1 s2.0 S0963996913004821 Main

Food Research International 54 (2013) 837851
Contents lists available at ScienceDirect
Food Research International

journal homepage: www.elsevier.com/locate/foodres
Co-extrusion encapsulation of canola oil with alginate: Effect of quercetin

addition to oil core and pectin addition to alginate shell on oil stability
Wei Wang, Geoffrey I.N. Waterhouse, Dongxiao Sun-Waterhouse
School of Chemical Sciences, The University of Auckland, Private Bag 92019, Auckland, New Zealand
a r t i c l e
i n f o
Article history:
Received 4 June 2013
Accepted 24 August 2013
Keywords:
Alginate
Alginatepectin mixtures
Canola oil
Co-extrusion encapsulation
Quercetin
a b s t r a c t
This study investigates the feasibility of encapsulating antioxidant-fortied canola oil via co-extrusion using
0.67% alginate (0.67%A), 1% alginate (1%A) or high methoxyl (HM) pectin-enhanced alginate (AP) as the
encapsulant. Results show that encapsulation conditions especially the coreshell ow rates and shell wall formulation, inuenced oil bead characteristics, core oil stability and retained phenolic content. Optical and scanning electron microscopy revealed that the 3 types of co-extruded oil beads were spherically shaped with the
AP beads having the largest bead size. All beads were physically and chemically robust, and remained intact
after being treated in acidied water at pH 3 for 2 h. Storage trials at 20 and 38 C for 30 or 60 days revealed
the interplay between shell formulations and storage conditions on oil primary and secondary oxidation, hydrolytic rancidity and total phenolic content of the encapsulated canola oils. Quercetin added to the oil core was
more effective than vitamin E or BHT in suppressing oil oxidation at 38 C. High performance liquid chromatography analyses indicated different decomposition pathways for quercetin in these beads during storage. FT-IR
studies conrmed the chemical composition and chemical stability of the 3 types of quercetin-containing oil
beads. 1%A and AP shells are both acceptable and comparable for preserving quercetin-containing canola oil,
with AP and 1%A being slightly better for 30 and 60 day storage periods, respectively. Thus, it is feasible and
benecial to deliver unsaturated oil and phenolic antioxidants in the form of pectin bre-enhanced alginate
microbeads.
2013 Elsevier Ltd. All rights reserved.
1. Introduction
Rising consumer preference for functional foods stimulates research
aimed at optimising the delivery of health-promoting substances like
dietary bres (DFs), polyphenols and unsaturated fatty acids (FAs) in dietary forms to consumers (Kumar & Goyal, 2008; Ratnayake et al., 2000;
Sherry et al., 2010; Warren et al., 2009). Unsaturated plant oils such as
canola oil are becoming an increasingly important part of the human
diet. However, these oils are highly susceptible to oxidation which if
left unchecked leads to rancidity and an unpleasant odour and taste.
Fortifying oils with antioxidants is a well-established approach for
improving oil stability against oxidation. Vitamin E has traditionally
been used for this purpose, although synthetic antioxidants such as butylated hydroxytoluene (BHT) are no longer in favour (Kahl & Kappus,
1993). Recently, more emphasis has been placed on phenolics as oilfortifying antioxidants due to their validated health benets and ability
to suppress lipid oxidation via donating hydrogen atoms to lipid peroxyl
radicals (Cheung, Szeto, & Benzie, 2007). Most phenolic antioxidants
Corresponding author. Tel.: +64 9 3737599x89024; fax: +64 9 3737422.

E-mail address: dx.sun-waterhouse@auckland.ac.nz (D. Sun-Waterhouse).
0963-9969/$ see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodres.2013.08.038
including quercetin can impart a bitter taste to foods (Jaeger, Axten,

