chapters Page No.

1. Biosurfactant: A Promising Alternative of Chemically
Synthesized Food Surfactant in Food Processing and
Environmental Applications 01-36
(Subhajit Ray and Amit Kumar Barman)
2. Molecular Aspects of Nutrition Molecular Gastronomy &
Related Techniques 37-48
(S. Sreeremya)
3. Sensory and Food Quality 49-75
(Kamaljit Kaur and Gursharan Kaur)
4. Prospects of Nano-Composites Containing Packaging on
Quality and Shelf Life of Fresh Fruits and Vegetables 77-90
(Janagam Venu Madhav, R. Swamy Sekhar and Shruti Sethi)
5. Machine Vision System: A Tool for Quality Inspection of
Food 91-112
(Monika Mathur and Raveena Kargwal)
6. Casein Micelle as a Nano Carrier System for Hydrophobic
Bio-Active Compounds 113-128
(Priti Saha and Rajesh Kumar Bajaj)
7. Novel Protein Foods: Alternative Sources of Protein for
Human Consumption 129-142
(Neelesh Kumar Maurya and Radha Kushwaha)
8. Potential of Functional Foods in Human Nutrition 143-168
(Mamta Thakur and H.W. Deshpande)
9. Nutrition and Behaviour 169-179
(Poonam Sharma)
 Chapter - 1
Biosurfactant: A Promising Alternative of Chemically
Synthesized Food Surfactant in Food Processing and
Environmental Applications

Authors
Subhajit Ray
Associate Professor, Department of Food Engineering &
Technology, Central Institute of Technology, Kokrajhar, Assam, India
Amit Kumar Barman
Assistant Professor, Department of Dairy Microbiology, West Bengal
University of Animal and Fishery Science, Mohanpur Campus, Nadia,
West Bengal, India

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Page | 2
 Chapter - 1
Biosurfactant: A Promising Alternative of Chemically
Synthesized Food Surfactant in Food Processing and
Environmental Applications
Subhajit Ray and Amit Kumar Barman

Abstract
Bio surfactants generally derived from microorganisms are amphiphilic
in nature consisting of the emulsification and surface active properties.
Interest in bio surfactants has increased considerably in recent years, as they
are potentially used in many commercial applications viz. petroleum,
pharmaceuticals, biomedical, and food processing industries. In this context
a strategy is required for the successive production and purification of bio
surfactant consisting of a series of steps. viz. isolation and identification of
bio surfactant producing strain, mutation for yield enhancement,
optimisation of process conditions e.g. fermentation time, temperature of
fermentation, inoculum age, inoculum size or volume (v/v), aeration rate,
agitation rate etc. in shake flask fermentation, selection of suitable media
with special emphasis on selection of carbon and nitrogen sources in similar
mode of fermentation, scale up in laboratory fermenter with the efficiency of
volumetric oxygen transfer coefficient (KLa), role of viscosity, cell growth
etc. and isolation and purification through solvent precipitation techniques
viz. acetone, ethanol, methanol or combination of two solvents etc. Due to
the aerobic nature of fermentation there is a vital role of dissolve oxygen
saturation, effective gas liquid interfacial area and specific oxygen uptake
rate (SOUR) as well as oxygen transfer rate (OTR) correlated with overall
mass transfer coefficient (KLa). The purified bio surfactant produced by
mutant strain of bacterial species both in the shake flask and laboratory scale
fermentation exhibits the reduced surface tension i.e. enhanced surface
activity. The surface property of the synthesized bio surfactant depends upon
the reduced surface tension value (dyne/cm).Now to establish the potency of
the bio surfactant, the bio surfactant activity at different concentration levels
are compared with synthetic surfactants and the highest surface activity was
found in microbiologically derived bio surfactant.

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 Keywords: Bio surfactant, surface tension, production, KLa purification,
application
Introduction
Bio surfactants are microbiologically derived surface active agents
produced by bacteria, yeast, mould etc. They are amphiphilic compounds
produced on living surfaces mostly on microbial cell surfaces or excreted
extra cellularly and contain hydrophilic and hydrophobic molecules that
confer the ability to accumulate between fluid phases, thus reducing surface
and interfacial tension at the surface and interface respectively1. Bio
surfactants are capable of lowering surface and interfacial tension effectively
and could be potential substitutes of widely used chemically synthesized
surfactants2. Microbial surfactants are promising substitute of chemically
synthesized surfactant and can be used for the food industry,
pharmaceuticals, cosmetics, specialty chemical industries, enhanced oil
recovery (EOR) and cleaning of oil spills by bioremediation. Several types
of Bio surfactant have been isolated and characterized including
lipopeptides, glycolipids, polysaccharides-protein complexes, phospholipids,
sorphorolipids, fatty acids and neutral lipids. Candida tropicalis 3, have been
shown to be potent producers of sorphorolipid type of bio surfactant.
Bacillomycin F. D, L, lichenysin, iturin, hallobacillin and plipastatin 4, 5, 6, 7
are lipopeptides bio surfactants produced by Bacillus strain. Production of
surfactin, a lipopeptides type bio surfactant was first reported for a strain of
Bacillus subtilis on 8. Though microbes have a great potential towards the
production of bio surfactant but it is always required to enhance the
productivity. In this context mutation is considered to be an effective tool.
Hence the basic procedure for classical mutation is to carry out the alteration
of genetic sequences so that potency of the microorganisms’ especially
bacterial strain can be achieved towards the productivity. Mutation can be
carried out by means of physical and chemical mutagens. Among the
chemical mutagens NTG (Nmethyl-N'-nitro-N-nitrosoguanidine) is one of
the effective mutagen. Application of NTG on Bacillus sp caused a good
increase of bio surfactant production capacity compared to potent strain9.
After obtaining mutated strain, it is always required to undergo for the
growth of microbial cell by means of shake flask fermentation as the primary
step followed by seed development in pilot plant scale and then to
commercial scale production. Therefore selection of the substrate in
particular carbohydrate and nitrogen sources are essential apart from other
mineral sources. However, hydrocarbons as a carbohydrate source are
commonly used as the substrate for the production of bio surfactants

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10
.Carbohydrates as a carbon and energy source are used for bio surfactant
production by Arthrobacter sp, Bacillus subtilis, Torulopsis bombicola and
Pseudomonas aeruginosa 11, 12, 13, 14. In general known bio surfactants are
synthesized from water-immiscible hydrocarbons; however Bacillus subtilis
strains are able to produce surfactants from water soluble substrates. Bio
surfactant production by Pseudomonas putida grown on BMS medium with
2% hexadecane as substrate further inoculated with 2ml from an overnight
culture on BMS with 2% glucose was reported15.Nitrogen sources were
found to affect the bio surfactant production by bacteria 16, 17, 18. Bio
surfactant production depends upon a suitable C: N ratio. However for the
mutant strain of Bacillus sp using different carbon sources as carbohydrates
and hydrocarbons and nitrogen sources in the fermentation medium and by
varying the concentrations of those sources can enable to formulate a
medium for the better production of bio surfactant. Production of the bio
surfactant from a particular organism may vary with the change of
environmental conditions for cell growth as well as fermentative
production19. Optimization of bio surfactant production through fermentation
route can be influenced by several environmental conditions viz. pH,
temperature of fermentation, time of fermentation, medium composition,
medium volume, inoculum size, aeration rate and agitation rate 20. Some
studies have been undertaken with the objective of optimizing surfactin
production by the alteration of variables such as medium composition,
agitator speed and aeration rate 21, 22. Rhamnolipid production by
Pseudomonas sp at a pH range from (6-6.5) was maximum and decreased
sharply at pH 723. The penta and disaccharide lipid production in Nocardia
corynebacteroides is unaffected in the pH range of (6.5-8) was shown24.
After optimisation of environmental conditions the production of bio
surfactant will be carried out in a laboratory scale fermenter with the
association of growth of the desired microorganism. Every fermentation
process depends upon the efficiency of the product formation based upon the
design economics of fermenter along with the optimisation of process
conditions. Now the function of a fermenter is to provide the environment
for the growth of organisms or cells under controlled conditions 25. The
biological transformation under the influence of microorganism in a suitable
environment takes place in such type of vessel. Shake flask fermentation do
not reflect very well the in terms of productivity in comparison to stirred and
aerated tank reactor as the oxygen transfer potential of a shake flask is in
general significantly less than that normally prevailing in an agitated/stirred
fermenter. So scale-up of laboratory fermenters for the development of
process parameters for bio surfactant production can be most effectively

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studied as the product uniformity is usually better in a stirred/agitated and
aerated fermenter than that in a shake flask fermentation 26, 27. It was reported
by28 that the fermentation processes were classified as product formation
with the synonymous growth rate, product formation associated with growth
and product formation not associated with growth. Now, when the second
type of fermentation is carried out in a batch process, it requires conditions
that allow the maximum utilization of substrate and it needs the adjustment
of pH, temperature, concentration of oxygen and nutrients in the bulk of the
media making assumption that these conditions prevail uniformly at the
surface of the cells. The cultivation of bacteria on the chief substrate glucose
to obtain bio surfactant production falls in this type of fermentation. In
fermentation, gas (oxygen) flow rate is subjected to variation by the control
system according to the growth phase of organism and its oxygen demand.
For the rapid oxygen supply a large area of contact between gas and liquid is
necessary which facilitate dissolution and turbulence and mixed the
dissolved gas into the bulk of fermentation. Therefore in aerated systems
oxygen supply and agitation systems are interrelated28.During the
fermentation process the transfer of oxygen from the gas phase into liquid
then into the cell phase is very important for microbial growth under aerobic
condition. For this purpose volumetric liquid phase oxygen transfer co-
efficient (KLa) is a significant factor for scale-up in aerobic fermentation. In
that fermenter equal volumetric gas transfer is not possible since the
superficial gas velocity through the vessel increases differently with the
scale. Moreover an increase in KLa may sometimes have an adverse effect
for the damage of organism in highly turbulent fermentation broth. So it is
necessary to optimize the air flow in the fermentation system and agitator
system in order to search a constant KLa value29.An increase in agitation
speed results in the reduction of bio surfactant yield due to the effect of shear
in Nocardia erythropolis 30.31,32. Bio surfactant recovery depends mainly on
its ionic strength, water solubility and location33.Trehalose lipids of
34, 35 36, 37
Mycobacterium sp , Arthrobacter paraffineus ,
trehalocorynomycolates and tetraesters of Rhodococcus erythropolis 14, 38,
mono,di and pentasccharide lipids of Arthrobacter paraffineus 37, 39,
Nocardia corynebacteroides 24, cellobiolipids of Ustilago sp 40, 41 and
rhamnolipid of Pseudomonas aeruginosa 41, 42, 43 are some well-known
examples of recovery of bio surfactant by solvent extraction. Bio surfactants
from Pseudomonas sp 44, 45, 46, Endomycopsis lipolytica 47, Candida
tropicalis and Debaromyces polymorphous 48 were best recovered by
acetone precipitation. Finally bio surfactant has a tremendous potential for
the purpose of commercial applications in different fields e.g. oil spill

Page | 6
management, oil industry, microbial enhanced oil recovery(MEOR),sludge
removal, crude oil transportation in pipe line, cleanup of oil container or
storage tank, microbial remediation of hydrocarbon, environmental
protection, medicinal applications e.g. as antibiotic, as immunological
adjuvants, anticancer agent, useful agent for gene delivery, antiviral agent
etc. apart from these applications in the area of metal removal, waste
treatment, mechanical dewatering of peat, bitumen release from tar sand,
ceramic processing industry, water treatment, promising food additives,
healthcare and cosmetic industries 49
Classification
Chemical surfactants are generally classified according to their
hydrophilic or lyophobic and lyophilic or hydrophobic character. However
microbial biosurfactants are categorize according to their biochemical
character and the producer strain. The structural backbone of some
biosurfactants with their respective producer strain50 are listed in the
following table (Table 1)
Table 1: Some Microbiologically Derived Surfactant

Type of
Microbe Structural Variety
Microorganism
Torulopsis bombicola Yeast Sorphorose lipid
Preudomonas acruginosa Bacteria Rhamnose lipid
Bacillus sublitis Bacteria Lipoprotein
Arthobacter paraffineus Bacteria Sucrose and Fructose lipids
Candida tropicalis Yeast Polysaccharide fatty acid complex
Coryncbacterium lepus Bacteria Corynomycolic and
Acinetobacter Fatty acids, mono-and di-glycerides,
Bacteria
calcoaceticus lipohetero- polysaecharides
Candida petrophilum Yeast Peptidolipids
Nocardia erythropolis Bacteria Neutral lipids
Corynebacterium
Bacteria Polysaecharide-Protein complex
hydrocurboclastus
Micrococcus sp Bacteria Glycolipids Phospholipids
Aminoacid-lipids
Bacillus sp Bacteria
Cellobiolipids

Some Important Types of Bio Surfactants

The surface active biomolecules are generally of complex lipids where
carbohydrates are combined with long chain aliphatic acids or
hydroxyaliphaticacids. Therefore it is chemically characterized as

Page | 7
glycolipids and again categorized into rhamnolipid, trehalolipids, and
sophorolipids. Now the structural variety and their synthesis will be
described below.
The specific chemical structure of glycolipid type bio surfactant will be
represented below (Figure1)

Fig 1: General structure of Glycolipid bio surfactant where sugar most commonly a
disaccharide containing glucose and or galactose and R-fatty acid components

Rhamnolipid Bio Surfactant

Rhamnolipid, in which one or two molecules of rhamnose are linked to
one or two molecules of ß-hydroxy decanoic acid are the best studied
glycolipid. Production of rhamnose containing glycolipid was first described
in Pseudomonas aeruginosa51. The structural moiety consists of L-
Rhamnosyl-L-rhamnosyl- -hydroxydecanoic acid with one and two
rhamnose units 52. The chemical configuration is represented below
(Figure2)

Fig 2: Structure of Rhamnolipid type Bio surfactant in which two rhamnose subunits
are linked to two -hydroxy-decanoic acids in side chain.

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Trehalolipids Bio Surfactant
There is several structural types of microbial trehalolipids bio
surfactants. Disacharide trehalose linked with C-6 and C-6’ to mycolic acids
is associated with most species of Mycobacterium, Nocardia, and
Corynebacterium 53. Mycolic acids are long chain,  - branched--hydroxy
fatty acids. Trehalose dimycolate produced by Rhodococcus erythropolis
(fig3) has been extensively studied54 and represented by figure 3.

Fig 3: Trehalose dimycolate from Rhodococcus erythropolis in which disaccharide

trehalose is linked to two long chain -branched--hydroxy fatty acids.

Sorphorolipids Bio Surfactant

Sorphorolipids are a mixture of at least six to nine different hydroxyl
fatty acid and consist of a dimeric carbohydrate sorphorose linked to a long
chain hydroxy fatty acid. Sorphorolipids are produced mainly by yeast such
as Torulopsis bombicola as indicated by figure 4 55, 56, 57. Torulopsis
petrophilum and Torulopsis apicola 58, 59. These sorphorolipids, which were
chemically identical to those produced by Torulopsis bombicola, did not
emulsify alkanes or vegetable oils. These results appear to contradict the
conventional belief that microbial emulsifiers and surfactants are produced
to facilitate the uptake of water-insoluble substrates. Although
sorphorolipids can lower surface and interfacial tension. They are not
effective emulsifying agents 55.

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 Fig 4: Sorphorolipid from Torulopsis bombicola in which dimeric sorphorose is
linked to a long chain (G8) hydroxy fatty acid

Lipopeptides and Lipoproteins Type Bio Surfactant

These classes of bio surfactants have a wide variety. A significant
development has been carried out in the area of lipid protein complex
category. Polymyxin dipeptide antibiotic produced by Bacillus brevis60 and
Bacillus polymixa 61 have shown significant surface tension. Pseudomonas
rubescens62 and Thiobacillus thioxidans 63 are able to produce ornithine
containing lipids. Gluconobacter cerinus IFO 3267 64 and Agarobacterium
tumefaciens IFO 3058 65 can be able to synthesise cerilipin, ornithine, taurine
containing lipid and lysine containing lipids respectively and simultaneously
exhibit excellent bio surfactant activity.It has been observed65 that the cyclic
lipopetide surfactin (Figure 5), synthesized by Bacillus subtilis ATCC 21332
is one of the most potent bio surfactants and reduces the surface tension from
72 to 27.4 m N/m at concentrations as low as 0.005%. The production of a
new lipopetide surfactant lichenysin A, by Bacillus licheniformis BAS-50
containing long chain β-hydroxy fatty acids was observed 66.

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 Fig 5: Structure of cyclic lipopetide surfactin produce by Bacillus subtilis

Isolation of Bio Surfactant Producing Strain

Microbial surfactants are usually extra cellular in nature. Different
bacterial strains produce different bio Surfactant with different bio
Surfactant activities. Most of the cases bio Surfactant producing bacteria are
isolated from soil or industrial wastewater. For example Nocardia species L-
417 isolated from soil sources possessed of the bio Surfactant activity
28mN/m. Bio Surfactant production by a new Pseudomonas putida strain
isolated from industrial wastewater possessed of the bio Surfactant activity
29mN/m 15. Screening of bacterial strains is essential for the selection of
most potent strain to achieve highest bio Surfactant activity.
Isolation of Different Bacterial Strains
Generally oil enriched soil contains a significant population of bacteria.
Hence bacterial isolate were collected from oil enriched soil samples of
different zones of petrol pumps. Count of bacterial strains or isolates was
examined by using plate and dilution techniques. The petridishes containing
different dilutions of sample were incubated at 300C for 24hr to obtain the
bacterial colonies.
Medium Composition
The medium for plate and slant culture was composed of 0.5% peptone,
0.3% beef extract) and 3% agar-agar powder at pH 7.0. The fermentation
medium for bio Surfactant production is formulated as C6H12O6, 0.1%
NaNO3, 0.05% MgSO4.7H2O, 0.001% FeSO4.7H2O, 0.1% KH2PO4, 0.025%
yeast extract, 0.01% CaCl2.2H2O at pH 7.0 19.
Production of Crude Bio Surfactant and Selection of the Most Potent
Bacterial Strain
Agar slant culture is prepared by aseptic transfer of bacterial colonies

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followed by incubation at 30ºC for 24hr. Incubated slant culture is
transferred to sterilized medium for inoculum development at 30ºC for 24hr
at 120 r.p.m shaker speed. Finally 0.5 ml of each inoculum was transferred
into 50ml of each fermentation medium for shake flask fermentation at 30ºC
for 7 days at 120 r.p.m shaker speed. After the completion of fermentation
bio Surfactant activity is measured and potent strain is selected.
Assay of Bio Surfactant Activity
After completion of fermentation the culture broth generally centrifuged
at 10,000r.p.m for 30mins. Bio Surfactant activity of the supernatant is
measured by ring surface tension meter. The unit of bio Surfactant activity
will be expressed in terms dyne/cm and author found as 52 dyne/cm 19.
Identification of Bio Surfactant Producing Strain
After isolation and selection of the bacterial strain, identification is an
important tusk. Identification not only characterize the morphological,
cultural and biochemical nature of the microbial strain but also whether the
microbial strain is toxic or non-toxic. Selection of toxic microbial strain is
not desirable because of its lack of fulfilling the objective of bio Surfactant
production. Therefore, identification of the selected potent bacterial strain is
carried out by performing various physical and biochemical tests67. In this
regard different media preparation for carrying out different biochemical
tests are required68. For the study of morphological, cultural and
physiological characteristics different physical and a series of biochemical
tests e.g. simple staining, gram staining, ammonia from arginine, protein
liquefaction, nitrate reduction test, catalase test, in dole test, starch
hydrolysis, synthesis of ammonia from urea, methylene blue milk test etc.
are performed so that the identification can be made and further study can be
progressed. In this regard the author identified the bio Surfactant producing
strain as Bacillus sp 19
Mutation of Identified Microbial Strain
Classically mutation is defined as the sudden and permanent change of
genetic sequence of a species. Basic procedure for classical mutation is to
subject a population of chosen strain to a mutagenic treatment with the
objective of obtaining a population of survivor organisms, which are
randomly genetically altered, from the parent population. Through mutation,
a large portion of population is killed but a high percentage of genetically
modified individuals are formed in the surviving population. Survivors are
then isolated and screened for the identification of the desired mutant.
Screening of a particular mutant may be either rational or random69. In the

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random (non selective) screen, all isolates are tested independently for the
character (selective) of interest. Rational screens are based upon available
and biochemical genetic knowledge of the system involved. Since screening
of mutant of identified bacterial strain Bacillus sp 19 had a subject of interest
on the performance of bio Surfactant production rather than studies of other
biochemical properties, the random screening procedure is considered.
Various physical mutagens e.g. UV ray, X ray, gamma ray etc. and chemical
mutagens e.g., methyl methane sulphonate (MMS), ethyl methane
sulphonate (EMS), ethyl ethane sulphonate, NTG etc. can be considered.
Among the chemical mutagens NTG (N-methyl-N/-nitro-N-
nitrosoguanidine) is one of the effective and potent mutagen. According to
recent report, application of NTG on Bacillus species caused a good increase
of bio surfactant production capacity compared to the parent strain70. For any
mutation it cannot be predicted that which dose of a particular mutagen and
which time of contact of the mutagen can be effective to fulfill the objective
of mutation. Therefore, it is necessary to apply different doses of chemical
mutagen with different time of contact of that mutagen with the organism
subjected to mutation. Mutation of the selected strain is carried out by using
chemical mutagen N-methyl-N'-nitro-N-nitro so guanidine at final
concentration ranging from (10-100) µg/ml in 10ml of 24hr culture broth and
incubated at 300C at 120r.p.m shaker speed and each isolate was tested for
bio Surfactant activity and author found as 28dyne/cm19.
Environmental Conditions for Bio Surfactant Production in Shake Flask
Fermentation
Production of the bio Surfactant from a particular organism may vary
with the variation of environmental as well as cultural conditions for cell
growth as well as fermentation conditions. Bacterial strains show different
growth phases in cultural media 71 and the growth factor of a particular
bacterial strain may affect the bio Surfactant production. The strains are
capable to produce bio Surfactant within a particular growth phase. So it is
necessary to optimize the age of inoculum to enhance the Bio Surfactant
production. When cell growth of a particular bacterial strain added in
different amounts in the fermentation medium may show a variation of
production of Bio Surfactant. An increase in amount of cell in the
fermentation medium may not enhance the productivity of the bio surfactant
due to the limitation of the nutrients. On the other hand lesser amount of cell
in the fermentation medium may also cause less bio surfactant production
since lesser number of cells are participating in the bio surfactant production.
So it is necessary to optimize the amount of cell growth to be added in the
fermentation medium for the bio surfactant production. The fermentation

Page | 13
process for bio surfactant production can be influenced by several
environmental as well as cultural factors e.g. period of fermentation,
temperature of fermentation, pH of the medium and volume of the
fermentation medium, volume of inoculum and age of inoculum. Therefore,
a successful fermentation usually requires an effective optimization of
process parameters so that a significant yield is obtained in terms of bio
surfactant production. It has been observed that the maximum bio surfactant
production is achieved at 300C, pH 7.0 on 7 days of fermentation. Optimum
volume of the medium, age of the inoculum and volume of the inoculum are
50ml, 24hr and 1% (v/v) respectively 19.
Selection of Carbon and Nitrogen Sources for Bio surfactant Production
For bio surfactant production, selection of nutrient i.e. specially carbon
and Nitrogen sources for the growth of a microbial strain as well as
production is an important factor. Microorganisms show a wide variation of
Nutrients that they can utilize for growth and bio surfactant production. So
the composition of the medium is a very important factor in determining the
yield of bio surfactant from any microbial Strain. Medium supporting the
higher growth of the organism does not always means the satisfactory
production of biological surface active agent. So for the growth of bacterial
species and production of bio surfactant, it is always required to select the
carbon source, nitrogen source and mineral source in the fermentation
medium. Now in order to optimize nutritional parameters different carbon
and nitrogen sources were used. For the selection of carbon sources different
carbohydrates viz. starch, lactose, arabinose, raffinose, xylose, galactose,
glucose etc. and hydrocarbons viz. purified kerosene as mixed hydrocarbon
(Figure 6), hexane, heptanes, octane, decane, dodecane and tridecane
respectively can be tested for their bio surfactant activities. For the selection
of nitrogen sources in the fermentation medium different simple viz. sodium
nitrate, ammonium chloride, ammonium nitrate, ammonium sulphate,
diammonium hydrogen phosphate, ammonium dihydrogen phosphate etc.
and complex nitrogen sources viz. peptone, tryptone, beef extract, malt
extract, yeast extract and molasses can be considered. In this study by
measuring the surface activity of the bio surfactant and cell mass
concentration, carbon and nitrogen sources were optimized. Now from these
studies the optimum C/N ratio was also selected. Superior carbon and
nitrogen sources for bio surfactant production are found as glucose (2%);
sodium nitrate (nitrogen level 16.47mg/100ml) and yeast extract (nitrogen
level 2.45mg/100ml) in combination respectively where the optimum C/N
ratio was found to be 0.12:1 19.

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 Fig 6: Treatment of Impure Kerosene for the conversion of pure kerosene

Process Conditions for Fermentative Production of Bio Surfactant in

Laboratory Scale Fermenter
The function of a fermenter is to provide the environment for the growth
of organisms or cells under controlled conditions72. The biological
transformation under the influence of microorganism in a suitable
environmental conditions e.g. temperature of fermentation, time of
fermentation, inoculum volume, inoculum age, substrate concentration,
volume of the medium, aeration rate (v. v. m, in case of aerobic
fermentation),agitation rate (r. p. m) etc. takes place in such type of vessel.
Every fermentation phenomenon generally follows the scale up pathway i.e.
from agar slant culture to shake flask fermentation to laboratory scale to pilot
plant scale and finally to commercial scale of fermentation. Moreover
inoculum development for a successful fermentation in a laboratory scale
fermenter is very important. In this regard the inoculum development
pathway will follow the aseptic transfer of inoculum from shake flask culture
to a series of seed vessels or tanks i.e. seed tank-I, seed tank-II, seed tank-III
etc. consisting of sterilized nutrient so that inoculum can be developed in a
optimum concentration thereby fulfil the efficient fermentation in large scale
fermenter for product synthesis. Now in comparison to oxygen transfer rate
(OTR), shake flask fermentation is less efficient than in an continuous stirred
tank fermenter (CSTF).During the fermentation process the transfer of
oxygen from the gas phase into liquid and then into the cell phase is very
important for microbial growth under aerobic condition where so many
resistances e.g. gas liquid interfacial resistance, liquid film resistance etc. are
important. For this purpose volumetric liquid phase oxygen transfer co-
efficient (KLa) which is the product of individual liquid film resistance and

Page | 15
gas liquid interfacial area is a significant factor for scale-up in aerobic
fermentation. In that fermenter equal volumetric gas transfer is not possible
since the superficial gas velocity through the vessel increases differently
with the scale. Moreover an increase in KLa may sometimes have an adverse
effect for the damage of organism in highly turbulent fermentation broth. So
it is necessary to optimize the air flow in the fermentation system and
agitator system in order to search a constant KLa value. Therefore
optimisation of process conditions including period of fermentation, agitator
speed, aeration rate and volume of the fermentation medium are carried out
and found as 5 days,150r.p.m,1.0v.v.m and 3liter.Moreover viscosity and
KLa determined under these conditions are found to be 129.76hr-1 and
0.037c.p 29.
Determination of KLa
The volumetric mass (oxygen) transfer co-efficient (KLa) in
fermentation system is determined by dynamic gassing out method73. In this
method most dissolved oxygen was replaced by nitrogen gas and then the
definite volume of air was supplied to the actively respiring culture and then
air supply was allowed to stop and the resulting fall in dissolved oxygen due
to respiration of organism was measured as a function of time to obtain the
rate of oxygen uptake by the total microbial population. As soon as the
dissolved oxygen reached the critical value, the aeration was resumed and
the change in D.O. concentration (dcL/dt) was measured from these data.
Such variation of D.O. concentration with time has shown in figure 7 and
figure 8 respectively. Now the value of concn of oxygen (CL) at a particular
point in the fermenter can be measured from the value of concentration of
oxygen (C*) in the liquid that would be in the equilibrium with the partial
pressure of oxygen in the air. The equilibrium concentration of oxygen C*
can be calculated from the mole fraction of gas x (oxygen) in solution as C*
= (32/18) x 106 x (x/1-x)……………………………… (1)
Again p = HX……………………. (2), where p is the partial pressure of
oxygen in air at a particular temperature and H is Henry’s Law constant at
that temperature. For that experiment which was carried out at 300C, the
values of P and H were taken from international critical tables. Hence from
different values of % of dissolved oxygen (D.0), the corresponding CL values
were obtained as CL = C* x % of D.O……… (3).
Oxygen uptake rate (OUR i.e. RX) can be measured from the slope of
(figure 7 i.e. fig 17). When CL is plotted against time in the non-gassing
situation i.e. when air is turned off. In that case the change in D.O.

Page | 16
 concentration (dCL/dt) = – rx …..…………………………………. (4),
Where r is the specific oxygen uptake rater (SOUR) and X is the cell
mass concentration. For the aeration period, the change in D.O.
concentration (dCL/dt) = KLa (C*-CL)-rx --- ……. (5)
The equation (5) can be rewritten as CL = -1/KLa [(dCL/dt) + rx] +
C*……………………6).
Now by plotting [(dCL/dt) + rx] against CL (figure 8 i.e. fig 18).) a
straight line can be obtained and from the reciprocal of slope of the line, the
value of KLa can be determined.

Fig 7: Dissolve oxygen concentration (CL) vs time (t) plot

Fig 8: CL vs (dCL/dt+rx) plot for the determination of KLa

Page | 17
 Fig 9: Schematic description of the direct measuring of OTR in bioprocess by the
classical dynamic technique74

Fig 10: Schematic description of the dynamic technique desorption–absorption of

oxygen for inert condition measurements74

Determination of Viscosity of Fermentation Broth

Viscosity of the fermentation broth in a dynamic fermentation system
was generally measured by Brookfield Synchroelectric Viscometer (Lab
Model) in centipoises (cp). Here viscosity was measured under different
fermentation conditions viz. variable aeration rate, variable agitation rate,
variable period of fermentation and changing volume of the medium inside
the fermenter. Finally the optimum viscosity was determined 29.
Determination of Cell Growth
Cell growth generally measured by separating the supernatant after
centrifugation at 10,000 r.p.m for 30 mints from fermentation broth. The cell

Page | 18
mass so obtained was dried in Aluminium cup at 600C overnight using a hot
air oven and then the weight of dry cell mass was estimated 29.
Recovery of bio Surfactant
After the completion of fermentation, it is important to recover the
product i.e. bio Surfactant from fermentation broth. Basically the product
recovery is based upon two general pathways viz. primary purification or
isolation and then purification or secondary purification. There are lots of
methods for the recovery of bio Surfactant viz. ammonium sulphate
precipitation technique, solvent precipitation viz. acetone, ethanol, methanol
etc., acid precipitation, thin layer chromatography (TLC), ion exchange
chromatography, isoelectric precipitation, adsorption etc. Different methods
of recovery is represented by table 2.
Table 2: Methods for the recovery of bio surfactants 50

Methods
Adsorption
Adsorption
Ion exchange Chromatography
Solvent extraction
Centrifugation
Acid Precipitation
Membrane Ultra filtration
Selective Crystallization
Ammonium Sulphate Precipitation
Organic Solvent extraction
Thin layer Chromatography
Dialysis
Lyophilisation
Iso-electric Focusing
Mechanisms
Wood absorption 75, 76
Polystyrene adsorption 77
Charge separation 79
Dissolves in organic solvent 78
Centrifugal Force 79
Low Ph 80
Micelles formation 80
Redissolution in organic solvent 81
Salting out 81
Dissolves in organic solvent 75
Relative flow difference 82
Difference in solute Concentration 83
Cryodesiccation 83
Charge difference 83

In a recent development continuous removal of bio surfactant during

fermentation by different techniques has increased the cell density in the
reactor and eliminated product inhibition, resulting in a several fold net
increase in bio surfactant yield 84, 85, 86. Other continuous recovery techniques
involving adsorption on Amberlite XAD-2 followed by purification and
freeze drying have also been reported to give 60% recovery with 90% purity
75
.Biosurfactant produced by Nocardia sp. L-417, were purified by
procedures that included ammonium sulphate fractionation, chilled acetone
and hexane treatment, silica-gel column chromatography and sephadex LH-
20 gel filtration 87. The recovery of bio surfactant was maximum by using

Page | 19
acetone as solvent in comparison to other solvent and it was 3.7217g/l. The
surface tension of the purified bio surfactant was 18 dyne/cm. So the activity
of the purified bio surfactant was increased by 1.5 fold in comparison to
crude bio surfactant 19.
Comparative Study with Synthetic Surfactant
The isolated and purified bio surfactant reduces the surface tension
effectively. Some commercially used chemical or synthetic surfactants were
also be able to reduce the surface tension. Different commercially used
chemical surfactants e.g. viscosin, Dodecylbenzene sulfonate (DBS), Linear
Alkyl benzene sulfonate (LABS), Tween 80, Span 20, Brij 35 were
considered in the present discussion. Lowering of surface tension by the
isolated and purified bio Surfactant produced from Bacillus sp took place
efficiently 19.The comparative study in context to surface tension of
microbiologically derived surfactant with synthetic surfactant is represented
by the following table (table 3)
Table 3: Comparative study of surface tension (dynes/cm) of bio surfactant with
synthetic surfactants at different concentrations

Concentrations Surface Tension

Name of the Surfactant
(g/l) (dyne/cm)
0.05 70
0.10 46
0.20 34
Viscosin 0.25 33
0.50 31
0.75 30
1.0 30
0.05 66
0.10 58
0.20 56
Dodecyl benzene sulfonate
0.25 54
(DBS)
0.50 52
0.75 50
1.0 50.5
0.05 62
0.10 52
Linear Alkyl benzene sulfonate
0.20 52
(LABS)
0.25 53
0.50 50

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 0.75 51
1.0 50
0.05 50
0.10 52
0.20 52
Tween 80 0.25 53
0.50 55
0.75 56
1.0 56.5
0.05 40
0.10 38
0.20 38
Span 20 0.25 37.5
0.50 38.0
0.75 40
1.0 40.5
0.05 45
0.10 42
0.20 40
Brij 35 0.25 40
0.50 38
0.75 40
1.0 41
0.05 72
0.10 70
0.20 68
Purified Bio Surfactant 0.25 62
0.50 30
0.75 26
1.0 18

In table 3, with some known synthetic surfactant in regard to their

capacity for reduction of surface tension of water at their different levels
(concentrations) it was observed that the bio surfactant has a much more
capacity for the reduction of surface tension of water upto 18 dynes/cm at its
concentration of 1.0g/l. Moreover, its aqueous solution has also capacity for
foam formation which was somewhat in fewer amounts as compared with
the synthetic or chemical surfactants.

