INCORPORATION OF PEARL MILLET AND FLAXSEED MILK FOR

THE FORTIFICATION OF SUGAR FREE ICE-CREAM USING

RESPONSE SURFACE METHODOLOGY

THESIS SUBMITTED IN PARTIAL FULFILMENT OF

THE REQUIREMENTS FOR THE DEGREE OF

Master of Science
IN

Food Science and Technology

Supervisor Submitted By
Dr. Amrita Poonia Prashasti Yadav

CENTRE OF FOOD SCIENCE AND TECHNOLOGY

INSTITUTE OF AGRICULTURAL SCIENCES
BANARAS HINDU UNIVERSITY
VARANASI-221005
INDIA

I.D. No. FST-15259 2018 Enrolment No. 379824

DEDICATED TO LORD KASHI
VISHWANATH

i
 Center of Food Science and Technology
Dr. Amrita Poonia Institute of Agricultural Sciences
Assistant Professor
Banaras Hindu University
Varanasi-221005, U.P, India
Center of Food Science and Technology
E-mail: dramritapoonia@gmail.com

Mob: +91- 9532030058

Ref. No . ………. Dated: ……………………………

CERTIFICATE
To,

The Registrar (Academic)

Banaras Hindu University, Varanasi - 221005 (U.P), INDIA
Varanasi-221005

Through: The Coordinator, Center of Food Science and Technology, Institute of

Agricultural Sciences, Banaras Hindu University.

Dear Sir,

I have great pleasure in forwarding the thesis entitled, “INCORPORATION OF

PEARL MILLET AND FLAXSEED MILK FOR THE FORTIFICATION OF SUGAR FREE ICE-
CREAM USING RESPONSE SURFACE METHODOLOGY” submitted by Ms. Prashasti
Yadav, I.D No. FST- 15259 in partial fulfillment of the requirements for the degree of
Master of Science in Food Science and Technology, Institute of Agricultural Sciences,
Banaras Hindu University, Varanasi and placing on record that she has completed the
requisite residential requirements as contained in the ordinance of the University.

I certify that the entire scheme of investigation, presented here in, was planned and
carried out solely by the candidate under my supervision and guidance. To the best of my
knowledge, the data presented in the thesis are genuine and original.

Thanking You

DR. ANIL KUMAR CHAUHAN DR. AMRITA POONIA

(COORDINATOR) (SUPERVISOR)

ii
INCORPORATION OF PEARL MILLET FLOUR AND FLAXSEED MILK FOR THE
FORTIFICATION OF SUGAR FREE ICE-CREAM USING RESPONSE SURFACE
METHODOLOGY

By

Ms. Prashasti Yadav

Thesis submitted in partial fulfilment of the requirements for the degree of

MASTER OF SCIENCE

(FOOD SCIENCE AND TECHNOLOGY)

FROM

CENTRE OF FOOD SCIENCE AND TECHNOLOGY,

INSTITUTE OF AGRICULTURAL SCIENCES,

BANARAS HINDU UNIVERSITY,

VARANASI,221005

2018

ID. No. FST-15259 Enrolment No. 379824

APPROVED BY ADVISORY COMMITTEE

Chairman : Dr. Amrita Poonia

Assistant Professor

Centre Of Food Science And Technology,

Institute Of Agricultural Sciences, B.H.U., Varanasi

Member : Prof. Anil Kumar Chauhan

Professor

Centre Of Food Science And Technology,

Institute Of Agricultural Sciences, B.H.U., Varanasi

Member : Dr. D. C. Rai

Professor

Dept.of Dairying and Animal Husbandry,

Institute Of Agricultural Sciences, B.H.U.

Varanasi

EXTERNAL EXAMINER:

iii
 Acknowledgments

I bow my head with great reverence in the pious feet of

Bharat Ratna Mahamana Pandit Madan Mohan Malviya Ji, a
man of great vision and founder of the Banaras Hindu
University, Varanasi, India, for his lifetime sacrifice and efforts
in establishing such a temple of learning for the cause of the
millions of students like me.

I bow my head in great reverence to the omnipotent and

omnipresent Lord ‘Kashi Vishwanath’ for their blessings and
true love towards me and to every creature resending on this
wonderful planet. Also, I'm highly obliged to this holy city
Varanasi which is situated on the banks of Maa Ganga.

It is my sublime privilege to express my deep sense of

reverence and gratitude to my supervisor Dr. Amrita Poonia,
Assistant Professor, Centre of Food Science and Technology,
Institute of Agricultural Sciences, Banaras Hindu University,
Varanasi for her inspiring and ingenious guidance, incisive and
articulate criticism, cordial discussion, unending zeal and
constant encouragement which assisted me to overcome every
problem that came in my way during the period of this
investigation and preparation of this manuscript. I shall always
remain deeply indebted to her for her affectionate glances and
generosity bestowed on me.

Words would fail to express my deep sense of gratitude to

explain the help, affection and constant encouragement
rendered by Prof. Anil Kumar Chauhan, Coordinator, Centre of
Food Science and Technology for providing the necessary
facilities during the present study.

I am also expressing my gratitude to the members of my

advisory committee Prof. Anil Kumar Chauhan, Coordinator,
Centre of Food Science and Technology and Dr. D. C. Rai,
Professor, Department of Dairying and Animal Husbandry,

iv
Institute of Agricultural Sciences for their constant
encouragement and for boosting my morale during
experimentation.

I owe an individual debt of gratitude to Asst. Professor, Dr.

A. D. Tripathi, Dr. Arvind, Er. D. S. Bunker, and I am thankful
to all the faculty members of Centre of Food Science and
Technology, Institute of Agricultural Sciences, B.H.U or their
invaluable cooperation and encouragement during my entire
course of the investigation. They had spent their valuable time
for me, whenever I needed.

I feel paucity of words to express my gratefulness and

warmest regard to all the non-teaching staff members especially
Mr. Amaresh Singh and Mr. Chandrashekhar for their timely
help and co-operation during my study.

I have no words to express my feelings about the

contribution of my respected father, Shri. Onkar Nath Yadav,
for his blessings, sacrifices, affection, encouragement, moral and
financial support throughout my life. I also express my heartfelt
regards to my loving mother, Smt. Padmawati Yadav whose
flawless love, blessings and guidance helped me to overcome all
the barriers in life. I also owe my heartfelt gratitude to my
maternal grandparents, Shri. Rajwant Singh and Smt. Badami
Devi for their love, blessings and constant support during my
stay in Varanasi.

This work would have been rather incomplete without

the moral support and constant inspiration of my beloved
brother Mr. Utkarsh Yadav.

I am also thankful to my cousins Eshita Singh, Atharv

Gaurav and Anushka Raj for their love and best wishes
throughout my life. I am also thankful to all my family
members along with relatives for their blessings and
encouragement without which, I could not have achieved my
task.

v
 I can’t refrain myself from expressing my deep debt of
gratitude to my friend Amit Yadav who always encouraged me
to keep my morale high. I am thankful to him for his moral
support, co-operation and priceless suggestions throughout my
studies.

It is difficult to express the gratitude I owe to all my

friends especially Priyanka Arya, Sarvjeet Kaur, Shubhum
Arora, Satya Prakash, Ranjeet, Anamika, Saumya Joshi, Ekta,
Satyam and all my batchmates for their love and affection
which always animated me to face the challenge.

Without the help of seniors, none can learn the lesson of

life. So, heartful and special gratitude to my seniors, Ms. Suman
Rai, Ms. Prachi Tyagi, Mrs. Sonika, Mrs. Poonam Yadav Mr.
Abhinay Shashank Jha and Mr. Mohammad Alsaebai.

Last but not the least, I record my sincere thanks to all

respectable people and teachers who helped and guided me but
could not find a separate mention. I still solicit their benediction
to proceed with every step of a perfect destined life.