Wohlers, & Sun-Waterhouse, 2009). In general, consumers are unwilling to compromise on the taste and avour of foods, so delivery of
high phenolic content foods and beverages to consumers is technically
challenging.
Co-extrusion is a microencapsulation method that is highly suitable
for preservation of oil through creating a shell wall barrier to enclose
an oil core, which itself can be fortied with an antioxidant (Fang
& Bhandaria, 2010; Sun-Waterhouse, Penin-Peyta, Wadhwa, &
Waterhouse, 2012; Sun-Waterhouse, Wadhwa, & Waterhouse, 2013;
Sun-Waterhouse, Zhou, Miskelly, Wibisono, & Wadhwa, 2011). Such
an encapsulation approach also ameliorates undesirable taste sensations such as bitterness of core substances through minimising contact
with oral taste receptors. The resultant antioxidant-fortied oil microbeads can be used either as nutritional supplements for direct consumption or as a food ingredient. The shell wall matrix inuences the size,
shape and integrity of microbeads, as well as encapsulation efciency
and stability of core substances (Fang & Bhandaria, 2010; SandovalCastilla, Lobato-Calleros, Garca-Galindo, Alvarez-Ramrez, & VernonCarter, 2010). Polysaccharides are widely used as encapsulants due
to their moderately low oxygen permeability (Sandoval-Castilla et al.,
2010; Williams & Phillips, 2000). Alginate gels formed in the presence
838
W. Wang et al. / Food Research International 54 (2013) 837851
of Ca2+ are frequently utilised as a physical barrier for microencapsulation because of their chemical stability, low toxicity and low immunogenicity (Liu et al., 2002). Pectins are health-promoting soluble bres and
often used to increase viscosity and gel strength of food products
(Thakur, Singh, & Handa, 1997). Blending different biopolymers has
proven to be a useful strategy to form shell walls with maximal protection and tailored controlled release properties for core substances (Islan,
de Verti, Marchetti, & Castro, 2012; Piculell, Bergfeldt, & Nilsson, 1995).
Adding pectins to alginate-based encapsulant formulations may confer
multiple benets: 1) enhanced protection on core substances and 2) increased DF consumption and product nutritional value (Fernandez,
2001). Pectins that are industrially extracted from apple pomace and
citrus peels are mostly in the form of high methoxyl (HM) pectins
(N50% degree of esterication, DE) (Daz-Rojas et al., 2004). Thus, it is
of interest to examine the combined use of alginate and HM pectins
for encapsulation of bioactive oils.
In this study, canola oil was selected as a model unsaturated plant oil
because of its high unsaturated FA content (i.e. monounsaturated FA
62.4%, polyunsaturated FA 31.3% with 61.6% C18:1, 21.7% C18:2n6
and 9.6% C18:3n3) (Przybylski, Mag, Eskin, & McDonald, 2005). The
effects of co-extrusion encapsulation conditions, i.e. coreshell feed
ow rates and bead hardening temperature and time were rst examined using the oil beads encapsulated by 1% alginate only (1%A). The optimal encapsulation conditions were then selected to produce canola oil
beads with or without an added antioxidant encapsulated by two alginate formulations (0.67% sodium alginate (0.67%A) or 1%A), as well as
by alginateHM pectin solution (AP). Quercetin was selected as a
model phenolic antioxidant for oil fortication because of its potent
scavenging activity and recognised health benets (Warren et al.,
2009). The effectiveness of quercetin in preserving the encapsulated
oil was compared to that of vitamin E or BHT over 60 days storages at
20 and 38 C. Optical and scanning electron microscopy techniques
were used to explore bead characteristics before and after treatment
at pH 3. FT-IR spectroscopy and high performance liquid chromatography (HPLC) are used to examine the compositional changes of the
encapsulated oil beads during the storage trials. From this work, we
hope to ascertain the best shell formulation and fortifying antioxidant
for the preservation of canola oil quality.
2. Materials and methods
2.1. Chemicals and materials
Canola oil (Simply, Malaysia) was purchased from Countdown in
Auckland, New Zealand. Sodium alginate (GRINSTED Alginate FD155,
viscosity 350550 mPas) was purchased from Danisco, Auckland,
New Zealand. HM pectin (classic AU 201 USP, degree of esterication
7276%) was purchased from Herbstreith & Fox KG (D-75305),
Neenburg, Switzerland. Quercetin, rutin, catechin, gallic acid, BHT,
-tocopherol (vitamin E), FolinCiocalteu's phenol reagent (2 N),
calcium chloride, active carbon, p-anisidine and tridecanoic acid were
purchased from Sigma-Aldrich Inc., St Louis, MO, USA. HPLC grade acetonitrile was purchased from J.T. Baker, Phillipsburg, NJ, USA. Glacial
acetic acid, absolute ethanol, chloroform, concentrated HCl (36%),
acetonitrile, n-hexane (95%), isooctane, methanol, potassium iodide,
potassium dichromate, potassium hydrogen phthalate, sodium hydroxide, sodium carbonate, sodium thiosulphate and sulfuric acid were
purchased from Ajax Finechem, Auckland, New Zealand. PEG 400
(carbowax) was purchased from AMCD Australia Ltd, Mulgrave,
Victoria, Australia. Ammonium chloride was purchased from May &
Baker, Dagenham, England. Iodine solution was purchased from
ACROS ORGANICS, New Jersey, USA. Starch soluble, sodium sulphite
and potassium chloride were purchased from AnalaR, BDH laboratory
chemicals, Poole, England. Cyclohexane (analytical grade) was purchased from Biolab (Aust) Ltd, Scoresby, Australia. Phenolphthalein indicator was purchased from Scharlau Chemie (Barcelona, Spain).
2.2. Preparation of encapsulated canola oil beads