Page | 21
Bio Surfactant in Food Processing and Environmental Applications
Promising Food Additives
Bio Surfactants also have several promising applications in the food
industry as food additives 50. Food additives are ingredient with no
nutritional value added to food to modify physical, chemical, biological, or
sensory characteristics during the manufacturing, processing, preparation,
treatment, packaging, storage, transport or handling. Lecithin and its
derivatives, fatty acid esters containing glycerol, sorbitan or ethylene glycol
and ethyoxylated derivatives of monoglycerides including a recently
synthesized oligopeptides88 are currently in use as emulsifiers in the food
Industries worldwide. A novel bio Emulsifier from Candida utilis has shown
potential use in salad dressing 89.Biosurfactants are potential candidates in
the search for functionally different products, as they meet the requirements
of functional food additives.
Bio Surfactant as Food Emulsifiers
Food emulsions and colloids are intricate systems consisting of several
components of food ingredients. Low molecular weight amphiphiles play
and essential role in the stability of such liquid emulsions as (beverages,
dressings, sauces and alcoholic emulsions and others). Food colloids
therefore mainly consist of blends of monomeric and polymeric
hydrocolloids and proteins. In many food industries emulsifiers are used in
order to obtain the right consistency and texture of food additives product
process. The capacity of surface-active compounds to modifying the
rheological characteristics of dough or the emulsification of partially broken
fat tissue explains the need for their usage application in bakery and meat
processing practices 90.A number of natural plant crop derived food
emulsifiers, such as lecithin and gum Arabic, have traditionally been
accepted by food industries. Cocoa powder is one of the most popular
ingredients in a number of dry mixes for desserts and drinks that are easy to
prepare and to store. Mayonnaise, butters, cream, margarine, salad dressing,
chocolates, and hotdogs are examples of foods processed from emulsions 91,
92
. In addition to the obvious role as surface and interfacial tension reducing
agents enhancing stabilization of emulsion formation, microbial bio
Surfactants have other important uses in food including; controlling texture
and shelf-life of starch-containing products, improving consistency and
texture of fat-based products, agglomeration of fat globules, stabilizing
aerated systems, and modifying rheological properties of wheat dough 93.In
bakery and ice cream formulations bio Surfactants have been reported to
improve consistency, delaying staling, and solubilising flavour oils and as fat

Page | 22
stabilizers and anti-spattering agents. Rhamnolipid in particular have been
used to enhance dough texture, stability, volume and conservation of bakery
products 94, 95.
Bio Surfactants as antioxidant agents
Bio Surfactants show some potential as antioxidant agents.
Mannosylerythritol lipids (MELs) are versatile bio Surfactants known for
their versatile interfacial and biochemical properties. A polysaccharide
emulsifier from Klebsiella sp was also shown to have potent inhibition of the
autooxidation of soybean oil 96.
Oils and Fats
Most oils and fats are used in the food industry, which generates great
quantities of wastes and so, their disposal is a growing problem. Candida
antarctica and Candida apicola synthesized surfactants (glycolipids) in a
cultivation medium supplemented with oil refinery waste, either with soap
stock (5-12% v/v) or post-refinery fatty acids (from 2 to 5% v/v). The
efficiency of glycolipids synthesis was from 7.3 to 13.4 g/L and from 6.6 to
10.5 g/L in the medium supplemented with soap stock and post-refinery fatty
acids, respectively 97. The meat processing industry is seeking new
applications for abundantly available, inexpensive animal’s fats.
Sophorolipid production by Candida bombicola was studied as a model of
fat utilization for bio Surfactant production. In a pH controlled fermenter,
120 g/L of sophorolipid was obtained and the cells at the end of fermentation
contained 37% of protein and 14% lipids 98.
Antiadhesive Agents
The involvement of bio Surfactants in microbial adhesion and
detachment from surfaces has been investigated. A surfactant released by
Streptococcus thermophilus has been used for fouling control of heat-
exchanger plates in pasteurizers as it retards the colonization of other
thermophilic strains of Streptococcus responsible for fouling 99. The use of
bio Surfactants released by Lactobacilli strains is very promising once these
microorganisms are naturally present in human flora and have also a
probiotic effect 100. Considering the interesting properties demonstrated by
bio Surfactants, future utilization as multipurpose ingredients, which exhibit
emulsifier, antiadhesive, and antimicrobial activities simultaneously and
thus, suitable for many food applications 92.
Environmental Applications
Due to good physicochemical properties, low toxicity and good

Page | 23
biodegradability bio Surfactants are widely applied in environmental
protection techniques, e.g. water and soil
Remediation, oil spills removal etc. Bio Surfactants turned out to
remove crude and model oils from sand columns or contaminated ground in
the washing process. The efficiency of bio Surfactant in removing crude oil
was comparable to those of synthetic surfactant and much higher than for
natural plant surfactant–saponin 101 and synthetic Tween 60 102. In the case of
removing hexadecane from sand bio Surfactant was much more efficient
than SDS and Tween 80103.Biosurfactants are also very effective in
enhancing of oils biodegradation. The addition of bio Surfactant produced by
Candida Antarctica to the fermentation process of n-undecane improved
degradation rate of petroleum hydrocarbons, while application of synthetic
surfactant Tween 40 and Span 80 didn’t show any improvement104.In
biodegradation of phenanthrene bio Surfactants produced by Pseudomonas
aeruginosaP-CG3 and ATCC 9027 strains were less effective than synthetic
Tween 80 and more effective than Triton X-100 in respect to enhancement
of the rate of biodegradation. However, they displayed the highest
phenanthrene solubilisation among all examined surfactants. The solubility
of phenanthrene in P-CG3 and ATCC 9027 bio Surfactants’ solutions at 10 x
CMC concentrations were 50 and 28 mg/l, respectively, while only 16 and
11 mg/l for Tween 80 and Triton X-100, respectively105.A review of
investigations on efficiency of bio Surfactants and synthetic surfactants in
enhancing biodegradation of polycyclic aromatic compounds (PAHs)
showed, that bio Surfactants display similar PAH bioavailability
enhancement as synthetic surfactant, however, are non-toxic to
microorganisms degrading pollutants 106.Summarizing, one can state that bio
Surfactants display a lot of advantages over chemically synthesized
surfactants. They are less toxic, highly effective and easily biodegradable,
what makes them environmentally friendly and proves their potential to
replace synthetic surfactants in many applications, not only of environmental
character. One of the possibility is exploiting their solubilising properties
and using in vegetable oils refining for removal of phospholipids.
Microbial Enhanced Oil Recovery (MEOR)
One of the potential applications of microbial bio Surfactants to be
recognized and to draw industrial attention was in microbial enhanced oil
recovery (MEOR). Surfactant based technology is being developed to
enhance recovery of the remaining 50%. A number of reviews exist in this
regard 107, 108. Bacillus licheniformis JF-2, an isolate from oilfield injection
water which, in addition to producing most effective bio Surfactants (CMC,

Page | 24
10 g.liter-1, interfacial tension of saline against decane lowered to 10-3
dyne.cm-1), has other properties such as being anaerobic, halo tolerant, and
thermo tolerant, makes bio Surfactants that are potentially useful for in situ
microbially enhanced oil recovery (MEOR) 109, 110, 111. It has been reported 112
that classical oil production technologies involving primary and secondary
can only partially recover the oil present in the field with an efficiency
estimated at 30-40% of the overall amount of oil available, such efficiencies
are expected to decrease with the gradual depletion of light crude resources
leaving the viscous crude oil, thus requiring the development of the
“tertiary” processes which aim at enhanced oil recovery. A report was also
came113 that the use of laboratory selected bio-surfactant producing
microorganism been injected into oil reservoir with resounding success.
These microbes are injected along with their nutrients into oil wells thus
their ability to grow and metabolize in these areas. Since most oil extraction
processes are carried out at high temperature, most organisms implicated
here are extremophiles. Another strategy is based on the concept that oil
reservoirs are inhabited by indigenous microbial communities however the
knowledge of such microbial ecosystems is limited due to difficulties in
collecting representative samples as well as carrying out in situ analysis.
Therefore, whether indigenous microorganisms are native or contaminants
exogenously introduced through water flooding, drilling or other oil well
operations is still to be confirmed as well as their metabolism and activities
established 113, 114.
Environmental Protection
Bio Surfactants are also useful in bioremediation of sites contaminated
with toxic heavy metal like uranium, cadmium, and lead and this aspect has
been very well reviewed recently 115. These observations suggest the
usefulness of natural microbial consortia and their products in solving
environmental related problems.
Metal Removal
There is an increased interest in using surfactants to complex metals
from industrial discharge and mining operations. Removal of lead from
wastewater using biosurfactant116 is an important commercial application.
Bio Surfactant in Metal Remediation
The addition of a bio surfactant could promote desorption of heavy
metals from its solid phases in two different approaches. The first approach
is through the complexation of the free form of metal ions residing in
solution. This would decreases the solution-phase activity of the metal and,

Page | 25
therefore, promotes desorption according to Le Chatelier's principle. The
second approach is through the accumulation of bio surfactants at the solid-
solution interface under the condition of reduced interfacial tension. This
would allow the direct contact between the bio surfactant and the sorbed
metal 117. Since the utilization of microorganisms and microbial products,
e.g., bio surfactants, in bioremediation of metal-contaminated soils shows
promising results, consequently, the development of remedial technologies
will require further study in several areas. For instance, soils contain
numerous cations that may compete with metal contaminants for the bio
surfactant complexation sites. Therefore, the selectivity of bio surfactants for
metals both in solution and in soil systems must be prior examined. The
mechanisms of heavy metal removal by bio surfactants consist of three main
steps: sorption and binding of the bio surfactant to the soil surface and also
to the metal; separation of the metal from the soil to the solution; and lastly
association of the heavy metal with micelles. Heavy metals are trapped
within the micelles through electrostatic interactions and can be easily
recovered through precipitation or membrane separation techniques118.
Conclusions
Bio surfactant has been proved as a suitable alternative of chemically
synthesized surface active molecules or surfactants. This is generally
produced from different species of microbial origin. The objective of the
current task is to produce bio surfactant from natural resources viz. oil
enriched earth sample by the active involvement of microbes. In this regard
among other microbial specimen, bacterial population plays an important
role in bio surfactant production. Here isolation, identification and mutation
of the potent strain by means of chemical mutagen are beneficial for higher
yield. In shake flask ferementation, the standardisation of environmental
conditions was carried out first and then suitable carbon source and nitrogen
sources were selected for further course of work. The scale up process helps
for the quantitative enhancement of production in laboratory scale fermenter.
A suitable comparative study about surface tension or surface activity
between microbiologically produced and purified surfactant and synthetic
surfactant at different concentrations will help to know the potentiality
towards surface active property. Finally the produced bio surfactant can be
able to utilize in so many areas including food processing and environmental
applications.
References
1. Karanth NGK, Deo PG. Veenanadig NK. Microbial production of bio

Page | 26
 surfactants and their importance. Current Science. 1999; 77:116-123.
2. Jiraporn T, Niran R, Takauki K, Mitsuru H, Tadayuki I, Manasaki M et
al. Production and characterization of bio surfactants from Bacillus
licheniformis F 2.2. Bioscience. Biotechnology Biochemistry. 2003;
67(6):1239-1244.
3. Sudha S, Kumanan R, Muthusamy K. Der Pharmacia Letter. 2010;
2(2):155-158.
4. Marion FD, Maget-Dana R. Europian Journal of Biochemistry, 1985,
153-158.
5. Grangemard I, Bonmatin JM, Bernillon J, Das BC, Peypoux F. Journal
of Antibiotics. 1999; 52:363.
6. Peypoux F, Guinand M, Michel G, Delcambe L, Das BC. Biochemistry.
1978; 17:3992-3994.
7. Nishikiori T, Naganawa H, Muraoka Y, Aoyagi T. Journal of
Antibiotics. 1986; 39:860-864.
8. Arima K, Kakinuma A, Tamura Surfactin G. a crystalline peptide lipid
surfactant produced by Bacillus subtilis: isolation, characterization and
its inhibition of fibrin clot formation. Biochemistry Biophysics.
Research Communication. 1968; 31:488-494.
9. Destin J, Robalin D, Thourt P. Biotechnology Letter. 1997; 19(2):105-
108.
10. Namir IAH, Wang J, Bozhong M. Identification of a Biosurfactant
Producing Strain: Bacillus subtilis HOB2. Protein & Peptide Letters.
2009; 16:7-13.
11. Suzuki T, Tanaka HH, Itoh S. Agricultural Biological Chemistry. 1974;
38:557.
12. Davis DA, Lynch HC, Varley J. Enzyme Microbiology Technology.
1999; 25:322-326.
13. Cooper DG, Paddock DA. Applied Environmental Microbiology. 1984;
47:173-178.
14. Guerra Santos LO, Kappeli O, Fiechter A. Growth and biosurfactant
production of a bacterium in continuous culture. Enzyme Microbiology
Technology. 1998; 24:322-326.
15. Tuleva KB, Ivanov RG, Christova EN. Bio surfactant production by a
New Pseudomonas putida strain. Z Naturforsch. 2002; 57C:356-360.

Page | 27
16. Ristau E, Wagner F. Formation of Novel anionic trehalose tetraesters
from Rhodococcus erythropolis under growth limiting conditions.
Biotechnology Letter. 1993; 5:95-100.
17. Duvnajale Z, Cooper DG, Kosaric N. Effect of nitrogen source on
surfactant production by Arthrobacter paraffineus ATCC 19558 in J.E.
Zajic, D.G. Cooper, T.R. Jack and N. Kosaric (Ed.) Microbial enhanced
oil recovery. Pennwell Books. Tulsa, Okla, 1998, 66-72.
18. Gerson DF, Zajic JE. Antonic van Leeunenhock, 1979, 45-81.
19. Ray S. Production of bio surfactant using an isolated bacterial strain of
Bacillus sp (m28). Journal of Microbiology and Biotechnology
Research. 2012a; 2(3):402-415.
20. Wagner F, Kim JS, Lang S, Li ZY, Marwed G, Metueovic U et al. Proc.
3rd. European. Congress of Biotechnology. Verlag Chemie Weinheim,
1984, 1-3.
21. Sen R, Swaminathan T. Applied Microbiology Biotechnology. 1997;
47:358-363.
22. Jenneman MJ, Mclnerney MJ, Knapp RM, Clark JB, Feero JM, Revus
DE et al. Developmental Industrial Microbiology. 1983; 24:485-488.
23. Guerra-Santos LH, Kappeli O, Fiechter A. Pseudomonas. Aeruginosa
bio surfactant production in continuous culture with glucose as carbon
source. Applied Environmental Microbiology. 1984; 48:301-305.
24. Powala M, Lang S, Wray V. Penta- and disaccharide lipid formation by
Nocardia corynebacteroides. Grown on n-alkanes. Applied
Microbiology Biotechnology. 1989; 31:473-479.
25. Solomons GL. Materials and Methods in Fermentation, Academic Press.
London, 1969, 42-44.
26. Diendoerfer FH, Gaden EL. Applied Microbiology. 1965; 3:253-256.
27. Gaden EL. Biotechnology and Bioengineering. 1962; 4:99-102.
28. Taguchi H, Humphrey AE. Dynamic Measurement of the volumetric
oxygen transfer coefficient in fermentation systems. Journal of
Fermentation Technology. 1966; 44:881-889.
29. Ray S. Optimization of process conditions for bio surfactant production
from mutant strain of Bacillus sp (m28) in a 5l laboratory fermenter,
Journal of Microbiology and Biotechnology Research. 2012b; 2(3):431-
439.

Page | 28
30. Margaritis A, Kennedy K, Zajic JE. Application of an airlift fermenter in
the production of bio surfactants. Developmental Industrial
Microbiology. 1980; 21:285-294.
31. Margaritis A, Zajic JE, Gerson DF. Production and surface active
properties of microbial surfactants. Biotechnology and Bioengineering.
1979; 21:1151-1162.
32. Margaritis A, Kennedy K, Zajic JE, Gerson DF. Bio surfactant
production by Nocardia erythropolis. Developmental Industrial
Microbiology. 1979; 20:623-630.
33. Desai JD, Banat IM. Microbiology and Molecular Biology Review.
1997; D:47-64.
34. Assetincau C, Asselineau J. Trehalose containing glycolipids. Progress
Chemistry Fats Lipids. 1978; 16:59-99.
35. Syldatk C, Wagner F. Production of bio surfactants. In N. Kosaric WL.
Cairns, and N.C.C. Gray (ed) Bio Surfactants and Biotechnology.
Marcel Dekker Incorporate, New York, 1987, 89-120.
36. Itoh S, Suzuki T. Fructose lipids of Arthrobacter, Corynebacteria,
Nocardia and Mycobacterium grown on fructose. Agriculture and
Biological Chemistry. 1974; 38:1443-1449.
37. Suzuki T, Tanaka H, Itoh S. Sucrose lipids of Arthrobacter,
Corynebacteria and Nocardia grown on sucrose. Agriculture and
Biological Chemistry. 1974; 38:557-563.
38. Kretschmer A, Bock H, Wagner F. Chemical and Physical
Characterization of interfacial-active lipids from Rhodococcus
erythropolis grown on n-alkane. Applied Environmental Microbiology.
1982; 44:864-870.
39. Li ZY, Lang S, Wagner F, Witte I, Wray V. Formation and
Identification of interfacial active Glycolipids from resting microbial
cells of Arthrobacter sp. and potential use in tertiary Oil recovery.
Applied Environmental Microbiology. 1984; 48:610-617.
40. Boothroyd B, Thern JA, Haskins RH. Biochemistry of the ustilaginales.
XII. Characterization of extracellular glycolipids produced by Ustilago
sp. Canadian Journal of Biochemistry and Physiology. 1956; 34:10-14.
41. Hiratsuka K, Nakahara T, Sano N, Yamada K. Formation of
Rhamnolipid by Pseudomonas aeruginosa: its function in hydrocarbons
fermentations. Agricultural Biological Chemistry. 1971; 35:686-692.

Page | 29
42. Desai AJ, Patel RM, Desai JP. Emulsifier production by Pseudomonas
fluorescens during the growth on hydrocarbons. Current Science. 1988;
57:500-501.
43. Yamaguchi M, Sato A, Yukuyams A. Microbial production of Sugar
lipids. Chemical Industry. 1976; 17:741-742.
44. Reddy PG, Singh HD, Pathak MG, Bhagat SD, Baruah JN. Isolation and
functional characterization of hydrocarbon emulsifying and solubilizing
factors produced by Pseudomonas sp. Biotechnology and
Bioengineering. 1983; 25:387-401.
45. Chameotra SS, Singh HD. Purification and Characterization of alkane
solubilizing factor produced by Pseudomonas PG-1. Journal of
Fermentation Bioengineering. 1990; 69:341-344.
46. Goswami P, Singh HD. Different modes of hydrocarbon uptake by two
different Pseudomonas sps. Biotechnology and Bioengineering. 1991;
37:1-11.
47. Roy PK, Singh HD, Bhagat SD, Baruah JN. Characterization of
hydrocarbon emulsification and solubilization occurring during the
growth of Endo mycopsis lipolytica on hydrocarbons. Biotechnology
and Bioengineering. 1979; 21: 955-974.
48. Singh M, Desai JD. Hydrocarbon emulsification by Candida tropicalis
and Debaromyces polymorphous. Indian Journal of Experimental
Biology. 1989; 27:224-226.
49. Ray Sources S. Characterization, synthesis and utilization of microbial
surface active biomolecules: A Review. The Journal of Bioprocess
Technology. 2014; 99:360-388.
50. Cooper DG, Zajic JE. Advances in Applied Microbiology. 1980;
26:229-232.
51. Jarvis FG, Hohnson MJ. A glycolipid produced by Pseudomonas
aeruginosa. Journal of American Chemical Society. 1949; 71:4124-
4126.
52. Syldatk C, Lang S, Wagner F. Chemical and Physical Characterization
of four interfacial-active rhamnolipids from Pseudomonas sp. DSM
2874 grow on n-alkanes. Z. Naturforsch. 1985; 40C:51-60.
53. Lang S, Wagner F. Structure and properties of bio Surfactants, In N.
Kosaric, W.I. Cairns, and N.C. Gray (Ed), Bio surfactants and
Biotechnology. Marcel Dekker, Inc, New York, N.Y, 1987, 21-47.

Page | 30
54. Rapp BH, Wray V, Wagner F. Formation, isolation and Characterization
of trehalose dimycolates from Rhodococcus erythropolis grown on n-
alkanes. Journal of General Microbiology. 1979; 115:491-503.
55. Cooper DG, Paddock DA. Production of a bio surfactant from
Torulopsis bombicola. Appl. Environ. Microbiol. 1984; 47:173-176.
56. Gebbert U, Lang S, Wagner F. Sorphorose lipid formation by resting
cells of Torulopsis bombicola. Biotechnology Letter. 1984; 6:225-230.
57. Inoue S, Itoh S. Sorphorolipids from Torulopsis bombcola as microbial
surfactants in alkane fermentation. Biotechnology Letter. 1982; 4:3-8.
58. Cooper DG, Paddock DA. Torulopsis petrophilum and surface activity,
Applied Environmental Microbiology. 1983; 46:1426-1429.
59. Tulloch P, Hill A, Spencer JFT. A new type of macrocyclic lactone from
Torulopsis apicola. Journal of Chemical Society Communication, 1967,
584-586.
60. Itoh S, Suzuki T. Effect of rhamnolipids on growth of Pseudomonas
aeruginosa mutant deficient in m-paraffin utilizing ability. Agriculture
and Biological Chemistry. 1972; 36:2233-2235.
61. Fujikawa K, Hayashi K, Suzuki T, Tsukamoto K. The chemical
structure of polymyxin E. The identities of polymyxin E1 with colistin
A and Polymyxin E2 with colistin B. Journal of Biological Chemistry.
1965; 57:226-227.
62. Yamane T. Enzyme technology for the lipid industry: An engineering
overview. Journal of American Oil Chemist Society. 1987; 64:1657-
1662.
63. Knoche HW, Shiveley JM. The Structure of an ornithine containing
lipid from Thiobacillus thiooxidans. Journal of Biological Chemistry.
1972; 24(7):170-178.
64. Kondo K, Tahara Y, Yamada Y. A new lysine containing lipid isolated
from Agarobacterium tumifaciens. Agricultue and Biological
Chemistry. 1976; 40:243-244.
65. Arima K, Kokinuma A, Tamura G. Surfactin, a crystalline peptide lipid
surfactant produced by Bacillus subtilis: Isolation, characterization and
its inhibition of fibrin formation. Biochemistry Biophysics Research
Communication. 1968; 31:488-494.
66. Fredrickson HI, Timmis KN, Yakimov MM. Effect of heterogeneity of

Page | 31
 hydrophobic moieties on surface activity of lichenysin A, a lipopeptide
biosurfactant from Bacillus licheniformis BAS 50. Biotechnology and
Applied Biochemistry. 1990; 23:13-18.
67. Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST. Bergey’s
Manual of Determinative Bacteriology, Williams and Wilkins,
Baetimore, 1994.
68. Harrigan WF, McCance ME. Laboratory Methods in Microbiology,
Academic Press, London, 1970, 51-57.
69. Sounders AV, Sounders RT. Microbial genetics applied to
biotechnology, Croom Helm, London, 1987, 212.
70. Destin J, Robalin D, Thourt P. Biotechnology Letter. 1997; 19(2):105.
71. Salle AJ. Text Book of Bacteriology, Edn 7,Tata McGraw Hill
Publishing Co. Ltd, New Delhi, 1974, 274.
72. Solomons GL. Materials and Methods in Fermentation, Academic Press.
London, 1969, 26-28.
73. Taguchi H, Humphrey AE. Dynamic Measurement of the volumetric
oxygen transfer coefficient in fermentation systems. Journal of
Fermentation Technology. 1966; 44:881-889.
74. Ochoa GF, Gomez E. Bioreactor scale-up and oxygen transfer rate in
microbial processes: An overview. Biotechnology Advances. 2009;
27:153-176.
75. Dubey KV. Adsorption-desorption process using wood based activated
carbon for recovery of bio surfactant from fermented distillery
wastewater. Biotechnology Progress. 2005; 21:860-867.
76. Berensmeier S, Franzrerb M, Heyd M, Kohnert A, Kirschhofer BG,
Nusser FM et al. Development and trends of bio surfactant analysis and
purification using rhamnolipids as an example. Annual Bio analysis
Chemistry. 2008; 39(1):579-159.
77. Reiling HE, Wyass UT, Guerra-Santos IH, Hirt R, Kappeli O, Fiechter
A. Pilot plant production of rhamnolipid bio surfactant by Pseudomonas
aeruginosa. Applied Environmental Microbiology. 1986; 51:985-989.
78. Kuyukina MS. Recovery of Rhodococcus bio surfactants using methyl
tertiary butyl ether extraction. Journal of Microbiology Methods. 2001;
46:149-156.
79. Nitschke M, Pastore GM. Cassava flour wastewater as a substrate for

Page | 32
 bio surfactant production. Applied Biochemistry and Biotechnology.
2003; 106:295-302.
80. Sen R, Swaminathan T. Response surface modelling and optimization to
elucidate the effects of inoculum age & size on surfactin production.
Biochemical Engineering Journal, 2004; 21:141-148.
81. Arun GB, Banat IM, Balu AC, Dhakrphalkar PK, Surekha KS. Methods
for investigating bio surfactants and bio emulsifiers: a review. Critical
Reviews in Biotechnology. 2010; 30(2), 1-18.
82. Priya T, Usharani G. Comparative Study for Bio surfactant Production
by using Bacillus subtilis and Pseudomonas aeruginosa. Botany
Research International. 2009; 12(4):284-287.
83. Banpurkar AG, Banat IM, Chopade BA, Dhakephalkar PK, Satpute SK.
Methods for investigating bio surfactants and bio emulsifiers: a review.
Critical Reviews in Biotechnology. 2010; 30(2):127-144.
84. Neu TR, Poralla K. Emulsifying agent from bacteria isolated during
screening for cells with hydrophobic surfaces. Applied Microbiology
Biotechnology. 1990; 32:521-525.
85. Cooper DG, MacDonald CR, Duff SJB, Kosaric N. Enhanced
production of surfactin from B. subtilis by continuous product removal
and metal cation additions. Applied Environmental Microbiology. 1981;
42:408-412.
86. Mattei GE, Rambeloarisoa E, Giusti G, Rontanu JF, Bertrand JC.
Fermentation procedure of a crude oil in continuous culture on sea-
water. Applied Microbiology Biotechnology. 1986; 23:302-304.
87. Kim HS, Lim JE, Lee OS, Lee DJ, Lee HT. Purification and
Characterization of bio surfactants from Nocardia sp. L-417.
Biotechnology and Applied Biochemistry. 2000; 31:244-253.
88. Bloomberg G. Designing proteins as emulsifiers. Lebensmittel
Technologies. 1991; 24:130-131.
89. Rockey J, Roller S, Shephord R, Sutherland IW. Novel bio emulsifier
from microorganisms for use in foods. Journal of Biotechnology. 1995;
40:207-217.
90. Kourkoutas Y, Banat IM. Bio surfactant production and application. In:
Pandey AP, editor, The Concise Encyclopaedia of Bio resource
Technology, Philadelphia, Haworth Reference Press, 2004, 505-515.

Page | 33
91. Banat IM, Makkar RS, Cameotra SS. Potential commercial applications
of microbial surfactants. Applied Microbiology and Biotechnology.
2000; 53:495-508.
92. Nitschke M, Costa SGVAO. Bio surfactants in food industry. Trends
Food Science and Technology. 2007; 18:252-259.
93. Marchant R, Banat IM. Bio surfactants: a sustainable replacement for
chemical surfactants? Biotechnology Letter. 2012; 34:1597-1605.
94. Kralova I, Sjeoblom J. Surfactants used in food industry: a review.
Journal of Dispersion Science and Technology. 2009; 30:1363-1383.
95. Van Haesendonck IPH, Vanzeveren ECA. Rhamnolipids in bakery
products. W.O. 2004/040984; International application patent (PCT),
Washington, 2004.
96. Kawaguchi K, Satomi K, Yokoyama M, Ishida Y. Antioxidative
properties of an extracellular polysaccharide produced by a bacterium
Klebsiella sp. isolated from river water. Nippon Suisan Gakkaishi. 1996;
42:243-251.
97. Bednarski W, Adamczak M, Tomasik J, Plaszczyk M. Application of oil
refinery waste in the biosynthesis of glycolipids by yeast. Bio resource
Technology. 2004; 95:15-18.
98. Deshpande M, Daniels L. Evaluation of sophorolipid bio surfactant
production by Candida bombicola using animal fat. Bio resource
Technology. 1995; 54:143-150.
99. Busscher HJ, Vander KBM, Vander MHC. Bio surfactants from ther
mophilic dairy streptococci and their potential role in the fouling control
of heat exchanger plates. Journal of Industrial Microbiology &
Biotechnology. 1996; 16(1):15-21.
100. Singh P, Cameotra SS. Potential applications of microbial surfactants in
biomedical sciences. Trends in Biotechnology. 2004; 22(3):142-146.

101. Urum K, Grigson S, Pekdemir T, McMenamy S. A comparison of the

efficiency of different surfactants for removal of crude oil from
contaminated soils. Chemosphere. 2006; 62(9):1403-1410.
102. Kuyukina MS, Ivshina IB, Gein SV, Baeva TA, Chereshnev VA. In
Vitro Immunomodulating Activity of Biosurfactant Glycolipid Complex
from Rhodococcus Ruber. Bulletin Experimental Biological Medicine.

Page | 34
 2007; 144(3):326-330.
103. Bai G, Brusseau ML, Miller RM. Bio surfactant-enhanced removal of
residual hydrocarbon from soil. Journal of Contaminant Hydrology.
1997; 25(1-2):157-170.
104. Hua Z, Chen J, Lun S, Wang X. Influence of bio surfactants produced
by Candida antarctica on surface properties of microorganism and
biodegradation of n-alkanes. Water Research. 2003; 37(17):4143-4150.
105. Wong JWC, Fang M, Zhao Z, Xing B. Effect of Surfactants on
Solubilisation and Degradation of Phenanthrene under Thermophilic
Conditions. Journal of Environmental Quality. 2003; 33(6):2015-2025.
106. Makkar RS, Rockne KJ. Comparison of synthetic surfactants and bio
surfactants in enhancing biodegradation of polycyclic aromatic
hydrocarbons. Environmental Toxicological Chemistry. 2003;
22(10):2280-2292.
107. Davis JB. The application of foaming for recovery of surfactin from B
subtilis ATCC21332. Enzyme Technology. 2001; 28:346-354.
108. Ron NSF. The Role of Microorganisms in the Recovery of Oil.
NSF/RANN 770201. U.S. Government Printing Office, Washington,
D.C, 1975.
109. Javajeri M, Jenneman GE, Knapp RM, Mcinnerney MJ. An aerobic
production of a bio surfactant by Bacillus licheniformis JF-2, Applied
and Enviornmental Microbiology. 1985; 50:698-700.
110. Clark JB, Feezo JM, Jenneman GE, Knapp RM, Menzle DE, Mclnerney
MJ et al. A halotolerant bio surfactant producing Bacillus sp potentially
useful for enhanced oil recovery. Development of Industrial
Microbiology. 1983; 24:484-492.
111. Carswell KS, Georgiou G, Lin SC, Sharma MM. Continuous production
of the lipopeptides bio surfactant of Bacillus licheniformis JF-2. Applied
Microbiology and Biotechnology. 1994; 41:281-285.

112. Banat IM, Perfumo A, Rancich I. Possibility and challenges for bio
surfactant uses in petroleum industry. Advances in Exploratory and
Medical Biology. 2010; 67(2):135-145.
113. Maggot M. Indigenous microbial communities in oil fields In Olivier B,
Maggot M. Edn, Petroleum microbiology Washington DC, ASM Press,

Page | 35
 2005, 21-33.
114. Caredda P, Franzetti A, La Colla P, Ruggeri C, Tamburim E. Surface
active compounds and their role in bacterial access to hydrocarbons in
Gordonia strains. Federation of European Microbiological Societies
Microbiology and Ecology. 2008; 63:238-248.
115. Miller RM, Zhang Y. Effect of rhamnolipid (bio surfactant) structure on
solubilisation and biodegradation of n-alkanes. Applied and
Environmental Microbiology. 1995; 61:2247-2251.
116. Kim J, Vipulanandan C. Removal of Lead from wastewater using a Bio
surfactant, University of Houston, Houston, Texas, 1998.
117. Olaniran AO, Balgobind A, Pillay B. Bioavailability of Heavy Metals in
Soil: Impact on Microbial Biodegradation of Organic Compounds and
Possible Improvement Strategies. International Journal of Molecular
Science. 2013; 14:10197-10228.
118. Santos D, Rufino R, Luna J. Bio surfactants: Multifunctional
Biomolecules of the 21st Century. International Journal of Molecular
Science. 2016; 17:401.

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 Chapter - 2
Molecular Aspects of Nutrition Molecular Gastronomy
& Related Techniques

Author
S. Sreeremya
Assistant Professor, Department of Biotechnology, Sree Narayana
Guru College, Coimbatore, Tamil Nadu, India

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Page | 38
 Chapter - 2
Molecular Aspects of Nutrition Molecular Gastronomy &
Related Techniques
S. Sreeremya

Abstract
Molecular gastronomy is the new direction of gastronomy mostly
carried out by idea of implementation of science in cooking. Many things
allied with this term are not quite clear and many have a wrong
understanding. This direction of gastronomy seeks innovation and
improvement of the existing situation, the basic aim of improving ways of
preparing meals, so that they have such taste as it should be in the optimal
case, every time. The idea of a practical molecular gastronomy in restaurants
and forming a sort of concoction of traditional and modern, artistic and
scientific technique to cooking is widespread throughout the world, but the
greatest concentration of such restaurants are located within the European
Union, where actually were created the first prototypes of such restaurants.
Spherification is another unique and novel technique which is mostly
implemented in European restraunts. Blenderized food is another technique
which is especially advised technique for patients, this is a simple and much
efficient technique where culinary practices are concerned.
Keywords: Molecular gastronomy, cooking, Science, Technique,
Blenderized food
Introduction
In the global scenario the obesity is one of the biggest problems of
modern man, a result of sedentary lifestyles and unbalanced diets imposed
by lifestyle. Standard restaurants' offer is based on the portions that typically
exceed nutritional requirements and the entry of such foods further
undermine the notion of a balanced diet. For these reasons there is a need for
rationalization and regular moderate intake of what is majorly needed.
Rationalization of nutrition is the key features of the new attitudes adopted
by molecular gastronomy, as well as the use of food as a whole (Adria et al.,
2006). The economic analysis of food deserts is comprised of four factors:
issues related to defining the relevant products, issues that seriously apply to

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consumers (the demand side), issues that mainly apply to food
Retailers (the supply side), and then the interactions of these factors (the
market).The issue of food desert can be sorted out by using the concoction of
science and technique in food (Sreeremya, 2017).
History
In 1988, a new scientific stream, molecular gastronomy, was defined as
‘looking for the mechanisms of phenomena occurring during dish
preparation and consumption’ (Adria et al., 2005). This new definition
delineates the opportunity to discuss the precise content of molecular
gastronomy and its connection with other existing fields of science. There
has always been much anarchy between science and technology when it
comes to food, including over exactly what food is(Ajinomoto,2008).
However, one has to recognize that human beings very seldom eat non-
transformed tissues or natural products; raw materials are transformed so that
chemical and physical changes determine the final elements of all food as
well as its ‘bioactivity’, a term which we propose to delineate the sensory
effects, nutritional value, eventual toxic effects, and so on, of the
Various compounds released by food systems. During food preparation,
plant or animal tissues are at least rinsed in water and cut, and most food are
thermally processed. For example, even for a simple carrot salad, which
requires no thermal processing, there is a big difference between the raw
product in the field and what is consumed - that is, grated carrots on a plate:
this is because cutting the tissue activates enzymatic reactions and because
compounds get transferred between the dressing and the plant tissue
(Barham, 2008). This analysis paves to the conclusion that reagents and
products of ‘culinary transformations’ (transformations performed in the
kitchen) should not both be called food. The specific transformation
occurring from the raw materials to the last prepared dish is worth assessing,
both for scientific and technological reasons. Making the difference between
science and technology is particularly important for molecular gastronomy
because of the confusion between science and cooking, and because the
public unduly fears ‘chemistry in the plate’ (Barham et al., 2010).
Five Major Goals of Molecular Gastronomy
Five goals of this new science: -
1) To collect and investigate old wives’ tales about cooking
2) To model and screen existing recipes
3) To introduce new tools, products, and methods to cooking

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 4) To invent new dishes availing knowledge from the previous three
aims
5) To use the appeal of food to promote science”.
Molecular gastronomy still creates uncertainty (Chaptal, 1890). The
chimerical food creations we witness today-fruit caviar, hot ice cream, and
foam sauces-are creative majorly kitchen applications of the science. The
many names given to capture this kitchen approach, ranging from molecular
gastronomy to culinary amalgamation, fundamentally refer to the same idea:
Applications of Food Science
While food science is part of molecular gastronomy, its main application
has been to cater safe and nutritious food for the masses in the most efficient
and economical manner possible. Food science is “the discipline in which
biology, physical sciences, and engineering are availed to assess the nature
of foods, the causes of their deterioration, and the principles underlying food
processing”. The application of food science-food technology-is defined as
“the application of food science to the selection, preservation, processing,
packaging, distribution, and use of safe, nutritious, and wholesome food”.
Neither the science nor the technology is progressed toward the study of
restaurant cooking processes or the creation of new dishes in the kitchen.
Molecular gastronomy has helped bring the tools and technology that are
commonplace in the food industry to the restaurant kitchen (Jacobs et al.,
2015).
For example, the technique of spherification, where round “pearls” of
gel with liquid centers (“caviar- Caviar is a scrumptious delicacy consisting
of salt-cured roe (Roe or hard roe is the fully ripe internal egg masses in the
ovaries, or the oozed external egg masses of fish and certain marine animals,
such as shrimp, scallop and sea urchins) of the Acipenseridae family. The
roe can be "fresh" (non-pasteurized) or pasteurized, with pasteurization
decreasing its culinary and economic value. Traditionally, the word caviar
refers only to roe from wild sturgeon in the Caspian Sea and Black Sea) are
synthesized by dropping a flavorful base mixed with sodium alginate into a
calcium chloride solution. Upon contact with the calcium ions, gelation
occurs from the outside in (Phan et al., 2008). The longer the pearls stays in
the calcium solution, the firmer and less liquid the centers become. While
this is a fairly new application in the kitchen, alginates are well established
as an ingredient in food manufacturing. The thickening and gelling
properties of alginate have long been availed in sauces and re-formed
products such as restructured fish fillets, onion rings, and herb-flavored
alginate gels used to stuff olives(Eunice et al.,1989).

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 Maltodextrin, a product of partial starch hydrolysis, is another crossover
from food science. An element availed to transform oil to powder in the
molecular kitchen, it is a staple ingredient in food processing availed to
disperse dry ingredients, stabilize high-fat ingredients, and a source of car-
bohydrate in high-energy drinks (Valverde, 2008).
Preparation and Cooking Techniques
Foams and airs, forms of foods that are certainly not new, were made
fashionable in fine dining. Lecithin is a staple element used to emulsify
sauces like mayonnaise and vinaigrettes. Soy lecithin, in particular, avoids
concerns of cholesterol when compared to the traditional use of egg yolk for
emulsifying power. The emulsifying characteristics of lecithin also make it a
popular ingredient to create airs and foams.
Molecular Gastronomy: FACT (Von Elbe et al., 2008)
Molecular gastronomy is therefore situated on the interface between
science and application.
The Scientific Aim is
 To research on recipes, cooking habits and cooking wisdom;
 To describe the chemical and physical processes that take place
during cooking.
Other Related Techniques
Spherification
Spherification is the phenomena of encapsulating a liquid into a sphere
that will rise in the mouth. This is achieved when a film of gel membrane is
formed around a liquid. A reaction that occurs between sodium alginate and
a calcium salt such as calcium lactate. Spheres can vary in size ranging from
small caviar beads to much larger ravioles. Carefully follow each recipe as
there are various spherification methods.
Sodium Alginate Dissolution
 To avoid clumping, the use of a hand blender is preferred as some
egg beaters are not efficient enough to properly dissolve the sodium
alginate.
 When not in contact with calcium, sodium alginate acts more like a
thickening agent, so when blended a lot of air will be trapped in the
solution. Allow the solution to rest so that the air bubbles escape
from the alginate bath.