Date: ……………………

Place: Varanasi (PRASHASTI YADAV)

vi
 Contents

LIST OF TABLES ..................................................................................................................................... viii

LIST OF FIGURES ..................................................................................................................................... ix

ABBREVIATIONS ...................................................................................................................................... x

INTRODUCTION ........................................................................................................................... 1

REVIEW AND LITERATURE ............................................................................................................ 5

MATERIALS AND METHODS ........................................................................................................ 12

RESULTS AND DISCUSSION ......................................................................................................... 22

SUMMARY AND CONCLUSIONS .................................................................................................. 38

BIBLIOGRAPHY........................................................................................................................... 40

APPENDICES………………………………………………………………………………………………………………………………43

vii
 LIST OF TABLES

Table 1.1 Nutritional Profile of Flax seeds

Table 2.1 Classification of ice-cream on the basis of FSSAI (2006)

Table 2.2 Approximate composition (percentage by weight) of some types of

Ice-cream

Table 3.1 Independent variables for optimization

Table 3.2 Texture analyzer settings for the analysis of Ice-Cream

Table 4.1 Experimental design for optimization of PMF fortified ice-cream

Table 4.2 Coefficient of Estimation of Coded Factors for different factors of

optimization of PMF fortified Ice-Cream

Table 4.3 Constraints fixed for optimization of levels

Table 4.4 Optimization of Pearl Millet and Flaxseed fortified ice-cream

Table 4.5 ANOVA on the predicted values for the different attributes of PMF
fortified ice-cream optimized by RSM with the actual product prepared
experimentally

Table 4.6 Proximate composition of CSM ice-cream

Table 4.7 Various physical parameters of optimized PMF ice-cream

Table4.8 Representing the volume of melted ice-cream

Table 4.9 Textural attributes of PMF fortified ice-cream

Table 4.10 Sensory Score for optimized Pearl Millet and Flaxseed fortified ice-
cream

viii
 LIST OF FIGURES

Fig. 3.1 Flowchart For Preparation Of Millet And Flaxseed Milk Fortified Ice-
Cream

Fig. 4.1 Response Surface Plots showing the effect of the composition of
ingredients on Colour and Appearance of Ice-cream

Fig. 4.2 Response Surface Plots showing the effect of the composition of
ingredients on Body and Texture of Ice-cream

Fig. 4.3 Response Surface Plots showing the effect of the composition of
ingredients on Flavour of Ice-cream

Fig. 4.4 Response Surface Plots showing the effect of the composition of
ingredients on Overall Acceptability of Ice-cream

Fig. 4.5 Pearl Millet and Flaxseed Milk fortified ice-cream

ix
 ABBREVIATIONS

% : Percent

ANOVA : Analysis of variance

AOAC : Association of Official Analytical Chemists

CCRD : Central Composite Rotatable Design

g : Gram

IU : International Unit

K cal : Kilo calories

mg : Mili grams

ml : Mili litres

̊C : Degree celcius

PMF : Pearl Millet and Flaxseed

RSM : Response Surface Methodology

SDF : Soluble Dietery Fibre

TPA : Texture Profile Analysis

TS : Total Solids

TSS : Total Soluble Solids

x
INTRODUCTION

1
 INTRODUCTION

Pearl millet (Pennisetum americanum) is a native of Africa and is successfully cultivated in

India. Among millets, bajra is the predominant crop in India, occupying, during 2004 an area
of 7.5 million hectares with a production of 4.6 million tonnes and a yield of 6.1 quintals per
hectare. (N. Shakuntala Manay and M. Shadaksharaswamy,2008).

The coarse grain contains 8-10 percent husk. The average chemical composition of bajra
grain is as follows: moisture, 12.4%; protein, 11.6%; fat, 5.0%; carbohydrate, 67.1%; fiber,
1.2%; and mineral matter, 2.7%. The mineral matter is rich in calcium, phosphorus, and iron.
More than 50% of the phosphorus is phytinwhich is a major factor for the poor digestibility
of the bajra grains. The protein content of bajra varies from 8.8 to 16.1 percent. The protein
contains a high proportion of prolamine, followed by globulins and albumins. Among the
amino acids, tryptophan content is high, and lysine content is average to low. The
carbohydrates consist mostly of starch with the smaller amount of sugars (1.2 percent),
peniosans and hemicelluloses. The starch is composed of 32.1 percent amylose and 67.9
percent amylopectin. The grains are rich in thiamine, riboflavin, and niacin. (N. Shakuntala
Manay and M. Shadaksharaswamy,2008).

Flaxseeds are known as Linum usitatissimumwith its species name meaning ‘‘most useful''.
That would describe the versatility and nutritional value of this tiny little seed. Flax seeds are
slightly larger than sesame seeds and have a hard shell that is smooth and shiny. Their color
ranges from deep amber to reddish brown, depending upon whether the flax is of golden or
brown variety. While whole flaxseeds feature a soft crunch, the nutrients in ground seeds are
more easily absorbed. The warm, earthy and subtly nutty flavor of flax seeds combined with
an abundance of omega-3 fatty acids makes them an increasingly popular addition to the diets
of many a health-conscious consumer. Whole and ground seeds and oils are available
throughout the year. (N. Shakuntala Manay and M. Shadaksharaswamy,2008).

Flaxseed oil is rich in alpha-linolenic acid, an omega-3 fat found in fish oils called
eicosapentaenoic acid or EPA. Alpha-linolenic acid or ALA, in addition to providing several
beneficial effects of its own, can be converted in the body to EPA, thus providing EPA’s
beneficial effects. For this conversion to readily take place, however, depends on the presence
and activity of enzyme called delta-6-destaurase, which, in some individuals, is less available
or less active in others. Also, the delta-6-deataurase function is inhibited in diabetes and by
the consumption of saturated fat and alcohol. For these reasons, higher amounts of ALA-rich
flax seed oil must be consumed to provide the same benefits as the omega-3 fats found in the
oil of cold water fish. (N. Shakuntala Manay and M. Shadaksharaswamy,2008).

Pearl Millet is a good source of dietary fibres and is a cheap source of various nutrients. This
enables it to find use in various dairy products. It is easily available in the local market and is
a cheaper source of various minerals and carbohydrates. Flaxseed is also a rich source of

2
protein and fats, especially omega-3 fatty acids. Therefore, its usage in the ice-cream helps to
fortify it with the alpha linolenic acid which has numerous beneficial effects on the human
body on consumption.

Table 1.1 Nutritional Profile of Flax seeds

Nutrient amount DV (%) World’s healthiest

food rating
Omega-3 fatty 3.51 g 140.4 Excellent
acids
Manganese 0.64 mg 32.0 Very good
Dietary fiber 5.41 g 21.6 Very good
Magnesium 70.14 mg 17.5 Good
Folate 53.86 mcg 13.5 Good
Copper 0.20 mg 10.0 Good
Phosphorus 96.49 mg 9.6 Good
Vitamin B-6 0.18 9.0 Good
(pyridoxine)
Source: (N. Shakuntala Manay and M. Shadaksharaswamy,2008).

Ice-cream is a frozen product obtained from cow or buffalo milk or a combination thereof or
from cream, and/or other milk product, with or without the addition of cane sugars, eggs, fruit
juices, fruits, preserved fruits, nuts, chocolate, edible flavors and permitted food colors. It
may contain permitted stabilizer and emulsifier not exceeding 0.5% by weight. The mixture
must be suitably heated before freezing. The product should contain not less than 10% milk
fat, 3.5% protein and 36% total solids. (FSSAI,2011).

Stabilizers are defined as substances which help to preserve emulsions. A small quantity of
stabilizers is used in food products. It has a very important role to play in food products.
Stabilizers are added in ice-cream to produce smoothness in body and texture, retard or
reduce ice crystals growth during storage and provide uniformity in the product and
resistance to melting. Stabilizer functions through their ability to form gel structures in water
or combine with water as water of hydration. (Sukumar De,2013).

An emulsifier is defined as a substance which helps to form emulsions. The values of

emulsifying agents in the manufacture of ice-cream lies mainly in the improved whipping
quality of the mix, the production of a drier ice-cream with a smoother body and texture in
their superior drawing qualities in the freezer, and the possibility of maintaining more precise
control over the various manufacturing processes. (Sukumar De,2013).

Ice-cream is a product commonly enjoyed by people of all ages due to its cooling effect. The
nutritive value of ice-cream is high, as it is a milk based dessert. However, it is made up of
fat, emulsifiers and sugar. As today’s consumers search for healthier food products with
functional appeal, low caloric density, high fibre content and the sensory qualities of

3
traditional products, there is an opening in the market for the development of new
formulations of ice-cream to comply with consumer demands. (Karaman et al., 2014).

Objectives:

 To study the sensory attributes of ice-cream on addition of pearl millet flour and flax
seed milk.
 To study the effect of fortification on the nutritional composition of ice-cream.

4
REVIEW AND LITERATURE

5
 REVIEW AND LITERATURE

2.1 Incorporation of cereals in ice-cream

I.J. Patel et al.,(2012) studied that Ragi (Finger millet) improves the nutritional value of ice
cream by enhancing the iron and fibre content. Caramel flavoured medium fat ice cream (6 %
fat) was prepared by addition of gelatinized malted ragi flour roasted in butter (MRB) @ 8 %,
9 % and 10 % by weight of mix and compared with control (C) i.e. vanilla ice cream
containing 10 % fat. The overall acceptability score of product prepared using 9 % MRB was
statistically (P>0.05) at par with the C, hence, it was selected. In the next part of the study,
ragi ice cream was prepared using 4 different flavours viz. vanilla, mango, chocolate and
caramel. Chocolate flavoured ragi ice cream was adjudged as best, followed by mango,
caramel and vanilla ice cream. The iron and fibre content of chocolate flavoured ragi ice
cream was found to be 12.8 ppm and 1.36 % respectively. vs. 1.5 ppm and 0.18 %
respectively in control (C). Heat shock treatment as well as storage up to 30 days had no
adverse effect on the sensory quality of the chocolate flavoured ragi ice cream. Incorporation
of finger millet in ice cream resulted in reduction in the amount of stabilizer used and
effectively functioned as fat replacer in ice cream.