2.2.1. Preparation of encapsulant formulations
A sodium alginate solution (1%w/w) was prepared by adding the required amount of Na-alginate to pre-boiled distilled water at 85 C (50%
of nal volume), mixed at 5000 rpm for 1 min using a Silverson L5T
high shear mixer (Silverson Machines Ltd., Chesham, Bucks, UK), and
then made up to volume with distilled water at 20 C. The resulting
solution was then subjected to further mixing using the Silverson high
shear mixer (X screen; 5000 rpm, 1 min). The alginate solution was
stored overnight at 4 C in a fridge (SRS 535NW, Samsung, Korea) and
used the following day to make beads. A 0.67%w/w alginate solution
was prepared in the same manner.
HM pectin solution (1%w/w) was prepared by slowly adding the required amount of HM apple pectin powder to distilled water of 80 C
with gentle stirring (speed at 3, 3001 magnetic stirring hotplate,
Germany). The resultant solution was gently homogenised using the
Silverson high shear mixer (X screen; 800 rpm, 1 min).
The alginatepectin (AP) solution was prepared by mixing carefully
the 1%w/w alginate solution with the 1%w/w pectin solution at a volume
ratio of 2:1, followed by gentle stirring with a magnetic stirrer (speed at
3 for 2 min) to obtain a homogenous shell solution.
2.2.2. Preparation of canola oil fortied with quercetin, vitamin E or BHT
Quercetin dihydrate (200 ppm, nal concentration in oil) was rst
mixed with PEG 400 (2%w/w, nal concentration in oil) in a beaker, to
which canola oil was slowly added. The beaker containing canola oil
and quercetin was then placed in an ice bath, and the mixture was
homogenised using the Silverson high shear mixer (X screen;
2000 rpm for 1 min followed by 5000 rpm for 2 min). Fortication of
canola oil with BHT or vitamin E was conducted in the same manner
but without the use of PEG. PEG was added as a plasticiser and surfactant to help blending quercetin with oil. After preliminary trials,
2%w/w was determined to be the minimal concentration needed to dissolve quercetin.
2.2.3. Encapsulated canola oil using co-extrusion technology
The canola oil with or without a fortifying antioxidant was encapsulated by an Inotech Encapsulator (Inotech Encapsulation AG, Dottikon,
Switzerland), following the method of Sun-Waterhouse, Zhou, et al.
(2011) with some modications. During encapsulation, the core uid
(200 mL of unfortied or fortied oil in a glass syringe), and the shell
uid (i.e. 200 mL of 0.67%A, 1%A or AP solution in a plastic syringe)
were simultaneously pumped into a mixing chamber to give a core
shell uid stream which are sprayed out through a nozzle (200 m)
under a vibration frequency of 1750 Hz and a voltage of 1.5 kV. On contact with a 3%/w/w CaCl2 solution (200 mL), regular-sized microbeads
formed through cross-linking. After hardening in this CaCl2 solution,
the beads were collected on a 50-m nylon mesh, washed with water
at 30 C, and transferred to 50 mL glass tubes and lled up to a volume
of 35 mL (wrapped with aluminium foil), and freeze dried using a
freeze dryer (Labconco, Total Lab Systems Ltd, New Zealand). Preliminary encapsulation trials were conducted using different combinations
of coreshell feed ow rates and encapsulated bead hardening
conditions Table 1: a core feed ow rate of 30 or 60 mL/h, a shell feed
ow rate of 200 and 250 mL/h, and hardening at room temperature
for 10 min or at 4 C for 2 h in 3% CaCl2 solution. The optimal encapsulation conditions were determined for preparing oil beads for the storage trials and pH treatment studies. The unencapsulated oil (control)
was prepared by placing the canola oil in the same glass syringe (as
the core uid), then subjecting the oil to the same encapsulation process treatment but in the absence of a shell uid in the plastic syringe,
and harvested by collection in the same beaker. The control oil was
thus exposed to the same light and heat exposures as the encapsulated
oils.
839
Table 1
Effect of encapsulation conditions on beads encapsulated using 1% alginate.
Encapsulation conditions
Bead parameters and oil loading efciency
Shell ow rate
(mL/h)
Core ow rate
(mL/h)
Hardening
time
Beads size
(m)
Wall thickness
(m)
Oil loading efciency