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Caviar Serving
 The spherification process availed to create caviar will not stop
after the pearls are rinsed. Membranes will get thicker and thicker
progressively until the beads are completely congealed. Therefore,
it is best to serve immediately to ensure that the interior of the
caviar remains liquid.
There is a rapid increase in the population which had resulted in
widening the horizons of various sectors, the major field which have prolific
focus is the food industry. Basically due to the nature of food ingredients,
these products contain bacteria, even when hospital kitchens follow clean
preparation procedures (Horst et al., 2007). When kept at room temperature
for feeding contamination by bacterial overgrowth is likely to occur.
Blenderized feeds must be delivered to the stomach efficiently, where the
acidic terrain helps limit, but not eliminate, bacterial growth. Contamination
can also result in patient infections that slow recovery and cause longer
hospital stays and prolifically higher costs of care (Carratu et al., 2005).
Importance of Blenderized Food in Diet
The blenderized liquid diet is often used as an intriguing step between a
full liquid diet and a regular diet. This diet comprises of fluids and foods
blenderized to a liquid form. It may be appropriate for individuals who have
chewing or swallowing problems. This diet must only be followed at the
recommendation of the physician (Henriques et al., 1999). After the
observed blenderized diet by the physician (Araújo et al., 2006). Physician
after assessing the result will suggest to return to your regular diet. If an
individual wishes follow regular blenderized diet after two week, physician
will suggest certain energy boosting supplements to be added (Von Atzingen
et al., 2007).
Blending Tips
1) To prepare a blenderized food a food processor will work better
than a blender. Food may blenderize much easily when cut into
small pieces before placing in a blender or food processor. Meats
blenderize more easily when they are warm.
2) Mixing equal portion of solids and liquids can liquefy most foods.
Fruits and vegetables do not need as much liquid since they are
naturally high in water.
3) Strain the blenderized food if it comprises lumps or chunks.
4) Blenderized foods may be stored up to 48 hours in the refrigerator.

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 5) Use of gravy (extracts), vegetable juices, cream soups, cheese or
tomato sauce, milk and juice rather than water is recommended
when blenderizing food.
Protocol to Blenderize Food: (Felício et al., 2012)
 Food should already be cooked or canned foods may be availed
 Cut solids into small cubes before adding in blender
 Blend for a second or two (pulse)
 Add a small portion of liquid and pulse several times
 Add more liquid and blend to get desired thickness
 Excellent results are obtained when you only blend small amounts
of food at a time (e.g., 1 cup/ 250 mL servings)
 Safety Measures:
 Wait until blender blades have completely stopped before removing
the lid
 After you blend a food, either serve it or refrigerate it
 Foods Storage Measures:
 Do not store blended food for longer than 3 days in the fridge
 Freeze in small portions for a quick meal. One can place
blenderized food into ice cube trays, freeze, then store in freezer
bags for convenient use
 Blended foods can be stored in the freezer for up to 1.5-3 months
 Thaw frozen food in the refrigerator (rather than on the counter)
Blenderized Tube Feeding Formulas
Preparation consisted of three stages: stage I: the raw foods (rice,
carrots, and ground beef) were cooked together in the same wok. The rice,
carrot, and ground beef proportions were calculated availing the cooking
factors, which were 2.5, 0.8, and 0.7, respectively. Beans cooked in a
pressure cooker for 30-32 minutes were added to the mixture. These foods
were placed in a pot containing 150mL of boiling water and cooked in low
heat (using the small burner of standard stoves) for approximately 15-20
minutes (Zaban et al., 2009). Stage II: once the raw foods were cooked, they
were added in a blender with half of the milk volume and the other
ingredients, and blended for three minutes at moderate speed (Lima et al.,
2015). The remainder of the milk was added, and the mixture was blended
again for another three minutes at medium speed. Stage III: the ready-to-eat

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formula was sieved three times through a fine mesh strainer, placed in a
sterile plastic container aseptically, and stored in the refrigerator (Sousa et
al., 2014).
The study blenderized enteral formula was mainly assessed according to
the Dietary Reference Intake (DRI) for men aged more than 51 years. To
assess the stability and homogeneity, the samples were added in a glass
beaker, refrigerated for one day, and inspected visually. Flow rate was
mainly assessed by counting how long it took for 200mL of the formula to
drip through the enteral feeding extension set (Medina et al., 2008). The
formula was stored in a specific 300mL plastic bag, which was kept on a
hook one meter above from the drip chamber. Drip time was mainly
measured on two occasions, at Time 0 (T0), immediately after formula
preparation, and at Time 3 (T3), after three hours of refrigeration at 4°C. The
formula was removed from the refrigerator and heated in a water bath for
five minutes, which is enough time for it to reach room temperature (25ºC).
Osmolality of the blenderized food products were determined by a
cryoscopic osmometer (Advanced Digmatic 3D2, Diversified Equipment
Company, Inc., Lorton, Virginia, United States), and pH was measured by a
digital pH meter (pH Meter Tec - 2 Tecnal, Piracicaba, São Paulo, Brazil),
properly calibrated. The major parameters analysed included were ash,
moisture, energy, proteins, lipids, crude fiber, vitamin C, calcium, iron,
magnesium, zinc, and phosphorus (Bréton, 2002). Carbohydrate content was
determined indirectly. All these tests were repeated three times.
The study nutrients were assessed by the following methods: protein by
the Kjeldhal method, availing the Jones factor and respecting ingredient
ratios to convert total nitrogen into crude protein; lipids by the Bligh-Dyer
method; crude fiber by the acid digestion method; energy by bomb
calorimetry; minerals by the inductively coupled plasma mass spectrometer
ELAN DRCII (Perkin Elmer SCIEX, Toronto, Ontario, Canada); vitamin C
by spectrophotometry using a wavelength of 510nm.
The Application-Oriented Aim is
 To use the knowledge about the physical and chemical processes of
cooking in order to develop novel cooking instruments and
ingredients,
 To design and invent typically new dishes with the help of the
acquired knowledge about food and cooking processes. In this way,
the cook no longer innovates by trial and error but on the basis of
scientific knowledge.

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 The last point concerns molecular gastronomical dishes. These are
dishes that are innovated by applying knowledge about molecular
gastronomy, thus providing us with a refreshed outlook on eating and
gastronomy (Jacobs et al., 2015).
Conclusion
Molecular Gastronomy encompasses modelling recipes, testing old
wives tales, inventing new dishes and new tools, methods and ingredients in
the kitchen. As any intellectual enterprise is improved when clearly
delineated, technological applications were finally excluded from itself and a
more precise. This paper gives a detailed information about several
techniques used in food and cooking with the influence of molecular
gastronomy.
References
1. Adria F, Blumenthal H, Keller T, McGee H. Statement on the ‘new
cookery. The Guardian. Dec. 10, 2006.
2. Adria F, Soler J, Adria A. El Bulli: “1998-2002”. Ecco Press, 2005.
3. Ajinomoto. Activa TG Transglutaminase. Www.activatg.com. Accessed
May 6, 2008.
4. Araújo EM, Menezes HC. Formulações com ali-mentos convencionais
para nutrição enteral ou oral. Ciênc Tecnol Aliment. 2006; 26(3):533-8.
5. Barham P. Molecular Gastronomy. Discovery Channel.
Www.discoverychannel.co.uk. Accessed April 28, 2008.
6. Barham Peter, Leif Skibsted H, Wender LP. Bredie, Michael Bom Frøst,
Per Møller, Jens Risbo, Pia Snitkjær, and Louise Mørch Mortensen.
Molecular Gastronomy: A New Emerging Scientific Discipline.
Chemical Reviews. 2010; 110(4):2313-2365.
7. Bréton JO, Ruesca PB, Laborda SG, Nogueras EF. Evaluación deun
programa de nutrición enteral domiciliaria. Endocrinol Nutr. 2002;
49(6):179-84.
8. Carratu E, Sanzini E. Sostanze biologicamente attive presenti negli
alimenti di origine vegetale. Ann Ist Super Sanitá. 2005; 41(1):7-16.
9. Chaptal JA. Essai sur le perfectionnement des arts chimiques en France;
Imprimerie nationale: Paris, 1890.
10. Eunice LC, Shuryo Crit N. Rev. Poultry Biol. 1989; 2:21-58.

Page | 46
11. Felício BA, Pinto ROM, Pinto NAVD, Silva FD. Food and nutritional
safety of hospitalized patients under treatment with enteral nutrition
therapy in the Jequitinhonha Valley, Brazil. Nutr Hosp. 2012;
27(6):2122-9.
12. Henriques GS, Rosado GP. Formulação de dietas enterais artesanais e
determinação da osmola-lidade pelo método crioscópico. Rev Nutr.
1999; 12(3):225-32.
13. Horst MA, Lajolo E. Biodisponibilidade de com-postos bioativos de
alimentos. In: Cozzolino SMF. Biodisponibilidade de nutrientes. 2ª ed.
São Paulo: Manole, 2007, 697-735.
14. Jacobs J, Zoran A. Hybrid Practice in the Kalahari: Design
Collaboration through Digital Tools and Hunter Gatherer Craft. The
33th international conference on Human factors in computing systems
(CHI '15). ACM, New York, NY, USA, 2015.
15. Lima VS, Souza FCA, Aguiar JPL, Yuyama LKO. Composição
nutricional de dieta enteral artesanal a partir de alimentos convencionais
do Município de Coari, Estado do Amazonas, Brasil. Rev Pan-Amaz
Saúde. 2015; 6(2):29-36.
16. Medina JM, Nascimento GGF, Oliveira MRM, Mar-ques MR.
Contaminação microbiológica de dietas enterais. Rev Bras Nutr Clin.
2008; 23(4):262-9.
17. Phan VA, Liao YC, Antille N, Sagalowicz L, Robert F, Godinot N.
Delayed volatile compound release properties of self-assembly
structures in emulsions. J. Agric. Food Chem. 2008; 56(3):1072-1077.
18. Sousa LRM, Ferreira SLR, Schieferdecker MEM. Physicochemical and
nutritional characteristics of handmade enteral diets. Nutr Hosp. 2014;
29(3):568-74.
19. Valverde J, This H. 1H NMR quantitative determination of
photosynthetic pigments from green beans (Phaseolus vulgaris L.). J.
Agric. Food Chem. 2008; 56(2):314-320.
20. Von Atzingen MC, Silva MEMP. Características físico químicas de
dietas enterais artesanais com hidrolisado protéico de carne. Alim Nutr.
2007; 18(2):183-9.
21. Von Elbe JH, Huang AS, Attoe EL, Nank WKJ. Agric. Food Chem.
1986; 34:51-54.

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22. Zaban ALRS, Novaes MRCG. Demographic, epidemiological and
nutritional profile of elders in home enteral nutritional therapy in
Distrito Federal, Brazil. Invest Clin. 2009; 50(3):347-57.

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 Chapter - 3
Sensory and Food Quality

Authors
Kamaljit Kaur
Department of Food Science and Technology, Punjab Agricultural
University, Ludhiana, Punjab, India
Gursharan Kaur
PG Department of Food Technology, Khalsa College, Amritsar, Punjab,
India

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Page | 50
 Chapter - 3
Sensory and Food Quality
Kamaljit Kaur and Gursharan Kaur

Abstract
Food quality and sensory analysis are important concepts. The selection
of food for consumption depends largely on food quality and sensory
analysis is performed to determine consumer preference/acceptability. In this
chapter food quality concept, attribute, sensory analysis procedures and
artificial senses are discussed. Both sensory and objective testing are
important in evaluating food quality.
Key words: Quality, Sensory analysis, Color, Flavor, Texture
Introduction
Many years ago, about 2500 years BC Mosaic and Egyptian laws had
provisions to prevent the contamination of meat. More than 2000 years ago,
India already had regulations prohibiting the adulteration of grains and
edible fats. The Roman government also provided state control over food
supplies to protect consumers against bad quality and fraud. Adamson [1]
reported that, when food was scarce it resulting in expected increase in
demand which made prevalence of fraudulent practices. The initiation of a
system of scientific management of quality can be traced back to the
teachings of people such as Frederick Taylor, who introduced the scientific
method when evaluating workplace processes involved in quality and
production. He advocated that each task should be standardized, workers
must be trained and job functions divided to maximize efficiency [2]. Walter
She wart is credited with the introduction of the control chart in 1924, that
can be used to monitor the variation of any process or quality factor [3]. The
control chart is instrumental in understanding the variability of a process,
when something is “out of control” or irregular and for optimizing a process
to improve economic efficiency and product quality.
Food quality may be defined as the “degree of excellence”, “customer
satisfaction”, “doing things right the first time” or “zero defects”. Quality of
food depends upon taste, appearance, nutritional quality as well as in

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bacteriological or keeping quality. Sensory analysis plays major role in
defining food quality. Sensory science is a multidisciplinary area comprising
measurement, interpretation and understanding of human responses to
product properties as perceived by the senses such as sight, smell, taste,
touch and hearing [4].
The quality of a given commodity is a combination of attributes that
contribute its acceptability. Acceptability in turn depends upon a set of
consumer expectations parameters for selecting a product. The choice of a
food product depends upon characteristics that differentiate it from others.
The definitions of quality are mentioned in Table 1. Food items are selected
on the basis of a person’s total assessment of its desirable sensory attributes.
Table 1: Some definitions of quality

Quality concepts Reference

Composite of characteristics/attributes which differs from
Kramer & Twig, 1983 [5]
unit to unit
Totality of features/characters of a product that bear on its
Amerine et al., 1965 [6]
ability to satisfy a given need
Uniformity, consistency and conformity to a given
Kramer & Twig, 1982 [7]
standard or specifications
A statement of what the user wants and what the
Gatchalian, 1989 [8]
manufacturers can provide
Fitness for use Juran, 1974 [9]
Quality control is doing things right the first time and
Harrington, 1986 [10]
every time

Aspects of Food Quality

Food quality has both subjective and objective aspects. Subjective
evaluation of food is done by human senses, which may cause drift and there
could be personal bias. Whereas, objective evaluation is evaluation of food
by using instruments and it includes physical, chemical and microbiological
aspects. Subjective measurement of food may include evaluation of food that
may utilize one or more of the different tests. Sensory evaluation can be seen
as a link between research and development, with a focus made on technical
aspects of food, consumer and marketing research on consumers behavior
and psychology. Objective test measures one particular attribute of a food
rather than overall quality of the product. Certain principle must be
emphasized when considering objective tests to evaluate the quality of food
products [4]:
 Objective evaluation of food involves instrumentation and use of

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 physical and chemical techniques instead of variable sensory organs
to evaluate food quality.
 The objective tests must be appropriate for the food product to be
tested. i.e. it must measure an attribute of the food that has a major
effect on quality.
 The objective test results should be correlated with sensory testing
of similar product to check the reliability index of quality of food.
 Objective tests include all types of instrumental analysis which
cover chemical, nutritional and microbiological properties of food.
 If the instruments are calibrated well, the results will be the absolute
measurement of a particular property of food.
 Objective testing of food must be correlated with sensory testing,
because no single test can measure overall acceptability of a
specific food.
 Objective tests are repeatable and are not subject to human
variation. If the equipment is properly maintained and used
correctly, it should give reliable results.
Quality Systems
A quality system is a mechanism that coordinates and maintains the
activities needed to ensure that the characteristics of products, processes or
services are within certain limits. A quality system involves every part of an
organization that directly or indirectly affects these activities. The quality
system is documented in a quality manual with associated documents that
specify procedures and standards.
There are three basic components of a quality system. These are quality
management, quality control and quality assurance.
Quality Management: Quality managements is the means of
implementing and carrying out quality policy. In quality management goal
planning, quality control and quality policies are performed. Responsibility
of quality management is to see that all quality goals and objectives are
implemented and that corrective actions have been achieved. Quality system
is periodically reviewed to ensure effectiveness and to identify and review
any deficiencies.
Quality Control: The term quality control can simply be defined as the
maintenance of specified finished product characteristics every time it is
manufactured. It encompasses all techniques and activities of an organization
that continuously monitor and improve the conformance of products,

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processes or services to specifications. It may also include the review of
processes and specifications and make recommendations for their
improvement. Quality control eliminates causes of unsatisfactory
performance by identifying and helping to eliminate the source of variation.
The objectives of a quality control program are to provide a system where
product meets design requirements and give feedback for corrective actions
and process improvement.
The basic tools of quality control are:
 Ingredient specifications
 Approved supplier list
 Product formulas
 Product standards (specifications)
 Manufacturing procedures
 Critical control point identification/sampling program
 In-process analysis, records and reporting packaging specifications
 Label specifications
 Cleaning and sanitizing program
 Good manufacturing practices (GMP) requirements
 Recall program
 Warehousing, shipping and receiving program
 Laboratory analysis
Quality Assurance: The term quality assurance describes all the
planned and systematic actions necessary to assure that a product or service
will satisfy the specified requirements. Quality assurance is a way of
preventing mistakes or defects in manufactured products and provides
confidence that quality requirements are fulfilled. The difference between
quality control and quality assurance is that “quality control has to do with
making quality what it should be and quality assurance has to do with
making sure quality is what it should be”. Table 2 summarizes the difference
between quality assurance and quality control.
Table 2: Difference between quality assurance and quality control

Quality Assurance Quality Control

Planned & systematic activities implemented in Activities focused on finding defects
quality system. So, that quality requirement for in specific deliverables.

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 a product or service will be fulfilled.
It is used to verify that the products
In general, it does not concern with the specific
are of acceptable quality that they
requirements of the product being developed.
are complete and correct.
It is process oriented It is product oriented
It makes sure the result of right things, in the It makes sure the result of what you
right way. have done & what you expected.
Oriented towards prevention Oriented towards detection.
Its goal is to improve test procedures and Its goal is to identify defect after a
processes so that defects do not occur when the product is developed and before it is
product is being developed. released.
It include process checklist, project audits and It includes inspection, deliverable
standard development peer review and the testing process.

Major Quality Attributes

Right from the initial stages of production to the time till the produce
reaches the consumer, the farmer has to combat many unfavourable
circumstances. Among these are pests, micro-organisms which infest the
farmland, foreign matter which may be dangerous or otherwise, poisonous
substances or impurities which get into products from materials used in
processing, micro-organisms and dirt introduced into the product through
unhygienic practices of the people who handle the produce, as well as loss of
quality that results from short-comings in storage practices. The quantified
measurement of quality characteristics is essential for the development of a
quality control programme.
These characteristics can be classified into two groups:
1) Physical attributes
2) Hidden attributes
1) Physical Attributes: Man accepts food products on the basis of
certain characteristics, which he defines and perceives with his
senses. These attributes are described in terms of sensations and are
sometimes referred to as quantitative or sensory qualities. They
include perceptions of appearance factors such as color, size, shape
and physical defects, kinesthetic factors like texture, viscosity,
consistency, finger feel, mouth feel and flavor factors or sensations
combining odour and taste.
2) Hidden Attributes: Unlike the physical characteristics, the hidden
attributes of food can neither be seen nor felt and are measurable
only by standard chemical or microbiological procedures. Some of

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 these attributes such as food items' nutritive content are positive and
need to be maintained. Other attributes are negative and if present
render food unsafe or unfit for human consumption. Figure 1
depicts the types of quality characteristics of foods.

Characteristics of
quality

Sensory Hidden Quantitative

Characteristics Characteristics

Fig 1: Different types of quality characteristics of foods

Methods for Determining Quality

Both sensory evaluation and objective evaluation of food quality are
essential in food industry in order to routinely monitor food quality and to
ensure that the foods being produced are acceptable to the customer. These
two methods of evaluation are complementary to each other.
Sensory testing is expensive and time consuming, because panellists are
required to test a single product in order for the results to be meaningful. On
the other hand, objective testing if efficient and, but after the initial purchase
of the needed equipment, also inexpensive. One can usually perform an
objective test on many samples in a day, whereas it may take a day to
perform a complete sensory test on line or two sample. Objective samples
give repeatable results, whereas sensory test may give variable results due to
variation of human responses and opinions.
Sensory evaluation gives a judgement of the overall acceptability of a
product, and objective method evaluate one aspect of food, and this may not
always be sufficient to determine whether the quality of product is
acceptable.
The true judge of the acceptability of a food product is a consumer.
Therefore, objective test must correlate with sensory tests to give a reliable
index of food quality.
Objective tests are essential for routine quality control of food products,
however, sensory evaluation of product is requirement of research and

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development. Only consumers can tell whether there is a perceivable
difference in a product when the formulation or packaging is changed, and
only consumers can determine whether a new product is acceptable or
preferred over another brand.
Broadly two Methods are used for Determination of Quality in Food
Industry as Shown Below
Subjective Method: In this method, individual is required to give his
opinion about qualitative/quantitative values. This method is also referred as
sensory method. It is by experience of the individual. Different subjective
methods are used for estimation.
Objective Methods: These are based on recognized standard scientific
tests to any sample of food. They represent the modern idea in quality
control because human element is excluded. Objective method is divided
into three groups:
a) Physical methods
b) Chemical methods
c) Microscopic methods
Physical Method: These are quickest methods and are used to measure
size, colour consistency, etc. Table 3 summarizes some physical methods to
check food quality.
Table 3: Physical methods to check quality of food products
Physical
Test Working
Factor
Measures differences in tri-stimulus
Colour difference meter
values Based on colour standards
Colour Munsell colour system
Measure light reflectance at different
Spectrophotometry
wavelengths
Flow through a capillary tube
A rotating cylinder is immerged in the
Ostwald Viscometer
fluid and stress measured.
Viscosity Rotating Spindle Falling
Measures the time required for a weight
weight
to fall through a tube containing the
sample.
Finger feel Mouth feel Test of firmness and softness
Texture value (texture Test of chewiness, fibrousness and
Texure
meter, Penetrometer) grittiness Indication of texture, firmness,
and Shear press tenderness and shear value.
Length, breath and
Shape and
diameter (Vernier Uniformity and classification.
Size
Callipers)

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Colour
Colour is one of the most widely measured product quality attributes in
postharvest handling and in the food processing research and industry. Apart
from differences in instrumentation, colour measurements are often based on
different colour indices even for the same product. Appearance is one of the
most important sensory quality attributes of fresh and processed food
products and their marketing [11, 12]. It is an all-inclusive term involving
size, shape, texture, mass, gloss, colour and others. The colour of food
surface is the first quality parameter evaluated by consumers, and it is
critical to product acceptance. Food appearance determined mostly by
surface colour is first sensation that the consumer perceives and uses as a
tool to either accept or reject food [13]. Visual appearance of the food
manifested as its colour has a strong influence on a consumer’s opinion
about the food quality [14, 15].
Gloss is a visual aspect of quality that depends on the ability of a surface
to reflect light [16]. Gloss on the outside of the whole fruit tends to be a
desirable attribute for whole fruits. Products that are freshly harvested often
have a bright, glossy surface, and this appearance factor can be greatly
reduced with weight loss and other postharvest handling conditions. Fresh
cut fruits and vegetables must appear to be fresh, generally indicated by the
brightness of colour and the absence of visual defects or drip. Sheen on the
outside of most cut fruits is preferred to a dried appearance. Colour and
appearance of the package can also influence the purchase decision.
Colour Systems (Colour Spaces) can be described by several colour
coordinate systems. Some of the most popular systems are RGB (red, green
and blue), which is used in colour video monitors; Hunter L a b,
Commission Internationale de l’Eclairage’s (CIE) L*a*b*, CIE XYZ, CIE
L*u*v*, CIE Yxy, and CIE LCH. These differ in the symmetry of the colour
space and in the coordinate system used to define points within that space.
According to CIE concepts, the human eye has three colour receptors- red,
green and blue and all colours are combinations of those. The amounts of
red, green and blue needed to form any particular colour are called the
tristimulus values and are denoted X, Y and Z, respectively. The most
commonly used notations are the CIE XYZ colour space devised in 1931 by
the International Commission on Illumination. The system is based on the
trichromatic principle, but instead of using real red, green and blue primaries
with their necessity for negative matching, it uses imaginary positive
primaries, X, Y and Z. It uses the chromaticity diagram to designate various
colours (Fig. 3). Primary Y, known as luminous reflectance or transmittance,

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contains the entire lightness stimulus. The application of the weighting to a
reflectance curve gives the tristimulus values, which are denoted by the
capital letters X, and Z. These values are then used to calculate the
chromaticity coordinates, designated by lowercase letters x (red), y (green)
and z (blue). The Hunter L a b developed in 1948 for photoelectric
measurement and the CIE L*a*b* colour space (Fig. 3) devised in 1976
provide more uniform colour differences in relation to human perception of
differences.

Fig 3: CIELAB colour space [17]

Viscosity
Viscosity is a direct measurement of a fluid’s quality. It is important to
be measured because a change in viscosity can indicate a fundamental
change in the material under test. Viscosity is often an extraordinarily
sensitive measurement of another property such as solids content and crystal
concentration. The viscosity characteristics of food products influence many
aspects of the fluid performance during processing (pumpability, droplet
breakup in spray drying, emulsion formation, flow into molds and
formability) and the quality of liquid products (texture, flavour release,
stability and appearance). Viscosity correlates with composition and
molecular distribution, and hence the characteristics of the material. By
determining viscosity, it is possible to quantify changes in flowing
compositions with respect to time. By monitoring the changes over time, one
can adjust the composition of the flowing media in order to achieve optimum
flowing and product characteristics. Viscosity describes a fluid’s internal
resistance to flow and may be thought as a measure of fluid friction. All real

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fluids (except super fluids) have some resistance to stress, but a fluid which
has no resistance to shear stress is known as an ideal fluid or viscid fluid.
The various types of viscometers can be classified according to the principle
on which they work. The most commonly used viscosity measurement
devices are capillary flow viscometers, orifice type viscometers, falling ball
viscometers and rotational viscometers.
Texture
Texture can be described as the properties of a foodstuff apprehended
both by the eyes, by the skin and muscle senses in the mouth, embracing
roughness, smoothness, graininess, and so forth. Texture deals with the sense
of feel with fingers and more precisely with mouth.
Classification of Textural Properties
Finger Feel
Firmness: Firmness is encountered by consumer while selecting a firm
apple or any other fruit or vegetable. It is measured physically by
compression forces.
Softness: It is also measured physically by compression forces e.g.,
peaches, plums etc.
Juiciness: In mature sweet corn juiciness is judged with the help of
thumb nail to test the ease and the amount of juice squeezed out of it. It is
measured physically by puncturing the fruit.
Mouth Feel
Chewiness: it is energy required to masticate the solid food to a state
ready for swallowing. It is sensed by resistance of the product to the
compression and shearing action of teeth.
Fibreness: it is sensed by the presence of an inedible residue remaining
in the mouth after mastication as well as resistance to cutting force of teeth.
Grittiness: it is sensed by presence of small grit particles which can be
stones or sands.
Mealiness: it is served by the coating of starch or other material with
adhesive properties over mouth tissues.
Stickiness: it is the property in which mouth feels the adhesiveness of
chewing gum.
Oiliness: it is sensed by mouth caused by oily product.

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Principles Involved in Measurement of Texture
All the sensation of texture is based on principle of resistance to force.
The force is applied in number of ways through the combination of two or
more methods/forces.
Compression: this refers to the squeezing together of test material so
that it still remain as a single individual unit but may occupy less volume.
The pressure tester falls under this category.
Shearing: it results from the application of force where test material is
separated into two or more parts with one part sliding beyond the other part.
Cutting: occurs when force is applied in such a way that the test
material is divided into two or more parts. So, that the two portions remain in
their original form in relation to each other.
Tensile Strength: it is application of force away from material rather
than towards the material i.e. when force is applied to pull the test material
apart from each other and this test is mostly done in textile industry. In food
industry it is used to check toughness of strings in the string beans.
The most important components of food texture are indicated in Table 4.
Table 4: Relation of kinesthetic characteristics or physical methods of application of
force

Sensory Relation Physical test Instrument or Procedure for measurement

Firmness Compression Pressuretester, Shear press
Softness Compression Pressuretester, Shear press
Puncture tester, Succlometer, Shear tester,
Juiciness Compression
Moisture tester
Tendrometer, Texturometer,Shearpress,
Chewiness Shear press
Specific gravity, Solids
Cutting,
Fibrousness Fibrometer, Shearpress, Fiber analysis
communiting
Grittiness Combination Sedimentation starch or germ analysis
Mealiness Tensile Strength Jelly strength, Pectin, Gum analysis
Stickiness Tensile Strength Jelly strength, Pectin, Gum analysis

Flavour
Flavour is conventionally regarded as a combination of sensations
derived from several distinct types of chemical stimuli. Tastes, detected by
receptors on the tongue and other oral surfaces, are in volatile chemical
stimuli that are carried in solution by saliva from the food to the receptors. It

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is now widely accepted that there are five basic tastes – sweet, salt, bitter,
acid, savoury (umami) – although this list is sometimes extended to include
other sensations. Odours (aromas) are volatile chemical stimuli detected by
receptors located in the olfactory epithelium in the nasal cavity. These are
transmitted to the receptors directly through sniffing (ortho nasal route), or
from the mouth during eating (retro nasal route). The odour response is
complex, with around 2500 odorous chemicals found in food [18]. A third
component of flavour, a chemical sense that stimulates the trigeminal nerves
is responsible for sensations such as burning and cooling. Trigeminal
sensations can arise from both chemicals in dissolved saliva, for example the
tingling sensation from carbonic acid in fizzy drinks, and from volatile
chemicals- pungent thiocyanates in mustard and horseradish.
Measurement of flavour components is influenced by the widely
differing volatilities of flavour-active chemicals. The relatively large number
of volatile chemicals contributing to flavour has been reflected in the wide
range of instrumental methods that are now commonly used for volatile
analysis, in particular gas chromatography/mass spectrometry systems [19].
More recently, considerable publicity has been given to the development of
so-called ‘electronic noses’, which are more correctly volatile sensors
operating on a pattern recognition basis. Although these are finding
numerous uses in other fields, relatively few routine uses have been recorded
within the food industry [20]. Measurement of taste chemicals has relied
predominantly on traditional methods of chemical analysis for salty and
acidic stimuli, with high-pressure liquid chromatography being used for less
volatile chemicals such as sugars.
Chemical Methods
These are standard food analysis methods and are used for quantitative
analysis of nutritive value like moisture, fat, protein, carbohydrates, fibre,
vitamins etc. Table 5 summarizes common chemical methods to check food
quality.
Table 5: Common chemical methods used for food products

Chemical factor Method Description

Moisture Drying Measure weight loss due to evaporation
Solids Hydrometer Concentration of dissolved solids
Total Soluble
Refractive index Measurement of TSS
Solids
Ascorbic Acid Dye method Measures Vitamin C content
Protein Kjeldahl method Total nitrogen determination

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 Carbohydrates Molisch general test Colour reaction with Napthol
Measures organic residues including
Fibre Acid-alkali extraction
cellulose and lignin
Burns at 550oC in Determines total ash by weight of
Ash/Minerals
Muffle Furnance residue after incineration
Dried sample extracted with
Fat Ether extraction
hexane/petroleum ether
Bioassys for each Vitamin analysis by analytical
Vitamins
vitamin measurement
pH and acidity pH meter and titration Measures alkalinity or acidity of samples
Chemical reaction with H2O2 or
Enzymes Catalase, peroxidise
indicators

Sensory Quality
Sensory quality is a scientific discipline used to evoke, measure, analyze
and interpret reactions to the characteristics of foods that are perceived
through the five senses of sight, smell, touch, taste and hearing.
As the definition depicts sensory evaluation implies the measurement
and evaluation of sensory properties of foods and other materials. Sensory
evaluation involves the analysis and interpretation of the responses by the
sensory professionals; i.e. the individual who provides the connection
between the internal world of product design technology and the external
world of market-place. This connection is essential so that the processing
and development specialists can anticipate the impact of product changes in
the market-place. Objectives of sensory evaluation are defined in the terms
of what sensory evaluation does and what it is capable of doing, as
mentioned below [21]:
 Provide useful and timely information and recommendations about
the sensory properties of product as requested.
 Provide quantitative information about the sensory characteristics of
all the food industries and competitive products.
 Maintain a pool of individuals qualified to participate in a wide
range of tests.
 Develop the methods that are for general use and are unique to
specific products.
 Develop procedures to relate sensory parameters with analytical
information for use in product research and quality.
 Maintain awareness of new developments in product evaluation and
their application to food industry.

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  Ensure that no product of the company fails because of a sensory
deficiency.
Sensory evaluation utilizes one or more of the five senses to evaluate
foods. Sensory evaluation is the study of reactions of the five senses: sight,
smell, hearing, taste and touch. Taste panels comprising groups of people
taste specific food samples under controlled conditions and evaluate them in
different ways depending upon the particular sensory test being conducted.
Sensory methods are used to determine:
 Whether foods differ in taste, odor, tenderness, juiciness, texture
etc.
 To what extent foods differ
 To determine consumer preferences
 To determine whether a certain food is acceptable to a specific
consumer group.
Aim of Sensory Evaluation
The aim of sensory evaluation is to determine the food quality
characteristics and the degree of compliance with the legal requirements and
consumer habits. The first and most important parameter of food is the
sensory characteristics. It is complex property, and it is an opinion about the
product itself, which cannot be replaced by any other method.
The aim of the sensory testing is to describe the product, to
distinguishing two or more products, to tell any differences between the
quality, its magnitude and direction. So the enjoyment is the sum of the
organoleptic characteristics.
Sensory analysis of food is performed with the human senses. Sensory
science is the study of the reactions of the five senses i.e. sight, hearing,
smell, taste and touch. It helps to know the characteristics of physical matter.
 Tongue and oral cavity
 Nose
 Skin touch
 Eye
 Ears
Panellists
The panellists are people who test the food and judge it. A panellist can

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be one person or several hundred. It depends on the type of the sensory
method. The panellists must be trained.
The panellists can be classified in three groups (trained panellists, semi
trained panellists, untrained panellists). In case of trained panellists 4-5
people is enough for the test. In case of semi trained panellist 10-20 people is
preferable. In case of untrained people, the largest number is the best.
Panellist should be of good health and should stop smoking, eating and
drinking before 30 minutes of the test. Factors affecting judge sensitivity:
 Health
 Smoking
 Memory
 Motivation
 The panellist mental fitness
Conducting Sensory Test
Testing Area: a special testing area is used so that obstruction of the
panel can be minimized.
 The testing area should have quite comfortable environment.
 Preferably room should be air conditioned and it is desirable that
the room should have controlled humidity.
 The testing should be such that it is easily reached by the panelist.
 Preparation area should be separated from the testing area.
 The panelist should not enter the preparation area as they might
gain information that would influence their judgment.
 Foreign colors from other food ingredients should be kept away
from testing room.
 Smoking should not be permitted at any time as it is recommended
by American Society for Testing and Material i.e. ASTM.
 A slight positive pressure should be maintained in the testing room
so that the odour from surrounding area will not enter testing area.
Testing Laboratory Consists of Three Separate Units
 Reception room: where the panel members meet the person in
charge of laboratory and get different type of sample to be tested.
 Sample preparation room: This is clean and well equipped area for
preparation and serving of samples.