Sivakumar et al.,(2017) conducted a study to develop a nutrient-rich foxtail millet

incorporated sugar-free ice cream. Foxtail millet is a good source of protein, dietary fibre and
low in carbohydrate content. Foxtail millet also has good thickening and water binding
properties Ice cream was prepared using foxtail millet at 2%,3% and 4% levels and sugar was
replaced by stevia calorie free sugar substitute at 3% level and subjected to sensory
evaluation. Various sensory parameters like flavour, body and texture, colour and
appearance, sweetness and overall acceptability were analysed for the samples S1, S2, S3 and
S4. It was found that the sample with 4% foxtail millet (S4) gained highest scores and was
liked the most when compared to other samples. The Statistical analysis of data revealed that
highly significant difference (P ≤0.01) was observed in flavour, body and texture, colour and
appearance and overall acceptability between samples.

2.2 Flax and flaxseed oil

Flaxseed is one of the oldest crops, having been cultivated since the beginning of civilization.
The Latin name of the flaxseed is Linum usitatissimum, which means “very useful”. Flax has
been used until 1990s principally for the fabrication of cloths (linen) and papers, while
flaxseed oil and its sub-products are used in animal feed formulation (Singh et al., 2011a).
There is a small difference in using the terms flaxseed and linseed. Flaxseed is used to
describe flax when consumed as food by humans; while linseed is used to describe flax when
it is used in the industry and feed purpose (Morris, 2008). In the last two decades, flaxseed
has been the focus of increased interest in the field of diet and disease research due to the
potential health benefits associated with some of its biologically active components. Flax
contains approximately 40% fat, 28% dietary fibres, 21% proteins, 4% ash and 6%
carbohydrates such as sugars, phenolic acids, lignans, hemicelluloses.(AOCS, 2000).

6
2.3 Ice-cream

Marshell et al., (2003) describe0d ice-cream as a frozen emulsion of dairy or non-dairy fat,
sugars, proteins, stabilizers, and flavours, which undergoes several manufacturing steps that
confers the desirable smoothness, and softness of ice-cream.

Stanley et al., (1996) defined ice-cream as a complex food foamed emulsion sustained by the
combination of several components; ice crystals, fat network, air bubbles and a highly
concentrated unfroze part.

Khanna, (2004) defined ice-cream as a complex food colloid made up of four phases of air
(50%), ice (20%), fat (5%) and matrix (25%).

2.3.1 Classification of ice-cream

There are various type of ice-cream available in market. They are classified on the basis of
their composition i.e. fat, total solids, sucrose and stabilizers/emulsifiers and per cent overrun
as classified under the following:

Table 2.1 Classification of ice-cream on the basis of FSSAI (2006)

Requirement Ice-cream Medium fat ice- Low fat ice-

cream cream
Total solids Not less than Not less than Not less than
36% 30% 26%
Wt/vol (g/l) Not less than 525 Not less than 475 Not less than 475
Milk fat Not less than More than 2.5% Not more than
10% but less than 10% 2.5%
Milk protein Not less than Not less than Not less than
3.5% 3.5% 3.5%

Table 2.2 Approximate composition (percentage by weight) of some types of Ice-cream

ConstituentsDairy ice- Ice milk Sherbet Ice lolly

cream
Milk Fat 10 4 2 0
Milk Solids 11 12 4 0
Not Fat
Added Sugar 14 13 22 22
Additives 0.4 0.6 0.4 0.2
% Overrun 100 85 50 0
Energy, 390 300 340 370
KJ/100 ml
Wastrel et al., (2006)

7
2.3.2 Composition and properties of ice-cream

Mehu, (1993) has suggested the main ingredients of ice-cream and they are milk, cream,
sugar, stabilizer, emulsifier and flavouring materials. The most of these ingredients in ice-
cream are multifunctional, contributing to different aspects of ice-cream manufactured
product quality and safety.

Marshall, (1991) and Goff et al., (1995) reported that m ilk fat is the main ingredient of ice
cream which forms the body and texture of ice-cream. Milk fat should be present in ice-
cream in range of 10%-20%.

Marshall and Arbuckle, (1996) have observed that the effect of milk fat in ice-cream is
dependent on several factors including the type and level of fat, the amount of non-fat milk
solids and sugar as well as variety of flavour compounds used in ice-cream. Milk fat also
affects the textural properties such as viscosity, tenderness, elasticity, emulsification, ice
crystallisation and other desirable attributes such as richness, smoothness.

Arbuckle, (1986) reported that the non-milk solid not fat contains approximately 55%
lactose, 37% proteins, 8% minerals and others such as vitamins, acids, enzymes. Legal limit
of non-milk solid not fat is in the range of 9%-12%. Emulsifier in ice-cream plays an
important function. Emulsifier has improved the meltdown resistance and smoother body
texture.

Keeney, (1982) reported that if emulsifiers are present in ice-cream then the overrun of ice-
cream is high, air is finely dispersed and foam structure is more stable.

2.4 Stabilisers

Maryam Bahramparvar and Mostafa Mazaheri Tehrani, (2011) has suggested that
stabilizers are substances that, are used in low levels in ice-cream mix, but have very
important functions, such as increase in viscosity of ice-cream mix, aeration improvement,
cryoprotection and control of melt down. A good stabilizer should be non-toxic, readily
disperse in the mix, not produce excessive viscosity or separation or foam in the mix, not clog
strainers and filters, provide ice-cream with desirable meltdown, be economical, and not
impart off flavours to the mix.

Keeney, (1982) studied the role of stabilizers and states that the basic role of stabilizer is to
reduce the amount of free water in the ice-cream mix by binding it as ‘’water of hydration’’,
or by immobilizing it within a gel structure. Also it is the ability of small percentage of
stabilizer to absorb and hold large amounts of bound water, which produces good body,
smooth texture, slow melt down and heat shock in the resultant product.

Marshall R.T. et al., (2003) reported that the amount and kind of stabilizer required in ice-
cream depend on its properties, mix composition and ingredients used; processing times,
temperatures and pressures; storage temperature and time; and many other factors. Usually
0.1-0.5% stabilizer is utilized in the ice-cream mix. Mixes high in fat or total solids (40%),
chocolate mixes or ultra-high temperature pasteurized mixes require less stabilizer than do

8
mixes that are low in total solids (37%), are high-temperature short-time (HTST) pasteurised,
or are to be stored for extended periods of time.

2.4.1 Common Stabilizers used in Ice-cream

Maryam Bahramparvam and Mostafa Mazaheri Tehrani, (2011) have studied the types
of stabilizer and its functions in ice-cream. Following stabilizers are studied.

Clarke C. (2004) has observed that guar gum is extracted from the seeds of a tropical
legume, Cyamoposis tetragonolba, called guar. It has been grown in India and Pakistan for
centuries and, for a short time and to a limited extent, in the United States. It is the least
expensive stabilizer and effectively decreases the undesirable effects of heat shock in ice-
cream. It readily disperses and does not cause excessive viscosity in the mix. Generally, 0.1-
0.2% is required in a mix a nd, therefore, this substance is considered to be a strong stabilizer.

Glicksman, (1986) reported that sodium carboxymethyl cellulose (CMC) chemically

modified natural gum is a linear, long-chain, water-soluble and anionic polysaccharide.
Purified sodium carboxymethyl cellulose is a white-to-cream coloured, tasteless, odourless,
free-flowing powder. CMC forms weak gels by itself but gels well in combination with
carrageenan, locust bean gum, or guar gum. It is a strong stabilizer and only 0.1-0.2% is
needed in a mix. It imparts body and chewiness to ice-cream.

Kilara A. (2008) gelatin is a relatively expensive stabiliser and is effective at concentrations

of 0.3-0.5%; however, it may not prevent the effects of heat shock. It is also not acceptable to
certain religious and vegetarian populations. The use of gelatin as a stabilizer produces thin
mixes that require a long aging period. Gelatin disperses easily and does not cause wheying
off or foaming.

2.4.2 Effects of Stabilizers on Rheological Properties of Ice-cream mix

E. Mahdian and R. Karazhian, (2013) had observed that the ice-cream mixes were
pseudoplastic fluids with apparent viscosity values decreasing with increasing shear rate. Mix
viscosity and consistency coefficient increased while flow behaviour index decreasing by use
of MPC (Milk Protein Concentrate) and inulin and as well with an increase in either type of
stabilizer level. Ice-cream hardness was not affected by type and amount of stabilizer but
overrun values decreased with increasing MPC, inulin and stabilizers’ level in the product
formulation.