(%)
200
30
60
30
60
30
60
30
60
RT, 10 min
350400
400450
350400
400450
450600
450600
450600
450600
6070
4060
6070
4060
7090
6080
7090
6080
65
62
68
65
62
59
63
60
250
4 C, 2 h
RT, 10 min
4 C, 2 h
1%ab
1%b
1%a
1%ab
1%b
1%b
1%ab
1%b
Breakage (%)
Before freeze drying
2
4
2
2
2
4
4
3
0.5%b
0.5%a
0.5%b
0.5%b
0.5%b
0.5%a
0.5%a
0.5%a
After freeze drying

5
8
4
6
5
7
6
6
0.5%bc
0.5%a
0.5%c
0.5%b
0.5%bc
0.5%ab
0.5%b
0.5%b
Note: RT = room temperature. Data are expressed as mean standard. acValues of the same column with different superscripts are signicantly different (p b 0.05).
2.2.4. Measurement of oil loading efciency and bead breakage

The oil loading efciency was evaluated using this equation:
Oil loading efficiency % TS=T 100%
where T is the total amount of the oil (g) in syringe used for preparing
beads, and S is the amount of surface oil (g) in the CaCl2 solution.
The bead breakage was measured before and after freeze drying. A
small amount of fresh encapsulated beads (initially suspended in a 3%
CaCl2 solution) was randomly withdrawn and transferred onto a
50 m nylon mesh. An aliquot (1 g) of the beads was accurately
weighed out, from which the broken beads were removed aided by optical microscopy. The remaining unbroken beads were accurately
weighed (Wunbroken). The bead breakage before freeze drying was dened using the following equation: Breakage before freeze drying % =
(1 Wunbroken) 100%.
A predetermined amount of each type of bead (30 beads) was
subjected to freeze drying. The number of the broken beads after freeze
drying was assessed by optical microscopy (Section 2.6). The weights
of 30 dried beads (Wdried) and the unbroken dried beads (Wdried,unbroken)
were measured. The bead breakage after freeze drying was dened as:
Breakage after freeze drying % = [(Wdried Wdried,unbroken) / Wdried]
100%. All determinations were run in triplicate and the mean values reported here.
2.3. Exposure quercetin-containing canola oil beads to acidic pH
An acidic aqueous solution was prepared by adding a 0.5% HCl solution (prepared from concentrated HCl of 36.8%w/w concentration) to
Milli-Q water until pH 3 (0.001 mol/L). The pH was monitored using a
pH metre (Schott Instrument pH Meter Lab 850, Xylem Inc, Germany).
Fresh quercetin-containing beads (initially suspended in the CaCl2 solution) were transferred onto a 50-m nylon mesh, and partially dried by
absorbing excess water with a paper towel held beneath the nylon
mesh. The partially dried beads (300 mg, accurately weighed) were immediately placed into the aqueous solution (5 mL) at pH 3 for 2 h (simulating the pH of acidic food matrix and the human stomach). Gentle
and regular shaking was applied during this pH treatment. After the
pH treatment, the wet beads were collected and subjected to optical microscopy examinations, with the residual aqueous solution subjected to
total phenolic content analysis by the FolinCiocalteu assay (to examine
if any quercetin and its derived products were released from the beads
during the pH treatment).
2.4. Storage trials
Unencapsulated canola oil and freeze dried encapsulated canola oil
beads were stored in sealed glass scintillation vials (20 mL) with the
same headspace ratio (3 mL). The bead-containing vials were ushed
with N2, tightly capped, wrapped with aluminium foil, and stored
(away from light) for 30 or 60 days at room temperature (in cooled