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  Test booth: where actual sensory evaluation of the samples are
carried out by panel members. Whenever samples with difference in
color are tested then color lights should not be used as they will
mask the color of samples. The samples should be served in
stainless steel, glass or bone china utensils.
Testing Setup
 The panelists are required to make an independent judgment during
sensory evaluation.
 In order to eliminate obstruction and prevent communication among
the panelist individual booth are used which can be even a simple
partitions on a table.
 The booth may be fairly simple or much elaborated.
 Many laboratories have a sink and tap built into each booth to
provide water for rinsing mouth. But it has also been noted that
these sinks cause sanitation and odour problem.
 Usually a switch in each booth is connected so that a panelist can be
signaled when he is ready for another sample.
 The panelist should be provided with comfortable chair or stools.
 Color of the booth should not influence the appearance of a product
being tested. An off white or light grey color is usually
recommended.
 Lighting should be uniform and must not influence the appearance
of the product to be tested. Type of light should be carefully chosen,
if colour and appearance are important factors to be judged. Since,
fluorescent light may distort color.
Testing Schedule: testing should be done when the panel members are
fresh. The test time is generally between 10-12 A.M. in the morning. Too
many samples should not be used as it causes fatigue and errors in the result.
Samples should not be more than 4-5 at a time.
Preparing Samples: A well equipped kitchen is the foremost
requirement with specialized equipment needed. The preparation area should
have:
 Good ventilation system for removing cooking odours.
 There should be sufficient counter space for serving samples.
 Preliminary testing is usually necessary to determine the methods of

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 preparation of product such as time and temperature of cooking,
amount of water required, time and speed of blending, type and size
of utensils being used is considered.
 It is often necessary to cut and dice and mix fresh product to get
representative samples.
 Panelists are influenced by irrelevant characteristics of a sample
because every effort is made to look the sample identical in all the
characteristics. It is sometime necessary to grind, dice, chop or
puree the sample to obtain uniformity but the best way is to judge
the sample in their normal state.
 Some workers uses colored container such as ruby red glass or
black lined cup to mask color difference.
 Samples are presented with 3-5 digit code and then identification of
samples are made by checking the sample randomly.
Techniques of Smelling and Tasting: for testing food products a
special technique is used to perceive aroma more clearly. Smelling is done
with short and rapid sequence of sniff, tasting of coffee or tea or fruit juices
is done by slurping one teaspoon of liquid is rolled on the tongue so that the
liquid reaches to all parts of the tongue where the taste buds are located.
Dilution and Carriers: foods are served in the way they are normally
consumed. Products such as hot sauces and spices must be diluted before
they are tasted. The spices can be mixed with white sauce and sugar syrup,
considering that the sauce or syrup should not blend the flavour of spice. The
use of carrier such as crackers for jam and frankfurters for ketchups adds to
the cost and effort and it is difficult to select an appropriate carriers. Carriers
are also a source of experimental errors because the proportion of product to
carrier may not be constant. The nature of product may sometimes gives the
necessity of carrier. E.g. pie filling shall be tested with pastry to see that how
it interact especially in terms of texture and icing shoud be tested on cake not
only to check flavour combination but also to see the icing handles.
The temperature also serves as important factor, generally the samples
are served at a temperature at which they are normally eaten but sometimes it
can be slightly warmer or cooler than normal stage. E.g. Edible oils are
evaluated at 44̊C i.e. bit higher than normal range, hot foods are served at
60-66̊ C and ice cream at -1 to 2̊C.
Utensils: they must not impart any taste or odour to the product.
Identical containers should be used during testing which is preferably

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colorless or white. Disposable dishes made from plastic or paper etc. For
odour testing stamped wine glasses covered with watch glasses are used.
Quantity of Sample: the sensory evaluation committee of ASTM
recommends that in discrimination test each panellist should receive at least
16 ml of liquid and 28g of solid. The panellist should receive constant
sample and enough samples to taste back and forth until they make decision.
In case of flavoured milk, a panellist is suggested to drink 192ml of the
product.
Rinsing: the panellists are provided with an agent for oral rinsing
between 2 samples. Generally, neutral water at room temperature is preferred
but warm water is used during testing of fatty foods. The time period
between samples should be constant.
Choosing and Training Panellist: The sensory quality particularly
flavour attributes are essential to be measured subjectively. The judgement is
done by experts. Hence, for this purpose a trained panel is being selected
because the sensory evaluation depends upon the objective precision and
reproducibility of the panellist. Interest, motivation, general attitude and
emotional state of panellist may be especially for inconsistent judgement.
Panellists are usually office bearer, plant or research staff.
Requirements for Ideal Panellist Are
 He should be able to discriminate easily between samples and
should be able to distinguish appreciable difference between taste
and smell.
 The panelist should be in good health and should abstain
themselves when suffering from conditions that might interface
with normal function of taste and smell. Emotional factors, interest
and motivation appears to be more important than the age or sex of
panelist. A panelist should avoid smoking, chewing gum, eating or
drinking for at least 30 min before testing.
 He should be experienced in a particular field.
 Willingness to spend time for sensory evaluation is required.
 He should present his interest in sensory analysis of sample since,
motivation of panelist affects his response. An interested panelist is
always more efficient. The panelist should be made to feel that
sensory evaluation is an important activity and their contribution is
important.
 The selected panelist should have ability to concentrate on the

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 samples and give proper conclusion and evaluate the production
with duplicate judgment.
 Panelist must be trained to disregard their personal preferences and
he should be made familiar with the testing methods that will be
used. All the panel members must know and agree upon the exact
meaning of each descriptive term used.
Different Types of Panels
Trained Panel: i.e. Laboratory panel: they are trained for specific food
product. They are aimed to find the difference in specific quality
characteristics of a given sample. The no. of member in a trained panel may
vary from 5-10.
Descriptive or Semi Trained Panel: this type of panel includes
technical people and their families who are normally familiar with quality of
different types of foods they are capable with few preliminary tests. Under
this above 25-30 panel members list is made to find the acceptability and
preference before a product is launched for consumer trials.
Consumer Panel: these panels are made up of untrained people chosen
at random places to present for which a product is being evaluated. A greater
number of panel members are required in case of consumers, which may
increase to 100 or more.
Sensory Test Methods
Discrimination/Difference Tests
Difference tests are the simplest test of the food product testing. They
are used to determine whether there is or not a difference between two
samples or one sample is preferred to another. This method is a routine
quality control and the effects of change is monitored in production. This test
is a good step to determine the complex sensory evaluation of the products.
Commonly Used Difference Tests:
Paired comparison: Two samples compared, one to be selected (1-tailed)
where preference is asked either sample can be correct (2-tailed)
Triangle Test: Only one response can be correct
Difference to Reference/Control: Duo-trio, multiple comparison
Ranking Procedures
In this test there are three or more samples (maximum 6 samples), which
are presented at the same time. They are coded, and there is no information
about the samples.

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Scaling Procedures
Scale is the instrument used by panellists to make explicit their
perception. Types of Scale: Verbal, Numerical, Line.
Descriptive Method/Test
This test is usually carried by a small number of highly trained
panellists. For this test, panellists are not simply asked whether they can
determine differences between two products but are asked to rate particular
aspects of the flavour of a particular product on scale. Flavor aspects vary
depending upon type of product being studied e.g. flavour notes in tea may
be bitter, smoky and tangy whereas flavour notes in yoghurt may be acid,
chalky, smooth and sweet. A descriptive flavour map or profile of a product
is thus developed. Any detectable changes in the product would result in
changes in the flavour map. The trainees required to be able to detect,
describe and quantify subtle changes in specific flavour notes is extensive. In
this test trained panellists act as analytical instruments and their evaluation
of product is not related to their like or dislike of a product.
Difference Test
This test is easy, it examines the perceivable difference between the two
samples. Such tests are useful where a company was changing the source of
one of its ingredients or substituting one ingredient for another. These tests
can also be used to see of the quality of a product changes over time or to
compare shelf life of a particular product packaged in different packaging
materials. Two of the most frequently used difference tests are the triangle
test and duo-trio test. Typical ballots for these tests are shown in Fig 4. In the
triangle test, each panellist is given three samples, two of which are alike and
is asked to taste the sample from left to right, cleansing their palate before
analysis of each sample. They circle the number on the ballot sheet that
corresponds to the sample they believe to be different. In duo-trio test, each
panellist is given a reference and two samples. Panellist is asked to taste the
reference first and then each sample, from left to right and circle on the
ballot the sample that is the same as reference. Statistics must be applied to
results to determine whether there is a significant difference among products.
Test# _______________ Panelist # _______________
Triangle Diffeence Test
Product_________________
Instructions: Proceed when you are ready. (Quietly as not to distract

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 others)
For Each Sample:
1) Take a bite of the cracker and a sip of water to rinse your mouth.
2) Two of the samples are the same and one is different. Circle the
Odd sample. If you cannot tell, guess.
________________ _________________ __________________
3) Describe the reason why the ODD sample is DIFFERENT. (Be
specific)

Test# _______________ Panelist # _______________

Duo-Trio Difference Test
Product_________________
Instructions: Proceed when you are ready. (Quietly as not to distract
others)
For Each Sample
1) Take a bite of the cracker and a sip of water to rinse your mouth.
2) CIRCLE the number of the sample which is THE SAME as the
reference R. If you cannot tell, guess.
R ____________________ ____________________
3) Why is R and the sample you choose the same?

Fig 4: Ballot for difference tests

Artificial Senses
With the developments in electronics and computer hardware
technology, the instruments that can mimic the human senses are being
developed. These analytical instruments replaces the simple sensory analysis
and are playing a major role in building the framework for development of
the instrumental methods for sensory evaluation. These instruments include
electronic tongue, electronic eye and electronic nose. The advantages
associated with use of these instruments are that they can be calibrated
consistently and can give results in the form of important data that is directly
correlated with quality and safety control. These instruments can safely be
used in case where the samples are unfit for human consumption [22].

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 E-nose or electronic nose is an instrument that is designed having wide
variety of heterogeneous gas sensors with partial specificity and a pattern
recognition system. These were constructed so as to distinguish and detect
between the complex odors or aroma components of a food sample. The
sensors are electrochemical in nature and their operation is based on
interactions among gaseous molecules and the coating materials of sensors.
A variety of sensor types have been developed in which three types of
materials are commonly used: metal oxides, conducting polymer composites
and intrinsically conducting polymers. These coating materials modulate the
electric current and this current can be easily detected using a transducer that
converts the modulation into a signal that can be easily recorded [23]. This
device has gained importance in the field of sensor technology from past few
decades mainly due to its wide applications in various industries. Recent
trends in use of e-nose can be contributed to advances in sensors,
improvements in materials, innovative software and progress in designs,
circuits and systems. The developments in sensor types and arrays for
construction of e-nose have been closely related with the expansion of
applications of e-nose [24].
E-tongue or electronic tongue is analytical tool designed to differentiate
the tastes of various chemical substances in liquid phase samples. An
electronic tongue can be used to identify, classify and depict the taste
components. E-tongues can be called as the wet counterparts of e-nose. The
non-specific array of sensors displays an output in the form of different
patterns for different taste-causing substances. This data is collected and is
treated statistically. Various kinds of chemical sensors are being used in the
designs of e-tongues. The selection of these sensors depends on chemical
nature of the food samples analyzed. An e-tongue is essentially composed of
three elements namely an automatic sampler, a set of chemical sensors with
varying selectivity and a suitable signal processing data [25, 26].
Computer vision or e-eye focuses on mimicking the effects of human
vision by electronic means and obtaining images. This method involves five
basic components namely illumination source, a camera, an image capture
board and a computer software and hardware. E-eye deals with visual
inspection of quality characteristics of food products by capturing,
processing and analyzing images. This technology is experiencing a
substantial growth in food industry due to its cost effectiveness, accuracy,
superior speed and consistency. Computer vision can be explained as
construction of detailed and explained description of the physical entity from
the captured images. Physical image sensor gathered these images and then

Page | 72
analyzed by use of computer hardware and software. E-eye has been
successfully used for quality analysis of meat, pizza, fish, bread, cheese, and
cereal grains quality [27].
References
1. Adamson Melitta Weiss. Food in medieval times, Greenwood
Publishing Group, 88 Post Road West, Westport, 2004, 64-67.
2. Knouse SB, Philips Carson P, Carson KW. Edwards Deming and
Frederick Winslow Taylor: a comparison of two leaders who shaped the
world’s view of management. International Journal of Public
Administration. 1993; 16:1621-1658.
3. Best M, Neuhauser D. Walter a Shewhart, 1924 and the Hawthorne
factory. Quality and Safety in Health Care. 2006; 15:142-143.
4. Singham P, Birwal P, Yadav BK. Importance of Objective and
Subjective Measurement of Food Quality and their Inter-relationship. J
Food Process Technol. 2015; 6:488. doi:10.4172/2157-7110.1000488.
5. Kramer A, Twigg BA. Quality control for the food industry:
Fundamentals. AVI Publishing Co., Westport, Conn, 1983, 3.
6. Amerine MA, Pangborn RM, Roessler EB. Principles of sensory
evaluation of food, Academic Press, New York and London, 1965.
7. Kramer A, Twigg BA. Quality control for the food industry:
Fundamentals. AVI Publishing Co., Westport, Conn, 1982, 2.
8. Gatchalian MM. Sensory evaluation methods: for quality assessment
and development. College of Home Economic, University of the
Philippines, Dilimon Quezon City, Philipines, 1989.
9. Juran JM, Jr FM, Gryna Bingham RS. Quality Control Handbook, 2nd
Ed., New York: Mc Graw-Hill, 1974.
10. Harrington JJ. Total Improvement Management-The Next Generation in
Performance Improvement, McGraw-Hill, New York, NY, 1986.
11. Costa C, Antonucci F, Pallottino F, Aguzzi J, Sun D, Menesatti P. Shape
analysis of agricultural products: a review of recent research advances
and potential application to computer vision. Food Bioproc Technol.
2011; 4(5):673-692.
12. Grossman RL, Wisenblit JZ. What we know about consumers’ color
choices. Journal of Marketing Practice: Applied Marketing Science.
1999; 5(3):78-88.

Page | 73
13. Leon K, Mery D, Pedreschi F, Leon J. Color measurement in L* a* b*
units from RGB digital images. Food Research International. 2006;
39(10):1084-1091.
14. Nisha P, Singhal RS, Pandit AB. Kinetic modelling of colour
degradation in tomato puree (Lycopersicon esculentum L.). Food
Bioproc Technol. 2011; 4:781-787.
15. Pereira AC, Reis MS, Saraiva PM. Quality control of food products
using image analysis and multivariate statistical tools. Industrial and
Engineering Chemistry Research. 2009; 48(2):988-998.
16. Mitcham B, Cantwell M, Kader A. Methods for determining quality of
fresh commodities. Perishables Handling Newsletter. 1996; 85:1-6.
17. Pathare PB, Opara UL, Al-Said FA J. Colour measurement and
analysis in fresh and processed foods: a review. Food Bioproc
Technol. 2013; 6(1):36-60.
18. Taylor AJ, Roberts DD. Flavor perception, Blackwell. Adamson Melitta
Weiss Food in medieval times, Greenwood Publishing Group, 88 Post
Road West, Westport, 2004; 64-67.
19. Kress-Rogers E. Instrumentation for food quality measurement. In
Instrumentation and sensors for the food industry, Ed. E. Kress-Rogers
and C. Brimelow, Woodhead Publishing Ltd, 2001.
20. Röck F, Barsan N, Weimar U. Electronic nose: Current status and future
trends. Chemical Reviews. 2008; 108:705-725.
21. Piggott JR. Sensory analysis of food, Elservier Applied science, London
and New York, 1988.
22. Elizabeth Baldwin A, Jinhe Bai, Anne Plotto, Sharon Dea. Electronic
Noses and Tongues: Applications for the Food and Pharmaceutical
Industries. Sensors. 2011; 11:4744-4766.
23. Rodríguez-Mendez ML. Electronic noses and tongues in food Science
(1sted.). London: Academic Press, 2016.
24. Alphus D. Wilson and Manuela Baietto. Applications and Advances in
Electronic-Nose Technologies. Sensors, 2009, 5099-48.
25. Miguel P, Laura EG. A 21st century technique for food control:
Electronic noses. Analytica Chimica Acta. 2009; 638:1-15.
26. Cordero C, Kiefl J, Schieberle P, Reichenbach SE, Bicchi C.
Comprehensive two-dimensional gas chromatography and food sensory

Page | 74
 properties: potential and challenges. Anal Bioanal Chem. 2015;
407(1):169-191.
27. Brosnan T, Sun DW. Improving quality inspection of food products by
computer vision. J Food Eng. 2004; 61:3-16.

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Page | 76
 Chapter - 4
Prospects of Nano-Composites Containing Packaging on
Quality and Shelf Life of Fresh Fruits and Vegetables

Authors
Janagam Venu Madhav
Division of Food Science and Postharvest Technology, IARI, New Delhi,
India
R. Swamy Sekhar
BCKV Mohanpur, IARI, West Bengal, India
Shruti Sethi
Division of Food Science and Postharvest Technology, IARI, New Delhi,
India

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Page | 78
 Chapter - 4
Prospects of Nano-Composites Containing Packaging on
Quality and Shelf Life of Fresh Fruits and Vegetables
Janagam Venu Madhav, R. Swamy Sekhar and Shruti Sethi

Introduction
In the horticultural sector of the world, India is the second largest
producer of fruits (81.2 million tons) with a global share of over 12% and
also second largest producer of vegetables (162.1 million tons) with a global
share of over 14%. In spite of all these achievements, about 20 to 30% of the
produce is lost annually due to lack of adequate infrastructure and less use of
modern postharvest technologies. Fresh fruits and vegetables have limited
shelf life ranging from a few hours to few weeks at ambient conditions
because of High respiration rate, high moisture content and bulky in nature.
Packaging Functions
Containment
The container must enclose the produce in convenient units for
handling and distribution. The produce should fit well inside the container,
with little wasted space. Small produce items that are spherical or oblong
(such as potatoes, onions, and apples) may be packaged efficiently utilizing a
variety of different package shapes and sizes. However, many produce items
such as berries, or soft fruit may require containers specially designed for
that item. Packages of produce commonly handled by hand are usually
limited to 50 pounds. Bulk packages moved by forklifts may weigh as much
as 1,200 pounds.
Protection
The package must protect the produce from mechanical damage and
poor environmental conditions during handling and distribution. To produce
buyers, torn, dented, or collapsed produce packages usually indicate lack of
care in handling the contents. Produce containers must be sturdy enough to
resist damage during packaging, storage, and transportation to market.
Because almost all produce packages are palletized, produce containers

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should have sufficient stacking strength to resist crushing in a low
temperature, high humidity environment.
Identification
The package must identify and provide useful information about the
produce. It is customary (and may be required in some cases) to provide
information such as the produce name, brand, size, grade, variety, net
weight, count, grower, shipper, and country of origin. It is also becoming
more common to find included on the package, nutritional information,
recipes, and other useful information directed specifically at the consumer.
New Technologies in Packaging
There are several new technologies such as modified atmospheric
packaging, active packaging, smart packaging and biodegradable packaging
but there are several advantages and drawbacks in different packaging
systems. For example starch has received considerable attention as a
biodegradable thermoplastic polymer. However, it has a poor performance
by itself because of its water sensitivity and limited mechanical properties.
The application of nanotechnology to these polymers may open new
possibilities for improving not only the properties but also the cost-price-
efficiency.
Nanotechnology: Nanotechnology is an emerging field that involves
the understanding, manufacture and manipulation of materials at the
molecular or atomic level. Nanotechnology deals with particles and
structures larger than 1nm but smaller than 100 nm. Because of their size,
nano- particles have proportionally larger surface area and consequently
more surface atoms than their micro scale counterpart.
Nanotechnology Applications in Food Packaging
 New food packaging materials with improved mechanical, barrier
and antimicrobial properties.
 Nano-composite: improved mechanical, moisture & gas barrier
properties.
 Antimicrobial and antifungal surface coatings with nano-particles
(silver, magnesium, zinc).
 Nano-biosensors: For pathogen & contaminant detection.
 Smart and active packaging systems.
 Improved tagging, monitoring and traceability.

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 Nano-Composites: The use of fillers which have at least one dimension
in the nanometric range (nanoparticles) produces polymer nano-composites
(Alexandre & Dubois, 2000).
Table 1: Types of Nano-Composites
S. No Nano Composite Application Mechanism
Reinforcement Moisture, gas barrier and
1 Clays
of packaging Improving mechanical strength
Improving the polymer elasticity,
Cellulose based
Reinforcement thermo mechanical
2 Nano-
of packaging properties and reducing water
Composites
sensitivity
Improves the tensile strength and
Carbon Nano- Reinforcement
3 thermal properties of the
Tubes of packaging
polymers
Decreased water absorption by
Reinforcement
4 Silicates starch, improved the Oxygen
of packaging
barrier, tensile strength.
Starch Reinforcement
5 Improved tensile strength
Nanocrystals of packaging
Improved tensile strength, water
Reinforcement barrier properties.
6 Chitin/chitosan of packaging, Increasing membrane permeability
Anti-microbial and causing rupture and leakage of
intracellular material of pathogen
Spoilage By sensing the change in gas
7 Metal oxides
sensors composition package
Adhesion to the cell surface,
Silver Nano-
8 Anti-microbial degrading lipopolysaccharides and
particles
forming ‘‘pits’’ in the membranes.
Zinc Oxide (ZnO) Disruption of cell membrane
9 Anti-microbial
Nano-particles integrity of pathogens
Peroxidation of the polyunsaturated
Anti-microbial,
10 Titania (TiO2) phospholipids of
O2 indicator
microbial cell membranes

Mechanisms Nano-Composite Packaging

Barrier Properties
The most widely known theories to explain the improved barrier
properties of polymer– clay nanocomposites are based on a theory developed
by Nielsen (1967), which focuses on a tortuous path around the clay plates,
forcing the gas permeant to travel a longer path to diffuse through the film.
The increase in path length is a function of the high aspect ratio of the clay
filler and the volume % of the filler in the composite.

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 Fig 1: Tortuous path of a permeant in a clay nano-composite

Antimicrobial Nano-Composites
The incorporation of antimicrobial compounds into food packaging
materials has received considerable attention. Films with antimicrobial
activity could help control the growth of pathogenic and spoilage
microorganisms. An antimicrobial nano composite film is particularly
desirable due to its acceptable structural integrity and barrier properties
imparted by the nano-composite matrix, and the antimicrobial properties
contributed by the natural antimicrobial agents impregnated within.
Antimicrobial property of silver nanoparticles (Ag-NPs): adhesion to the cell
surface, degrading lipopolysaccharides and forming ‘‘pits’’ in the
membranes, largely increasing permeability, penetration inside bacterial cell,
damaging DNA and releasing antimicrobial Ag+ ions by Ag-NPs dissolution.
Ethylene Scavenging Property of TiO2
TiO2 decomposes the ethylene into CO2 and H2O in presence of light

Fig 2: TiO2 reaction with ethylene in presence of light

Chitin/Chitosan Nanoparticles
The possible antimicrobial mechanisms are interactions between

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positively charged chitosan and negatively charged cell membranes,
increasing membrane permeability and eventually causing rupture and
leakage of intracellular material and chitosan also causes chelation of trace
metals by chitosan, inhibiting enzyme activities and, in fungal cells,
penetration through the cell wall and membranes to bind DNA and inhibit
RNA synthesis.
Detection of Gases Produced by Food Spoilage
Food spoilage is caused by microorganisms, whose metabolism
produces gases which can be detected by conducting polymer nano
composites (CPC) or metal oxides, which can be used for quantification
and/or identification of microorganisms based on their gas emissions.
Sensors based on CPC consist on conducting particles embedded into an
insulating polymer matrix.
O2 Indicators
SnO2 as a photosensitizer in a colorimetric O2 indicator comprising a
sacrificial electron donor (glycerol), a redox dye (methylene blue – MB), and
an encapsulating polymer (hydrox yethyl cellulose). Exposure to UV-B light
led to activation (photo bleaching) of the indicator and photo reduction of
MB by the SnO2 nanoparticles. The color of the films varied according to O2
exposure, bleached when not exposed, and blue upon exposed.
Preparation of Nano-Composite Packaging
Nano-composite packaging can be prepared by several methods such as
melt-blow method, extrusion and even as edible coating by using plasticizer.
Few methods are given below
Hu et al., (2011) prepared a novel nano composite-based packaging
(NCP) by blending polyethylene (PE) with nano-Ag, nano-TiO2 and
montmorillonite by following method The low-density polyethylene (LDPE)
was used as matrix material (Translucency, Melt flow index 2.2 g/10 min,
density 0.92 kg/m3, softening point 95°C). The nano-powders (35 wt. % of
nano-silver, 40 wt. % of nano-TiO2 and 25 wt. % of montmorillonite) in the
range of 40-80nm. Firstly a PE-nano composite master batch containing 30
wt. % of the commercial nano-powder, 56 wt. % of PE granule and 14 wt. %
of cross-link reagent KH-570 were immingled in uniformity through a high-
speed mixer for 1 h. After air cooling, they were extruded to PE nano-
composite master batch using a twin-screw extruder with a screw diameter
of 22 mm, a screw length/diameter ratio of 42 and a screw speed of 600 rpm.
In the second extrusion step, 1.5 kg of master batch and 38.5 kg of PE

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granule were immingled for 30 min. Subsequently the compounds were
blown into a film of 50μm thickness via a plastic extruder. After cooling,
films of 50 μm thickness were used to make bags of 20×22 cm2 using a heat
sealer. Polyethylene bags of the same thickness and size without nano-
composite master-batch nano-powder served as controls.
An et al., (2008) prepared the silver nanoparticles-PVP nano-composite
coating by following method. AgNO3, NaBH4 and PVP were dissolved in
deionized water to form aqueous solution of AgNO3 (0.1 M), NaBH4 (0.01
M) and PVP (0.01 M), respectively. The aqueous solutions of PVP (0.01M)
and NaBH4 (0.01 M) were mixed at a volume ratio of 1:1. About 500 ml of
this solution was transferred to a beaker, and agitated with a magnetic stirrer
before adding the AgNO3 (0.1M) solution. Upon addition of silver nitrate
drop by drop, the colorless solution of NaBH4-PVP slowly changed from
yellow to pale brown indicating the formation of silver nanoparticles.
Glycerol was added to the solution of silver nanoparticles. as a plasticizer
(1.5/100 ml) and the solution was stirred over a magnetic stirrer for 15 min
under magnetic stirring at ambient temperature. The final practical silver
concentration 0.06 mg l-1 for coating was used.
Effect of Nano Composite Packaging On Different Quality Parameters
of Fruits and Vegetables
Hu et al., (2011) studied the effect of nano composite-based packaging
on postharvest quality of ethylene-treated kiwifruit (Actinidia deliciosa)
during cold storage. A novel nano composite-based packaging (NCP) was
prepared by blending polyethylene (PE) with nano-Ag, nano-TiO2 and
montmorillonite by and they reported the effects of NCP on the quality
parameters of ethylene-treated mature kiwifruit were investigated during the
42 d of storage at 4 °C. The results showed that adding nanoparticles to the
PE significantly decreased the oxygen, water vapour permeability and
longitudinal strength, and inhibited spore germination. The weight loss,
softening, color variation and soluble solid content of kiwifruit were
significantly inhibited by 22.67%, 124.84%, 23.46% and 14.42%
respectively, which indicated that NCP could delay the ripening of kiwifruit.
However, ascorbic acid and total phenols contents in NCP-treated fruit were
increased compared with the controls. Additionally, kiwifruit in NCP
exhibited 57.44% lower headspace ethylene concentration, 29.44% for
malondialdehyde (MDA), lower polyphenol oxidase (PPO) activity and
higher peroxidase (POD) activity than the controls. These results suggest
that NCP may be a useful technique to reduce fruit decay and maintain
quality in kiwifruits during postharvest storage.

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 Li et al., (2009) reported the effect of a novel nano-packing material on
preservation quality of Chinese jujube (Ziziphus jujuba Mill. var. inermis
(Bunge) Rehd) during room temperature storage. The nano-packing material
with lower relative humidity, oxygen transmission rate and high longitudinal
strength (2.05 g/m2 24 h, 12.56 cm3/m2 24 h 0.1 MPa and 40.16 MPa,
respectively) was synthesized by blending polyethylene with nano-powder
(nano-Ag, kaolin, anatase TiO2, rutile TiO2). The results showed that the
nano-packing material had a quite beneficial effect on physicochemical and
sensory quality compared with normal packing material. After 12-day
storage, fruit softening, weight loss, browning and climatic evolution of
nano-packing were significantly inhibited. Meanwhile, the contents of
titrable acid and ascorbic acid were decreased to 0.21%, 251 mg/100 g, for
nano-packing and 0.15%, 198 mg/100 g, for normal packing; The contents of
total soluble sugar, reducing sugar, total soluble solids and malondialdehyde
were increased to 28.4%, 5.2%, 19.5% and 98.9 µmol/g for nano-packing
and 30.0%, 6.3%, 23.1% and 149 µmol/g for normal packing. Therefore, the
nano-packing could be applied for preservation of Chinese jujube to expand
its shelf life and improve preservation quality.
Yang et al (2010) prepared a novel nano-packing material with lower
relative humidity, oxygen transmission rate and high longitudinal strength
was synthesized by blending polyethylene with nano-powder (nano-Ag,
kaolin, anatase TiO2, rutile TiO2), and investigated its effect on preservation
quality of strawberry fruits (Fragaria ananassa Duch. cv Fengxiang) during
storage at 4oC and reported that nano-packaging was able to maintain the
sensory, physicochemical, and physiological quality of strawberry fruits at a
higher level compared with the normal packing (polyethylene bags). After a
12-d storage, decreases in the contents of total soluble solids, titra table
acidity, and ascorbic acid of nano-packing were significantly inhibited.
Meanwhile, decay rate, anthocyanin, and malondialdehyde contents were
decreased to 16.7%, 26.3 mg/100g, 66.3 μmol/g for nano-packing and
26.8%, 31.9 mg/100g, 75.4 μmol/g for normal packing; poly phenoloxidase
(PPO) and pyrogallol peroxidase (POD) activities were significantly lower in
nano-packing than the control. These data indicated that the nano-packaging
might provide an attractive alternative to improve preservation quality of the
strawberry fruits during extended storage.
Ram et al., (2013) investigated the Effect of ZnO Nano Particle
Containing Packaging on Shelf Life of Fresh Nagpur Mandarin (Citrus
reticulata Blanco) Segments. The ZnO Nano composites film used for the
study was characterized by 40 nm ZnO nano particle embedded in 60 μm

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thickness poly film with tensile strength of 3.35±0.50 kg longitudinally and
1.67±0.27 kg transversely average breaking load which was provided by
CIRCOT, Mumbai and they they reported that packaging containing ZnO
nano particle was suitable to maintain the microbial load below the threshold
population limit (6 log CFU/ml) till 30 days of storage. The segments had
better color in ZnO nano packaging after 30 days of storage. The total
soluble solids (TSS), vitamin-C and carotenoid contents were found better in
ZnO nano packaging than that of LDPE packaging. Relatively lesser value of
browning was recorded in ZnO nanoparticle containing film (0.62mg/l). The
limonin content was recorded to be higher than the threshold limit in both
the packaging. The changes in segments juice reducing sugar; non reducing
sugar and total sugar were found to be at higher side in LDPE packaging
than that of nano packaging. Study revealed that segments in nano packaging
had 30 days of shelf life with better quality under refrigerated storage (4oC)
temperature.
An et al., (2008) evaluated the effect of silver nanoparticles-PVP
coating on weight loss, ascorbic acid, total chlorophyll, crude fiber, color,
firmness and microbial qualities of asparagus spears stored at 2 and 10oC.
Asparagus samples were first sanitized with 100mg l-1 sodium hypo chloride
solution for 15 min. They were then immersed in coating solution containing
silver nanoparticles for 3 min at room temperature. During 25-day storage at
2 or 10oC, the coated asparagus demonstrated lower weight loss, greener
color and tender texture compared with the control samples. The growth of
microorganism was significantly hindered by the coating. Based on
comprehensive comparison and evaluation they suggested that asparagus
spears coated by silver nanoparticles could be kept in good quality for 25
days at 2oC and for 20 days at 10oC.
Health Risk of Nano Materials
Our bodies’ defensive mechanisms are not as effective at removing
nanoparticles from our lungs, gastro-intestinal tract and organs, as they are
with larger particles. Nanoparticles are also more adhesive than larger
particles to surfaces within our bodies. As a result of these factors and their
very small size, nanoparticles are much more likely to be taken up into our
cells and tissues than are larger particles. Numerous in vivo experiments
using rats and mice have demonstrated gastrointestinal uptake of
nanoparticles and small microparticles. Pathological examination of human
tissues also suggests ingestion and translocation of microparticles up to
20μm in size. A growing body of evidence demonstrates that some
manufactured nanoparticles will be more toxic per unit of mass than larger

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particles of the same chemical composition. For example titanium dioxide is
considered to be biologically inert in bulk form and is widely used as a food
additive. However in vitro experiments show that as a nanoparticle or
particle up to a few hundred nanometres in size, titanium dioxide damages
DNA, disrupts the function of cells, interferes with the defence activities of
immune cells and, by adsorbing fragments of bacteria and ‘smuggling’ them
across the gastro-intestinal tract, can provoke inflammation. A single high
oral dose of titanium dioxide nanoparticles caused significant lesions in the
kidneys and livers of female mice. The potential for ingested non degradable
nanoparticles to cause long term pathological effects in addition to short-
term toxicity is of great concern. A small number of clinical studies suggest
that non-degradable nanoparticles and small microparticles which do not
provoke an acute toxic response can accumulate in our bodies and over time
result in the development of ‘nano pathologies’, for example granulomas,
lesions (areas of damaged cells or tissue), cancer or blood clots.
Table 2: Toxicity of Nanomaterials used in Packaging

Nanomaterial Size Experimental evidence of toxicity

Destroyed DNA (in vitro; Donaldson et al.,
20 nm
1996)
30 nm mix of rutile Produce free radicals in brain immune cells
Titanium dioxide
and anatase forms (in vitro; Long et al., 2006)
25 nm, 80 nm and accumulated in liver, spleen, kidneys and
155 nm lung tissues (in vivo; Wang et al., 2007)
15 nm, 100 nm and Highly toxic to rat liver cells, brain cells (in
Silver
ionic form vitro; Hussain et al., 2006)
Toxic to human cells even at very low
19 nm
concentrations (in vitro; Brunner et al., 2006)
Zinc oxide
Damage liver, spleen and pancreas in mice
20 nm and 120 nm
(in vivo; Wang et al., 2007)
Inhibit cell growth, neurodegenerative
Silicon dioxide 50nm and 70nm disorders (in vitro; Chen and von Mickecz,
2005)

Regulatory Aspects of Nanotechnology

There are currently no special regulations for the application or
utilization of nanotechnology in foods in the United States, and although
recommendations for special regulations in the European Union (EU) have
been made, laws have yet to be changed. The U.S. Food and Drug
Administration (FDA) states that it regulates “products, not technologies,”
and anticipates that many products of nanotechnology will fall under the
jurisdiction of multiple centers within FDA and will therefore be regulated

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by the Office of Combination Products. FDA regulates on a product-by-
product basis and stresses that many products that are currently regulated
produce particles in the nano-size range. FDA says that “particle size is not
the issue” and stresses that new materials, regardless of the technology used
to create them, will be subject to the standard battery of safety tests
(http://www.fda.gov/nanotechnology/regulation.html). In contrast to the
FDA view on particle size, a recent report by the Institute of Food Science
and Technology (IFST)-a United Kingdom-based independent professional
qualifying body for food scientists and technologists-states that size matters
and recommends that nanoparticles be treated as new, potentially harmful
materials until testing proves their safety. None the less, the European
Commission intends to use existing food laws with respect to food products
derived through nanotechnology, where applicable, but acknowledges that
the technology will likely require modifications to the law. The European
Commission plans to use a case-by-case approach to risk assessment.
Recommendations by the Royal Society and the Royal Academy of
Engineering, commissioned by the UK government to assess the potential
impact of nanotechnology, included a call for identification of the use of
nanoparticles in ingredient lists. The UK government agreed that this was
necessary for consumers to make informed decisions and that modifications
to current labeling requirements would be necessary. The IFST suggested
that when nanoparticles are used as food additives, the conventional E-
numbering system for labeling be used along with the subscript “n” (IFST
2006). The UK government also agreed to consult with EU partners
regarding the IFST report’s recommendation that nanoparticle ingredients be
subjected to a full safety assessment before use in consumer products.
Future Thrust
Greater focus to elucidate the definite mechanism of nano-packaging
functions during storage to facilitate the application of nano-technology in a
broader range in the future. There are numerous data gaps that need to be
filled in order to demonstrate product safety to a wary public such as Nano-
material migration through polymer films, the interaction of nano-material
biomolecules and cellular components and the interrelationships between
nano-particle characteristics and toxicity.
References
1. Alexandre M, Dubois PP. Material science and engineering, Part R,
2000, 28.
2. An J, Zhanga M, Wang S, Tang J. Physical, chemical and

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 microbiological changes in stored green asparagus spears as affected by
coating of silver nanoparticles-PVP. LWT- Food Science and
Technology. 2008; 41:1100-1107.
3. Brunner TJ, Wick P, Manser P, Spohn P, Grass RN, Limbach LK et al.
In vitro cytotoxicity of oxide nanoparticles: comparison to asbestos,
silica, and effects of particle solubility. Environ. Sci. Technol. 2006;
40:4374-4381.
4. Chen M, Von Mikecz A. Formation of nucleoplasmic protein
aggregatesimpairs nuclear function in response to SiO2 nanoparticles.
Experiment Cell Res. 2005; 305:51-62.
5. Donaldson K, Beswick PH, Gilmour PS. Free radical activity associated
with the surface of particles: a unifying factor in determining biological
activity. Toxicol Lett. 1996; 88:293-298
6. Hu Q, Fang Y, Yang Y, Ma Y, Zhao L. Effect of nanocomposite-based
packaging on postharvest quality of ethylene-treated kiwifruit (Actinidia
deliciosa) during cold storage. Food Research International. 2010;
44:1589-1596.
7. Hussain SM, Javorina MK, Schrand AM, Duhart HM, Ali SF, Schlager
JJ. The interaction of manganese nanoparticles with PC-12 cells induces
dopamine depletion. Toxicol. Sci. 2006; 92:456-463.
8. Li H, Li F, Wang Y, Sheng J, Xin Z, Zhao L et al. Effect of nano-
packing on preservation quality of Chinese jujube (Ziziphus jujuba Mill.
var. inermis (Bunge) Rehd). Food Chemistry. 2009; 114:547-552.
9. Long TC, Saleh N, Tilton RD, Lowry GV, Veronesi B. Titanium
dioxide (P25) produces reactive oxygen species in immortalized brain
microglia (BV2): implications for nanoparticle neurotoxicity. Environ
Sci Technol. 2006; 40:4346-4352.
10. Ram L, Kumar D, Vigneshwaran N, Khewle A. Effect of ZnO nano
particle containing packaging on shelf life of fresh Nagpur mandarin
(Citrus reticulata Blanco) segments. Journal of Biological and Chemical
Research. 2013; 30(2):381-386.
11. Wang J, Zhou G, Chen C, Yu H, Wang T, Ma Y. Acute toxicity and bio
distribution of different sized titanium dioxide particles in mice after
oral administration. Toxicology letters. 2007; 168:176-185.
12. Yang FM, Li HM, Li F, Xin ZH, Zhao LY, Zheng YH, Hu QH. Effect
of Nano-Packing on Preservation Quality of Fresh Strawberry (Fragaria

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ananassa Duch. Cv. Fengxiang) during Storage at 4 ◦C. Journal of Food
Science. 2010; 75(3):236-240.