Maryam Bahramparvar & Mostafa Mazaheri Tehrani, (2011) reported that the viscosity,
which is one of the most important rheological properties of ice-cream mix and the unfrozen
portion of ice-cream, is influenced by mix composition (mainly stabilizer and protein), type
and quality of the ingredients, processing and handling of the mix, concentration (total solid
content) and temperature. The viscosity of ice-cream mix is set through mix composition
particularly stabilizer content and level. Although it is generally understood that mix
viscosity is important to impart desirable qualities of ice-cream, the specific rheological
parameters required are not well understood. Generally, as the viscosity increases, the

9
resistance to melting and the smoothness of texture increases, but the rate of whipping
decreases.

Dogan and Kayacier, (2007) investigated the effects of ageing on the rheological properties
of kahramanmaras-type ice-cream mix. By evaluating n, K and apparent viscosity values,
they suggested that 24 hours of ageing at 0̊ C would be a proper ageing time for the ice-
cream mix. After 24 hours, K and apparent viscosity reached the highest values, whereas the
n reached the lowest value. However, it has been indicated for ice-cream mixes containing
stabilizers that about 4 hours of ageing is sufficient.

2.4.3 Effect of Stabilizers on Volume Increase (Overrun)

Qamar Abbas Syed and Muhammad Shoaib Ullah Shah, (2016) called ice-cream, an
aerated product and defined as frozen texture. More ice-cream quantity is one responsibility
of stabilizer, bring about from beginning to end more viscosity and persisting the air
molecules. The quantity of air in product is central since it affects superiority and income
saves for also since of authorised principles that should be gained. Moreover, the air cubicle
organization has confirmed to be the chief agent affecting melting, and the flow characters in
the frozen condition, this is linked to creaminess. Slighter air cells perk up the superiority
concerning these three factors.

Hartel, (1998) observed the impacts of unit operations on expression of air cells. Alteration
in stabilizer that consisted of 13% carrageenan, 37% guar gum, and 31% carboxymethyl
cellulose (CMC) did not affect freeze thawing. Adding up stabilizer, but less air, small room
volume compared to an analogous product ended devoid of stabilizer. Modifications in air
chamber volume can be frankly contributed to modifications in flow characteristics of the
product.

2.4.4 Effect of stabilizers on melting rate of ice-cream

Melting rate is of important parameter for the consumers especially when the ice-cream is in
the cone. Wildmoser et al., (2005) observed that the slow meltdown, slow serum drainage,
good shape retention, and slower foam collapse are some of the desired important quality
parameters of ice-cream. A fast and slow melting of ice-cream is the indication of defective
ice-cream.

Muse, M.R. et al., (2004) has suggested that as the ice-cream melts, heat transfers from the
warm air surrounding the product into the ice-cream to melt the ice crystals. Initially the ice
melts at the exterior of the ice-cream and there is a local cooling effect. The water from the
melting ice must diffuse into the viscous unfrozen serum phase, and this diluted solution then
flows downwards (due to gravity) through the structural elements (destabilized fat globules,
air cells and remaining ice crystals) to drip.

2.4.5 Effect of stabilizer on sensory qualities of ice-cream

Numerous studies have been carried out to find the relationship between viscosity and
sensory properties.

10
Maryam Bahramparvar & Mostafa Mazaheri Tehrani, (2011) reported that viscosity of
an ice-cream mix was also highly correlated with the overall acceptability of ice-creams
containing guar, gelatin, acacia and sodium alginate.

There is a relationship between stabiliser concentration and viscosity of ice-cream mix in

most cases with body and texture. Viscosity of an ice-cream mix was also highly correlated
with overall acceptability of ice-cream.

Soukoulis et al., (2008) furnished important information for the correlation of objective and
sensory properties and discrimination of stabilizing systems based on quality criteria, using
principal components and cluster analysis of instrumental and sensory data. In this research,
hydrocolloid type and content significantly affected vanilla flavour release. Higher the
hydrocolloids, better is the vanilla flavour perception.

2.5 Emulsifiers

Sukumar De, (2013) defined emulsifier as a substance which helps to form emulsions.
Emulsifiers improved the whipping quality of the mix and produce a drier ice-cream with a
smoother body and texture.

Sukumar De, (2013) reported that two types of emulsifiers are used in the manufacturing of
ice-creams. One is mono and diglycerides derived by the chemical reaction of naturally
occurring glycerides and other in polyoxyethelene derivatives of hexahydric alcohols, glycol
and glycol esters. Emulsifiers have little effect on the acidity, pH or viscosity of the ice-
cream mix. Whipping time decreases when emulsifiers are incorporated in the ice-cream. It
also decreases the melting rate of the finished ice-cream. It produces small air cells which
result in smoother ice-cream.

From above studies, we can say that emulsifiers and stabilizers both play an important role in
the preparation of ice-cream.

Karaman et al., (2004) had observed that ice-cream is a product which is liked by all age
groups of people because of its cooling effects.

11
MATERIALS AND METHODS

12
 MATERIALS AND METHODS

Different materials used and methods followed during the period of investigation are
described in this chapter.

3.1 Plan of work

This study entitled as ‘‘Incorporation of pearl millet and flaxseed milk for the fortification of
ice-cream using Response Surface Methodology’’ was conducted in the laboratory of Food
Science and Technology, Institute of Agricultural Sciences, Banaras Hindu University,
Varanasi during 2017-2018.

3.2 Materials

3.2.1 Flax Seeds

Flax seeds were purchased from the local market.

3.2.2 Pearl Millet Seeds

Pearl Millet seeds were purchased from the local market.

3.2.3 Milk

Packaged Amul full cream milk was purchased from Amul outlet of Varanasi.

3.2.4 Cream

Packaged Amul table cream was purchased from Amul outlet in Varanasi.

3.2.5 Skimmed Milk Powder

Nestle everyday dairy whitener was purchased from local market in Varanasi.

3.2.6 Artificial Sweetener

Normal Sugar free Natura Diet Sugar was purchased from the local market.

3.2.7 Glassware and Metal Ware

All the glassware and metal ware that were used during the research were of high quality.

3.2.8 Chemicals

A R Grade Chemicals and certain food grade chemicals were used during research work.

3.2.9 Basic laboratory requirement

All glassware, metal ware and containers used for the investigation were of good quality and
intact. The basic requirements included glassware like test tubes, petri plates, glass rods,
beakers, conical flasks, volumetric flasks, flat bottom flasks, pipette, burette, reagent bottles
etc. Other requirements included test tube stands, micropipettes etc.

13
3.2.10 Experimental Setup

The process physical and chemical parameters were analysed using electronic balance, digital
pH meter, refractometer, laminar air flow, autoclave, hot air oven, centrifuge, deep freezer.

3.3 Preparation of Millet incorporated ice-cream

3.3.1 Optimisation of ingredients for ice-cream by using Response Surface Methodology

Methodology:

The optimization in the preparation of ice-cream was done with the help of Response
Surface Methodology using Minitab 17 software. The parameters taken into consideration
were milk, cream and pearl millet flour. Keeping in view the objectives of the investigation,
the plan of work is discussed as under.

The levels of independent parameters are presented in Table 3.1

Table 3.1 Independent variables for optimization

Variables Lower limit Upper limit

Milk 150 ml 170 ml

Cream 70 g 90 g

Pearl millet flour 12 g 18 g

As responses sensory attributes (colour and appearance, body and texture, flavour and overall
acceptability was selected. On the basis of these responses, the effect of the three independent
variables was evaluated. A combination of 20 trials (experiments) was generated in the
Minitab 17 software using Central Composite Rotatable Design (CCRD) during
investigation.

3.3.2 Process Optimization

Numerical optimization technique of the Minitab 17 was used for simultaneous

optimization of the multiple responses. The desired goals for each factor and responses was
chosen, wherein the goals were acceptable to either the factors or the responses. The possible
goals or constraints presented were: Maximise, Minimize, Target and none (for responses
only) and set to an exact value (for factors only). In order to search a solution maximizing
multiple responses, the goals were combined into an overall composite function called the
desirability function. Desirability is an objective function that ranges from 0 to 1. The
Minitab software seeks to maximize the function. The goal seeking begins at a random start
point and proceeds up a steepest slope to a maximum. To compare the effect of milk, cream
and pearl millet flour on sensory attributes, various analyses were carried out.

14
3.4 Methodology

3.4.1 Extraction of Flax seed milk

10 gram of seed samples were taken in a beaker. Soaked them in 100 ml portable drinking
water for 1 hour. Ground it in a blender and strained it to get fresh and clean milk.
Pasteurized the milk at 80̊ C for 10 minutes and cooled the milk.

3.4.2 Preparation of Pearl Millet flour

Took 100 grams of whole, fresh, clean pearl millet grains. The grains should not be from the
older produce. Ground them into the grinder to obtain fine powder. Sieved it to remove the
husk from the flour.