room, 20 2 C) or at 38 1 C (in an incubator oven, MIR 162,
Sanyo Electric Co., Ltd, Osaka, Japan). Two subsamples from each vial
were withdrawn on Day 0, 30 or 60. A portion of each bead subsample
was used for microscopic, physical and chemical analyses. The
remaining freeze dried beads in the vials were ushed with N2 and
kept in a freezer (20 C) for later use.
2.5. Water activity of the freeze dried beads
The water activity (aw) of the freeze dried canola oil beads was measured at 25 C using a dew point water activity metre (AquaLab 4TE,
Decagon Devices Inc., Pullman, WA, USA). For each type of bead, duplicate samples were collected from two separate bead production batches
and triplicate measurements were performed on each of the duplicate
samples.
2.6. Microscopic examination of encapsulated oil beads
The appearance, size and wall thickness of fresh encapsulated
canola oil beads (in one drop of 3% CaCl2 solution) were examined by
optical microscopy with a Nikon Eclipse E600 microscope (Nikon
Corporation, Chiyoda-ku, Tokyo, Japan) equipped with a Nikon Coolpix
995 3.34 mega pixel camera (10, Nikon corporation, Chiyoda-ku,
Tokyo, Japan).
A small amount of fresh encapsulated beads (initially suspended in a
3% CaCl2 solution) was transferred using a plastic dropper onto a 50 m
nylon mesh, partially dried (through using a paper towel beneath the
nylon mesh to absorb excess water), and then placed directly on the
sample stage of an environmental scanning electron microscope
(ESEM) (FEI-Quanta-200, FEI, Hillsboro, OR, USA). The images were
taken at an electron gun accelerating voltage of 10 kV at a sample temperature of 2 C and a relative humidity of 95%. Ten beads per bead production batch (for two production batches) were examined, and three
pictures per bead production batch were taken.
2.7. Fourier transform infrared (FT-IR) spectroscopy
The bead ingredients (i.e. canola oil, quercetin, alginate, HM pectin
and water), as well as the freeze dried quercetin-containing oil encapsulated beads were examined by FT-IR spectroscopy. FT-IR absorbance
spectra were collected over the range 4000650 cm1 (4 cm1 resolution, 32 accumulated scans) using a Perkin Elmer Spectrum 100 FT-IR
spectrometer equipped with a universal ATR sampling attachment and
standard optical geometry. A sample background was collected in air
before sample analyses. Each ingredient or bead sample was placed
directly onto the ATR crystal at room temperature under dim light
(in room with no sunlight and direct light but reduced bulb lighting
from 5 m away). Absorbance spectra were normalised using SigmaPlot
(version 11.0).
840
2.8. Oil stability evaluation
2.9. High performance liquid chromatography (HPLC) phenolic proling
2.8.1. Extraction of canola oil from the encapsulated beads and extraction of
phenolics from the obtained oil
The extraction of canola oil from encapsulated beads followed the
method of Sun-Waterhouse et al. (2012) with some modications.
Freeze dried beads (2 g) were rst ground using a mortar and pestle,
to which 20 mL of KCl solution (0.88% w/v) was added. After the suspension was mixed using a MT19 vortex mixer (Chiltern International,
Slough, UK, operating at the speed of 4), chloroform (20 mL) and methanol (20 mL) were added. The resultant mixture was homogenised
using a homogeniser (12,000 rpm for 2 min; CH 6005 model, Luzern,
Switzerland). The chloroform layer was then collected, and the extraction step was repeated. The two collected chloroform layers were combined, evaporated using a Labconco RapidVap Concentrator (40 min at
40 C &10 kPa; Model 79100-01, Labconco Corp., Kansas City, Missouri,
USA) under N2, dried in the Ultra-Low Cold Trap Centrivap Centrifugal
Concentrator (Model 78100-01, Labconco Corp., Kansas City, MO) at
40 C for 4 h under vacuum (Sun-Waterhouse, Thakorlal, & Zhou,
2011; Sun-Waterhouse, Zhou, et al., 2011; Sun-Waterhouse et al.,
2012), and stored in a freezer (80 C) until analysis.
The extraction of the phenolics in the extracted oils followed the
method of Sun-Waterhouse, Zhou, et al. (2011) with some modications.
An aliquot of extracted oil (0.5 g) was dissolved in n-hexane (1.25 mL),
to which aqueous methanol (80% v/v, 0.5 mL) was added. The mixture
was vigorously mixed for 2 min using the MT19 vortex mixer at speed
4, centrifuged at 3000 rpm for 3 min (Centrifuge 5702, Eppendorf,
Hamburg, Germany), and the methanolic phase was collected and transferred to a vial wrapped with aluminium foil. This methanolic extraction
was repeated three times. The combined methanolic extracts were dried
in the Ultra-Low Cold Trap Centrivap Concentrator at 40 C for 4 h
under vacuum, and stored in a freezer (80 C) until analysis.
An analytical HPLC (HP Agilent 1100 series, Agilent Technologies,