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 Chapter - 5
Machine Vision System: A Tool for Quality Inspection of
Food

Authors
Monika Mathur
Center of Food Science and Technology, CCSHAU, Hisar, Haryana,
India
Raveena Kargwal
Department of Processing and Food Engineering, COAE & T, CCSHAU,
Hisar, Haryana, India

Page | 91
Page | 92
 Chapter - 5
Machine Vision System: A Tool for Quality Inspection of
Food
Monika Mathur and Raveena Kargwal

Quality investigation of food and Agricultural products create are

troublesome and work concentrated. At the same time, with expanded
desires for food results of high caliber and wellbeing measures, the
requirement for exact, quick and target quality assurance of these attributes
in food items keeps on developing. Number of motorized preparing lines has
been created for business generation. In any case, the visual investigation
and quality control is finished by human eyes. Human examination is a
moderate procedure, has poor repeatability and result shifts from individual
to individual. Machine vision framework (MVS) is a picture
handling/examination procedure which is utilized for target assessment of
value parameters. Its speed and exactness fulfill regularly expanding creation
and quality prerequisites, henceforth helping in the advancement of
completely computerized forms.
Large scale manufacturing of food was likewise connected with two
noteworthy issues. The first is the decrease in food quality, and the second
one is the "waste" issue related with handling and planning activities. The
wastage by and large is an immediate outcome of the quality issue, where the
quality decrease achieves unaccepted cutoff points. Henceforth, is the
requirement for quality examination and affirmation instruments to be
introduced in the generation lines of such mass food preparing and creating
plants.
MVS can perform many functions simultaneously in a food processing
line such as segregation by species, by size, by visual quality attributes,
determination of composition, volume, measurement of shape parameters,
and quantification of the meat colour, automated portioning and detection of
defects. MVS system is also being used in food industry for the detection of
defects in apples, oranges, olives, cherries etc., sorting of potatoes, online
monitoring of baking conditions, measurement of browning in chips. It is
also being used for checking ripening stages of banana, tomato, cherries etc.

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Also it can be used to classify different varieties of cereal grains and check
their adulteration. Recent developments in MVS and supporting technologies
has resulted in general acceptance of the feasibility and profitability of
implementing visual inspecting systems in quality assurance operations.
Quality control is becoming increasingly more important in today’s
industry. Food quality is the quality characteristics of food that is acceptable
to consumers. This includes external factors as appearance (size, shape,
color, gloss, and consistency), texture, and flavour; factors such as federal
grade standards (e.g. of eggs) and internal (chemical, physical, microbial).
Customers demand cheap but high quality products. Many consumers also
rely on manufacturing and processing standards, particularly to know what
ingredients are present, due to dietary, nutritional requirements
(kosher, halal, vegetarian), or medical conditions (e.g., diabetes, or allergies)
Food quality also deals with product traceability, (e.g., of ingredient,
and packaging suppliers), should a recall of the product be required. It also
deals with labeling issues to ensure there is correct ingredient and nutritional
information. So, the ability to manufacture high-quality products consistently
is the basis for success in the highly competitive food industry. For an
efficient and successful production, manufacturers must rely on a robust
quality control system. These systems can prevent product rejection by
detecting all defective products and simultaneously avoid false defect
detections. Also, an automated quality control system integrated into the
manufacturing process can detect production deviations early and thus avoid
waste. Quality has been the subject of a large number of studies (Shew felt &
Bruckner, 2000).
Quality can be analyzed by subjective and objective means. Compared
to the traditional quality control tasks performed by humans, automated
quality control systems offer various advantages, including the ability to
work in hazardous environments 24 hours a day, and in some tasks perform
quicker measurements with higher accuracy and consistency than humans.
Consequently Francis (1980) found that human perception could be easily
fooled. Together with the high labour costs, inconsistency and variability
associated with human inspection accentuates the need for objective
measurements systems. Additionally, when the automated quality control
system is computer-controlled, the results of monitoring can easily be
integrated into the manufacturer’s statistical process control. Machine vision
quality control systems play a growing role in modern manufacturing quality
control systems. One of the reasons explaining this growth is that quick,
accurate noncontact measurement and complex feature analysis can now be

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performed by low-cost machine vision systems. So, Machine vision defined
by the machine vision association of society “The use of devices for optical
non-contact sensing to automatically receive and interpret an image of a real
scene in order to obtain information and/or control machine or processes”.
Another definition Machine vision is the technology to replace or
complement manual inspections and measurements with digital cameras and
image processing. The technology is used in a variety of different industries
to automate the production, increase production speed and yield, and to
improve product quality. However, the speed, reliability and cost savings
inherent to process automation have provided sufficient impetus to the food
industry to pursue aggressively the automatic, on-line control of all plant
operations.
Innovation of new technology often undergoes four phases. The first is a
research or discovery phase, in which new knowledge results in advances in
the technology. Phase two is the early commercialization phase, in which a
small market for the technology develops. Phase three represents the
emergence of niche-specific applications of the technology. The last phase is
that of widespread proliferation. A key characteristic of phase four is that the
technology is transparent to users, that is, those who use it do not have to
understand its inner workings. Based on this classification, we can clearly
say that machine vision technology is in phase three, and steadily marching
towards becoming a phase-four technology within the next decade or so. In
concern of history, in 1950 – Two dimensional imaging for statistical pattern
recognition development. In1960’s – Roberts begins studying 3D machine
vision. 1970s –MIT’s artificial intelligence lab opens a machine vision
course. 1980’s- new theories and new concepts emerging; optical character
recognition systems were used in various industrial applications. 1990’s-
highly advanced and improved versions of MVS are developed and cost of
MVS starts dropping. Sonka et al. (1999) noted that it aims to duplicate the
effect of human vision by electronically perceiving and understanding an
image, and provides suitably rapid, economic, consistent and objective
assessment. Zuech et al. 2000 studied that the use of machine vision system
devices for optical, non-contact sensing to automatically receive and
interpret the image of a real scene in order to obtain information and/or
control machines or process image. Today – 3D vision systems that can scan
products running at high speeds are becoming affordable and system do
everything from thermal imaging to slop measurement.
In general, a machine vision sensor is used to measure some aspect of
the results of the manufacturing process (e.g. shape, size, texture, location)

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that is indicative of the accuracy, efficiency or quality of the process. The
measured parameter can then be used as feedback in a real-time control loop
that optimizes the manufacturing process through variations in process
parameters (speed, temperature, flow rate, etc.) Computer vision applications
currently range from vision-guided robot assembly to inspection tasks. Most
practical applications can be broken down into six general categories. First is
gauging: performing precise dimensional measurements, second is
verification: qualitatively ensuring that one or more desired features are
present and/or undesired features are absent, third one is flaw detection:
finding and discriminating unwanted features of unknown size, location and
shape Fourth is identification: determining the identity of an object from
symbols, including alphanumeric characters. Recognition: determining the
identity of an object from observed features of the object is fifth and sixth is
locating: determining the location and orientation of an object.
The food industry continues to be among the fastest growing segments
for machine vision applications. It now ranks among the top ten industries
using machine vision technology. Most initial vision systems are isolated
batch-type operations that target a specific task. However, in order for vision
systems to be a mainstay of the food processing industry, they must integrate
into the overall system design and provide on-line, continuous and automatic
control capabilities. Much progress has been made in this direction over the
past five years. Currently, several commercial vendors offer automatic
vision-based quality-evaluation systems for the food industry. Despite their
initial cost, such systems are becoming increasingly popular.
Recently automatic inspection systems, mainly based on camera––
computer technology have been investigated for the sensory analysis of
agricultural and food products. Machine vision has proven to be successful
for objective measurement of various agricultural (He, Yang, Xue, & Geng,
1998; Li & Wang, 1999) and food products (Sun, 2000; Wang & Sun, 2001).
Machine vision includes the capturing, processing and analyzing
images, facilitating the objective and nondestructive assessment of visual
quality characteristics in food products (Timmermans, 1998). The potential
of Machine vision in the food industry has long been recognized (Tillett,
1990) and the food industry is now ranked among the top 10 industries using
this technology (Gunasekaran, 1996). Recent advances in hardware and
software have aided in this expansion by providing low cost powerful
solutions, leading to more studies on the development of Machine vision
systems in the food industry (Locht, Thomsen, & Mikkelsen, 1997; Sun,
2000). There is increasing evidence that machine vision is being adopted at

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commercial level (Locht et al., 1997). Computer vision has long been
recognized as a potential technique for the guidance or control of agricultural
and food processes (Tillett 1990). Therefore, over the past 20 years,
extensive studies have been carried out, thus generating many publications.
Components of Computer Vision
Machine vision systems commonly used in agricultural applications
acquire reflectance, transmittance, or fluorescence images of the agricultural
materials under UV, VIS, or NIR illumination. Generally it consists five
basic components: illumination, a camera, an image capture board (frame
grabber or digitizer), computer hardware and software as shown in Fig. 1
(Wang & Sun 2002a).
Image Illumination
Illumination systems are the light sources. The light focuses on the
materials (especially when used). Lighting type, location and colour quality
play an important role in bringing out a clear image of the object (Narendra
and Hareesh, 2010).
The light range can be in the UV (200-400 nm), VIS (400-700 nm), or
NIR (700-2500 nm). There are also applications in thermal imaging (above
2500 nm) for agricultural products. When radiation from the lighting system
illuminates an object, it is transmitted through, reflected, or absorbed. These
phenomena are referred to as optical properties. The absorbed light can also
be re-emitted (fluorescence), usually at longer wavelengths. A number of
compounds emit fluorescence in the VIS region of the spectrum when
excited with UV radiation. The optical properties and fluorescence emission
from the object are integrated functions of the angle and wavelength of the
incident light and chemical and physical composition of the object. So, the
lighting system, a critical part of a controlled machine vision system, must
be carefully designed. The ultimate purpose of lighting design is to provide a
consistent scene eliminate the appearance of variations, and yield
appropriate, application-specific lighting. Proper selection of lighting
sources (incandescent, fluorescent, halogen, Xenon, LED), lighting
arrangements (backlighting, front lighting, side lighting, structured lighting,
ring lighting), and lighting geometry (point lighting, diffuse lighting,
collimated lighting) is the “key to value” (Zuech 2004). Primary factors that
influence the selection is whether the object under inspection is: 1) flat or
curved; 2) absorbing, transmissive or reflective; and 3) the nature of the
feature to be imaged in comparison with the background. For instance,
backlighting is usually used for detecting objects that are translucent, such as

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hatching eggs (Das and Evans 1992) or used to measure the geometric
dimensions of obscured object, such as measuring the shoot height or root
diameters of pine to estimate the pine seedlings (Wilhoit et al. 1994). The
side light scatter can be used to determine the cellular granularity (Jain et al.
1991). Structured light can be used to form a 3-D shape of apple surfaces for
stem/calyx detection (Yang 1993). Moreover, controlled lighting design
sometimes acts as an active sensing means that can “create” new
information. Because of a controlled lighting system design, intelligent
image processing technology applied to a machine vision system is normally
simplified and can achieve high accuracy. Major Key to a successful
machine vision application is to start with a good contrast, repeatable image
that is not affected by ambient light or the surroundings. A functional block
diagram of basic machine vision system has been given in Fig. 1(b).
Image Acquisition
With advances in digital cameras, the camera and the image capture
system generally merge into a single device. This device communicates with
the computer via cables (e.g., USB or Fire wire), or by wireless means.
There can be three light detection sensors in the camera, dedicated to each
primary colour (Red, Green, Blue), or one sensor can be selectively used to
handle all the three primary colors. The software can control the camera
settings, the timing of image acquisition, the light source and can analyze the
image to extract desired features to make decisions. These may include non-
contact sensing, measuring object shape and dimensions, detecting product
defects; providing process control feedback alerting production line
operators for in process system failures; and providing product quality
statistics (Sarkar, 1991; Sun, 2004; Balaban et al., 2005). Image capturing is
an application program that enables users to upload pictures from digital
cameras or scanners which are either connected directly to the computer or
to network. A system consisting of a high-pixel resolution CCD (charge
coupled device) chip and associated hardware is the most common method
for generating digital images. However, because digital images are
inherently monochrome, or black and white, other hardware and software are
needed to generate color images. An electronic system that can classify fruits
by mass, by colour, by diameter, by bruising, shape and size should be
composed of a camera, a lens, a light source, a filter, a PC and image
processing software (Sarkar and Wolfe 1985; Hahn and Sanchez 2000). Lino
et al. (2008) used electronic systems composed of CCD camera and PC for
image capturing in quality evaluation of tomatoes and lemons. Recently
Kanali et al. (1998) investigated the feasibility of using a charge simulation

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method (CSM) algorithm to process primary image features for three
dimensional shapes recognition. However, only 2- dimensional (2D) data is
needed for grading, classification, and analysis of most agricultural images.
3- Dimensional may be needed for information on structure or added details.
Sonka et al. (1999) have developed 3-D vision technique from a series of 2-
D images to derive a geometric description. The imaging unit consists of a
high performance 12-bit digital CCD camera and spectral range 290–1,100
nm with the peak quantum efficiency of 60% at 500 nm were used for
quality evaluation of pickling cucumbers using hyperspectral reflectance and
transmittance imaging by Ariana and Lu (2008).

Fig: A Principle components of machine vision system (Pundit et al. 2005) (b)
Functional block diagram of basic machine vision system (Singh et al. 2004)

Frame Grabber
The process of converting pictorial images into numerical form is called
digitisation. In this process, an image is divided into a two dimensional grid
of small regions containing picture elements defined as pixels by using a
vision processor board called a digitiser or frame grabber. There are

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numerous types of analogue to digital converters (ADC) but for real time
analyses a special type is required, this is known as a ‘flash’ ADC. Such
‘flash’ devices require only nanoseconds to produce a result with 50–200
mega samples processed per second (Davies 1997). Selection of the frame
grabber is based on camera output, spatial and grey level resolutions
required, and the processing capability of the processor board itself
(Gunasekaran and Ding 1994).
Image Processing and Analysis
Image analysis can provide a wide range of information about a product
from a single image in a fraction of a second, making it possible to analyze
products as they pass on a conveyor belt (Storbeck and Daan, 2001). Image
processing/analysis involve a series of steps, which can be broadly divided
into three levels: low level processing, intermediate level processing and
high level processing (Sun, 2000).
Low Level Processing
Low level processing includes image acquisition and pre-processing.
Image acquisition is the transfer of the electronic signal from the sensing
device into a numeric form. The machine vision system, basically, comprises
two main processes: (1) Image processing and (2) Image analysis (Zuech et
al. 2000). Image preprocessing refers to the initial processing of the raw
image data for correction of geometric distortions, removal of noise, grey
level correction and correction for blurring. Averaging and Gaussian filters
are often used for noise reduction with their operation causing a smoothing
in the image but having the effect of blurring edges.
Intermediate Level Processing
Intermediate level processing involves image segmentation, image
representation and description. Image segmentation is one of the most
important steps in the entire image processing technique as subsequent
extracted data are highly dependent on the accuracy of this operation. Its
main aim is to divide an image into regions that have a strong correlation
with objects or areas of interest.
High Level Processing
High level processing involves recognition and interpretation, typically
using statistical classifiers or multilayer neural networks of the region of
interest. These steps provide the information necessary for the
process/machine control for quality sorting and grading. Statistical
procedures from basic image statistics such as mean, standard deviation and

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variance to more complex measurement such as principle component
analysis can be used to extract features from digital images. Once image
features are identified, the next step is feature classification. (Minz, 2013).
Working Principle of Machine Vision: Principle of working involves
four basic steps:
1. Imaging: Take an image.
2. Processing and Analysis: Analyze the image to obtain a result.
3. Communication: Send the result to the system in control of the
process.
4. Action: Take action depending on the vision system's result.
Machine vision is the construction of explicit and meaningful
descriptions of physical objects from images. Images are acquired with a
physical image sensor and dedicated computing hardware and software are
used to analyses the images with the objective of performing a predefined
visual task.

Fig 2: Different levels in the image processing process (Sum, 2000).

Machine vision system analyze image and produce result on the basis of
different attributes of product: shape, size, Texture and Appearance. Size of
the product is an important parameter to measure e.g. On many food
production lines, jamming of packaging machines due to oversized products
is one of the major problems. 2 D size of a product is a relatively
straightforward parameter to measure if the edges of the product are clearly
defined against the background. Use of 3-D techniques with structured
lighting or laser lines. it is also possible to determine the height information
and hence the volume of the product. Measurement of size and shape can
also be used in the grading and separation of products. For example eggs,
potatoes, chickens, and so on.

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Shape and Size
Many food products contain separate distinct features, for example, the
items in a complete set meal as provided by airlines, or the different
components on a pizza (tomatoes, olives, peppers, etc). Région labelling
operator will shade the objects according to the order in which they are
found during a conventional raster scan of the input image.The objects could
be shaded according to their areas, i.e. the biggest one becomes the brightest
(or darkest). To isolate groups of similar objects in an image for analysis:
The segmentation process would rely on the structural and colour properties
of these objects. Figure illustrated how certain features such as the cherries
and the crimped pastry ring in a packet of bakewell cakes could be enhanced
by using their colour attributes.
Appearance of Product
 It is very difficult to quantify. For example, what constitutes a good
loaf shape? And when does the appearance of a pizza become
unattractive? By extracting and analyzing certain individual
features, it is possible to form a quantitative description of the
appearance of a food product in a limited way. Surface defects are
much easier to inspect and are based on detecting an abrupt change
in the characteristics of the surface.

Fig 4.19: Measurement using machine vision: (a) original image of croissant,
(b) after thresholding, the centroid, the angle of least moment of inertia and the
maximum object width are found.

Fig 4.11: Using RGB informaation to enhance certain features of a product: (a) the
red channel showing the redness of the scene.

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 For example, if a baked product
is detected to be overcooked by
a vision system, feedback
control can instantaneously
reduce the temperature of the
oven whilst a feed forward loop
can operate a reject mechanism.
Swift and automated reaction to

Fig 21.16: Group of eight biscuit images serving any defects means that machine,
as a reference of eight baking time: Group 1 (8 man and material utilization are
min), Group 2 (10 min) Group 3 (12min), group all improved.
4 (14 min), Group 5 (16 min), Group 6 (18 min),
Group 7 (20 min), Group and Group 8 (24 min)
(Nashat and Abdullah).

Texture
Two main approaches for recognizing and classifying textures,
structural method: considers the texture to be composed of primitive
elements arranged in a repeated manner, statistical method,: describes a
texture in terms of the distribution of pixel values in the image. In food
products, the former approach is better suited to textures containing
relatively large primitives which can be readily identified and to textures
where the pattern is regular. For example, tea leaves and decorations on
cake.
Non-visual Imaging Techniques
When defects cannot be detected by normal visual means, the use of
other sensing methods yielded extremely better results. For example X-ray
imaging can be processed to identify the presence of foreign bodies. X-ray
imaging is also useful in the analysis of the internal texture of the food
product (Chan, 1988). Ultraviolet light Nuclear magnetic resonance: it is
possible to reveal the texture and composition of foods by using NMR.

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Fig 4.23: Inspectng decoration on a cake: (a) original image, (b) enhanced image, (c)
isolating pattern decoration, (d) pattern with a break

Fig 4.24: Crack detection: (a) original image, (b) result of morphotogical oprator to
remove creck, (c) subtracting ((a) from (b)). (d) crack is found, (e) creck
superrimposed on original image

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Fig 21.10: Group of muffin images captured from a single batch after baking: top left
is undercooked, top right is optimally cooked, bottom left is slightly overcooked, and
bottom right is a substantially overcooked batch (McConnel and Blau, 1995)

Applications
 Assessment of fruits & vegetables
 For grain classification and quality evaluation
 Meat and meat products
 Evaluation of Beverages
 Evaluation of bakery/ Snacks/ Confectionaries Products
 Processed Food Products
 Automatic Process Monitoring
 Packaging/Product Integrity
 Identification of Foreign Objects

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 Basic comparison between imaging techniques
Ultrasound X-rays Gamma-rays Ultraviolet Infrared
Low density material
Packages food wwith no with low water Similar to Xrays but
Application to Fluorecent analysis of
air content, and high content, foreign body denser materials may Thermal imaging of food
food stuff certain foodstuffs
water content detection and texture be inspected
analysis
Real time imaging Real time imaging Real time imaging Real time imaging Inspection rate approx 5
Speed
possible possible possible possible samples per sec.
At high intensities eye
Safety requirement None Considerable Considerable None
damage or skin cancer
Temperature must Image intensifier
Effect of ultraviolet on
Additional system remain constant and converts to visible Environmental temp must
Similar to X-rays the food stuff must be
requirement coupling medium is spectrum. System remain relatively constant
assessed
required must be caged in lead
Low flexible transducer only distance or type
system easily to
must be made of source may be Suitable only for certain Good flexibility and
Flexibility different imaging
specifically for material altered to change materials recalibration is simple
applications
to be imaged imaging properties
Gel or water interface Source cannot be
between transducer switched off, creating
Other advantages Safe when switched Colour vision necessary Camera examined limited
required. Transducer not storage and transport
and disadvantages off for fluorescent analysis to 0.5 degrees sensitivity
well suited to industrial complications. Beam
environ met can’t be directed

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Assessment of Fruits and Vegetables
 Narendra and Hareesh (2010) reported variety classification, defects
detection and segmentation, identification of stems and calyxes and
sugar content prediction in apples. Also investigated the use of
computer vision in sorting fresh strawberries based on size and
shape, shape characteristics of papaya, tomato color, size, shape and
abnormalities with inner quality, inspect and grading of
mushrooms, separation method for grading of potatoes, color of
lemon. Marakeby et al., (2013) also studied on fast quality
inspection of food products using computer vision. Leemans et al.
(1998) investigated the defect segmentation of ‘Golden Delicious’
apples using machine vision. Hahn and Sanchez (2000) classifies
chilli by three different width sizes, by means of a photodiode
scanner, was built they reported that the accuracy on the necrosis
detection and width classification was of 96.3 and 87%,
respectively. Arjenaki et al. (2013) sorting the tomato online based
on shape, maturity, size, and surface defects using machine vision.
Silvia (2011) reported quality assessment of blueberries by
computer vision. Sadegaonkar (2013) evaluated the quality
Inspection and grading of mangoes by computer vision & image
analysis. Arefi, (2011) The algorithm was able to identify ripened
tomato by high accuracy in different lighting conditions of
greenhouse.

Ripeness level is determined by computing the ratio of number of red

pixels to the number of pixels of the berry object: (A) original image, (B)
berry object, and (C) red pixels.

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 RGB image for (A) unripe, (B) ripe, and (C) overripe strawberries. The
cropped small square for texture analysis is depicted at the bottom left corner
of each image. (D) Extraction of GLCM at different directions (0, 45, 90, or
135 degree) and distance (D) for each pixel in the cropped square image.
For Grain Classification and Quality Evaluation
Quality inspection of cereal grains and pulses like rice, corn, wheat,
gram, beans, etc., can be performed based on size (length/width) and color
quantification of samples. Narendra and Hareesh (2010) used machine vision
to identify different variety of wheat and to discriminate wheat from non-
wheat components, developed a machine vision algorithm for corn kernel
mechanical and mould damage measurement and whiteness of corn has been
measured by on line computer vision approach, measuring the degree of
milling of rice. Arefi et al. 2011, four varieties of wheat were identified by
integrating machine vision and artificial neural network using Matlab
software. Lai et al.1986 suggested some pattern recognition techniques for
identifying and classifying cereal grains. Luo et al. 1999 used a color
machine vision system to identify damaged kernels in wheat. S. Majumdar
and D. S. Jayas 1999 analysed and Classify Bulk Samples of Cereal Grains
using Machine Vision system. M. A. Shahin and S.J Symons 2003 lentil type
identification using machine vision
Quality and Safety Evaluation of Meat, Poultry and Fish Products
Machine vision application in meat industry can be grouped as:
determination of composition, fat/muscle ratio, measurement and evaluation
of size and volume, measurement of shape parameters, quantification of the
outside or meat color, and detection of defects during quality evaluation.
Moisture and fat content of meat has been correlated with the colour (Chen
et al., 2002)
Process Monitoring and Control: Vision systems can also have great
potential in other areas
Monitoring individual processes to ensure that the correct action is taken
(cutters, depositors, mixers, etc.) also used for controlling electromechanical
devices for manufacture an assembly (lights, robot arms, etc.). Checking the
general factory environment is also very well performed by MVS.
Processed Food Products
Visual features such as color and size indicate the quality of many
prepared consumer foods. Patel et al. (2012) and Sun (2000) investigated
this in research on pizza in which pizza topping percentage and distribution

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were extracted from pizza images. To avoid the misguideness of quality
assessment by visual based human perception, computer vision has been
widely used in the assessment of confectionary products so far. For example,
if a baked product is detected to be overcooked by a vision system, feedback
control can instantaneously reduce the temperature of the oven whilst a feed
forward loop can operate a reject mechanism. Swift and automated reaction
to any defects means that machine, man and material utilization are all
improved.
Role of MVS in Packaging/Product Integrity
Used in product integrity verification to provide a comprehensive
solution for any particular packaging line monitoring. It verify the shape of a
container or the correct position of a label and its registration (Zeuch, 2002).
Pattern recognition in order to verify the correct label. No spill onto the outer
walls that the label is not torn or wrinkled, or has no folded comers, etc.
Strickland (2000) reported the use of digital imaging technology for the
automatic monitoring of dry sugar granules and powders. This system
provides particle size data to production line operators for process control
and product quality improvement.
Role of MVS in Identification of Foreign Objects
Detection of foreign objects and contaminants in food: A machine vision
system is much more effective than human observers that can automatically
detect foreign matter along the way. It continuously watches the product
stream and doesn’t become distracted. Can freeze motion on a relatively
high-speed belt, and its resolution can be specifically tailored for the
observation task
The efficiency of accuracy of machine vision system for detecting
defects is very good. Below is a table which gives summery about the
percentage of accuracy of detecting defects.
Summary of Machine Vision Applications For Food And Agricultural Produce
Product type Product Application Accuracy
Defects detection --
Classification 78%
Apple
Grading 95%
Fruits Oranges
Classification 93%
Strawberries
Quality evaluation 87%
Sorting 94%
Potato Classification 84%
vegetables Chilli Classification 87%
Lemon Classification and sorting ----

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 Classification 94%
Weed identification -----
Wheat
disease infection 97%
Corn
Cereals Size 73–90%
Rice
Whiteness ------
Wheat, barley, oats, rye
Grading 91%
Classification 98%

Advantages of Machine Vision System

It Generate precise & descriptive data, quite quick and objective. It
reducing tedious human involvement and as compared to human efficiency it
is more consistent, efficient and cost effective. Also it automating many
labour intensive processes making process easy and quick, consistent. It is a
very important non-destructive and undisturbing technique for quality
evaluation. It is quite robust and competitively priced sensing technique. It
gives permanent record, allowing further analysis later
Disadvantages of Machine Vision System
There are very few disadvantages related to MVS. One of which is
object identification being considerably more difficult in unstructured
scenes, and need of artificial lighting needed for dim or dark conditions. But
with the recent advancements in technology, these disadvantages can be
overcome.
Economic Consideration in Using Machine Vision
It saves energy, resulting from increased reliability of output, ultimately
proved to be more economical as compared to manual handling of products.
It helps the manufacturer in maintaining the competitive position. Feedback
from the vision system may be used to control the manufacturing process,
thus improving product quality and reducing waste. Faults can be recognized
and corrected quickly, saving cost of wastage. Benefits from adding
increased flexibility to the automation process. Automatic record keeping
and reporting of production trends.
Future Applications
It will be very cost effective in the future due to their low development
costs. MVS constructed, capable of performing a variety of inspection and
monitoring tasks. It would include a variety of different imaging methods
and a flexible lighting arrangement as well as robot arms for pick and place
tasks and non-visual sensors for more specific tasks. The food industry is
one sector that could reap huge benefits from machine vision techniques.
Indeed, inspection and monitoring of such products and their production

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processes will no doubt provide many interesting challenges for machine
vision systems for many years to come.
Conclusion
It has potential application for automatic food quality evaluation which
is quite useful for Indian food and beverage industry in today's quality
conscious and competitive world. A framework for machine vision system
embedded with the emerging artificial intelligent techniques for developing
advanced vision systems for better efficiency/precision. Various aspect of
food quality such as product quality and safety; classification and sorting;
and process automation have much need in future for implementation of
machine vision system.
References
1. Abdullah MZ, Abdul-Aziz S, Dos-Mohamed AM. Quality inspection of
bakery products using color based machine vision system. J Food Qual.
2000; 23:39-50.
2. Blasco J, Alexiose N, Molto E. Machine vision system for automatic
quality grading of fruit. Biosystems Engineering, London. 2003;
85(4):415-423.
3. Chan JP, Batchelor BG. Machine vision for the food industry', food
process monitoring systems. uk: Springer Science Business Media
Dordrecht, 1993, 58-100.
4. Cubero S, Aleixos N, Moltó E, Sanchis JG, Blasco J. Advances in
machine vision applications for automatic inspection and quality
evaluation of fruits and vegetables. Food Bioprocess Technol. 2011;
4:487-504.
5. Da-Wen Sun. Computer Vision Technology for Food Quality
Evaluation, London: academic press. Second edn, 2016, 1-660.
6. Leemans V, Magein H, Destain MF. On-line fruit grading according to
their external quality using machine vision, Bio systems engineering.
2002; 83(4):397-404.
7. McConnel RK, Blau HH. Color classification of non-uniform baked and
roasted foods. In: Proceedings of the Food Processing Automation
Conference IV, American Society of Agricultural Engineers, St. Joseph,
MI, USA, 1995, 40.
8. Nashat S, Abdullah MZ. Multi-class colour inspection of baked foods
featuring support vector machine and Wilk’s k analysis. J Food Eng.

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 2010; 101:370-380.
9. Patel K, Kar A, Jha SN, Khan MA. Machine vision system: a tool for
quality inspection of food and agricultural products, J Food Sci Technol.
2012; 2:123-141.
10. Saini Mandeep, Singh Jagjit, Prakash Neelam R. Analysis of wheat
grain varietees using image processing-a review. Int. J Sci. & Res. 2014;
3(6):490-495.
11. Sonka M, Hlavac V, Boyle R. Image processing, analysis, and machine
vision. Chapter 14. PWS publishing, California, 1999.
12. Brosnan T, Sun D. Inspection and grading of agricultural and food
products by computer vision systems-a review. Computers and
Electronics in Agriculture. 2002; 36:193-213.
13. Zuech N. Understanding and Applying Machine Vision, Vision Systems
International Yardley, Pennsylvania, Marcel Dekker, 2nd eds, Inc, 2000,
1-336.

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 Chapter - 6
Casein Micelle as a Nano Carrier System for
Hydrophobic Bio-Active Compounds

Authors
Priti Saha
Ph.D. Scholar, Dairy Chemistry Division, National Dairy Research
Institute, Karnal, Haryana, India
Rajesh Kumar Bajaj
Principal Scientist, Dairy Chemistry Division, National Dairy Research
Institute, Karnal, Haryana, India

Page | 113
Page | 114
 Chapter - 6
Casein Micelle as a Nano Carrier System for Hydrophobic
Bio-Active Compounds
Priti Saha and Rajesh Kumar Bajaj

Abstract
The superiority of milk protein is not only considered in terms of
nutritional point of view but also it contributes different functional properties
to dairy, food and nutraceutical applications. Casein is the main milk protein,
mainly present as micellar form, hence able to form supra molecular structure
to encapsulate different bio-active hydrophobic molecules of interest. Casein
also able to binds different bio-molecule by hydrophobic and electrostatic
interactions. Apart from reassemble caseins which able to play a superior role
as nano carrier system for bioactive hydrophobic molecules, but also some
researchers are focusing for use of native casein as encapsulate material in
encapsulation method.
Keywords: Casein micelle, nano carrier, supra-molecular structure,
hydrophobic bio-active compound, encapsulation
Introduction
Milk is a complex biological material with superior quality of proteins,
lipid and minerals. The main function of milk is to supply nutrients such as
essential amino acids required for the growth of the newborn. Bovine milk
contain near about 3.5% milk protein and casein is the main and unique protein
present in milk. Casein contains 94% of proteins and 6% of inorganic
constituents like calcium phosphate. There are functional properties of
different dairy products like solubility, foaming and emulsification depends
on the casein. Apart from those functional properties, the caseins can be
applied as a potent carrier system for hydrophobic bioactive substances to
provide food with additional health benefits.
For preservation of the beneficiary activities of difference bioactive
molecule, encapsulation is a very useful approach. Casein contributes various
functional properties to bind or encapsulate molecules, such as chemically
binding hydrophobic molecules, surface activity, aggregation, gelation, and

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interaction with other polymers. However, every food system has a specific
matrix that consists of technology (Florian and Schmitt, 2015), hence casein
is a natural nano carrier system to provide systematic protection of different
bioactive compounds specially hydrophobic bioactive molecules.
Structure of Casein Micelles
Casein is the unique protein and almost 80% of the total protein of milk
(Brunner, 1977). There are mainly four casein fractions viz: αs1, αs2, β and κ
caseins in the proportion of 3:0.8:3:1, respectively (Fox, 2003). The αs1, αs2
and β caseins are highly phosphorylated (containing phosphorus) and κ-
caseins are glycosylated (containing sugar residues). The αs1, αs2, β and κ
caseins are phosphorylated at specific serine residues and contain 8, 9-12, 5
and 1 phosphates respectively (Horne, 1998). 95% of caseins present in milk
associate together into colloidal particles, termed as casein micelles. These
micelles are intricate particles that consist of 94% casein, 6% inorganic
constituents like calcium, phosphate, magnesium, citrate (Fox, 2003).
There are mainly four casein micelle structures have been proposed,
including coat-core models, subunit (sub-micelles), internal structure models
(Phadungath, 2005) and nano-cluster models (Dalgleish, 2011). All of these
models are in consensus regarding the presence of κ-casein on the micelle
surface and CCP clusters in the core of the micelle. Recently emphasis has
been placed on nanocluster model where bridging between phosphoserine
groups of caseins by calcium and phosphate, as well as hydrophobic
interactions between caseins resulting in association.
What is Encapsulation?
Encapsulation is the process of surrounding substances (coat) within
another substance (core) yielding capsules like structure (Anonymous, 2018).
The aims of bioactive compounds for encapsulations are: enhancing entrapped
compounds stability against oxidation processes, modifying physicochemical
properties of bioactive compounds in order to better integrate these
compounds during the processing and manufacturing, controlling their
release. Thus, the bioactive component would be kept as fully functional.
Also, this technology may provide barriers between sensitive bioactive
materials and the environment, and thus, to allow taste and aroma
differentiation, mask bad tasting or smelling, stabilize food ingredients or
increase their bioavailability. Encapsulation was originally introduced in the
area of biotechnology to make production-processes more efficient as the
matrix around the cells allows for rapid and efficient separation of the
producer cells and the metabolites. Such technologies, developed

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approximately 60 years ago, are of significant interest to the pharmaceutical
sector (especially for drug and vaccine delivery). This technology is also
relevant in food industry. There is a multitude of possible benefits of
encapsulated ingredients in the food industry. Therefore encapsulation aims to
preserve stability of the bioactive compounds during processing and storage
and to prevent undesirable interactions with food matrix.
Native Caseins as an Effective Encapsulation Material for Bio-Active
Components
The structure (secondary, tertiary and quaternary) of casein makes it
unique from other protein, this unique feature of milk protein helps to interact
between themselves or with other polymers (Florian and Schmitt, 2015).
The caseins are held together in the micelle by hydrophobic interactions
and by bridging of calcium-phosphate nano clusters therefore caseins are self-
assembled and spontaneously agglomerate (Florian and Schmitt, 2015).
Studies proposed that casein molecules are able to interact with other molecule
through various interactions, therefore casein act as a nano-carrier for various
bioactive components. According to Shapira et al. (2010) that hydrophobic
and electrostatic interactions were main force to entrap bioactive hydrophobic
molecule. Those authors suggested that hydrophobic and electrostatic
interactions provide casein micelles to act as nano carriers of different
beneficial moieties like, calcium and phosphate to the neonate. Shapira et al.
(2010) also explained that due to action of hydrophobic forces between caseins
within a casein micelle, act as sponge-like structure with porous surface
consisting of internal cavities connected to each other by channels. These
unique properties help to protect and transport specific molecule
(hydrophobic) of interest.
Tavares et al. (2014) reported that the ability of “native” casein micelles,
for formation of supra molecular structure make it a well-known as bioactive
molecule carrier. In a different study Portnaya et al. (2008) suggested that
highly amphiphillic nature of β-casein, which is highly calcium sensitive phos
phoprotein. Thus it exhibit self-association under different conditions in
aqueous solution to form micelles of size 7.3 to 13.5 nm hydrophobic probes
that able to hold vitamin D, vitamin A and sucrose esters. Zana (2005)
proposed the hydrophobic interaction of β-casein in solution prevents its self-
compaction and formation of globular structures. Such block copolymers are
more effective in entrapping hydrophobic molecules. Another mechanism
proposed by Creamer et al. (1977) that β-casein moves in and out of the casein
micelles with fluctuations of temperature, hence this characteristic provides

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more opportunities for hydrophobic probes to associate with casein micelles
in the place of removed β-caseins. The quaternary structure of the casein
micelles also promotes the incorporation of hydrophobic molecules.
Re-Assembled Caseins as an Effective Encapsulation Material for Bio-
Active Components
Casein micelles can be reconstructed by mixing a source of casein
together with phosphate, calcium and citrate (Knoop, Knoop, & Wiechen,
1979) and it is called Re-assembled Caseins. Sodium caseinate is a readily
available source of casein that has been used for this purpose (Zimet et al.,
2011). O’Kennedy (2006) reported that production of sodium caseinate begins
by lowering the pH of milk to 4.6 that causes caseins to aggregate. Then the
liquid (whey) is removed from the aggregated casein, and followed by
addition of alkali such as sodium hydroxide to the aggregated casein. This
author also reported that the resulting product was readily dispersable in water.
Re-assembled micelles (rCN) are significantly affected by changes in pH,
temperature, and concentrations of the system components during the process
of formation (Aoki et al., 1996). Additionally, altering the composition of re-
assembled casein micelles has an influence on micelle properties. Modifying
the amount of αs-casein, β-casein, κ-casein, colloidal phosphate, or citrate
present in micelles induces changes in size and stability (Schmidt, 1982). Re-
assembled casein with αs and β caseins, sodium caseinates and
phosphocaseins have been extensively used to study the interactions of caseins
with hydrophobic probes.
Advantages of Casein Micelles as an Encapsulated Material
According to Tavares et al. (2014) the main advantages of the use of
casein micelles for the encapsulation of hydrophobic compounds are: i) direct
incorporation of micellated hydrophobic substances into final products occurs;
ii) high dermal penetration and intestinal release of micellated active
ingredients; iii) increased bioavailability of the active compounds consumed
as casein nanoparticles; iv) casein micelles are very stable in comparison with
emulsions or liposomes; v) the micellation of active substances opens up
completely new application possibilities for difficult or unprocessable
substances; vi) cost reduction and environmental protection.
Application of Casein for Encapsulation of Bio-Active Components
The utilization of casein micelles as a vehicle for hydrophobic bio-active
components has been investigated thoroughly in the past few years. A
systematic brief review of casein micelles for encapsulation of bio-active
components has been given below.