3.4.3 Preparation of ice-cream

3.4.3.1 Materials

 Milk- Milk was taken from the local market. Amul full cream milk was used.
 Cream- Amul fresh cream was procured from the local market.
 Sweetener- Sugar free natura diet sugar was procured from the local market.
 Skimmed Milk powder- Nestle Everyday dairy whitener was used for this purpose.
 Flaxseed milk
 Pearl millet flour
 Vanilla essence
 Sodium alginate
 Guar gum
3.4.3.2 Methodology for preparation of ice-cream

20 trials of ice-cream were run as per the suggestions by the software. A basic method of
preparation was followed. 150 ml of milk was taken in a pan. 70 g of cream was whipped and
added to the milk. 20 ml of pasteurised flaxseed milk was added to it. 20 g of skimmed milk
powder was added to increase the solids content of the ice-cream. 18g Pearl millet flour was
added to the mixture and whipped. The mixture was whipped to mix all the ingredients
together along with artificial sweetener and stabilizers. The mixture was then kept for aging
for 2 hours. The aged mixture was whipped to incorporate air into the mix and then kept for
freezing in deep freezer for overnight.

15
 Fig.3.1 FLOWCHART FOR PREPARATION OF MILLET AND FLAXSEED MILK
FORTIFIED ICE-CREAM

Took 150 ml of milk in a pan

Added 70g of cream to it and whipped

Added 18g of pearl millet flour to it an d whipped

Added 5g artificial sweetener to the mix and 20g of SMP and whipped

Added 20 ml flaxssed milk, 1g sodium alginate and 1g of guar gum and whipped

Kept the mixture for aging for 2 hours

Added a few drops of vanilla essence to the mix and whipped it to incorporate air into the mix

Transferred the mixture to ice-cream cups and froze it to -4̊ C

3.5 Analysis of optimized ice-cream

3.5.1 Moisture Content: (AOAC,2000)

Procedure:

2g of sample was taken in a clean, dried and previously weighed petriplate and it was placed
in the hot air oven at 105̊ C for 2 hours. The petriplate was removed and immediately
placed in a cooling dessicator. It was allowed to cool and weighed.

Calculation:
𝑊3− 𝑊1
% Moisture Content = x 100
𝑊2− 𝑊1

Where, W1 = Weight of empty petriplate

W2 = Weight of petriplate + sample

W3 = Weight of petriplate + dried sample

16
3.5.2 Protein Content Estimation

Principle:

In the Kjeldahl method, the protein and other organic food components are digested
with concentrated sulphuric acid in the presence of a catalyst (1:5 cupric sulphate: sodium
sulphate). The total nitrogen is converted to ammonium sulphate. The digest is diluted with
water. Alkali (NaOH) is added to neutralise the H2SO4. The ammonia formed is distilled in
H2SO4 solution containing methyl red indicator.

Digestion:

2g of sample was weighed in Kjeldahl digestion flask and 15g of the digestion mixture
was added (Na2SO4/K2SO4 + 1g CuSO4) was added followed by 25 ml conc. H2SO4. The
tubes were carefully placed in the digestion chamber of the Kelplus apparatus and the process
of digestion was carried out. The content was boiled vigorously until the solution appeared to
be transparent/ clear. Heating was continued to 2-3 hours.

Distillation and Titration:

The digested sample was taken in a conical flask filled with 25 ml of 4% boric acid
neutralized with a mixture of methyl red and bromocresol blue in the ratio of 5:7. This was
placed in the distillation chamber of Kel-plus. The sample was diluted and alkali was added
till the sample changed to brown colour. Kel-plus was allowed to run for 10 minutes, after
which the conical flask was taken out of the distillation chamber and was immediately titrated
against 0.1N HCl. The titre value was noted.

Calculations:

% Protein = TV (s-b) x 14.01 x N of HCl x 100

Weight of sample x 1000

3.5.3 Ash Content: (AOAC,2000)

Procedure:

2g of homogenised sample was measured accurately (up to 4 decimal points) and

taken in a previously weighed, clean dry crucible. The crucible was placed on a hot plate till
the sample charred and the smoking discontinued. The crucible with the charred sample was
then transferred to a muffle furnace where ashing was carried out at 550̊ C for 4 1 hour. The
muffle furnace was turned off and the sample was removed after 10 minutes and placed in the
dessicator. The weight of the crucible with ash was taken when the crucible had cooled down.

17
Calculations:
𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑎𝑠ℎ
% Ash Content = x 100
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒

3.5.4 Fat content: (IS: 1224, 1958)

Ice-cream fat was determined by Gerber method using butyrometer. For this, 10
ml of conc. Sulphuric acid (Sp. Gr. 1.820-1.824 at 27̊ C) was transferred into a milk
butyrometer using automatic tilt measure taking due care as not to wet the neck of the
butyrometer. The thoroughly mixed sample of ice-cream was added slowly using 10.75ml
pipette followed by 1ml of amyl alcohol (Sp. Gr. 0.803- 0.805) using an automatic tilt
measure. The mouth of the butyrometer was firmly stoppered and contents were shaken well
by applying gentle swirling motion until most of the organic constituents were dissolved. The
contents were warmed in a water bath (maintained at 70̊ C±2̊ C) for 5 minutes. The
butyrometer after wiping it dry, was subsequently loaded in a Gerber’s centrifuge for 5
minutes at 1200 rpm. Again the butyrometer was transferred to the water bath (70̊ C) for 3
minutes. Before recording the reading, the fat column was adjusted in such a way that it
makes readable.

3.5.5 Total carbohydrate content:

The total carbohydrate content was determined as a difference of sum of protein,

ash, moisture and fat content from 100mg of sample, as per Serrem et al.,(2011).

Total carbohydrate = 100 – (Moisture + Protein + Ash + Fat)

Where moisture, protein, ash and fat content are expressed in grams per 100g of
sample.

3.5.6 Titratable Acidity: (AOAC 2000)

One gram of sample was taken in a standard flask and volume was made up to 100 ml. 10 ml
of this was taken into the conical flask and phenolphthalein indicator was added. It was
titrated against 0.1N NaOH till an end point of pink was obtained.

Calculations:

% Titratable acidity = Titre value x N of NaOH x Dilution x 90 x 100

(as citric acid) Weight of sample x Volume of aliquot x 1000

3.5.7 pH Determination

The pH of controlled sample and optimized ice-cream was measured using a digital pH
meter according to the standard of the International Dairy Federation (1992). Each sample
was mixed thoroughly and the pH was noted.

18
3.5.8 Total Solids: (AOAC, 1990)

Procedure:

5g sample was weighed into a previously weighed moisture dish. It was first evaporated
to dryness over a steam bath and was dried in an oven at 105̊ C for 3 hours. It was cooled in
a desiccator and weighed. The weighed sample was returned to the oven for final drying.
After that, it was cooled and weighed at an hourly interval until no further difference in the
weight was observed (this is, constant weight is observed). The moisture content was
calculated as a percentage ratio of the weight of moisture content, to that of the weight of
sample analysed.

Calculations:
100 𝑋 𝑤2−𝑤3
% MC =
𝑤2−𝑤1

Where MC = Moisture Content

W1 = Weight of empty moisture dish

W2 = Weight of empty dish + sample before drying

W3 = Weight of sample + sample after drying to constant weight

% dry matter (total solids) = 100 - % MC

3.5.9 Solids Not Fat (SNF)

This was estimated by difference as the solids not fat (SNF). It was calculated as
shown below % Solid Not Fat = % T.S. - % Fat, where T.S. = Total Solids

3.5.10 Overrun percentage (Marshall and Arbuckle, 1996)

Procedure:

Overrun is the amount of air incorporated in the frozen ice-cream. It is expressed by the
percentage increase in volume that the initial ice-cream mix undergoes during freezing/
whipping.

This was calculated by comparing the weight of a known volume of ice-cream (M2) to
the weight of the same volume of unfrozen ice-cream mix (M1).

Calculations:

Overrun (%) = (M1-M2)/M2 x 100

Where,

19
 M1 is the weight of ice-cream

M2 is the weight of ice-cream that had the same volume as that of the mix.

3.5.11 Melting Rate

Procedure:

A 50 gram sample of controlled and optimized ice-cream was placed in a Buchner funnel
on the top of the flask and was allowed to melt at room temperature (24̊C) for 30 minutes.
After this time, dipped volume was weighed and melting resistance was obtained using the
following equations.

Calculations:
𝐴1−𝐴2
Melting Rate = 𝐴1
x 100

Where,

A1 is the weight of the initial sample

A2 is the weight of the melted sample

3.5.12 Viscosity:

Procedure:

Calibrate the instrument using auto zeroing mode of the viscometer. In the manual
mode select the spindle and set the speed of the spindle. Insert the spindle in the spindle
holder of the viscometer. Immerse the spindle in the beaker containing the sample at 25̊ C.
Switch on the motor. Note the consistent value in centipoises as shown on monitor.

3.5.13 Textural Analysis

Textural attributes of the ice-cream such as cohesiveness, gumminess and springiness,

were analysed by Texture Profile Analyser (TPA) with the Texture analyser Model TA-TXT
Plus.