USA) equipped with a G1313A autosampler, a G1322A degasser, a
G1311A Quat pump, a G1316A column compartment and a Phenomenex
Luna C18 column (4.6 250 nm, 5 m particle size), was used to analyse the phenolics including quercetin in the encapsulated canola oil
beads (Phani, Vinaykumar, Umamaheswara rao, & Sindhuja, 2010). The
mobile phases consist of (A) 1% acetic acid (45.0%), (B) acetonitrile
(15.0%) and (C) methanol (40.0%). The ow rate was 1.0 mL/min and injection volume was 20 L with a total run time of 30 min. The wavelength detection range was 200 to 540 nm.
The phenolics extracts (~1 g) obtained in Section 2.8.1 were constituted in aqueous methanol (80%v/v, 0.5 mL), vortexed and centrifuged
(3000 rpm for 3 min). The resultant supernatant was ltered through a
0.45 m lter (Nylon Membranes; Supelco, Bellefonte, PA, USA) and
then transferred to brown vials for HPLC analysis. The undissolved quercetin was recovered from the obtained residues by centrifugation and
ltration. Individual phenolics were identied based on their retention
time and absorbance maximum (max). The concentrations of nonavonoid phenolics present at 280 nm and avonoids present at
370 nm were estimated using a catechin or quercetin external standard,
respectively. Quercetin was quantied at 370 nm using internal standard rutin hydrate (25 ppm). The concentration of quercetin was calculated using this equation: Concentrationquercetin = (Peak Areaquercetin
Concentrationrutin) / Peak Arearutin.
2.8.2. Peroxide value (PV) determination

The PVs of the oils were determined (AOCS, 1998a) following the
procedures of Sun-Waterhouse, Thakorlal, et al. (2011), and expressed
as milliequivalent (meq) per kg of oil.
2.8.3. p-Anisidine value (p-AV) determination
The p-AVs of the oils were determined (AOCS, 1998b), following
the procedures of Sun-Waterhouse, Thakorlal, et al. (2011). A spectrophotometer (SpectraMax Plus 384, MDS Analytical Technologies,
Hawthorn, Australia) was used to measure the absorbance at 350 nm.
2.8.4. Totox value calculation
Totox values were calculated from the PVs and p-AVs of the oil samples using the equation of Totox value = 2PV + p-AV (O'Connor, Lal, &
Eyres, 2007).
2.8.5. Free fatty acid (FFA) determination
FFAs were determined using the direct titration method of AOCS
(2000) and following the procedures of Sun-Waterhouse, Thakorlal,
et al. (2011). FFA content was expressed as g FFA as oleic acid per
100 g oil.
2.8.6. Iodine value (IV) determination
IV was determined using the AOCS Ofcial Method (1997) and
expressed as g I2/100 g oil.
2.8.7. Analysis of total extractable phenolic content
The total extracted phenolic content (TEPC) was determined by the
FolinCiocalteu assay (Singleton, Orthofer, & Lamuela-Ravento, 1997),
and expressed as mg gallic acid equivalents (GalE) per kg oil.
2.10. Statistical analysis

All chemical analysis data are expressed as mean standard deviation of three replicates. Analyses were carried out using MINITAB 15
(Minitab Inc., Pennsylvania, USA) statistical software with one-way
ANOVA followed by Tukey's multiple comparison test at p b 0.05.
3. Results and discussion
3.1. Effect of encapsulation conditions on bead characteristics, oil loading
efciency and breakage percentage of the 1% alginate canola oil beads
The effects of co-extrusion encapsulation conditions, in particular
coreshell feed ow rates and bead hardening temperature and time,
were rst examined using oil beads encapsulated with 1% alginate
only (1%A). Optical microscopy examinations (Fig. 1AD) revealed
that all the 1%A beads prepared using different coreshell feed ow
rates and hardening conditions were spherically shaped which is desirable as it minimises the surface area to volume ratio of the beads (http://
en.wikipedia.org/wiki/Surface_tension) and hence reduces O2 permeation through the shell to the core oil. Table 1 summaries the bead
size, wall thickness, oil loading efciency and breakage percentage of
the alginate encapsulated beads that were prepared using a core feed
ow rate of 30 or 60 mL/h, and a shell feed ow rate of 200 and
250 mL/h, and hardening at room temperature for 10 min or at 4 C
for 2 h in 3% CaCl2 solution. These conditions were selected based on results of our preliminary trials, and yielded individual intact beads.
The feed ow rates of core and shell solutions and the hardening time
of encapsulated beads all inuenced the characteristics of oil beads. The
effect of hardening conditions on bead hardening was consistent with
the rapid gelation and cross-linking of calcium-alginate to form the
egg-box junction during co-extrusion encapsulation (Li, Yun, Xing, Liu,
& Xu, 2011). At the same shell ow rate, higher core ow rates led to a
larger bead size but a thinner wall thickness. This is understandable as
within a given time period, a higher core ow rate gives a greater
amount of core uid relative to a xed amount of shell uid. The alginate
beads obtained at the high shell ow rate (250 mL/h) yield inconsistent
bead sizes (i.e. a wider bead size distribution). At the same core ow rate,
the bead size and wall thickness increased at higher shell ow rates, and
A, 30-200 mL/h
841
B, 30-250 mL/h
100 m
100 m
100 m
100 m
C, 60-200 mL/h
D, 60-250 mL/h
Fig. 1. Optical micrographs (magnication 10) of fresh canola oil beads encapsulated with 1% alginate, which were prepared using different coreshell feed ow rates (A, 30200 mL/h;
B, 30250 mL/h; C, 60200 mL/h; D, 60250 mL/h), but the same hardening conditions: at 4 C for 2 h in 3% CaCl2 solution.
the increased shell ow rate led to the presence of small individual