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Curcumin
Curcumin, a plant derived lipid soluble compound, has antioxidant,
antitumerogenic and anti-inflammatory properties (Aggarwal et al., 2007).
Curcumin was found to hydrophobically interact with non-polar regions of
αs1-casein (at pH 7.4, 27°C) at two binding sites with varying affinities.
Esmaili et al. (2011) used self-assembled -casein to encapsulate curcumin.
In the presence of 10 g /L -casein above the protein critical micelle
concentration, the solubility of curcumin was increased at least 2500 times.
Simultaneously, the cytotoxicity of curcumin against human leukemia cells
and its antioxidant activity slightly increased after encapsulation. The authors
also proposed to use -casein as a nano vehicle for this hydrophobic
components. Diacetyl curcumin was also shown to associate by hydrophobic
interactions with β-casein micelles at a single site in the core of the micelle
with relatively high affinity (Mehranfar et al., 2013).
Polyphenols
Tea polyphenols which include catechin (C), epicatechin (EC),
epigallocatechin (EGC) and epigallocatechin gallate (EGCG). Tea
polyphenols have been known to interact with proline-rich proteins like
salivary proteins and caseins (Fox, 2001). Yuksel et al., (2010) reported that
milk proteins, especially caseins, bind to green tea flavonoids by their decrease
in hydrophobicity upon binding to these hydrophobic polyphenols.
Glycosylated caseins obtained by maillard reaction were also used to
encapsulate and retain EGCG without aggregation or fusion during storage
(Xue et al., 2014). Bohin et al. (2012) used micellization of bovine -casein
at 25 °C and pH 7.0 for the encapsulation of phenolic components such as
epigallocatechin-3-gallate (EGCG).
Sahlan and Pramadewi (2012) tested low pH and a rennet hydrolysis
casein fraction from cow milk that able to encapsulate flavonoids. The specific
treatment recommended by those authors produce casein nanoparticles with a
mean diameter of 109 nm, close to the diameter of native casein micelles.
These authors also reported that these casein nanoparticles possessed an
encapsulation efficiency of about 42%, corresponding to an encapsulation of
almost 1.0 mg flavonoid per gram of casein. Casein micelles were also tested
for their ability to stabilize minerals in supplemented foods. Other plant
polyphenols have also been studied in association with their interaction with
caseins.
Resveratrol, is an another plant polyphenol those interactions with caseins
have been studied. The number of resveratrol molecules binding with αs-

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casein and β-casein were 1.2 and 1.14, respectively (Bourassa et al., 2013).
Acharya et al. (2013) reported that sodium caseinate micelles mainly by
hydrogen bonding and to a lesser extent by hydrophobic interactions able to
bind with this type of polyphenols. Genistein is another plant polyphenol with
high antioxidative activity, which is found to bind to αs-casein and β-casein
(Bourassa et al., 2013).
Iron and Calcium
Raouche et al. (2009) used a reversible acidification process by
carbonation to enhance the iron content in the casein colloidal fraction of
skimmed milk. The process of acidification by carbonation probably reduced
competition between iron and calcium ions for casein phosphoseryl sites. The
interaction between calcium and the casein phosphoserine residues is known
to be affected by both pH and ionic strength: increasing ionic strength and
decreasing pH reduce the affinity of casein for calcium.
In contrast, the interaction between iron and the casein phosphoserine
residues is less affected by pH change because iron binding involves
electrostatic interaction and coordination links. Consequently, calcium was
partially removed from the casein micelles during carbonation, allowing its
substitution by iron atoms. Supplementation with iron modified the properties
of the casein micelles by decreasing the zeta potential, hydration and thermal
stability of the casein micelles; in addition, it slowed down the enzymatic
coagulation kinetic of casein micelles (Raouche, et al., 2009). Compared to
whey proteins, caseins exhibit a higher ability to stabilize iron (Sugiarto et al.,
2009). The ability of casein micelles to encapsulate iron under magnetic field
was reported by Sangeetha and Philip (2012). They produced stable iron oxide
nanoparticle casein nano-complexes that can reversibly self-assemble after
application of an external magnetic field.
Vitamin
Mohan et al. (2013) reported that unmodified casein micelles to bind
hydrophobic molecules such as vitamin A. Florian and Schmitt (2015),
reported that casein micelles have a high affinity towards vitamin A, however
other milk protein fractions not able to bind with vitamin A. Semo et al. (2007)
applied the re-assembly of caseinate to encapsulate vitamin D2. The re-
assembly process was triggered by successive additions of a mixture of
potassium hydrogen phosphate and calcium chloride, dropped into a solution
of sodium caseinate and tri-potassium phosphate. The authors obtained casein
micelle-like structures with a diameter of about 156 nm, which allowed an
efficient encapsulation of vitamin D2 of approximately 27%. In this way,

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vitamin D2 was efficiently protected against photochemical degradation.
A similar re-assembly process was used to form casein micelle-like
structures from sodium caseinate to encapsulate vitamin D3 (Haham et al.,
2012). The obtained supra-molecular structures were further homogenized at
a pressure of 155 MPa and the protective effect against thermal and oxidative
degradation of the vitamin. Penalva et al. (2015) demonstrated that casein
nanoparticles could be used as carriers for folic acid. The nanoparticles
displayed a size of approximately 150 nm and could carry 25 μg folic acid per
mg nanoparticle. Bioavailability of folic acid was increased significantly.
Docosahexanoic Acid (DHA)
Docosahexanoic acid (DHA) is a fatty acid that binds at 3 to 4 sites of a
protein molecule in reconstituted sodium caseinate micelles with and without
addition of calcium and phosphate ions (Zimet et al., 2011). The DHA-loaded
casein nanoparticles had a diameter of approximately 300 nm and showed
good colloidal stability. They reported that oxidative stability of DHA during
cold storage (4°C/16 days) and heat treatment (74°C/20 seconds) improved by
encapsulation in re-assembled micelles. Similarly, -casein micelles were
used to encapsulate a hydrophobic drug for the treatment of gastric cancer
(Shapira et al., 2010).
Processing Conditions Affecting Hydrophobic Binding of Casein Micelles
with Bioactive Components
According to Ahmad et al. (2008) that hydrophobic and electrostatic
interactions, hydrogen bonding and presence of calcium phosphate linked to
casein molecules are the forces responsible for the structure and the stability
of casein micelle. Cleavage of these by changing different processing
conditions like heating, pressure treatment and cooling have been reported to
have impact on the binding of hydrophobic probes by caseins and casein
micelles.
Effect of High Pressure Treatments on Casein Micelles during
Encapsulation
A number of studies have investigated the effect of different types of
pressure treatments including high hydrostatic pressure (HHP) and high
pressure homogenization (HPH), ultra-high pressure homogenization
(UHPH).
HHP involves static high pressure and was found to increase the binding
and loading capacity of curcumin (Yazdi and Corredig, 2012) and vitamin D2
(Menendez-Aguirre et al., 2011) in casein micelles. The rearrangements

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occurring within the casein micelles on application of pressure (>200 MPa)
favor binding to hydrophobic probes. Calcium and phosphate ions are also
known to contribute to this high pressure effect.
HP treatment at pressures <200 MPa has little effect on casein micelle
size, but treatment at >250 MPa causes considerable disruption of casein
micelles in raw skim milk (Needs et al. 2000) and reconstituted skim milk
powder (RSMP; Gaucheron et al., 1997). HP induced disruption of casein
micelles causes changes in the optical properties of milk, such as lightness and
turbidity (Johnston et al., 1992). It has been suggested that disruption of casein
micelles under pressure is due to solubilisation of colloidal calcium phosphate,
as well as dissociation of hydrophobic and electrostatic interaction (Needs et
al., 2000).
Apart from the spontaneous binding of α-tocopherol acetate to
phosphocasein dispersion in water (1:1 ratio), more binding was observed at
HPH pressures ≥ 200 MPa. This was also obvious from the 1.5 times higher
binding efficiency at 250 MPa and 3 times higher at 300 MPa compared to
control without treatment (Chevalier-Lucia et al., 2011). This is attributed to
the changes in micelle assembly during HPH by the dissociation and re-
association to form smaller compact structures that spontaneously entrap
hydrophobic molecules during its formation. Similar observations were
reported by another group indicating a 1.5 to 2 fold increase in the binding
efficiency of curcumin to native phosphocasein suspended in simulated milk
ultrafiltrate (SMUF) at 300 MPa HPH pressure (Benzaria et al., 2013).
Pressure below 200 MPa (Semo et al., 2007) and presence of small quantities
of ethanol (Chevalier-Lucia et al., 2011) did not modify or change the size of
the casein micelles. The increment of binding efficiency is due to the increase
in hydrophobic interactions within the casein micelles. This is further proven
by the increase in binding of α-tocopherol acetate to casein micelles at 34°C
compared to that at 14°C. The hydrophobic interactions are known to
strengthen at higher temperatures and this can play a role in the changes
occurring in casein micelles during HPH which induces a short heating period.
Ultra-high pressure (UHP) involves the application of pressures from 100
to 1000 MPa. UHP utilization in food products was initiated during the early
1980s and is a nonthermal pasteurization method. Sandra and Dalgleish (2005)
reported that during ultra-high pressure (more than 200MPa) homogenization
(UHPH) processing casein micelles size reduced and disruption of casein
occurred. The UHPH caused some of the micellar k- and S caseins to be
more soluble, possibly because of disruption of hydrophobic interactions, or
the effects of mechanical shear, on the proteins nearest the micellar surface.

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These authors also reported that UHPH was able to modify casein micelle
structural properties.
Effect of Ultra-Sonication on Casein Micelles during Encapsulation
Ultra sound has been shown to be a potential alternative to thermal
treatment for reducing microbial activity (Piyasena et al., 2003), an effective
method for homogenizing milk fat, and a means of improving the heat stability
of whey protein concentrates. Few studies found on the effect of ultra-sonic
sound on casein micelles. Madadlou et al. (2009) reported that the average
size of reassembled casein micelles could be reduced by exposure to
ultrasound (35 kHz frequency) for 6 h, provided the pH was above 8. In
another study involving true CN micelles, a decrease in particle size resulting
from ultrasonication was observed (Nguyen and Anema, 2010). Very recently
Chandrapala et al. (2012) found no effect on the intregrity of casein micelles
applying ultrasound 20 kHz frequency for 60 min.
Conclusion
Caseins, the unique phosphoprotein and almost 80% of the total protein
of milk. In milk, about 90% of the casein associated with different minerals as
a colloidal particle, hence termed as casein micelles. The structure of casein
makes it unique from other protein, this unique feature of milk protein helps
to interact between themselves or with other polymers and act as natural
carrier for bioactive hydrophobic molecules like curcumin, polyphenols,
vitamin, DHA etc. The supra-molecular structure, hydrophobic and electro-
static interaction etc. helps to bind casein with different bioactive compounds.
Most of these studies of encapsulation efficiency of caseins were mainly based
on cow milk but till date this type of studies are limited on buffalo milk. Owing
that the different compositional changes of buffalo milk casein more research
are needed so that buffalo milk could be used as a nano carrier for bioactive
compounds.
References
1. Acharya DP, Sanguansri L, Augustin MA. Binding of resveratrol with
sodium caseinate in aqueous solutions. Food chemistry. 2013;
141(2):1050-1054.
2. Aggarwal BB, Sundaram C, Malani N, Ichikawa H. Curcumin: the Indian
solid gold. In The molecular targets and therapeutic uses of curcumin in
health and disease. Springer, Boston, MA, 2007, 1-75.
3. Ahmad SI, Gaucher F, Rousseau E, Beaucher M, Piot J, Grongnet F, et
al. Effects of acidification on physicochemical characteristics of buffalo

Page | 123
 milk: A comparison with cow’s milk. Food Chemistry. 2008; 106:11-17.
4. Anonymous. www.google.co.in.ENCAPCULATION. (Last access 1st
June 2018), 2018.
5. Aoki T, Uehara T, Yonemasu A, El-Din MZ. Response surface analyses
of the effects of calcium and phosphate on the formation and properties
of casein micelles in artificial micelle systems. Journal of agricultural and
food chemistry. 1996; 44(5):1230-1234.
6. Benzaria A, Maresca M, Taieb N, Dumay E. Interaction of curcumin with
phosphocasein micelles processed or not by dynamic high-pressure. Food
chemistry. 2013; 138(4):2327-2337.
7. Bohin MC, Vincken JP, Van Der Hijden HT, Gruppen H. Efficacy of food
proteins as carriers for flavonoids. Journal of agricultural and food
chemistry. 2012; 60(16):4136-4143.
8. Bourassa P, Bariyanga J, Tajmir-Riahi HA. Binding sites of resveratrol,
genistein, and curcumin with milk α-and β-caseins. The Journal of
Physical Chemistry B. 2013; 117(5):1287-1295.
9. Brunner JR. Milk Proteins, in Food Proteins. (Eds Whitaker JR. and
Tannenbaum SR.) Westport, Conn. Avi Publishing Co. Inc. Google
Scholar, 1977, 175-208.
10. Chandrapala J, Martin GJO, Zisu B, Kentish SE, Ashokkumar M. The
effect of ultrasound on casein micelle integrity. Journal of dairy science.
2012; 95(12):6882-6890.
11. Chevalier-Lucia D, Blayo C, Gràcia-Julià A, Picart-Palmade L, Dumay
E. Processing of phosphocasein dispersions by dynamic high pressure:
Effects on the dispersion physico-chemical characteristics and the binding
of α-tocopherol acetate to casein micelles. Innovative food science &
emerging technologies. 2011; 12(4):416-425.
12. Creamer LK, Berry GP, Mills OE. Study of the dissociation of beta casein
from the bovine casein micelle at low temperature. New Zealand Journal
of Dairy Science and Technology, 1977.
13. Dalgleish DG. On the structural models of bovine casein micelles-review
and possible improvements. Soft matter. 2011; 7(6):2265-2272.
14. Esmaili M, Ghaffari SM, Moosavi-Movahedi Z, Atri MS, Sharifizadeh
A, Farhadi M et al. Beta casein-micelle as a nano vehicle for solubility
enhancement of curcumin; food industry application. LWT-food science
and technology. 2011; 44(10):2166-2172.

Page | 124
15. Fox PF. Milk proteins as food ingredients. International Journal of Dairy
Technology. 2001; 54(2):41-55.
16. Fox PF. Milk proteins: general and historical aspects. In Advanced Dairy
Chemistry-1 Proteins Springer, Boston, MA, 2003, 1-48.
17. Florian K, Schmitt JJ. Review: Milk Proteins as Nanocarrier Systems for
Hydrophobic Nutraceuticals. Journal of Food Science. 2015; 80:2361-
2367.
18. Gaucheron F, Famelart MH, Mariette F, Raulot K, Michela F, Le Graeta
Y. Combined effects of temperature and high-pressure treatments on
physicochemical characteristics of skim milk. Food Chemistry. 1997;
59(3):439-447.
19. Haham M, Ish-Shalom S, Nodelman M, Duek I, Segal E, Kustanovich M
et al. Stability and bioavailability of vitamin D nanoencapsulated in
casein micelles. Food & function. 2012; 3(7):737-744.
20. Holt C, De Kruif CG, Tuinier R, Timmins PA. Substructure of bovine
casein micelles by small-angle X-ray and neutron scattering. Colloids and
Surfaces A: Physicochemical and Engineering Aspects. 2003; 213(2-
3):275-284.
21. Horne DS. Casein Interactions: Casting Light on the Black Boxes, the
Structure in Dairy Products, 1998.
22. Johnston DE, Austin BA, Murphy RJ. Effects of high hydrostatic pressure
on milk. Milchwissenschaft. 1992; 47:760-763.
23. Knoop AM, Knoop E, Wiechen A. Sub-structure of synthetic casein
micelles. Journal of Dairy Research. 1979; 46(02):347-350.
24. Madadlou A, Mousavi ME, Emam-djomeh Z, Ehsani M, Sheehan D.
Sonodisruption of re-assembled casein micelles at different pH
values. Ultrasonics Sonochemistry. 2009; 16(5):644-648.
25. Mehranfar F, Bordbar AK, Fani N, Keyhanfar M. Binding analysis for
interaction of diacetylcurcumin with β-casein nanoparticles by using
fluorescence spectroscopy and molecular docking calculations.
Spectrochimica Acta Part A: Molecular and Bio molecular Spectroscopy.
2013; 115:629-635.
26. Menendez-Aguirre O, Stuetz W, Grune T, Kessler A, Weiss J, Hinrichs
J. High pressure-assisted encapsulation of vitamin D2 in reassembled
casein micelles. High Pressure Research. 2011; 31(1):265-274.

Page | 125
27. Mohan MS, Jurat-Fuentes JL, Harte F. Binding of vitamin A by casein
micelles in commercial skim milk. Journal of Dairy Science. 2013;
96(2):790-798.
28. Needs EC, Stenning RA, Gill AL, Ferragut V, Rich GT. High-pressure
treatment of milk: effects on casein micelle structure and on enzymic
coagulation. Journal of Dairy Research. 2000; 67(1):31-42.
29. Nguyen NH, Anema SG. Effect of ultrasonication on the properties of
skim milk used in the formation of acid gels. Innovative food science &
emerging technologies. 2010; 11(4):616-622.
30. O’Kennedy BT, Mounsey JS, Murphy F, Duggan E, Kelly PM. Factors
affecting the acid gelation of sodium caseinate. International dairy
journal, 2006; 16(10):1132-1141.
31. Penalva R, Esparza I, Agüeros M, Gonzalez-Navarro CJ, Gonzalez-
Ferrero C, Irache JM. Casein nanoparticles as carriers for the oral delivery
of folic acid. Food hydrocolloids. 2015; 44:399-406.
32. Phadungath C. Casein micelle structure: a concise
review. Songklanakarin Journal of Science and Technology. 2005;
27(1):201-212.
33. Piyasena P, Mohareb E, McKellar RC. Inactivation of microbes using
ultrasound: a review. International journal of food microbiology. 2003;
87(3):207-216.
34. Portnaya I, Ben-Shoshan E, Cogan U, Khalfin R, Fass D, Ramon O et al.
Self-assembly of bovine β-casein below the isoelectric pH. Journal of
agricultural and food chemistry. 2008; 56(6):2192-2198.
35. Raouche S, Dobenesque M, Bot A, Lagaude A, Marchesseau S. Casein
micelles as a vehicle for iron fortification of foods. European Food
Research and Technology. 2009; 229(6):929-935.
36. Sahlan MUHAMAD, Pramadewi INDRIANTI. Nanoencapsulation of the
flavonoids isolated from Phaleria macrocarpa leaf by casein
micelle. International Journal of Pharmaceutical Bio Science. 2012;
3:472-478.
37. Sandra S, Dalgleish DG. Effects of ultra-high-pressure homogenization
and heating on structural properties of casein micelles in reconstituted
skim milk powder. International Dairy Journal, 2005; 15(11):1095-1104.
38. Sangeetha J, Philip J. The interaction, stability and response to an external
stimulus of iron oxide nanoparticle-casein nanocomplexes. Colloids and

Page | 126
 Surfaces A: Physicochemical and Engineering Aspects. 2012; 406:52-60.
39. Schmidt DG. Association of caseins and casein micelle structure.
In Developments in Dairy Chemistry-1-Proteins (Ed. Fox, P. F.), in
press, London: Applied Science Publishers Ltd Google Scholar, 1982.
40. Semo E, Kesselman E, Danino D, Livney YD. Casein micelle as a natural
nano-capsular vehicle for nutraceuticals. Food hydrocolloids. 2007; 21(5-
6):936-942.
41. Shapira A, Assaraf YG, Livney YD. Beta-casein nanovehicles for oral
delivery of chemotherapeutic drugs. Nanomedicine: Nanotechnology,
Biology and Medicine. 2010; 6(1):119-126.
42. Sneharani AH, Singh SA, Appu Rao AG. Interaction of αS1-casein with
curcumin and its biological implications. Journal of agricultural and food
chemistry. 2009; 57(21):10386-10391.
43. Sugiarto M, Ye A, Singh H. Characterisation of binding of iron to sodium
caseinate and whey protein isolate. Food Chemistry. 2009; 114(3):1007-
1013.
44. Swaisgood HE. Chemistry of the caseins. In advanced dairy chemistry-1
Proteins. Springer, Boston, MA, 2003, 139-201.
45. Tavares GM, Croguennec T, Carvalho AF, Bouhallab S. Milk proteins as
encapsulation devices and delivery vehicles: applications and
trends. Trends in food science & technology. 2014; 37(1):5-20.
46. Walstra P. On the stability of casein micelles1. Journal of Dairy Science.
1990; 73(8):1965-1979.
47. Xue J, Tan C, Zhang X, Feng B, Xia S. Fabrication of epigallocatechin-
3-gallate nanocarrier based on glycosylated casein: Stability and
interaction mechanism. Journal of agricultural and food chemistry. 2014;
62(20):4677-4684.
48. Yazdi SR, Corredig M. Heating of milk alters the binding of curcumin to
casein micelles. A fluorescence spectroscopy study. Food chemistry.
2012; 132(3):1143-1149.
49. Yuksel Z, Avci E, Erdem YK. Characterization of binding interactions
between green tea flavanoids and milk proteins. Food Chemistry. 2010;
121(2):450-456.
50. Zana R. Dynamics in micellar solutions of amphiphilic block copolymers.
Dynamics of Surfactant Self assemblies. New York: CRC Press, Taylor

Page | 127
 & Francis Group, 2005, 161-231.
51. Zimet P, Rosenberg D, Livney YD. Re-assembled casein micelles and
casein nanoparticles as nano-vehicles for ω-3 polyunsaturated fatty
acids. Food Hydrocolloids. 2011; 25(5):1270-1276.

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 Chapter - 7
Novel Protein Foods: Alternative Sources of Protein for
Human Consumption

Authors
Neelesh Kumar Maurya
Institute of Home Science, Bundelkhand University, Jhansi, Uttar
Pradesh, India
Radha Kushwaha
Centre of Food Technology, University of Allahabad, Allahabad, Uttar
Pradesh, India

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Page | 130
 Chapter - 7
Novel Protein Foods: Alternative Sources of Protein for
Human Consumption
Neelesh Kumar Maurya and Radha Kushwaha

Abstract
Proteins are a major macronutrient of the human diet needed for
survival. Its crucial function in nourishment is to provide sufficient amounts
of amino acids to the body as these amino acids work as anaplerotic
substrates in the building block of the body. As the growth of population
increases continuously, the demand for protein also increases over the next
decades, and it is very important to search alternative sources of protein for
human consumption. The present food industrialists aim to develop a
cheaper, protein rich that have almost essential amino acids with highest
bioavailability and more convenient food products. Single cell protein from
algae and fungi, leaf protein extract and many insects could be an alternative
of protein, because they have almost all the essential amino acids required
for the human body for the survival.
Introduction
Proteins are a vital nutrient element of the human diet needed for
survival. Its crucial function in nourishment is to provide sufficient amounts
of amino acids to the body as these amino acids work as the building block
of the body. The quality of protein also known as the nutritive value of the
product, which is mainly, depends on its amino acid content and its
physiological application after digestion, absorption. Metabolism of amino
acids is determined by the number of amino acids used in protein synthesis.
Accessibility of amino acids varies with the protein source, processing
methods, and interaction with other components of the food like fat, minerals
etc.
As the growth of population increases continuously, the demand for
protein also increases over the next decades, and it is very important to
search alternative sources of protein for human consumption. Proteins that
are not currently used as animal feed, and proteins that are currently used as

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animal feed modified and improved for human consumption. The problem of
obtaining sufficient protein is further compounded because the mean daily
protein requirement for consumers also increased. Major conventional
sources of protein in the diet in developing and developed countries are
cereals, meat, Pulses, milk and dairy, fish, seafood, oil crops, vegetables,
starchy roots, eggs, offals, fruit; but they will not be sufficient enough in
next decades. So it’s very important to search the alternative sources of good
quality of protein for human consumption.
Some novel protein sources like insects, algae, duckweed, microbial
protein, my coprotein, leaf protein, and rapeseed are expected to enter in
food market as replacers for animal‐derived proteins. A wide variety of
vegetarian alternatives is also available on the market like seitan, tofu, soy
meat, tempeh, Quorn and meatless based on lupins, canola. However, food
safety aspects of these alternative sources of protein are not well‐known.
The aspire of this article is to review the state of the art on the safety of
major novel protein sources for food production, in particular insects,
rapeseed, duckweed, microalgae, and seaweed.
Various Sources of Protein are as Follows

1. Mycoprotein
Mycoprotein is a form of single-cell protein, also known as fungal
protein. Fungi have been influencing human affairs for thousands of years,
whether as a direct food source, as a medicine, or in a food. Fungi have
provided food for man, primarily in the form of fruit bodies of
basidiomycetes and/or ascomycetes. The process of bringing mycoprotein
into the marketplace began in 1985. The nutritional value of mycoprotein, is
found to be comparable with eggs in amino acid composition, but contained
no cholesterol and had a substantial fiber content. Mushrooms are one of the
most common fungi which can be used as the source of protein as they have
the high protein content, usually around 20-30% by dry weight.

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 Table 1: Protein content of mushrooms (Kalac, 2009)

S. No Species of Mushrooms Protein Content (%)

1. Button Mushrooms 56.3
2. Porcini Mushrooms 26.5
3. Portobello Mushrooms 25
4. Enoki Mushrooms 22
5. Oyster Mushrooms 19
6. Hericium erinaceus 22
7. Shiitake Mushrooms 28
9. Morchella esculenta, 32.7

Another fungus Fusarium venenatum strain PTA-2684 also used as

protein source. It has 42% protein. Quorn’M is a meat substitute product
originating in the UK, which is composed of Fusarium with mixed with egg
albumen and vegan is the product of dried fungus with potato protein where
egg albumen and potato work as binding agents. The PDCAAS for
mycoprotein is 0.99, better than beef at 0.92.

Fig 1: Mushroom varieties

The essential amino acid of mycoprotein and other foods are given
below in table 2.

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 Table 2: Essential Amino Acid Content of mycoprotein and other protein foods (g
amino acids per 100g)

Essential Amino Cow’s Soy

Mycoprotein Egg Beef Peanuts Wheat
Acids Milk Isolate
Histidine 0.35 0.09 0.30 0.66 0.6 0.65 0.32
Isoleucine 0.52 0.20 0.68 0.87 1.1 0.91 0.53
Leucine 0.86 0.32 1.10 1.53 1.8 1.67 0.93
Lysine 0.83 0.26 0.90 1.60 1.4 0.92 0.30
Methionine 0.21 0.08 0.39 0.50 0.3 0.32 0.22
Phenylalanine 0.49 0.16 0.66 0.76 1.1 1.30 0.68
Tryptophan 0.16 0.05 0.16 0.22 0.3 0.25 0.18
Threonine 0.55 0.15 0.60 0.84 0.8 0.88 0.37
Valine 0.62 0.22 0.76 0.94 1.1 1.08 0.59

2. Single Cell Protein

The shortage of protein-rich food and global survival of millions of
people have forced researchers to look for alternative sources of protein that
can replace traditional and other expensive sources like soymeal or fishmeal.
Therefore, the center of attention has shifted towards microbes as food
sources for consumption as single cell protein (SCP). The term ‘single cell
protein’ was given in 1968 at the Massachusetts Institute of Technology by
replacing microbial protein and petroprotein (Mateles and Tannenbaum,
1968; Tannenbaum and Wang, 1975). Currently, SCP is being produced
from many species of microorganisms like algae, fungi, and bacteria. It is
suitable to use fungi and bacteria for the production of SCP when grown on
economical waste products. Their quick growth and high protein content
have made them the major sources of SCP.
2.1 Sources
Algae, fungi, and bacteria are the main sources of SCP. They are as
follows-
Algae as the Source of SCP: Caulerpa rocemosa, Chlorella salina CU-
1(28), Chlorella spp., Chlorella spp. (M109, M121, M122, M138, M150),
Dunaliella Laminaria, Sargassum, Spirulina maxima, Spirulina spp.,
Gracilaria changgi (Fig 2).
Algae also contain 40–60% protein, 7% mineral salts, chlorophyll, bile
pigments, and fiber.

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Fig 2: A. Chlorella; B. Laminaria; C. Sargassum; D. Spirulina; E. Brevibacterium;
F. Pseudomonas fluorescens: G. Fusarium.

Fungi as Source of SCP: Aspergillus niger, Sporotrichum

pulverulentum, Candida krusei SO1& Saccharomyces spp., Candida
tropicalis ceppo, Chrysonilia sitophilia, Fusarium graminearum,
Paecilomyces variolii, Penicillium cyclopium, Penicillium roqueforti,
Penicillium camemberti, Pichia pastoris, Saccharomyces cereviceae,
Schwanniomyces occidentalis, Scytalidium acidophilum, Trichoderma
reesei, Kluyveromyces marxianus, White rot, Yeast.
Bacteria as the Source of SCP: Brevibacterium spp., Cellulomonas
spp., Methanomonas methanica, Methylophilus methanotrophus,
Pseudomonas fluorescens, Rhodopseudomonas gelatinosus, Streptomyces
spp.
Bacterial SCP has high protein content and certain essential amino
acids. The crude protein content is around 80% of the total dry weight. The
essential amino acid compositions of different Lactobacillus bacteria are
comparable to the FAO reference protein and SCP from other sources.
Although algae are very nutritious there are some limitations which are
somewhat unfit for human consumption. The most important is the algal cell
wall which cannot be digested by humans because of lack of the cellulase
enzyme. To overcome this problem algal wall must be digested before the

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final product is eaten. The fungus produces many allergic reactions, diseases,
and liver cancer in humans as they have mycotoxins in certain fungal species
especially Aspergillus parasiticus and A. flavus. Hence, it is a requirement
that mycotoxins are eliminated before fungal SCP is consumed.
Table 3: Comparison of SCP production from algae, fungi, and bacteria (Singh,
1998.)

Fungi
Parameter Algae Bacteria Fungi (Filamentous)
(Yeast)
Lower than bacteria and
Growth rate Low Highest Quite high
yeast
Light, carbon Wide range
Mostly
Substrate dioxide or Wide range except carbon
Lignocellulosics
inorganic sample dioxide
pH range Up to 11 5–7 5–7 3–8
Cultivation Ponds, Bioreactors Bioreactors Bioreactors Bioreactors
Contamination Precautions Least if pH is less
High and serious Low
risks needed than 5
S-containing
Low Deficient Deficient Low
amino acids
Nucleic acid
– Required Required Required
removal
Endotoxins from
Mycotoxins in many
Toxin – gram-negative –
Species
bacteria

The chemical composition of any SCP product must be characterized

clearly in terms of percentage protein, type of amino acids, nucleic acid,
lipids, fats, toxins, and vitamins.
3. Leaf Protein Extract
Now a day’s researcher processed study to extract protein from various
fresh leaves of plants. The protein content of the dried product from the
sources is about 50-70%. The major problem face during leaf protein
production its keeping quality, shelf life, acceptability, and taste. the
concentrated form of the proteins extracted from the leaves of plants known
as Leaf protein concentrate (LPC). Leaf protein concentrate (LPC) are very
nutritious food made by separating mechanically indigestible fiber and
soluble anti-nutrients from much of the protein, vitamins, and minerals in
certain fresh green plant leaves. They are rich in β-carotene, iron, and high-
quality protein so it will be very effective in fighting malnutrition. The
amino acid profile of leaf protein concentrate indicates that it is nutritionally
superior to most cereal and legume seed proteins; it can also compare
positively with animal proteins except egg and milk.

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 Protein content of LPC was found to be higher in the leaves in
Achyranthes aspera (19.9%), Aerva javanica (31.9%), Pulicaria angustifolia
(21.2%), Sesbania sesban (15.2%), Solanum nigrum (24.6%), Tephrosia
purpurea (32.3%), Withania somnifera (22.9%). LPCs are also rich in
polyphenols. Ribulose-1, 5-bisphosphate carboxylase oxygenase, is an
enzyme generally known as RuBisCO. It catalyzes the first major step of
carbon fixation, for the production of glucose with atmospheric carbon
dioxide, water, and sunlight. RuBisCO can make up to 50% of the total
amount of the protein fraction of green plants. It is found inside the mobile
phase of the chloroplast.
4. Insects
Edible insects contain high-quality protein, vitamins and amino acids for
humans. Insects, a traditional food in many parts of the world, are highly
nutritious and especially rich in proteins and thus represent a potential food
and protein source. Insects, a traditional food in many parts of the world, are
highly nutritious and especially rich in proteins and these represent a
potential food and protein source. The ethnic people of India also consume
insects as food, eating insects known as entomophagy. Figure 3 shows
various insects which can be used as the source of protein and their protein
content in table 4.
In this context, edible insects are an important and promising food
resource to be developed in the near future. Indeed, even though insects are
considered a nonconventional food and often a low-prestige food by
occidental cultures, they form an integral part of the daily diet of many
ethnic groups in the world. Insects are eaten in their different developmental
stages, including eggs, larvae and pupae, and even the adults. However, most
insects are eaten in immature stages when the exoskeleton is reduced and
soft. Insects generally have a crunchy consistency, often enhanced by frying
or roasting, so when they are ingested salivation and mastication occurs
which produces a sensation of satisfaction. Insects are cooked, roasted, fried
or boiled and they are generally eaten in combination with other food items.
They can preserve and store during periods of abundance, when insect
collection occurs, them for later consumption.