20
3.5.13.1 Test Settings:

Table 3.2 Texture analyser settings for the analysis of Ice-Cream

Mode Measure force in compression

Option Return to start

Pre-test Speed 2mm/s

Test speed 2mm/s

Post speed 5mm/s

Distance 80mm

Trigger type Auto-10g

Tare mode Auto

3.5.14 Sensory Analysis:

The optimised millet and flaxseed milk fortified ice-cream was evaluated for its sensory
characteristics by a semi-trained panel consisting of 10 judges drawn from the Centre of Food
Science and Technology. The panelists were asked to mark their scores according to the 9
points hedonic scale. Sensory evaluation of the sample was performed at a room temperature
of 37̊ C.

3.6 Statistical Analysis

Data obtained during the investigation were suitably analysed by employing statistical
designs viz., Central Composite Rotatable Design. Response Surface Methodology was also
used to optimize the various parameters such as sensory attributes of ice-cream.

21
RESULTS AND DISCUSSION

22
 RESULTS AND DISCUSSION

For the optimization of Pearl Millet and Flaxseed Milk fortified ice-cream, RSM based on
CCRD was used. 20 trials of the experiment were conducted and presented in Table 4.1. The
response study of sensory values (colour and appearance, body and texture, flavour, overall
acceptability) was performed.

Table 4.1 Experimental design for optimization of PMF fortified ice-cream

Factors Response

Milk (ml) Cream (ml) Pearl Millet Colour and Body and Flavour Overall
Flour(g) appearance Texture Acceptability

160 80 20.04 7.5 7.5 8 7.66

170 70 12 8.3 8.8 7.5 7.93

170 90 18 7.5 7.55 8 7.68

143.18 80 15 6.55 7.5 8 7.35

160 96.81 15 7.66 7.55 7.88 7.69

160 80 9.95 7 7.6 8 7.53

160 80 15 7.6 7.6 8 7.73

150 70 18 8.55 8.7 8.6 8.61

170 70 18 7.55 8.3 7.5 7.78

160 63.18 15 8 7.6 8 7.88

170 90 12 8 7.5 7.9 7.78

160 80 15 7.8 8 7.8 7.86

150 90 12 7.6 7.5 7.8 7.63

150 70 12 7.5 6.4 6.8 6.9

176.81 80 15 7.5 7.6 8 7.7

150 90 18 7 7.66 7.55 7.4

160 80 15 8.5 7.55 7.6 7.88

160 80 15 8.5 7.55 7.6 7.88

160 80 15 8.5 7.55 7.6 7.88

160 80 15 8.5 7.55 7.6 7.88

23
 Table 4.2 Coefficient of Estimation of Coded Factors for different factors of
optimization of PMF fortified Ice-Cream

Factors Coefficients

Colour and Body and Flavour Overall

Appearance Texture Acceptability

A- Milk (ml) 0.618 0.0921 0.0110 0.0907

B- Cream (ml) -0.174 -0.0933 0.0475 -0.0754

C- PMF (g) 0.003 0.1934 0.1208 0.1046

A2 -0.336 0.0017 0.0447 -0.0956

B2 -0.051 0.0105 0.0235 -0.0037

C2 -0.256 0.0017 0.0447 -0.0708

AB 0.138 -0.164 0.119 0.0312

AC -0.213 -0.264 -0.181 -0.2187

BC -0.175 -0.299 -0.244 -0.2387

4.1 Effect of variables on sensory properties of PMF fortified ice-cream

4.1.1 Effect on colour and appearance

It is an important property for any food materials that increase the consumer's
aesthetic appeal. The model for the sensory score of pearl millet and flaxseed fortified ice-
cream could be explained by the equation given as:

Colour and appearance = -95.1+ 1.087A-0.068B+2.45C-0.00336A2-0.00051B2-

0.0284C2+0.00137AB-0.00708AC-0.00583BC

24
Where, A = Milk

B = Cream

C = Pearl Millet Flour

Here, A, B and C are coded terms for three variables, i.e., milk, cream and pearl millet
flour respectively. The coefficient of determination (R2) was 0.6283 indicating that 62.83%
of the variability in the response could be explained by the model.

The ‘‘Pred. R-square’’ of 0.00 was in reasonable agreement with the ‘‘Adj. R-
square’’ of 0.2937. This model can be used to navigate the design space. The model of
polynomial equation fitted well and lack of fit was insignificant.

The coefficient of estimation of pearl millet and flaxseed milk fortified ice-cream
(Table 4.2) shows that the level of cream had a negative impact on colour and appearance of
the ice-cream but the level of milk and pearl millet flour had a positive effect on the score of
colour and appearance.

Fig. 4.1(a) shows the response surface plot for colour and appearance as influenced by
the level of milk and pearl millet flour. From the Fig. 4.1(a) it can be concluded that with the
increase in the level of pearl millet flour, the colour and appearance increases, but the
increase in the level of milk has significantly no effect on colour and appearance.

Fig. 4.1(b) shows the response surface plot for colour and appearance as influenced
by the level of cream and pearl millet flour. From the Fig. 4.1(b) it can be concluded that with
the increase in the level of pearl millet flour, the colour and appearance increases, but the
increase in the level of cream has significantly no effect on colour and appearance.

Fig. 4.1(c) shows the response surface plot for colour and appearance as influenced by the
level of cream and milk. From the Fig. 4.1(b) it can be concluded that with the increase in the
level of milk, the colour and appearance increases, but the increase in the level of cream has
significantly no effect on colour and appearance.

(a)

25
 (b)

(c)

Fig. 4.1 Response Surface Plots showing the effect of the composition of ingredients on
Colour and Appearance of Ice-cream

(a) Milk and Pearl Millet Flour

(b) Cream and Pearl Millet Flour
(c) Milk and Cream

26
4.1.2 Effect on Body and Texture

The data fitted the following quadratic model equation:

Body and Texture = - 46.9 + 0.267A + 0.385B + 2.262C + 0.000017A2 + 0.000105B2 +

0.0002C2 - 0.00164AB - 0.00879AC - 0.00996BC

The coefficient of determination (R2) was 0.6397 for the quadratic model. The
‘‘Pred. R-square’’ of 0.00 was in reasonable agreement with the ‘‘Adj. R-square’’ of 0.3155.
This model can be used to navigate the design space. The model of polynomial equation
fitted well and lack of fit was insignificant. The F-value of the model was 1.97.

The coefficient of estimation of pearl millet and flaxseed milk fortified ice-
cream (Table 4.2) shows that the level of cream had a negative impact on body and texture of
the ice-cream but the level of milk and pearl millet flour had a positive effect on the score of
body and texture.

Fig. 4.2(a) shows the response surface plot for body and texture as influenced by
the level of milk and pearl millet flour. From the Fig. 4.2(a) it can be concluded that with the
increase in the level of pearl millet flour, the body and texture increases, but the increase in
the level of milk has significantly no effect on body and texture.

Fig. 4.2(b) shows the response surface plot for body and texture as influenced by
the level of cream and pearl millet flour. From the Fig. 4.2(b) it can be concluded that with
the increase in the level of both milk and pearl millet flour, the body and texture increases.

Fig. 4.2(c) shows the response surface plot for body and texture as influenced by
the level of milk and cream. From the Fig. 4.2(c) it can be concluded that with the increase in
the level of cream, the body and texture increases, but the increase in the level of milk has
significantly no effect on body and texture.

27
 (a)

(b)

(c)

Fig. 4.2 Response Surface Plots showing the effect of the composition of ingredients on
Body and Texture of Ice-cream

(a) Milk and Pearl Millet Flour

(b) Cream and Pearl Millet Flour
(c) Milk and Cream

28
4.1.3 Effect on Flavour

The data fitted the following quadratic model equation-

Flavour = 11.6-0.146A-0.101B+1.508C+0.000447A2+0.000235B2+0.0050C2+

0.00119AB-0.00604AC-0.00812BC

The coefficient of determination (R2) was 0.4876 for the quadratic model.
The ‘‘Pred. R-square’’ of 0.00 was in reasonable agreement with the ‘‘Adj. R-square’’ of
0.0264. This model can be used to navigate the design space. The model of polynomial
equation fitted well and lack of fit was insignificant. The F-value of the model was 1.06.

The coefficient of estimation of pearl millet and flaxseed milk

fortified ice-cream (Table 4.2) shows that the level of all the three ingredients namely, cream,
milk and pearl millet flour had a positive effect on the score of flavour.

Fig. 4.3(a) shows the response surface plot for flavour as influenced
by the level of milk and pearl millet flour. From the Fig. 4.3(a) it can be concluded that with
the increase in the level of pearl millet flour, the flavour increases, but the increase in the
level of milk has significantly no effect on flavour.

Fig. 4.3(b) shows the response surface plot for flavour as influenced by the
level of cream and pearl millet flour. From the Fig. 4.3(b) it can be concluded that with the
increase in the level of pearl millet flour, the flavour increases, but the increase in the level of
cream has significantly no effect on flavour.

Fig. 4.3(c) shows the response surface plot for flavour as influenced by the level
of milk and pearl millet flour. From the Fig. 4.3(c) it can be concluded that with the increase
in the level of milk, the flavour increases, but the increase in the level of cream has
significantly no effect on flavour.