spherical features in the shell (which are likely to be oil droplets or
possibly air pockets) (Fig. 1B and D). The alginate beads obtained at
the core ow rate of 60 mL/h largely tended to be present as twins
or triplets and suspended as clusters in the CaCl2 solution, with uneven
shell wall thickness (Fig. 1C and D). In general, these ndings agreed
with the results of Whelehan and Marison (2011) who reported that
bead wall thickness was controlled by the ow rate ratio of shell to
core solutions. Room temperature was selected as it has advantages relating to energy cost-effectiveness. 4 C was selected as an alternative
to RT. Our preliminary trials showed that the minimal hardening time
at RT or 4 C was 10 min and 2 h, respectively. When the same core
ow rate and a 200 mL/h shell ow rate were used, bead hardening at
4 C for 2 h appeared to be more benecial (i.e. higher oil loading efciency) than at room temperature for 10 min. Hardening time was
found to have no effect on bead size, a result which was also observed
by Lotpour, Mirzaeei, and Maghsoodi (2012). Longer hardening times
are expected to lead to a higher degree of cross-linking (RosasLedesma, Leon-Rubio, Alarcon, Morinigo, & Balebona, 2012; Smrdel,
Bogataj, & Mrhar, 2008). Deladino, Anbinder, Navarro, and Martino
(2008) showed that 15 min hardening facilitated bigger beads with
complete cross-linking and maximum mechanical strength. The highest
oil loading efciency was 68 1%, and the lowest breakage percentages
before and after freeze drying were 2 0.5% and 4 0.5%, respectively
(Table 1). The 30200 coreshell ow rate in combination with 2 h
hardening at 4 C appeared to be the best conditions for oil encapsulation by alginate, because of the resultant high oil loading efciency and
low breakage percentages before and after freeze drying. Therefore,
this set of process parameters was selected as the optimal conditions
and used for all other encapsulation studies.
The breakage of some beads during freeze drying was likely due to
the fast sublimation of frozen water from the alginate gel matrix,
resulting in the formation of various pores without having sufcient
time to shrink (Smrdel et al., 2008). The amount of surface oil detected
on the freeze dried beads was low and constant for the three types of encapsulated beads in this study i.e. 0.30.4 g surface oil/10 g dried beads.
The presence of large amounts of oil on the surface of encapsulated
beads (surface oil) is undesirable, since the surface oil not only deteriorates rapidly causing off-avour but also affects the wettability and
dispersability of encapsulated beads (Drusch & Mannino, 2009). The
degree of surface wrinkling caused by freeze drying the beads was
dependent on the encapsulation parameters i.e. core and shell feed
ow rates and bead hardening conditions. Most severe wrinkling was
observed for the beads produced at a core ow rate of 60 mL/h, a shell
ow rate of 200 mL/h and hardened at room temperature for 10 min.
3.2. Appearance, size and wall thickness of the quercetin-containing
canola oil beads
The freeze dried quercetin-containing canola oil beads that were
encapsulated by 0.67%A, 1%A or AP under the optimal encapsulation
conditions selected in Section 3.1. (i.e. 30200 coreshell ow rates
and 2 h hardening at 4 C) had water activities of 0.135 0.001,
0.352 0.001, and 0.460 0.001, respectively. The water activity
values suggest that the freeze dried AP beads might have a slightly
higher risk of microbial spoilage compared to the 2 types of alginate
only beads (Sun-Waterhouse et al., 2013).
The optical and ESEM micrographs (Fig. 2) conrm the structured
integrity of all 3 types of co-extruded canola oil beads. The ESEM micrographs (Fig. 2, 1st row) show that the 0.67%A (magnication 400),
842
A 0.67% Alginate
B 1% Alginate
200 m
200 m
Before pH treatment
Before pH treatment
C Alginate-pectin
300 m
Before pH treatment
100 m
100 m
100 m
After pH 3 for 2 h
After pH 3 for 2 h
After pH 3 for 2 h
100 m
100 m
100 m
Fig. 2. ESEM (1st row, before pH treatment; magnication 400 for alginate alone beads and 300 for alginatepectin beads) and optical microscopy images (magnication 10, before,
2nd row, and after, 3rd row, the treatment at pH 3 for 2 h) of fresh (wet) canola oil beads prepared with shell formulations: A) 0.67% alginate, B) 1% alginate, and C) alginate + High
methoxyl (HM) pectin.
1%A (magnication 400) and AP (magnication 300) oil beads had