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Fig 3: A. Macrotermes bellicosus; B. Macrotermes notalensis; C. Brachytrypes spp.;
D. Zonocerus variegates; E. Analeptes trifasciata; F. Anaphe infracta; G. Anaphe
recticulata; H. Anaphe venata; I. Cirina forda; J. Apis mellifera; K. Oryctes boas; L.
Rhynchophorus phoenicis; M. Blattodea; N. Leafcutter ants; O. Honeypot ants; P.
Dung Beetle; Q. Waxworm; R. Termites (Redford & Dorea 1984)
From total nitrogen content of insects, approximately 93% is unbound.
The biological quality of insect protein is good, having chemical scores from
amino acid profiles, compared to the WHO/FAO/UNU pattern, that range
from 10% to 96%. The calcium, iron, and potassium contents are higher than
those of most food products of vegetable and animal origin. Insects are also
rich in vitamins of the B group such as niacin, riboflavin, and thiamin. Thus,
insects have the potential to improve people's diet by significantly
contributing to their protein intake and by reducing deficiencies of minerals
and vitamins. Hemiptera (true bugs) being low in isoleucine, lysine,
phenylalanine + tyrosine, and valine and the order Diptera (flies) being low
in leucine and cysteine, in general, most edible insects satisfactorily provide
the essential amino acids required for human nutrition according to the
WHO. High amino acid values have been found for phenylalanine + tyrosine
in some species, and some insects are rich in tryptophan, lysine, and
threonine.
Table 4: Protein content of insects (Banjo et al., 2006)

S. No. Insects Protein content (% dry basis)

1. Macrotermes bellicosus 20.4
2. Macrotermes notalensis 22.1
3. Brachytrypes spp. 6.25
4. Cytacanthacris aeruginosus 12.1

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 unicolor
5. Zonocerus variegates 26.8
6. Analeptes trifasciata 29.62
7. Anaphe infracta 20.0
8. Anaphe recticulata 23.0
9. Anaphe venata 25.7
10. Cirina forda 20.2
11. Apis mellifera 21.0
12. Oryctes boas 26.0
13. Rhynchophorus phoenicis 28.42
14. Cockroaches (Blattodea) 57.30
15. Beetles (Coleoptera) 40.69
16. Flies (Diptera) 49.48
17. Beetles (Hemiptera) 48.33
18. Bees, wasps, ants (Hymenoptera) 46.47
19. Termites (Isoptera) 35.34
20. Caterpillars (Lepidoptera) 45.38
21. Dragonflies (Odonata) 55.23
22. Grashoppers, locusts, crickets 61.32
(Orthoptera

This review showed that several insects which are known as pests also
having high nutritional qualities. All the non-toxic insects are indeed a good
source of protein and other nutrients for human consumption, therefore,
should be encouraged. Insects are already being used traditionally in the diet
in the different culture. They can be reared for larger production, therefore;
their nutritional value can be utilized in various products and could be
industrialized.
Conclusion
There are various non-general sources of protein and other nutrients
which are being used by the certain group of culture and community like
fungi, algae, leaf protein extract and the large variety of insects. These are
providing the high quality of proteins and supplements (minerals and
vitamins) even when dried. Therefore it could be the best area for the
production of proteins and minerals that could be incorporated into various
products to make it more nutritious. So, this is recommended to explore
these areas for the better results and make it possible reduce the malnutrition

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among the consumers. The major problem for the use of these as food
include legal restrictions and it is supposed to lack consumer acceptance can
also a barrier. Both issues could be defeat with the help of scientific
exploration on beneficial properties and also on safety risks as well as the
development of effective safety measures. Production of high-quality non
general protein source based food products also requires studies on the
impact of farming conditions and processing methods on nutritional quality
and functional properties.
References
1. Banjo AD, Lawal OA, Songonuga EA. The nutritional value of fourteen
species of edible insects in south western Nigeria. African Journal of
Biotechnology. 2006; 5(3):298-301.
2. Betschart AA, Kinsella JE. Influence of storage on composition, amino
acid content, and solubility of soybean leaf protein concentrate. Journal
of agricultural and food chemistry. 1974; 22(1):116-123.
3. Bhalla TC, Sharma NN, Sharma M. Production of metabolites,
industrial enzymes, amino acid, organic acids, antibiotics, vitamins and
single cell proteins, 2007.
4. Boland MJ, Rae AN, Vereijken JM, Meuwissen MP, Fischer AR, Van
Boekel MA et al. The future supply of animal-derived protein for human
consumption. Trends in Food Science & Technology. 2013; 29(1):62-
73.
5. Chanda S, Chakrabarti S. Plant origin liquid waste: a resource for single
cell protein production yeast, 1996.
6. Erdman MD, Bergen WG, Reddy CA. Amino acid profiles and
presumptive nutritional assessment of single-cell protein from certain
lactobacilli. Applied and environmental Microbiology. 1977; 33(4):901-
905.
7. FAO. Vitamin and mineral requirements in human nutrition: Report of a
joint FAO/WHO expert consultation, Bangkok, Thailand, 1998, 2004,
21-30. http://www.who.int/nutrition/publications/micronutrients/924154
6123/en/.
8. Kalač P. Chemical composition and nutritional value of European
species of wild growing mushrooms: A review. Food chemistry. 2009;
113(1):9-16.
9. Kovač DJ, Simeunović JB, Babić OB, Mišan AČ, Milovanović IL.

Page | 140
 Algae in food and feed. Food Feed Res. 2013; 40(1):21-31.
10. Matassa S, Boon N, Pikaar I, Verstraete W. Microbial protein: future
sustainable food supply route with low environmental
footprint. Microbial biotechnology. 2016; 9(5):568-575.
11. Maurya NK, Arya P. Amaranthus grain nutritional benefits: A review.
Journal of Pharmacognosy and Phytochemistry. 2018; 7(2):2258-2262.
12. Miller SA, Dwyer JT. Evaluating the safety and nutritional value of
mycoprotein. Food technology. 2001; 55(7):42-47.
13. Möller A. L-tryptophan production from anthranilic acid by amino acid
auxotrophic mutants of Candida utilis. Process biochemistry. 1994;
29(7):521-527.
14. Nasseri AT, Rasoul-Amini S, Morowvat MH, Ghasemi Y. Single cell
protein: production and process. American Journal of food technology.
2011; 6(2):103-116.
15. Ramos‐Elorduy J. Insects: a sustainable source of food?. Ecology of
food and nutrition. 1997; 36(2-4):247-276.
16. Rathore M. Leaf protein concentrate as food supplement from arid zone
plants. Journal of dietary supplements. 2010; 7(2):97-103.
17. Ramos-Elorduy J. Los Insectos Como Fuente de Proteínas en el Futuro.
Edit. Limusa 2 edic, 1982, 149.
18. Ramos-Elorduy J, Morales J, Pino J, Nieto Z, Contenido de Tiamina.
Riboflavina y Niacina en algunos insectos comestibles de México. Rev.
Tecn. Alim. 1988b; 22:76-81.
19. Redford KH, Dorea JG. The nutritional value of invertebrates with
emphasis on ants and termites as food for mammals. Journal of Zoology.
1984; 203(3):385-395.
20. Rumpold BA, Schlüter OK. Nutritional composition and safety aspects
of edible insects. Molecular nutrition & food research. 2013; 57(5):802-
823.
21. Rumpold BA, Schlüter O. Insect-based protein sources and their
potential for human consumption: Nutritional composition and
processing. Animal Frontiers. 2015; 5(2):20-24.
22. Singh BD. Biotechnology. New Delhi: Kalyani Publishers, 1998, 498-
510.
23. Spolaore P, Joannis-Cassan C, Duran E, Isambert A. Commercial

Page | 141
 applications of microalgae. Journal of bioscience and bioengineering.
2006; 101(2):87-96.
24. Tee RD, Gordon DJ, Welch JA, Taylor AN. Investigation of possible
adverse allergic reactions to mycoprotein (‘Quorn’). Clinical &
Experimental Allergy. 1993; 23(4):257-260.
25. Trehan K. Biotechnology. New Delhi: Wiley Eastern Limited, 1993, 79-
88.
26. Vashista BR. Botany for degree students-Algae. New Delhi: S. Chand
and Co. Ltd, 1989, 503-14.
27. WHO. Protein and amino acid requirements in human nutrition: Report
of a joint FAO/ WHO/ UNU expert consultation. WHO technical report
series, 2007. http://whqlibdoc.who.int/trs/who_trs_935_eng.pdf.
28. Wiebe MG. Quorn TM Myco-protein-Overview of a successful fungal
product. Mycologist. 2004; 18(1):17-20.

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 Chapter - 8
Potential of Functional Foods in Human Nutrition

Authors
Mamta Thakur
Ph.D. Research Scholar, Department of Food Engineering and
Technology, Sant Longowal Institute of Engineering and
Technology, Longowal, Punjab, India.
H.W. Deshpande
Head and Professor, Department of Food and Industrial Microbiology,
College of Food Technology, Vasantrao Naik Marathwada Krishi
Vidyapeeth, Parbhani, Maharashtra, India.

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Page | 144
 Chapter - 8
Potential of Functional Foods in Human Nutrition
Mamta Thakur and H.W. Deshpande

Abstract
Changing food habits, lifestyle and increasing disposable income of
consumers these days strongly influence the food demand shifting more
towards the need of healthy nutrients enriched foods. The concept of
functional foods is not recent and they must be related to the health benefits
in addition to providing the basic nutrition. Numerous functional or bioactive
components found in the food include carotenoids, dietary fibre, flavonoids,
isothiocyanates, phenolic acids, phytoestrogens, plant sterols,
probiotics/prebiotics/synbiotics and soy protein. This chapter discusses about
the potential of functional foods and components in human nutrition as well
as in mitigating the health issues. Indeed, the functional foods lower the risk
of obesity, osteoporosis, cancer, cardiovascular and gastrointestinal diseases,
aging symptoms, diabetes, dental issues, allergies, etc. because they possess
the anti-arrhythmic, anti-inflammatory, antioxidant, anti-hypertensive,
immune-modulatory, antimicrobial, hypocholesterolemic, anti-obesity, and
anti-cancerous characteristics. The evidences for such health benefits from
functional foods are based on several animal, clinical and epidemiological
studies. However, in developing nations, the recognition of functional foods
and components is a great issue thus influencing the nutrition security in
these countries.
Key words: Functional foods, functional components, human nutrition,
carotenoids, dietary fibers, flavonoids, fatty acids, isothiocyanates and
glucosinolates, phenolic acids, phytoestrogens, plant sterols and stanols,
probiotics/prebiotics/synbiotics, soy protein.
Introduction
Nowadays the consumers are aware of the relationship between health
and food that has led to completely different perspective of food from the
last few decades. The consumers demand is moving from the taste, price,
and convenience to health and wellness that has developed a novel

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interesting area of food and nutrition sciences, known as “functional foods”.
Functional foods are those which when consumed provide health benefits
beyond the traditional nutritional effects 1. Here, the traditional nutrition is
considered only due to the consumption of vitamins and minerals which are
important to reduce or eliminate or prevent the classical nutrient deficiency
disease. For example, scurvy is being corrected by vit C consumption.
Therefore, the ascorbic acid in citrus correcting scurvy is not considered as
functional food. On the other hand, the soy protein which is not essential to
health but related with the alleviation of cardiovascular disease is a
functional food. The Japanese government in 1980s introduced the term
“functional foods” that later on received the legal status with the
establishment of “Foods for Specified Health Use” (FOSHU) regulatory
system 2. Initially, around 300 products received the FOSHU status in Japan
on July 2002 and from then, several other products have also been granted
the FOSHU status. The functional foods appear alike the conventional foods
and also consumed like regular diet. They have the potential to benefit the
health on their daily-basis consumption at effective levels thus furnishing the
necessary amount of fats, proteins, vitamins, carbohydrates, etc. required for
healthy survival. Nutraceuticals which are the isolated component or part of
food offering the medicinal or health benefits, including prevention and
treatment of disease can be obtained from functional foods and marketed in
the form of solutions, capsules, powders, tablets, or potion that are not
related to the food. The nutraceuticals when incorporated back to the food, it
becomes a functional food.
The regular consumption of functional foods maintains the micro-flora
health and reduces the chronic diseases caused among the masses due to the
modern lifestyles and longer life. The food is considered functional only
when it contains any bioactive components like ω-3 fatty acids,
isothiocyanates, oligosaccharides, polyphenols, inulin, dietary fiber,
prebiotics and probiotics, carotenoids, soy protein, etc. 3 Extensive work has
been conducted worldwide to study the health benefits linked with bioactive
components of commercially available functional foods. The distinguished
functional foods include fruit and vegetables, herbs, whole grains, energy
bars, beverages, medicinal plants, yoghurt, and many more which have the
potential to improve satiety too. The whole grains including the oats, maize,
wheat, barley, rye, etc. are the concentrated source of B-complex vitamins,
trace minerals, dietary fibbers, and other functional components like lignans,
folates, polyphenols, etc. thus possessing the antioxidant and anti-
carcinogenic effects. Similarly, the fruits and vegetables contain the

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vitamins, minerals, flavonoids, carotenoids, simple sugars, organic acids, etc.
which have potent anti-inflammatory, antioxidant, and chemo-preventive
characteristics to prevent the risk of chronic diseases like cardiovascular,
obesity, and diabetes 4.
The impact of functional foods on nutritional status of people is the
most prevalent research area in the nutrition sciences these days. Numerous
factors are responsible for this including the relationship between the dietary
constituents and health benefits, drastic growth of demand about health and
wellness products in the market, consumer self-care phenomenon and
suitable legislation environment. Therefore, a strong need is felt to
understand the nutritional significance of functional foods to take correct diet
for being healthy. Keeping this in view, this chapter deals with knowledge
about several functional foods, bioactive constituents present in them and
their role in maintaining the human health.
Classification of Functional Foods
Some foods contain inherent bioactive components while other food can
be modified to make them functional. The functional foods can be produced
using the fortification of product or genetic engineering to improve or
modify the nutrient content. The application of biotechnology permits the
modification of genetic blueprint of animals and plants so that the harmful
compounds can be decreased and nutritional components can be enhanced.
Likewise, several nutritional components can be incorporated into the food
like vitamins and mineral, phytochemicals, dietary fiber, fat substitutes, ω-3
fatty acids, etc.
According to American Dietetic Association (ADA), the functional
foods can be classified into the following four groups 5:
 Conventional Foods: As the name suggests, they are the food
which have not been modified and are consumed in their natural
state. For example, the whole fruits and vegetables which are rich in
phytochemicals and other healthy nutrients.
 Modified Foods: They are improved either by fortifying or
enriching the food with nutrients or other bioactive components.
For example, the folic acid enriched breads, calcium added in
orange juice, and margarine containing the plant sterols. The
incorporation of herbs like guarana and ginseng to energy drinks
also considers under this group.
 Medical Foods: The foods that are formulated to reduce the disease

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 or any medical condition for which characteristic nutritional
requirements are important. These foods must be consumed or
administered under the physician’s examination. They are designed
specifically to meet the nutritional needs of specific disease. For
example, Axona-developed by Accera (USA) to treat the
Alzheimer’s disease by providing necessary nutrients and
Lofenalac-developed by Mead Johnson Nutrition to treat the
pehylketonuria.
 Foods for Special Dietary Use: These are almost similar to the
medical foods, however, commercially available. There is no need
to consume such foods under the supervision of physician. For
example, lactose-free dairy products, infant foods gluten-free foods,
low-fat product, etc. These foods meet the particular dietary
requirement of people because of their health problems like obesity,
celiac disease, lactose intolerance, etc.
Functional Food: Sources and Nutritional Significance
Several evidences from epidemiological, in-vivo, in-vitro, and clinical
studies demonstrated the reduction of chronic diseases by consuming the
plant-based diet such as fruits, vegetables, herbs, spices, whole grains, and
others. Although the vast number of naturally occurring health-enhancing
substances are of plant origin, the increasing evidences also supports the
improvement of human health through the intake of animal based food.
Numerous functional components are found in animal foods which have
biological importance. The dairy products contain the calcium, probiotics,
whey proteins and whey peptides, fish provide mainly the ω-3 fatty acids,
beef supplies the conjugated linoleic acid, eggs are rich in sphingolipids and
several conditionally-essential nutrients like α-lipoic acid, coenzyme Q10, L-
carnitine, choline and taurine are widely diffused in animal products.
Among plant based functional foods, the oat products containing the
soluble fiber β-glucan lower the total and low density lipoprotein (LDL)
cholesterol thus reducing the coronary heart disease. Soybean is considered
to have the preventive and therapeutic contribution in cholesterol reduction,
cardiovascular disease, cancer, osteoporosis, and the alleviation of
menopausal symptoms on account of many anticarcinogens such as protease
inhibitors, phytosterols, saponins, phenolic acids, phytic acid, and
isoflavones present in them. Flaxseed contains the most (57%) of the ω-3
fatty acid, α-linolenic acid in addition to the lignans. Tomatoes are rich in
lycopene, the primary carotenoid which has role in cancer risk reduction.

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Garlic possesses the medicinal properties including cancer chemopreventive,
antibiotic, anti-hypertensive, and cholesterol-lowering properties whereas the
broccoli and other cruciferous vegetables are associated with the decreased
risk of cancer due to glucosinolates. Further, the oranges, lemons, limes, and
grapefruits are a essential source of important nutrients as vitamin C, folate,
and fiber, but they contain the phytochemicals responsible for the anticancer
activity. Tea particularly green tea contains the polyphenols mainly catechins
associated with the reduced risk of cancer and cardiovascular diseases.
Moreover, the wine, particularly red wine, can reduce the risk of
cardiovascular diseases due to its high phenolic content compared to white
wine because of adding the grape skins into fermenting grape juice during
production.
Functional Components and Human Nutrition
The functional components naturally-found in food have significant role
in the human diet and provides the nourishment to our body. These
components improve our health by keeping the system in perfect working
condition and reduce the prevalence of several diseases including the GI
inflammation, cardiovascular, aging, development disorders, etc. The
important functional components of foods and their contribution in human
nutrition for improvement of health are described in detail as follows:
1. Carotenoids
Carotenoids are the family of fat-soluble pigmented compounds which
are synthesized by microorganisms and plants excluding animals. They are
found mainly in plants, flowers, fruits and vegetables, algae, some yeasts and
molds, photosynthetic bacteria, and sometimes non-photosynthetic bacteria
too. However, in human dietary system, the carotenoids can be taken from
fruits and vegetables especially green leafy vegetables which contribute to
their yellow, orange and red colors. The major carotenoids compounds found
in food include α-, β- and ɣ-carotene, lycopene, β-cryptoxanthin, zeaxanthin,
lutein, xanthohylls, astaxanthin and capsanthin which are significant
precursors of vitamin A 6.
No carotenoids is considered as an essential nutrient (except lutein and
zeaxanthin- the conditionally essential nutrient for ocular disease) and
therefore not indulged in the important metabolic pathways and generation
of a specific deficiency or disease. However, they are inversely associated
with the risk of cardiovascular diseases, some cancers and mainly
degenerative eye diseases 7-9 as shown in Table 1. It is said that the human
intestine absorbs only 5% carotenoids of the whole while more than 50%

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belong to the micellar solution. The heat treatment disrupts the cell wall and
loosens the bond thus improving the carotenoids accessibility. Other factors
determining the bioavailability of carotenoids include the nutritional status,
genetic factors, infection, drug (sulfonamides/aspirin) consumption, level of
vitamin E, etc. 10. The potent antioxidant properties of carotenoids are
mainly responsible for several health properties due to the prevention of
cellular DNA and other molecules from free-radical induced damage. The
carotenoids absorb the singlet oxygen (1O2) and other reactive oxygen
species (ROS) which would minimize the uncontrolled generation and
related increase of ROS concentration thus leading to the reduction of
oxidative stress – important factor in the pathogenic processes of several
diseases in the body. Carotenoids can also play a great role in human health
through the following mechanisms 11:
i) modulation of growth factors
ii) immune system stimulation
iii) modulation of intracellular signaling pathways
iv) modulation of several receptors molecules
v) regulation of cell cycle and apoptosis
vi) cell differentiation
Further, α-and β-carotene and β-cryptoxanthin can convert to vitamin A
in the body thus beneficial in disease prevention.
Table 1: Several studies demonstrating the role of carotenoids in human health

Carotenoids Study Type Results References

Affected tumor necrosis factor-α induced
inflammation thus reducing the generation
Lycopene and of ROS and nitrotyrosine, showing the Di Tomo et
In-vitro
β-carotene carotenoids’ anti-inflammatory activity al. 12
and the protective effects of carotenoid-
rich diets against CVD risk
Inhibited the strain-induced ET-1
expression through suppression of ROS
generation through reduction of p22(phox)
mRNA expression and NAD(P)H oxidase
activity; inhibited strain-induced ET-1
Sung et al.
Lycopene In-vitro secretion by reducing ROS-mediated 13
extracellular signal-regulated kinase
(ERK) phosphorylation; enhanced heme
oxygenase-1 (HO-1) gene
expression showing beneficial effects on
cardiovascular system.

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 α-carotene Michaud et
- Reduced the risk of lung cancer
and lycopene al. 14
Reduction of class A scavenger receptor
and CD36 expression in the protein and
mRNA levels; decrease of MMP-1, -2, -3,
-9, -12, and -14 activity and expression;
Kishimoto
Astaxanthin In-vitro suppression of mRNA expression of tumor
et al. 15
necrosis factor-alpha, interleukin-1beta,
interleukin-6, inducible nitric oxide
synthase, and cyclooxygenase-2;
inhibition of macrophages activation
Protected the macula from damage by blue
light, improve visual acuity and scavenge
Lutein and Abdel-Aal
- harmful reactive oxygen species; reduced
zeaxanthin et al. 16
risk of age-related macular degeneration
and cataracts
lutein,
zeaxanthin, β-
Epidemiologi Protection against the early Dwyer et
cryptoxanthin
cal atherosclerosis. al. 17
and α-
carotene
Inhibited expression and production of
inflammatory mediators; blocked the
intracellular accumulation of ROS;
β-carotene In-vitro demonstration of anti-inflammatory Bai et al. 18
activity by acting as a potential inhibitor
for redox-based NF-kappaB activation due
to its antioxidant activity
Reduced the body weight, abdominal
adipose tissue weight, and serum lipid
β- concentrations; significant repression of
Takayanagi
Cryptoxanthi Mice adipocyte hypertrophy; repressed the 19
n inflammatory cytokine secretion and
improves the lipid metabolism and the
energy consumption
Lycopene, Plasma lycopene, lutein and beta-carotene
Epidemiologi Karppi et
lutein and β- were the most powerful antioxidants and
cal al. 20
carotene there was decreased LDL-oxidation

2. Dietary Fibers
No doubt dietary fiber is not a nutrient, but essential to prevent the
occurrence of diseases mainly associated to bowel like constipation,
appendicitis, colon and rectum cancer, diverticular disease, piles, etc. The
dietary fibre, also known as roughage refers to the group of plant derived
polysaccharides which are edible but can’t be digested and absorbed in the
small intestine and passes into the large intestine unchanged 21. The examples

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of dietary fibre include the non-starch polysaccharides (gums, cellulose,
hemicellulose, and pectins), oligosaccharides (inulin, raffinose, etc.), lignin,
and other related plant substances like suberin, waxes, etc. This definition
also includes the resistant starch as dietary fibre because of its resistance to
digestion in GI tract. The major sources of dietary fibre include the fruits
(apple, pear, orange, strawberries, etc.), vegetables (onion, green beans,
brussel sprouts, peas, etc.), pulses (chickpeas, lentils, beans, etc.) and whole-
grains like oat, all cereals bran, whole and mixed grain breads, etc. Generally
dietary fibre is classified into soluble and insoluble fibre on the basis of
solubility and each category has definite physiological benefits to the body
summarized in Table 2.
Table 2: Classification of dietary fibre and potential health benefits

Dietary
Example (s) Health Benefits
Fibre
1. Reduction of bowel transit time,
2. Bulking capacity improve the
Insoluble laxation
Lignin, cellulose, and hemicellulose
fibre 3. Act as food to the intestinal
microflora mainly probiotic species
for their growth
1. Regulation of glucose levels in
Gums, pectins, β-glucans, the blood
Soluble
oligosaccharides, and other non- 2. Prolong the gastric emptying
fibre
cellulosic polysaccharides 3. Reduce the serum cholesterol
levels
Adapted and modified from Li and Komarek 22

The consumption of each type of dietary fibre and its relation to

physiological benefits has been extensively investigated (Table 3).
Numerous studies revealed the reduced risk of cardiovascular diseases 23,
colorectal cancer 24, stroke 25, type-2 diabetes 26 and weight gain 27 with the
adequate intake of dietary fibre. It is also significant for the immune system
regulation by controlling the GI health and fibre-microbiota interactions 28.
The health benefits of dietary fibre have been linked with its functions in the
small intestine as per the recent investigations 29. The soluble fibre may
result in enhanced viscosity due to the high molecular-weight biopolymers
which modifies the gastric emptying providing the satisfaction and fullness
feeling with the sensing and release of nutrients in duodenum. Contrarily, the
insoluble fibre interacts with the colonic microbiota thus releasing the
several by-products like short chain fatty acids on fermentation 30. This
contributes significantly to the composition and metabolic activity of
microbiome thus improving the intestinal health and immune system,

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consequently the ability to resist some chronic diseases.
Table 3: Summary of types, sources and physiological benefits of dietary fibre

Dietary Fibre Sources Physiological Benefits

Lowering of blood lipid; reduced
β-glucan and oat bran Oats and barley
blood glucose response
Cellulose Plant foods Laxation
Chitosan or chitin Fungi or shellfish Lowering of blood lipid
Lowering of blood lipid; reduced
Guar gum Guar bean
blood glucose response
Lowering of blood lipid; laxation;
Fructooligosaccharide (inulin Jerusalem artichoke
gut health; modulation of
and oligofructose) or chicory root
microbiota
Dairy products and Modulation of microbiota and
Galactooligosaccharide
legume extract immune system; gut health
Lowering of blood lipid; reduced
Pectin Plant materials
blood glucose response
Lowering of blood lipid; reduced
Psyllium Psyllium husk
blood glucose response; laxation
Lowering of blood lipid; reduced
Resistant dextrins Corn and wheat
blood glucose response
Reduced blood glucose response;
Resistant starch Plant materials
laxation; gut health
Reduced blood glucose response;
Soluble corn fibre Corn
laxation; gut health
Wheat bran Wheat Laxation
Adapted and modified from Clemens et al. 31

3. Flavonoids
Flavonoids-a class of polyphenols are basically the secondary plant
metabolites which are extensively distributed in the plant kingdom and
known for mainly blue, purple and red anthocyanin pigments in the plant
tissues 32. Out of more than 4000 flavonoid species, there are about six major
flavonoids groups (Fig. 1) including the flavones, flavonols, flavanols,
flavanones, anthocyanidins and isoflavonoids as per their chemical structure
33
. They are mainly found in nuts, fruits, vegetables, and beverages including
wine and tea. According to a study, people usually consume 1-2 g flavonoids
per day in a normal diet. The flavonoids are the excellent source of natural
antioxidant in human diet which is based on the functional-OH groups in
their structure that scavenges the free radicals or chelates the metal ions to
minimize the radical generation thus reducing the occurrence of oxidative
stress and many other diseases 34. They are typically more potent antioxidant

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than vitamins C and E. They can inhibit the injury caused by free radicals
through following mechanisms:
1. Direct scavenging of ROS
2. Antioxidant enzymes activation
3. Metal chelating activity
4. Reducing α-tocopheryl radicals
5. Inhibiting oxidases
6. Mitigating the oxidative stress caused due to NO
7. Increasing the uric acid levels
8. Enhancing the antioxidant characteristics of low-molecular
antioxidants

Fig 1: Classification of flavonoids with examples and associated health benefits

The flavonoids are considered as one of the safest drugs due to their
therapeutic application instead of being smaller organic compounds. The
flavonoids possess several biological activities like cytotoxic, anti-
inflammatory, cystostatic, anti-viral and anti-cancer properties on account of
its free radicals scavenging activity, chelation and interaction with
biomolecules like regulatory enzymes. They are very effective antimicrobial
substances against several micro-organisms due to their capacity to disrupt
the microbial membranes and inactivate the microbial adhesins, enzymes and
cell transport proteins 35. Each flavonoid are associated with numerous
positive physiological effects (Fig. 1).
Most potent flavonoids like flavones and catechins are employed to

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protect the body from adverse effects of oxidative stress. The occurrence of
cancer, liver and cardiovascular diseases can be prevented due to the
protective effects of quercitin, morin, myricetin, etc. The oxidative stress that
may lead to cardiovascular diseases can be decreased by flavonoids thus
attributing to the beneficial health effects. Flavonoids may cause the
inhibition of cell cycle to proliferate the lymphoid cells to produce the
growth inhibitory effects on several malignant tumor cells which reduces the
risk of human breast cancer, ovarian and prostate cancer 36. In addition to
this, the flavonoids are as enzyme inhibitor, immune modulators or skin care
and wound-healing agents.
4. Fatty Acids
The fatty acids – the building blocks of fats and oils are typically the
carboxylic acid containing a long-aliphatic chain either saturated or
unsaturated. The fats act as a medium for the absorption of fat-soluble
vitamins and indispensable in the survival, growth and development of
human during the embryonic development and initial growth just after birth
– on through infancy and childhood. Thus the fatty acids are significant in
the human development, health and disease. The high consumption of fat are
associated with the heart disease, obesity, and type II diabetes mellitus in
rich population while the poor nutritional status of children in non-affluent
groups are partially due to the diet low in fat. Fatty acids influence the cell
membrane structure, regulate the gene transcription, and function as cytokine
precursors in addition to the energy sources thus affecting human health.
Among fatty acids, the families of omega-3 (ω-3 or ω-3 like (α-linolenic
acid, eicosapentaenoic acid, and docosahexaenoic acid) and omega-6 (ω -6
or n-6 like linoleic acid and arachidonic acid) fatty acids are comprised of
polyunsaturated fatty acids containing 18-22 carbons. Also, α-linolenic acid
and linoleic acid are the essential fatty acid as they can’t be synthesized by
human body and therefore, must be provided from diet. Foods such as flax
seeds, walnuts, canola oil, fatty fish (tuna, salmon and herring), leafy greens,
etc. have large concentration of ω-3 fatty acids whereas the ω-6 fatty acids
are obtained mainly from vegetable oils like sunflower, corn, and soybean 37.
During metabolism, 0.2-5% of α-linolenic acid (ALA) can be converted into
eicosapentaenoic acid (EPA) and nearly 1% can change into the
docosahexaenoic acid (DHA) 38 and EPA and DHA are the precursors of
eicosanoids which have antithrombotic, anti-inflammatory, anti-arrythmic,
and vasodilatory characteristics whereas the linoleic acid is converted to
arachidonic acid (AA) - a precursor to a group of eicosanoids possessing the

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pro-inflammatory and prothrombotic properties.
The adequate quantity of ω-6 fatty acids also aid in the improvement of
health however, its high concentration may lead to the hyper-immune
responses which hinders the normal working of ω-3 fatty acids and linked
with adverse effects like chronic cardiovascular diseases and inflammatory
responses. Therefore, there should be a balance between the ω-6 and ω-3
ratio (1:1) in the healthy diet due to the opposing effects of ω-6 and ω-3 fatty
acids. The western diets contain the ω-6: ω-3 varying from 15:1-17:1 which
promotes the many chronic diseases which must be reduced for better health.
On the other hand, higher consumption of saturated fatty acids and trans-
fatty acids are attributed to the cardiovascular diseases, therefore their intake
should also be lowered. The trans-fatty acids are also considered as
carcinogenic in nature and their consumption has been declared as unsafe by
Food and Drug Administration (FDA).
The ω-3 fatty acids are the most important to cardiovascular, brain,
central nervous system and eye health and also during pregnancy and
childhood. Increased concentration of ω-3 fatty acids in blood plasma is
linked with the decreased risk of neurodegenerative diseases like
Alzheimer’s disease, mental disorders like schizophrenia and depression 39-
41
. Ω-3 fatty acids are reduce the growth of several inflammatory diseases
such as from inflammatory bowel disease to diseases of skin and joints, to
other auto-immune diseases like multiple sclerosis and lupus42.
1. Isothiocyanates and Glucosinolates
Isothiocyanates (R–N=C=S, ITCs) are the volatile substances which are
the hydrolysis product of glucosinolates – S- containing substances in the
cruciferous vegetables under the action of myrosinase, an enzyme released
on the damage to the plant tissues due to cutting or chewing. The ITCs are
usually stored in the form of thioglucoside conjugates (or glucosinolates) in
the cruciferous vegetables like broccoli, cauliflower, kale, mustard, and
cabbage. In addition to the breakdown of glucosinolates using myrosinase,
the intestinal microflora can also synthesize ITCs43. When glucosinolates are
present as sequestered in the sub-cellular compartments of plant, they are
chemically stable but biologically inactive and their breakdown forms the
several bioactive compounds including ITCs which can be absorbed from the
small bowel and colon44. Each glucosinolate can generate a different
isothiocyanate, for instance, the broccoli is a good source of glucoraphanin-
glucosinolate precursor of sulforaphane, and sinigrin - glucosinolate
precursor of allyl-ITCs.

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 Several animal and epidemiological studies showed that the cruciferous
vegetables rich in glucosinolates and ITCs are associated with the better
health, antioxidant and anti-inflammatory properties, and reduction of
cardiovascular disease, cancer incidence and progression. ITCs inhibit the
numerous pathogens at low concentrations thus presenting them as the
promising antimicrobial substances. In-vitro studies revealed the ITCs as
strong inducers of Phase II enzymes which enhances the metabolism and
detoxification of carcinogens, inhibit mitosis and stimulate apoptosis in
human tumour cells thus increasing the potential to block the DNA damage
and inhibit the growth of tumour cells. Epidemiological evidence suggests
that the glucosinolate breakdown products may protect gastrointestinal tract,
lung, breast and colon cancer 45-47.
6. Phenolic Acids
Phenolic acids are the important sub-category of important secondary
plant metabolites called as “phenolics”, extensively found in the plants. All
the 8000 naturally-existing phenolic compounds contains a common
structural feature - a phenol and when phenol is attached to carboxylic acid,
then phenolics are referred to as phenolic acids which provide protection
against the oxidative damage diseases such as stroke, coronary heart disease,
type 2 diabetes and cancers. The common example of these aromatic
secondary metabolites includes the caffeic, p-coumaric, vanillic, chlorogenic,
ferulic, gentisic, syringic and protocatechuic acid. Phenolic acids are the
strong antioxidants based on the reactivity of phenol unit containing -OH
group through the radical scavenging via electron donation, H-atom
donation, and singlet oxygen (1O2) quenching 48. After favonoids, phenolic
acids are the major sub-category (1/3rd) of phenolics present in teas, fruits
and vegetables, grains, coffees, red wine, and spices and consumed 25 mg/g
in a day based on the diet 49.
Phenolic acids are associated with the blocking of AP-1 transcriptional
activity due to the fact that AP-1 is an activator protein used in process
which controls the cell differentiation, inflammation, and proliferation 50.
Caffeic acid is related to the inhibition of leukotrienes biosynthesis, involved
in the asthma, immune-regulation diseases, and several allergic reactions. It
also possesses antitumor activity against the colon carcinogenesis and its
derivatives exhibit the strong inhibition of human immunodeficiency virus
type 1 (HIV-1) integrase which catalyzes the integration of viral DNA into
the host chromatin51. Certain foods rich in phenolic acids have been shown
the antibacterial effect and also reduce the risk of hypertension, but evidence

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from epidemiological studies focused on dietary phenolic acid intake is
scarce.
7. Phytoestrogens
Phytoestrogens are the non-steroidal and biologically-active
phytochemicals which possesses the chemical structure alike to estradiol - an
endogenous estrogen on the account of which it may attach to estrogen
receptors in numerous cells and pose estrogenic or anti-estrogenic effects.
They are found mainly in the whole grains, peas, seeds, clover and legumes
among which most-abundant food sources is soyabean and soyabean based
products. The classification of phytoestrogens includes the isoflavones
(genistein and daidzein), flavonoids (already discussed), lignans
(enterolactone and enterodiol), stilbenes (resveratrol and piceatannol),
terpenoids (limonene, menthol, citral, camphor, pinene, eugenol, thymol, and
geraniol) and coumestans (coumestrol) 52.
Phytoestrogens provide the protection against menopausal symptoms,
hyperlipidemia, cardiovascular and cognitive diseases, breat and uterus
cancers, osteoporosis, obesity, and several kinds of chronic renal diseases53-
54
. The health benefits of phytoestrogens are based on its competition with
17β-estradiol in order to bind the intranuclear estrogen receptor protein for
the modulation of gene transcription due to the similarities of its structure
with endogenous estrogens. Phytoestrogens binds more strongly to estrogen
receptor-β however; it can bind to estrogen receptors α too and both the
receptors are expressed in several tissues including adipose. Further, they
can exhibit positive health effects through potent antioxidative activity and
the non-estrogen receptor-mediated mechanisms by preventing the
enzymatic activity of DNA topoisomerase I (EC 5.99.1.2) and DNA
topoisomerase II (EC 5.99.1.3), protein tyrosine kinases, and ribosomal S6
kinase which are significant for the cell-signaling mechanisms, cell
proliferation, and differentiation 55.
8. Plant Sterols and Stanols
Plant sterols and stanols are the functional components in plant with
similar chemical structure and biological functions like that of cholesterol.
Basically, plant sterols are the steroid alkaloids which contain surplus
methyl, ethyl group or double bond than cholesterol whereas the stanols are
5α-saturated derivatives of plant sterols as they don’t carry any double bonds
in the sterol ring structure. Most common examples of plant sterols include
campesterol, sitosterol, and stigmasterol and plant stanols include the
sitostanol and campestanol56. The major dietary sources of plant sterols are

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vegetable oils, seeds, fruits and vegetables, spreads and margarines, nuts,
and grains products while the plant stanols are mostly found in the cereals,
mainly wheat and rye. The intake of plant sterols varies from 160–400
mg/day whereas the plant stanols are consumed in fewer amounts (17–
24 mg/day). Plant sterols are less absorbed (2–5%) than the cholesterol
(60%) in humans 57.
From 1950s, it is known that the absorption of cholesterol in the
intestine is interfered by plant sterols and stanols thus exhibiting cholesterol-
lowering effect due to the increase of ABC-transporter expression by them.
The consumption of plant sterols or stanols (2.5 g) on daily-basis may reduce
the serum cholesterol levels up to 10%. They are also found to reduce the
oxidative stress and inflammation. They exhibit the hypocholesterolemic
effects based on the interference of biliary and dietary cholesterol from the
intestinal tract. Beyond this, plant sterols and stanols demonstrate the anti-
cancer, anti-atherosclerosis, anti-inflammation, and anti-oxidation action 58-
59
.
9. Probiotics, Prebiotics and Synbiotics
Today, the consumers are being more health conscious leading to the
concept of probiotics, prebiotics and synbiotics in their diet for constant and
long-term health benefits. Generally, the probiotics refer to the live
microorganisms whose consumption in the adequate amounts; provide the
health benefits to host and prebiotics are the non-digestible foods (mainly
oligosaccharides) which stimulate the growth of probiotics in addition to
modify the their growth and activity while the synbiotics exhibiting the
synergism effect are the combination of prebiotic ingredients and probiotic
bacteria 60. Probiotics ostensibly fulfill their definition through a variety of
somewhat disparate and overlapping mechanisms. These include: regulation
of intestinal microbial homeostasis, antimicrobial activity, immuno-
modulation and pathogen exclusion. The safety, functional, and
technological characteristics of probiotics are very important while selecting
the microorganisms. The safety considerations of probiotics includes the
antibiotic resistance profiles, infectivity in immune-compromised animal
models, toxin production, hemolytic activity, metabolic activities (D-lactate,
bile salt de-conjugation), genetic and pathological side effects and
epidemiological surveillance of adverse incidents in consumers (post
market). Functional properties include the viability and stability of cells,
persistence activity in GI tract, species and strain characteristics, daily dose,
fermentation technology, storage conditions, and availability of prebiotics 61-
62
.