29
 (a)

(b)

(c)

Fig. 4.3 Response Surface Plots showing the effect of the composition of ingredients on
Flavour of Ice-cream

(a) Milk and Pearl Millet Flour

(b) Cream and Pearl Millet Flour
(c) Milk and Cream

30
4.1.4 Effect on Overall Acceptability

The data fitted the following quadratic model equation-

Overall Acceptability = -43.0+0.399A+0.068B+2.074C-0.000956A2-0.000037B2-

0.00787C2+0.000312AB-0.00729AC-0.00796BC

The coefficient of determination (R2) was 0.7036 for the quadratic model. The ‘‘Pred. R-
square’’ of 0.00 was in reasonable agreement with the ‘‘Adj. R-square’’ of 0.4369. This
model can be used to navigate the design space. The model of polynomial equation fitted well
and lack of fit was insignificant. The F-value of the model was 2.64.

The coefficient of estimation of pearl millet and flaxseed milk fortified ice-cream
(Table 4.2) shows that the level of all the three ingredients namely, cream, milk and pearl
millet flour had a positive effect on the score of overall acceptability.

Fig. 4.4 showed that the response surface plots for overall acceptability of PMF
fortified ice-cream by the levels of milk, cream and pearl millet flour. From Fig. 4.4 it can be
observed that the sensory score for overall acceptability is increased with the increase in the
levels of all the three ingredients.

31
 (a)

(b)

(c)

Fig. 4.4 Response Surface Plots showing the effect of the composition of ingredients on
Overall Acceptability of Ice-cream

(a) Milk and Pearl Millet Flour

(b) Cream and Pearl Millet Flour
(c) Milk and Cream

32
4.2 Optimization of the solution and their validations

To achieve the best combination for optimization of factors involved in the

preparation of ice-cream, the optimization command of the DOE of Minitab 17was used. The
optimizing conditions for all the attributes are as follows (Table 4.3). The method adopted for
the process optimization was based on numerical method. The constraints have been listed
below:

Table 4.3 Constraints fixed for optimization of levels

Response Goal Lower Target Upper Weight Importance

Colour and Maximum 6.55 8.55 1 1
Appearance
Flavour Maximum 6.80 8.60 1 1
Body and Maximum 6.40 8.70 1 1
Texture
Overall Maximum 6.90 8.61 1 1
Acceptability

Table 4.4 shows the optimized recipe condition as predicted by the RSM
software. The solution has been tabulated along with the desirability of each response.

Table 4.4 Optimization of Pearl Millet and Flaxseed fortified ice-cream

Constraints Optimized Desirability

Values

Factors Milk 154.734 -

Cream 63.1821 -

Pearl Millet Flour 18.9242 -

Responses Colour and 8.40447 0.92723

Appearance

Body and Texture 8.71684 1

Flavour 8.70345 1

Overall Acceptability 8.61004 1

Composite - - 0.9813
Desirability

33
4.3 Analysis of Optimized Product

Pearl Millet and Flaxseed fortified ice-cream was prepared according to the optimised
formulations as suggested by the RSM software and it was analysed for different
physicochemical, sensory and textural attributes. The data obtained has been tabulated in
Table 4.5. The predicted values were compared with the observed ones for the validation of
the formulation using one way ANOVA (Analysis of Variance). The table reveals that the
observed values were no significant, different from the predicted values concerning all the
properties of the product.

Table 4.5 ANOVA on the predicted values for the different attributes of PMF fortified
ice-cream optimized by RSM with the actual product prepared experimentally

Attributes Expected Observed P-value

Optimized Optimized
Attributes Attributes

Colour and 8.41 8.24±0.007 0.604

Appearance
Body and Texture 8.72 8.45±0.015 0.446
Flavour 8.70 8.37±0.005 0.435
Overall 8.61 8.84±0.015 0.303
Acceptability

4.4 Proximate Analysis of PMF fortified ice-cream

4.4.1 Proximate Composition of PMF fortified ice-cream

The proximate composition of PMF fortified ice-cream is given in Table 4.6

Table 4.6 Proximate composition of CSM ice-cream

Constituent Amount (g/100g of the sample)

Moisture 48.89±0.06
Protein 3.87±0.04
Fat 12±0.02
Carbohydrate 29.87±0.09
Ash 0.94±0.03
The no. of samples were 3.

34
 Table 4.7 Various physical parameters of optimized PMF ice-cream

Parameters Values
pH 6.8 ±0.03
Total Soluble Solids (TSS) 58̊ brix
SNF 29% ±0.05
Total Solids (TS) 42% ±0.04
Titratable Acidity 0.20 ±0.08
Overrun% 60 ±0.03

4.4.2 pH of Optimised PMF fortified ice-cream

pH of the PMF fortified ice-cream was found to be 6.8±0.03.pH of ice-cream should be 6.8
(De,1991). Thus, our finding was also significant as compared to De,1991.

4.4.3 Titratable acidity of PMF fortified ice-cream

Titratable acidity of PMF fortified ice-cream was found to be 0.20±0.08. Titratable

acidity of ice-cream should be 0.25% of lactic acid. (De, 1991).

4.4.4 Total solids content of PMF fortified ice-cream

The total solids content of PMF ice-cream was found to be 42%.The total solids
content of ice-cream should not be less than 36%. (De, 1991). The observed value matched
with the standard.

4.4.5 Total Soluble Solids (AOAC,1990)

Total soluble solids of PMF fortified ice-cream were found to be 58̊ brix.

4.4.6 Solids Not Fat (SNF)

SNF content of PMF fortified ice-cream was found to be 29%±0.05.Umelo M.C. et

al., (2014) reported that ice-cream recipe low in solids results in foam collapse and loss of
overrun and excessive shrinkage can result from partial melting at too high a freezer storage
temperature.

4.4.7 Overrun percentage of PMF fortified ice-cream

Overrun % of PMF fortified ice-cream was found to be 60%±0.03.Ozdemir et al.,

(2008) observed the Overrun% of ice-cream should be in the range of 80-100%. Whereas our
findings were lower than the standard overrun %. Chang and Hartel, (2002) reported that
the possible reason for getting a lower overrun value is due to the inconsistency during the
whipping process which is caused by the limitation of equipment.Guven et al., (2003);
Ozdemir et al., (2003); Dervisoglu et al., (2005)also agreed with our findings.

35
4.4.8 Melting Point of PMF ice-cream

The melting point of PMF ice-cream was noted at the interval of every 10 minutes. The
results are shown in Table 4.7. Muse and Hartel, (2004) observed that the melt-down rate of
ice-cream is affected by many factors, including the amount of air incorporated, the nature of
ice crystals and the network of fat globules formed during freezing.

Table4.8 Representing the volume of melted ice-cream

Time (min) Volume collected (ml)

0 0
10 10
20 22
30 38
40 48
50 60

4.4.9 Textural Analysis of Optimised PMF ice-cream

Textural attributes such as cohesiveness, springiness, gumminess were measured for

optimised PMF ice-cream. The results have been tabulated in Table 4.9.

Table 4.9 Textural attributes of PMF fortified ice-cream

Parameters Cohesiveness Springiness Gumminess

Values 0.03±0.04 44.62±0.05 0.89±0.03

Mean ± SEM

The texture analysis was done for the frozen sample. If the temperature is increased and the
ice-cream starts melting, the results would vary and that may cause an error.

4.4.10 Sensory analysis of optimized PMF fortified ice-cream

Sensory attributes such as colour and appearance, body and texture, flavour and
overall acceptability were measured for optimized PMF fortified ice-cream. Results found is
shown in the Table 4.10

36
Table 4.10 Sensory Score for optimised Pearl Millet and Flaxseed fortified ice-cream

Parameters Colour and Body and Flavour Overall

Appearance Texture Acceptability

Score 8.7 ± 0.04 8.8 ± 0.03 9.08 ± 0.03 9 ± 0.02

Figure 4.5: Pearl Millet and Flaxseed Milk incorporated sugar free ice-cream

37
SUMMARY AND
CONCLUSIONS

38
 SUMMARY AND CONCLUSION

The present study is titled, ‘‘Incorporation of Pearl Millet Flour and Flaxseed
Milk for the fortification of sugar free ice-cream using Response Surface
Methodology’’.

The study has been undertaken to verify the properties of Pearl Millet as an
ingredient to Ice-cream and also to fortify it with omega-3 fatty acids. The ice-cream is
sweetened by using artificial sweetener and therefore, can be consumed by the people with
diabetes.

5.1 Evaluation of potentiality of Pearl Millet flour and Flaxseed Milk in the preparation
of ice-cream

 The recipe was optimized by the addition of Pearl Millet flour at the rate of 18 grams
using Response Surface Methodology.
 The Flaxseed milk was added at the rate of 20 ml.
 Other ingredients like milk and cream were also optimised.
 The optimised values of ingredients as determined by the software are Milk-150ml,
Cream-70ml and Pearl Millet flour-18g.
 The desirability of the prepared recipe at the optimal level of ingredients as suggested
by the software gave the best product in terms of sensory evaluation factors.