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 Numerous research works have been carried out in the past few years to
identify the probiotics and prebiotics, their characterization, mechanisms,
potential as symbiotic, and contribution in the prevention and management
of several diseases. The lactic acid bacteria (LAB) and Bifidobacteria species
having “Generally recognized as safe” (GRAS) status are the most common
commercial starter cultures in addition to several other cultures63 (Table 4).
The promising prebiotic ingredients include the disaccharides,
oligosaccharides, and non-starch polysaccharides such as pectins, cellulose,
hemicellulose, fructooligosaccharides, gums, inulins, lactitol,
galactooligosaccharides, isomaltooligosaccharides, xylooligosaccharides,
lactulose, etc64.
Table 4: List of probiotic microorganisms significant in human nutrition

Lactobacillus Bifidobacterium Other LAB Non-LAB

Spp. Spp.
L. casei B. infantis Streptococcus Saccharomyces
thermophillus boulardi
L. rhamnosus B. animalis Propionibacterium Bacillus clausii
freudenreichii
L. reuteri B. brevis Leuconostoc dextranicum Bacillus cereus
L. johnsonii B. lactis Lactococcus cremoriss Clostridium
botyricum
L. helveticus B. adolescentis Lactococcus lactis Escherichia coli
Nissle 1917
L. lactis B. longum Pediococcus acidilactici
L. plantarum B. bifidum Enterococcus fraecium
L. paracasei
L. bulgaricus
L. johnsonii
L. fermentum
L. acidophilus
L. brevis
Being the complex microbial ecosystem, the intestinal microorganisms
have synbiotic co-evolution towards their host in addition to being the
commensal. The major functions of probiotics include the production of
nutrients to host, prevention from pathogens and boosting the immune
system. The daily consumption of probiotics therefore attain, restore, and
maintain the favourable ecosystem balance and the micro-flora in the GI
tract, indispensable for the better health of host. The consumption of
probiotics, prebiotics, or synbiotics which can be derived from raw fruits and
vegetables, fermented dairy products or pickles, pharmaceutical formulas

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and functional food on a regular basis is favourable for the health of
intestinal microbiota. Several clinical studies revealed the significance of
probiotic microorganisms in the prevention of irritable bowel syndrome
(IBS), obesity, non-alcoholic fatty liver disease, inflammatory bowel disease
(IBD), insulin resistance syndrome, diarrhoea, type 2 diabetes, allergic
diseases and elimination of Helicobacter in addition to boosting the immune
system (immune-modulation). Scientific investigations demonstrated the
advantages of prophylactic use of probiotics to treat different cancers and
their associated side effects. Prebiotics instead of stimulating the indigenous
gut bacteria growth, have the potential to modify the gut microbiota,
however, such alterations are possible only at individual strains and species
level. Several reports support the positive effects of prebiotics on human
health. On the other hand, the combination of probiotic and prebiotic
properties, the synbiotics ensures a superior health effect, than the probiotic
or prebiotic action alone 65-66.
10. Soy Protein
Soy proteins- constituting 40% soybeans are the cheapest source of
dietary proteins and considered analogous to animal protein in nutritional
composition containing almost all amino acids except cysteine and
methionine. Soybeans mainly contain two storage proteins viz. β-conglycinin
and glycinin. Animal studies reported the similar biological value of soy
protein and animal proteins (casein) on enrichment with methionine.
Numerous kinds of edible soy proteins are available in market like soy
protein concentrates, protein isolates, full-fat and defatted flours and grits
among which protein content varies from 90% in protein isolates to 40%
protein in whole ground soybeans. It is also present in traditional soy based
foods and foods or supplements containing soy protein ingredients.
Soy proteins supported the human health needs across the lifespan as a
source of lean, cholesterol-free and lactose-free protein. Their intake is
related to the reduced risk of breast, bladder, prostate, lungs, colon and liver
cancer in addition to preventing the initiation, promotion, and progression of
cancer. The soy proteins also help in the reduction of hypercholesterolemia
and aging symptoms 67.
Conclusion
Numerous investigations revealed the indispensable part of functional
foods in the human nutrition which contain the bioactive components from
plant or animal sources thus improving the health. Functional foods contain
bioactive substance in small amount which have biological importance.

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Many of bioactive components like ω-3 and ω-6 polyunsaturated fatty acids,
carotenoids, probiotics/prebiotics, phytoestrogens, dietary fibre, soy proteins,
etc. are however not a magical spell for poor health habits. Further, there is a
great need of research work based on diet-health relationships to substantiate
the probable health benefits of functional foods.
References
1. Hasler CM. Functional Foods: Benefits, Concerns and Challenges-A
Position Paper from the American Council on Science and Health. The
Journal of Nutrition. 2002; 132(12):3772-3781.
2. Prado FC, Parada JL, Pandey A, Soccol CR. Trends in non‐dairy
probiotic beverages. Food Research International. 2008; 41:111-23.
3. Jankovic I, Sybesma W, Phothirath P, Ananta E, Mercenier A.
Application of probiotics in food products-challenges and new
approaches. Current Opinions in Biotechnology. 2010; 21:175-181.
4. Shahidi F. Functional Foods: Their Role in Health Promotion and
Disease Prevention. Journal of Food Science. 2004; 69(5):R146-R149.
5. Hardy G. Nutraceuticals and Functional Foods: Introduction and
Meaning. Nutrition. 2000; 16:688-698.
6. Rao AV, Rao LG. Carotenoids and human health. Pharmacological
Research. 2007; 55:207-216.
7. Kantoff P. Prevention, Complementary Therapies, and New Scientific
Developments in the Field of Prostate Cancer. Reviews in Urology.
2006; 8(2):S9-S14.
8. Mozaffarieh M, Sacu S, Wedrich A. The Role of the Carotenoids,
Lutein and Zeaxanthin, in Protecting Against Age-related Macular
Degeneration: A Review Based on Controversial Evidence. Nutrition
Journal, 2003; 2:20.
9. Sesso HD, Liu S, Gaziano JM, Buring JE. Dietary Lycopene, Tomato-
Based Food Products and Cardiovascular Disease in Women. Journal of
Nutrition. 2003; 133:2336-2341.
10. Fernandez-Garcia E, Carvajal-Lerida I, Jaren-Galan M, Garrido-
Fernandez J, Perez-Galvez A, Hornero-Mendez D. Carotenoids
bioavailability from foods: From plant pigments to efficient biological
activities. Food Research International. 2012; 46:438-450.
11. Palozza P, Serini S, Ameruso M, Verdecchia S. Modulation of

Page | 162
 Intracellular Signaling Pathways by Carotenoids. In: Britton G, Liaaen-
Jensen S, Pfander H (eds.) Carotenoids: Nutrition and Health. 5,
Birkhauser: Basel, Switzerland, 2009, 211-235.
12. Di Tomo P, Canali R, Ciavardelli D, Di Silvestre S, De Marco A,
Giardinelli A et al. beta-Carotene and lycopene affect endothelial
response to TNF-alpha reducing nitro-oxidative stress and interaction
with monocytes. Molecular Nutrition and Food Research. 2012; 56:217-
227.
13. Sung LC, Chao HH, Chen CH, Tsai JC, Liu JC, Hong HJ et al.
Lycopene inhibits cyclic strain-induced endothelin-1 expression through
the suppression of reactive oxygen species generation and induction of
heme oxygenase-1 in human umbilical vein endothelial cells. Clinical
and Experimental Pharmacology and Physiology. 2015; 42:632-639.
14. Michaud DS, Feskanich D, Rimm EB, Colditz GA, Speizer FE, Willett
WC et al. Intake of specific carotenoids and risk of lung cancer in 2
prospective US cohorts, The American Journal of Clinical Nutrition.
2000; 72(4):990-997.
15. Kishimoto Y, Tani M, Uto-Kondo H, Iizuka M, Saita E, Sone H et al.
Astaxanthin suppresses scavenger receptor expression and matrix
metalloproteinase activity in macrophages. European Journal of
Nutrition. 2010; 49:119-126.
16. Abdel-Aal E-SM, Akhtar H, Zaheer K, Ali R. Dietary Sources of Lutein
and Zeaxanthin Carotenoids and Their Role in Eye Health. Nutrients.
2013; 5(4):1169-1185.
17. Dwyer JH, Paul-Labrador MJ, Fan J, Shircore AM, Merz CN, Dwyer
KM. Progression of carotid intima-media thickness and plasma
antioxidants: the Los Angeles Atherosclerosis Study. Arteriosclerosis,
Thrombosis, and Vascular Biology. 2004; 24:313-319.
18. Bai SK, Lee SJ, Na HJ, Ha KS, Han JA, Lee H et al. beta-Carotene
inhibits inflammatory gene expression in lipopolysaccharidestimulated
macrophages by suppressing redox-based NF-kappaB activation.
Experimental and Molecular Medicine. 2005; 37:323-334.
19. Takayanagi K. Prevention of Adiposity by the Oral Administration of β-
Cryptoxanthin. Frontiers in Neurology. 2011; 2:67.
20. Karppi J, Nurmi T, Kurl S, Rissanen TH, Nyyssönen K. Lycopene,
lutein and beta-carotene as determinants of LDL conjugated dienes in

Page | 163
 serum. Atherosclerosis. 2010; 209:565-572.
21. Van Der Kamp JW. Dietary Fiber-Bio-active Carbohydrates for Food
and Feed. Wageningen Academic Publishers, Wageningen, the
Netherlands, 2004.
22. Li YO, Komarek AR. Dietary fibre basics: Health, nutrition, analysis,
and applications. Food Quality and Safety. 2017; 1:47-59.
23. Threapleton DE, Greenwood DC, Evans CE. Dietary fiber intake and
risk of cardiovascular disease: systematic review and meta-analysis.
BMJ British Medical Journal, 2013; 347:68-79.
24. Dahm CC, Keogh RH, Spencer EA, Greenwood DC, Key TJ, Fentiman
IS et al. Dietary fiber and colorectal cancer risk: a nested case-control
study using food diaries. Journal of National Cancer Institute. 2010;
102:614-626.
25. Zhang A, Xu G, Liu D, Zhu W, Fan X, Liu X. Dietary fiber
consumption and risk of stroke. European Journal of Epidemiology.
2013; 28:119-130.
26. Yao B, Fang H, Xu W. Dietary fiber intake and risk of type 2 diabetes: a
dose response analysis of prospective studies. European Journal of
Epidemiology. 2014; 29:79-88.
27. Li SS, Kendall CW, De Souza RJ, Jayalath VH, Cozma AI, Ha V et al.
Dietary pulses, satiety and food intake: a systematic review and meta-
analysis of acute feeding trials. Obesity. 2014; 22:1773-1780.
28. Dong H, Sargent LJ, Chatzidiakou Y, Saunders C, Harkness L,
Bordenave N et al. Orange pomace fiber increases a composite scoring
of subjective ratings of hunger and fullness in healthy adults. Appetite.
2016; 107:478-485.
29. Mackie A, Bajka B, Rigby N. Roles for dietary fiber in the upper GI
tract: the importance of viscosity. Food Research International. 2016;
88:234-238.
30. Simpson HL, Campbell BJ. Review article: dietary fiber-microbiota
interactions. Alimentary Pharmacology and Therapeutics. 2015; 42:158-
179.
31. Clemens R, Kranz S, Mobley AR, Nicklas TA, Raimondi MP,
Rodriguez JC et al. Filling America’s fiber intake gap: summary of a
roundtable to probe realistic solutions with a focus on grain-based foods.

Page | 164
 Journal of Nutrition. 2012; 142:S1390-S1401.
32. Winkel-Shirley B. Flavonoid biosynthesis: a colorful model for
genetics, biochemistry, cell biology and biotechnology. Plant
Physiology. 2001; 126:485-493.
33. Hoensch HP, Oertel R. Emerging role of bioflavonoids in
gastroenterology: especially their effects on intestinal neoplasia. World
Journal of Gastrointestinal Oncology. 2011; 3(5):71-4.
34. Leopoldini M, Russo N, Chiodo S, Toscano M. Iron chelation by the
powerful antioxidant flavonoid quercetin. Journal of Agricultural and
Food Chemistry. 2006; 54(17):6343-6351.
35. Cushnie TPT, Lamb AJ. Antimicrobial activity of flavonoids.
International Journal of Antimicrobial Agents. 2005; 26:343-356
36. Tiwari SC, Husain N. Biological activities and role of flavonoids in
human health-A review. Indian Journal of Scientific Research. 2017;
12(2):193-196.
37. Martins MB, Suaiden AS, Piotto RF, Barbosa M. Properties of Omega-3
Polyunsaturated Fatty Acids Obtained of Fish Oil and Flaxseed Oil.
Revista do Instituto de Ciências da Saúde. 2008; 26:153-156.
38. Brenna JT, Salem JN, Sinclair AJ, Cunnane SC. Alpha-Linolenic Acid
Supplementation and Conversion to n-3 Long-Chain Polyunsaturated
Fatty Acids in Humans. Prostaglandins, Leukotrienes and Essential
Fatty Acids. 2009; 80:85-91.
39. Schaefer EJ, Bongard V, Beiser AS, Lamon-Fava S, Robins SJ, Au R et
al. Plasma phosphatidylcholine docosahexaenoic acid content and risk
of dementia and Alzheimer disease: The Framingham Heart Study.
Archives of Neurology. 2006; 63:1545-50.
40. McNamara RK, Jandacek R, Rider T, Tso P, Hahn CG, Richtand NM et
al. Abnormalities in the fatty acid composition of the postmortem
orbitofrontal cortex of schizophrenic patients: Gender differences and
partial normalization with antipsychotic medications. Schizophrenia
Research. 2007; 91:37-50.
41. Sanchez-Villegas A, Henríquez P, Figueiras A, Lahortiga F, Molero P,
Toledoet E. A longitudinal analysis of diet quality scores and the risk of
incident depression in the SUN Project. European Journal of Nutrition.
2007; 46:337-46.

Page | 165
42. Simopoulos AP. Omega-3 fatty acids in inflammation and autoimmune
diseases. Journal of the American College of Nutrition. 2002; 21:495-
505.
43. Fahey JW, Wehage SL, Holtzclaw WD, Kensler TW, Egner PA,
Shapiro TA et al. Protection of humans by plant glucosinolates:
efficiency of conversion of glucosinolates to isothiocyanates by the
gastrointestinal micro flora. Cancer Prevention Research. 2012; 5:603-
611.
44. Johnson IT. Glucosinolates in the human diet. Bioavailability and
implications for health. Phytochemistry Reviews. 2002; 1:183-188.
45. Zhao B, Seow A, Lee EJ, Poh WT, Teh M, Eng P et al. Dietary
isothiocyanates, glutathione S-transferase-M1, -T1 polymorphisms and
lung cancer risk among Chinese women in Singapore. Cancer
Epidemiology, Biomarkers and Prevention. 2001; 10:1063-1067.
46. Fowke JH, Chung FL, Jin F, Qi D, Cai Q, Conaway C et al. Urinary
isothiocyanate levels, brassica, and human breast cancer. Cancer
Research. 2003; 63:3980-3986.
47. Seow A, Yuan JM, Sun CL, Van Den Berg D, Lee HP, Yu MC. Dietary
isothiocyanates, glutathione S-transferase polymorphisms and colorectal
cancer risk in the Singapore Chinese health study. Carcinogenesis. 2002;
23:2055-2061.
48. Yanishlieva N, Gordon M. Antioxidants in food. Journal of Agricultural
and Food Chemistry. 2001; 52:2391.
49. Shahidi F, Naczk M. Phenolics in food and nutraceuticals: Sources,
applications and health effects. CRC Press, Boca Raton, FL, 2004.
50. Maggi-Capeyron MF, Ceballos P, Cristol JP, Delbosc S, Le Doucen C,
Pons M et al. Wine phenolic antioxidants inhibit ap-1 transcriptional
activity. Journal of Agricultural and Food Chemistry. 2001; 49:56-46.
51. Falsaperlaa M, Morgiab G, Tartaronec A, Arditoc R, Romano G.
Support ellagic acid therapy in patients with hormone refractory prostate
cancer (HRPC) on standard chemotherapy using vinorelbine and
estramustine phosphate. European Urology. 2005; 47(4):449-54.
52. Prakash D, Gupta C. Role of phytoestrogens as nutraceuticals in human
health. Pharmacology online. 2011; 1:510-523.
53. Lissin LW, Cooke JP. Phytoestrogens and cardiovascular health. Journal

Page | 166
 of the American College of Cardiology. 2000; 35:1403-10.
54. Velasquez MT, Bhathena SJ. Dietary phytoestrogens: A possible role in
renal disease protection. American Journal of Kidney Diseases. 2001;
37:1056-68.
55. Bhathena SJ, Velasquez MT. Beneficial role of dietary phytoestrogens
in obesity and diabetes, The American Journal of Clinical Nutrition.
2002; 76(6):1191-1201.
56. Moreau R, Whitaker B, Hicks K. Phytosterols, phyto stanols, and their
conjugates in foods: Structural diversity, quantitative analysis, and
health-promoting uses. Progress in Lipid Research. 2002; 41:457.
57. Piironen V, Lampi AM. Occurrence and levels of phytosterols in foods.
In Dutta PC (Ed.). Phytosterols as functional food components and
nutraceuticals. Marcel Dekker, New York, 2004, 1-32.
58. Bouic PJ. The role of phytosterols and phytosterolins in immune
modulation: A review of the past 10 years. Current Opinion in Clinical
Nutrition and Metabolic Care. 2001; 4:471-475.
59. Awad AB, Roy R, Fink CS. Beta-sitosterol, a plant sterol, induces
apoptosis and activates key caspases in MDA-MB-231 human breast
cancer cells. Oncology Reports. 2003; 10:497-500.
60. Markowiak P, Śliżewska K. Effects of Probiotics, Prebiotics, and
Synbiotics on Human Health. Nutrients. 2017; 9(9):1021.
61. Sandholm TM, Myllarinena P, Crittenden R, Mogensen G, Fonden R,
Saarela M. Technological challenges for future probiotic foods.
International Dairy Journal, 2002; 12:173-182.
62. Ranadheera RDCS, Baines SK, Adams MC. Importance of food in
probiotic efficacy. Food Research International. 2010; 43:1-7.
63. Thakur M, Satwadhar PN, Deshpande HW. Exploiting Fruits and
Vegetables for Development of Probiotic Products. Trends in
Biosciences. 2015; 8(22):6039-6046.
64. Crittenden R, Playne MJ. Prebiotics. In Lee YK, Salminen S. (eds).
Handbook of Probiotics and Prebiotics. John Wiley & Sons Inc.,
Hoboken, NJ, 2009, 535-561.
65. Shah NP. Functional cultures and health benefits. International Dairy
Journal, 2007; 17:1262-1277.
66. Saad N, Delattre C, Urdaci M, Schmitter JM, Bressollier P. An

Page | 167
 overview of the last advances in probiotic and prebiotic field. LWT-
Food Science and Technology. 2013; 50:1-16.
67. Hassan SM. Soybean, Nutrition and Health. In: El-Shemy H. (ed.)
Soybean. Intech Open, 2013. DOI: 10.5772/54545.
https://www.intechopen.com/books/soybean-bio-active-compounds/
soybean-nutrition-and-health. 18 June, 2018.

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 Chapter - 9
Nutrition and Behaviour

Author
Poonam Sharma
Associate Professor in the Divisin of Food Science and Technology of
Sher-e-Kashmir University of Agricultural Sciences and Technology of
Kashmir, Shalimar Campus, Srinagar, Jammu and Kashmir, India

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 Chapter - 9
Nutrition and Behaviour
Poonam Sharma

Abstract
Food and behaviour is a big topic nowadays and there is an insatiable
desire to find connections between what we eat and what we do. Social
media, various books, magazines, newspaper, articles and television and
radio programs may be one of the potent sources for increasing interest
among people and motivating strategies necessary to change their dietary
behaviours. Consumption of dietary supplements, antioxidants and nutrients
can reduce the risk of progression of age-related problems, nutritional
deficiency disorders and body building is considered useful. It is therefore
essential to design more effective dietary education and dissemination
methods for public awareness on nutrition and behaviour.
Key words: Nutrition, behaviour, malnutrition, neurotransmitter,
intervention
Introduction
Nutrition is defined as the act of process of nourishing or being
nourished, specifically the sum of processes by which an animal of plant
takes in and utilizes food substances. It is the science that interprets the
interaction of nutrients and other substances in food in relation to
maintenance, growth, reproduction, health and disease of an organism. It
includes food intake, absorption, assimilation, biosynthesis, catabolism and
excretion.
Psychology is defined as the study of mind and behaviour in relation to
a particular field of knowledge or activity. The nutritional psychology is the
study of mind and behaviour in relation to the process of taking in and
utilizing food. It studies how cognitive choices such as meal decision,
influence nutrition, psychological health and overall health of human beings.
It may be applied to numerous different fields including psychology,
dietetics, nutrition and marketing.
Nutritional behaviour is the sum total of planned, spontaneous or

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habitual action of individuals or social groups to procure, prepare and
consume food as well as those actions related to storage and clearance. In
this context, the term nutritional behaviour not only refers to influencing
factors but also to health, environmental, social and economic implications
along the entire product chain from farmer to consumer.
Recently food has been a source of technological development.
Nutritional psychologists have studied the public’s perception of food
technology such as genetic modification or additives that may extend the
shelf life of food. Nutritional labels on food products to provide consumers
with the necessary information to make educated decisions about the foods
that they have purchased and nutritional psychologists have done research on
how influential these labels are on how consumers choose what foods to buy.
Nutrition psychology has many implications not only related to how and
what people eat on day today basis but also in which and why they diet and
exercise. Fad diets are also popular in today’s society and they usually play
heavily on potential customer’s ideals about what they should weigh of look
alike. Various setting such as schools, workplaces, supermarkets, primary
care and community based studies have been used by the nutritionists in
order to identify what works for a particular group of people and strategies
are required to change a behaviour in different age groups. Campaigns that
incorporate tailored advice that include practical solutions as well as
environmental change are likely to succeed in facilitating dietary change
behaviour.
Factors that Influence Food Choice and Behaviour
The key driver for eating is of course hunger but what we choose to eat
is not determined solely by physiological or nutritional needs. Some of the
other factors that influence food choice and behaviour include:
Biological: Our physiological needs provide the basic determinants of
food choice. Humans need energy and nutrients in order to survive and will
response to the feeling of hunger and satiety. The central nervous system is
involved in controlling the balance between hunger appetite, stimulation and
food intake. Palatability is proportional to the pleasure someone experience
when eating a particular food. It is dependent on the sensory properties of the
food such as taste, smell, texture and appearance.
Economic: Household income and the cost of the food is an important
factor influencing food choices especially for low income consumers. There
is no doubt that the cost of food is primary determinant of food choice and
dependent on person’s income and socio-economic status. Accessibility to

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shops is important influencing food choice, which is dependent on resources
such as transport and geographical location.
Physical: Studies indicate that the level of education can influence
dietary behaviour during adulthood thus it is important to convey accurate
and consistent messages through media on food packages and health
professionals.
Social: Social influences on food intake refer to impact that one or more
persons have on the eating behaviour of others. Food choice is influenced by
social factors because attitude and habits develop through interaction with
others. What people eat is formed and constrained by circumstances that are
essentially social and cultural. Poor diets can result in under nutrition and
over nutrition and problems are faced in society requiring different level of
expertise and methods of intervention. Attitude, beliefs and knowledge about
food is widely recognized as significant in food decisions. Moreover, healthy
food choices at home and away from home also need to be more readily
available for the people.
Psychological: Psychological stress is a common feature of modern life
and can modify behaviour that affects health by making unhealthy food
choice. The influence of stress on food choice is a complex not least because
of various types of stress one can experience. The effect of stress on food
intake depends upon the individual, the stressor and circumstances. In
general some people eat more and some eat less than normal when
experiencing stress. Depressed mood appears to influence the severity of
food cravings.
Food choices factors also vary according to life stage vary from or group
of people to the next. From an early age, taste and familiarity influence
behaviour towards food. Thus one type of intervention to modify food choice
behaviour will not suit all population groups. Rather, interventions need to
be geared towards different groups of the population with consideration to
many factors influencing their decision on food choice.
Nutrition and Behaviour Relationships: Background
Since ancient times, people have consistently believed that the food they
eat has powerful effects on their behaviour. Some foods have been blamed
for both physical and mental ill heath, and others have been valued for their
curative or magic properties. Beliefs about food play an important role in our
efforts to maintain health and happiness and influence scientific inquiry on
the role of nutrition and behaviour in human life.

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 Direct Effects of Nutrition on Behaviour: Nutrition is usually taken
to be important for physical health but mental health-brain health in its
widest sense must be considered as equally important. A diet lacking
essential nutrients or containing too many ingredients that are detrimental in
excess is likely to have adverse consequences for brain function and thus
mental health and behaviour. It is widely agreed that a balanced diet is
required to support physical health and also beneficial for the healthy
functioning of the brain.
Food and Neurotransmitter: The food we eat affects chemical
composition of our brain and alter our behaviour. Nutrition affects cognitive
possibilities, including alertness and the production or release of
neurotransmitters, the chemical messengers that carry information from one
nerve cell to another. Foods are made of more than one nutrient and their
interaction is going to affect the production and release of neurotransmitters.
Neural impulses are largely resulting from sodium-potassium exchange but
numerous others such as complex carbohydrates, amino acids(tryptophan
and tyrosine),fatty acids particularly omega3 fatty acids affect permeability
of cell membrane, neurotransmitters metabolism and glial cells. The delicate
brain chemical balance is somewhat controlled by the blood-brain barrier.
Still brain remains highly susceptible to changes in body chemistry resulting
from nutrient intake and deficiency. The direct connection between nutrition,
brain function and behaviour exists without a doubt. It can be seen through
brain’s capability of receiving, storing and integrating sensory information
while initiating and controlling motor responses. These functions correspond
to mental activities and form the basis for our brain development and
susceptibility to diseases Banjari et al, (2014).
Certain foods contain precursors or starting material for some
neurotransmitters. If a diet is deficient, the brain will not be able to produce
some neurotransmitters. Neurological and mental disorders may occur when
this balance is upset. Examples of neurotransmitter precursors include
aspartic acid used to make aspartate, the building blocks of protein found in
peanuts, potatoes eggs and grains. Choline used to make acetylcholine which
affects both the peripheral nervous system and central nervous system and is
the only neurotransmitter used in the motor division of the somatic nervous
system associated with the voluntary control of body movements via skeletal
muscles mainly found in eggs, liver and soybeans. Glutamic acid used to
make glutamate which plays a key role in long term potentiation and is
important for learning and memory found in flour and potatoes.
Phenylalanine used to make dopamine, a single organic chemical released by

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nerve cells to send signals to other nerve cells mainly found in beets, soya
beans, almonds, eggs, meats and grains. Tryptophan used to make serotonin
found in gastrointestinal tract, platelets and central nervous system. They act
as many function in the brain such as regulation of mood, appetite and sleep.
Serotonin also has cognitive functions including memory and learning. The
food sources are eggs, meat, skim milk, bananas, yogurt, milk and cheese.
Tyrosine used to make norepinephrine which functions as the
neurotransmitter released from the sympathetic neurons affecting the heart.
As stress hormone norepinephrine affects parts of the brain where attention
and fight of flight senses are controlled mainly found in milk, meat and
legumes.
Long Term Effects of Chronic and Acute under Nutrition: Diet can
affect cognitive ability and behaviour in children and adolescents. Nutrient
composition and meal pattern can exert immediate or long term beneficial or
adverse effects. Beneficial effects mainly result from the correction of poor
nutritional status. For example, thiamine treatment reverses aggressiveness
in thiamine deficient adolescents. Deleterious behavioural effects have been
suggested for example sucrose and additives were once suspected to induce
hyperactivity but these effects have not been confirmed by rigorous
investigations. In spite of potent biological mechanisms that protect brain
activity from disruption some cognitive functions appear sensitive to short
term variations of fuel (glucose) availability in certain brain areas. A glucose
load, for example, facilitates mental performance particularly on demanding
long duration tasks. Even intelligent scores can be improved by
micronutrient supplementation in children and adolescents with very poor
dietary status.
Nutritional behaviour is framed by biological, anthropological,
economic, psychological, socio-cultural, and home economics and is shaped
by individual situation and from a public point of view it is associated with
preventable cases of various diseases. The diet of an organism is what it eats,
which is largely determined by the availability and palatability of foods. For
humans, a healthy diet includes preparation of food and storage methods that
preserve nutrients from oxidation, heat or leaching and that reduce the risk of
foodborne diseases. In humans an unhealthy diet can cause deficiency related
diseases such as blindness, anaemia, scurvy, preterm birth still birth and
cretinism or nutrient excess health-threatening conditions such as obesity
and metabolic syndrome and some common chronic systemic diseases as
cardio vascular diseases like diabetes and osteoporosis. Under nutrition can
lead to wasting in acute cases and stunting of marasmus in chronic cases of
malnutrition.

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Changing Nutrition and Behaviour: Successful Intervention
Dietary intervention is not easy because it requires alteration in habits
that have been built over a life time. Proper nutrition and health are closely
interrelated throughout life, but probably the highest importance is expressed
in the first years of life.
Impact of Protein Energy Malnutrition on Behaviour: Inadequate
nutrition causes lower cognitive development, reduced attention and
concentration and reduces performance in later life. Also, foetal
programming in utero should not be neglected for its proven influence on the
later development of a child. As nicely illustrated by Vanhees, et al. (2014)
we are what we eat and so are our children. Their extensive view on
epigenetic studies clearly illustrates the importance of balanced diet of both
mother and father. Besides macronutrient composition of the diet (high fat
diets, protein restricted diets, diet high in carbohydrates) intake of specific
micronutrients, especially those involved in one carbon metabolism(folic
acid, vitamin B2,B6 and B12) day by day shows more potential in
programming offspring’s epigenome. At birth the brain reaches 70% size
and 25% weight of an adult brain. In the subsequent period are created new
nerve cells (neurons) that travel to their final destination Brain changes
throughout life. Brain is a very dynamic organ, showing high plasticity. Due
to the characteristic altering our diet in terms of having a balanced nutrition
without any deficiency or over nutrition can preserve our brain from
deterioration. For example one study showed that high dose supplementation
with folic acid during early pregnancy shows association with increased
neurodevelopment resulting in enhanced vocabulary development,
communication skills and verbal comprehension at 18 months of age
(Chatzi, et al. 2012). Similar findings have been shown for boosting
cognitive performance and intake of iron (after correcting iron deficiency
anaemia. We can boost our mood by retaining available neurotransmitters in
the gap between nerve cells as long as possible and it seems possible but yet
to be tested that expression of foods in art can also serve to improve mood.
Regulation of three key neurotransmitters responsible for mood (dopamine,
noradrenaline and serotonin) by modulating food intake impacts durability of
their stimulation of nerve cells thus impacts mood and behaviour.
Caffeine and Dietary Sugar: A study of 8000 people has shown that
people who consume chocolate live longer compared to those who never eat
chocolate. Positive effect of a chocolate lies in its flavonoid content as they
reduce the amount of low-density lipoprotein (LDL) cholesterol and reduce
blood pressure. They also show the potential to slowdown the growth of

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cancer cells. Poletti et al., (2012). Overconsumption of sugar alone can
negatively impact young people’s mental health. In a sample of 5,498 youth
aged 15-16 in Oslo Norway researchers found a strong association between
mental health problems. In a related study researchers found a positive
association between consumption of sugary soft drinks and sweet foods and
risks for suicidal behaviours among adolescents aged 12-19, Jiangsu
Province, China.
Stress and Mental Functioning: Emotional eating-the habit of eating
to cope with stress boredom or negative feelings rather than to satisfy the
hunger is a behaviour that can lead to overeating and weight gain. Study
investigated the effects of a nutrition education program on dietary
behaviour and nutrition knowledge among second grade and third grade
students participating in 8 week nutrition education program. Results
indicated that the intervention effectively improved dietary behaviour and
increased knowledge.
Food Additives and Behaviour: The food additives and poor diet could
help explain poor school performance, criminal behaviour, alcoholism, and
the growing number of Alzheimer’s patients.
Obesity and Eating Disorders: Obesity represent a major health
problem and increases the risk of various diseases and is associated with a
range of adverse psychological consequences. As a result weight reduction
has become a preoccupation for many people and growing popularity of diet
centres, gymnasium, clubs and health professionals. Eating disorders like
anorexia nervosa and bulimia are mostly observed in young women due to
our culture’s recent obsession with thinness and a stigma against obesity.
Dietary Supplements and Brain Behaviour: Dietary supplements are
becoming a fast lucrative industry. There is a striking escalation in dietary
supplements believing that vitamin and minerals will prevent or delay or
prevent variety of diseases from common cold to arthritis and cancer, delay
the aging process and alleviate feelings of fatigue, and generally improve
psychological wellbeing. People take these supplements to make sure for
curing the treatment of diseases like Alzheimer’s and claims that foods and
supplements may reduce the risk and proved beneficial.
Behavioural Modification of Diet and Behaviour: Eating a healthy
diet that includes lot of fresh, locally grown organic vegetables and fruits.
Take a high quality omega-3 supplement such as fish oil. Avoid sugar and
grains including fruit juices, breads, white rice, pasta and potatoes. Avoid
anything with high fructose from syrup. Avoid all artificial sweeteners and

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limit monosodium glutamate intake by avoiding most processed foods.
Conclusion
Within the past decade there has been a veritable explosion of interest in
the relationship between food and nutritional behaviour. Food and behaviour
is a big topic nowadays and there is an insatiable desire to find connections
between what we eat and what we do. Social media, various books,
magazines, newspaper, articles and television and radio programs may be
one of the potent sources for increasing awareness in nutrition and
behaviour. When we make healthy food choices we tend to feel better and
when we eat heavy, sugary foods we can feel tired or sluggish. Studies have
shown that food can promote proper functioning of the brain. In order to
improve our mental abilities concentration memory and vigilance proper
nutrition is of great importance. By affecting neurotransmitters, substances
that activate different regions of the brain actively participate in the creation
of nerve impulses and thereby regulate our mental abilities emotions and
behaviour. Cognitive performance and maintenance of mental health
especially among elderly may be improved with proper diet consisting of
complex carbohydrates, poly-saturated fatty acids especially omega3 fatty
acids protein and specific foods containing specific nutrients like flavonoids.
In addition mood and concentration as well alertness can be affected by
moderate consumption of chocolate and caffeine beverages. Keeping in mind
the risk factors for loss of mental abilities by proper nutrition we can
potentially prevent or delay neurodegenerative changes in the brain
including Parkinson’s and Alzheimer’s disease. Designing nutrition
education programs that are likely to change dietary behaviour is a complex
task. Once mediating variables are identified and prioritized, strategies for
changing them can be developed. Nutritionists are encouraged to creatively
apply the behavioural nutrition principles to their education programs.
References
1. Bellisle F. Effects of diet on behaviour and cognition in children British
J Nutrition review article Oct. 2004; 92(2):227-32
2. Paoletti R, Poli A, Conti A. Chocolate and health, Springer, Milan,
2012.
3. Wenk GL. Your brain on food. How chemicals control your thoughts
and feelings. Oxford University press, New York, 2010.
4. Chatzi L, Papadopoulou E, Koutra K. Effect of high doses of folic acid
supplementation in early pregnancy on child neurodevelopment at 18

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 months of age; the mother child cohort Rhea study in Crete, Greece.
Public Health Nutr. 2012; 15(9):1728-1736.
5. Banjari Ines, Vuoje Ivana, Mandic Milena L. Brain food: how nutrition
alters our mood and behaviour in Hrana u zdravlju I bolesti, znanstveno
strucini casopis za nutricionizan I dijetetika. 2014; 3(1):13-21.
6. Vanhees K, Vonhogen IGC, Van Schooten FJ. You are what you eat and
so are your children: the impact of micronutrients on the epigenetic
programming of offspring. Cell Mol. Life Sci. 2014; 71:271-285.
7. Worobey John, Tepper Beverly J, Kanarek Robin B, Anci Krist DE.
Nutrition and behaviour-A multidisciplinary approach CABI
Publication, 2015.
8. Preedy Victor R, Watson Ronald Ross, Martin Collin R. Handbook of
Behaviour, Food and Nutrition-1st edition, Springer Science-Business
Media, LLC, 2011.
9. Robin B, kanarek Robin. Marks Kaufman Nutrition and behaviour
Springer, Boston MA, 1991.
10. Ban TA. Magavitamin therapy in Schizophrenia. In Nutrition and
behaviour, ed Miller SA, Philadelphia; Frankiin Institute, 1981, 247-
253.

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