EPILOGUE

The present study justifies the use of Pearl Millet flour and Flaxseed milk to be
used as an ingredient in ice-cream. This suggests that both these ingredients can be added to
the ice-cream without hampering the physical properties of ice-cream. Moreover, the
Flaxseed milk naturally increases the fat content of ice-cream, hence reducing the amount of
cream to be added to the ice-cream.

Pearl Millet flour having functional properties, especially being rich in fibres
can play a key role in the preparation of numerous products such as instant pudding, dairy,
confectionery, beverages and baked products.

39
BIBLIOGRAPHY

40
 BIBLIOGRAPHY

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A.O.A.C., (2000). Official methods of analysis(17th edition). Gaithersburg, MD: Association

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Bahramparvar, M., & Mazaheri Tehrani, M. (2011). Application and functions of stabilizers
in ice-cream. Food reviews international, 27(4), 389-407

Chang, Y., & Hartel, R. W. (2002), Development of air cells in a batch ice-cream freezer.
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Clarke, C. (2004). The science of ice-cream; the royal society of chemistry: Cambridge, UK.

Dervisoglu, M., Yazici, F., & Aydemir, O. (2005). The effect of soy protein concentrate
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Dogan, M., & Kayacier, A. (2007). The effect of aging at low temperature on the rheological

Properties of Kahramanmaras-type ice-cream mix. International Journal of PharmTech

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Hegenbert, S. (1996) The ice-cream evolution. Food Prod. Des. 6(7);29-24

IS: 1224-1958 (part I)- 1977. Determination of fat by the Gerber method of milk. Indian
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Karaman, S., Toker, O. S., Yuksel, F., Cam, M., Kayacier, A., & Dogan, M. (2014).
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Kilara, A., & Chandan, R. C. (2008). Ice-cream and Frozen Desserts (pp. 357-386). Wiley-
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Marshell, R. T., & Goff, D. (2003.). Formulationg and manufacturing ice-cream and other
frozen desserts. Food Technology.

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Marshall, R. T., & W.S. Arbuckle. (1996). Ice-cream. 5th Edn., Chapman and Hall, New
York, USA.,Pages., 368

Marshall, R.T., H.D. Goff and R.W. Hartel. (2003). Ice-cream. 6th Edn., Springer Science and
Business Media, London, USA., ISBN-13: 9780306477003, Pages: 371

Mehu, (1993) G.G. Evaluation of effects of several fat replacers on selected physical and
sensory properties of non fat frozen desserts. Master’s Thesis, University of
Missouri-Columbia,67

Muse, M.R., & Hartel, R.W. (2004). Ice-cream structural elements that affect melting rate
and hardness. Journal of Dairy Science, 87(1), 1-10.

Soukoulis, C., Chandrinos, I., & Tzia, C. (2008). Study of the functionality of selected
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42
APPENDICES

43
ANNEXURE 1: SENSORY EVALUATION PERFORMA FOR THE EVALUATION
OF PEARL MILLET AND FLAXSEED INCORPORATED ICE-CREAM

SENSORY EVALUATION OF MILLET BASED ICE CREAM

Name:- ______________________________ Date:- ____________Time:- ____________

Dear Sir/Madam,

You are receiving two samples of millet incorporated ice cream. Kindly evaluate the samples
on the below mentioned attributes, based on the nine point hedonic scale as follows:-

9- Like extremely

8- Like very much

7-Like moderately

6-Like slightly

5-Neither like nor dislike

4-Dislike slightly

3-Dislike moderately

2-Dislike very much

1-Dislike extremely

COLOUR TEXTURE FLAVOUR MELTNG OVERALL

QUALITY ACCEPTABILITY

SAMPLE 1

SAMPLE 2

Comments:-

Signature

44
ANNEXURE 2: ANOVA TABLE FOR THE ASSESSMENT OF COLOUR AND
APPEARANCE

Source DF Adj SS Adj MS F-Value P-Value

Model 9 3.90129 0.43348 1.88 0.170

Linear 3 0.79862 0.26621 1.15 0.375

Milk 1 0.38658 0.38658 1.67 0.225

Cream 1 0.41192 0.41192 1.78 0.211

Pearl millet flour 1 0.00012 0.00012 0.00 0.982

Square 3 2.34517 0.78172 3.39 0.062

Milk*Milk 1 1.62308 1.62308 7.03 0.024

cream*cream 1 0.03746 0.03746 0.16 0.696

pearl millet flour 1 0.94481 0.94481 4.09 0.071

*pearl millet flour

2-Way Interaction 3 0.75750 0.25250 1.09 0.396

milk*cream 1 0.15125 0.15125 0.66 0.437

milk*pearl millet flour 1 0.36125 0.36125 1.57 0.239

cream*pearl millet flour 1 0.24500 0.24500 1.06 0.327

Error 10 2.30821 0.23082

Lack-of-Fit 5 1.43487 0.28697 1.64 0.300

Pure Error 5 0.87333 0.17467

Total 19 6.20950

45
 ANNEXURE 3: ANOVA TABLE FOR THE EFFECT OF CONCENTRATION
OF INGREDIENTS ON BODY AND TEXTURE

Source DF Adj SS Adj MS F-Value P-Value

Model 9 2.23245 0.248050 1.97 0.152

Linear 3 0.74582 0.248606 1.98 0.181

milk 1 0.11591 0.115914 0.92 0.360

cream 1 0.11886 0.118864 0.95 0.354

pearl millet 1 0.51104 0.511041 4.06 0.071

flour

Square 3 0.00160 0.000532 0.00 1.000

milk*milk 1 0.00004 0.000040 0.00 0.986

cream*cream 1 0.00159 0.001587 0.01 0.913

pearl millet 1 0.00004 0.000040 0.00 0.986

flour *pearl millet flour

2-Way 3 1.48504 0.495013 3.94 0.043

Interaction

milk*cream 1 0.21451 0.214513 1.71 0.221

milk 1 0.55651 0.556512 4.43 0.062

*pearl millet flour

Cream 1 0.71401 0.714013 5.68 0.038

*pearl millet flour

Error 10 1.25727 0.125727

Lack-of-Fit 5 1.09393 0.218787 6.70 0.029

Pure Error 5 0.16333 0.032667

Total 19 3.48972

46
ANNEXURE 4: ANOVA TABLE FOR THE EFFECT OF CONCENTRATION
OF INGREDIENTS ON FLAVOUR

Source DF Adj SS Adj MS F-Value P-Value

Model 9 1.13861 0.126512 1.06 0.462

Linear 3 0.23176 0.077254 0.65 0.603

milk 1 0.00165 0.001648 0.01 0.909

cream 1 0.03076 0.030764 0.26 0.623

pearl millet flour 1 0.19935 0.199350 1.67 0.226

Square 3 0.05591 0.018637 0.16 0.924

milk*milk 1 0.02881 0.028813 0.24 0.634

cream*cream 1 0.00796 0.007959 0.07 0.802

pearl millet flour 1 0.02881 0.028813 0.24 0.634

*pearl millet flour

2-Way Interaction 3 0.85094 0.283646 2.37 0.132

milk*cream 1 0.11281 0.112812 0.94 0.354

milk 1 0.26281 0.262813 2.20 0.169

*pearl millet flour

Cream 1 0.47531 0.475313 3.97 0.074

*pearl millet flour

Error 10 1.19664 0.119664

Lack-of-Fit 5 1.05664 0.211329 7.55 0.022

Pure Error 5 0.14000 0.028000

Total 19 2.33526

47
ANNEXURE 5: ANOVA TABLE FOR THE EFFECT OF CONCENTRATION
OF INGREDIENTS ON OVERALL ACCEPTABILITY

Source DF Adj SS Adj MS F-Value P-Value

Model 9 1.37434 0.152704 2.64 0.073

Linear 3 0.33940 0.113134 1.95 0.185

milk 1 0.11234 0.112339 1.94 0.194

cream 1 0.07761 0.077613 1.34 0.274

pearl millet flour 1 0.14945 0.149448 2.58 0.139

Square 3 0.18830 0.062766 1.08 0.400

milk*milk 1 0.13164 0.131641 2.27 0.162

cream*cream 1 0.00019 0.000192 0.00 0.955

pearl millet flour 1 0.07229 0.072292 1.25 0.290

*pearl millet flour

2-Way Interaction 3 0.84664 0.282212 4.88 0.024

milk*cream 1 0.00781 0.007812 0.13 0.721

milk*pearl millet flour 1 0.38281 0.382812 6.61 0.028

cream*pearl millet flour 1 0.45601 0.456012 7.88 0.019

Error 10 0.57884 0.057884

Lack-of-Fit 5 0.56076 0.112151 31.01 0.001

Pure Error 5 0.01808 0.003617

Total 19 1.95317

48