regularly spherical shape and smooth surfaces. Optical microscopy
conrmed that all the 3 types of oil beads were in spherical shape
after encapsulation and before pH treatments (Fig. 2, 2nd row). The
average diameter of whole bead and oil core were: 492 15 and
286 8 m for 0.67%A beads, 348 11 and 208 6 m for 1%A
beads, and 533 16 and 367 9 m for AP beads, respectively. The
AP beads had the largest size. The average wall thickness decreased
in the order 0.67%A beads (~103 m) N AP beads (~83 m) N 1%A
beads (~70 m). A lower concentration of alginate resulted in signicantly larger beads, although the relative size of bead to core remained
almost constant.
When alginate is used alone, an increase of alginate concentration
would allow more Ca2+ binding sites and a greater number of alginate
strands held together in the egg-box structure within the bead shell,
thus resulting in a higher degree of cross-linking and a more rigid and
compact matrix (Liu et al., 2002; Mandal, Kumar, Krishnamoorthy, &
Basu, 2010). Accordingly, an elevated alginate concentration increased
the viscosity of alginate gel and subsequently a slower diffusion of core
substances, such as small polar phenolic compounds that were potentially derived from quercetin decomposition at pH like 3 (see
Section 3.3), through the alginate egg-box cavities to the outer solution
(Liu et al., 2002; Mandal et al., 2010). When the shell was AP mixture,
the actual concentration of alginate in AP was 0.67% (as a result of
mixing 1% alginate solution with 1% pectin solution at a 2:1 volume
ratio). Thus, it is not surprising that the shell wall thickness of the AP
beads ranked in between those of the 0.67%A and 1%A beads. Biopolymer
composition was reported to inuence bead diameter and roundness
(Sandoval-Castilla et al., 2010). Based on microscopy observations, the
AP combination was suitable for encapsulation. Alginate and pectin
are both polyuronates, and the Ca2+ binding mechanism occurred in

both alginate and the unmethoxylated pectins in HM pectin (although
N50% methoxylated pectins in HM pectin did not gel relying on Ca2+
but sugar or acid) (Siew & Williams, 2005; Whistler & BeMiller, 1997).
Madziva, Kailasapathy, and Phillips (2005) reported that AP encapsulated beads possessed greater encapsulation efciency, but less spherical
shape than the alginate encapsulated beads. Islan et al. (2012) reported
that incorporating 1% HM pectin into 2% alginate resulted in less rigid
structure of microspheres than those prepared using alginate only.
Thus, it is expected that the HM pectin component inuences the microstructure and network density of encapsulated beads (Morris & Chilvers,
1984; Sandoval-Castilla et al., 2010).
3.3. Effect of acidic treatment on the quercetin-containing oil beads
The integrity of the quercetin-containing 0.67%A, 1%A and AP beads
was retained after the acidic treatment at pH 3 for 2 h (Fig. 2, 3rd row),
suggesting that these bead shells were all chemically and structurally
robust. The size of these beads, however, decreased signicantly after
the pH treatment: The 1%A beads had the smallest bead size reduction
(12.9%), whilst the 0.67%A and AP beads exhibited comparable size reduction (37.6% and 29.1%, respectively). This shrinkage enabled a more
compact molecular arrangement within the shell network. Previous
studies have reported that alginate beads tended to shrink in highly
acidic media (Rayment et al., 2009), due to either strong hydrogen interactions or intermolecular lactonisation (Tataru, Popa, & Desbrieres,
2011). In this study, the oil beads did not rupture at low pH.
The total phenolics (including quercetin and its derived products)
released from the beads into the acidic water were evaluated by
the FolinCiocalteu assay (Table 2), and decreased in the order

Table 2
Total phenolic content of the aqueous solutions obtained following the treatments of
quercetin-containing oil beads at pH 3 for 2 h.
Sample
Total phenolic content

(g gallic acid equivalent per mL
solution or mg beads)
Water solution for pH treatment

0.67% Alginate
1% Alginate
Alginate-pectin
0.175
0.238
0.213
0.207
0.001c
0.001a
0.001b
0.001bc
Note: Data are expressed as mean standard. acvalues of the same column with
different superscripts are signicantly different (p b 0.05).
0.67%A N 1%A N AP beads. The release characteristics of bead shells are

strongly dependent on their permeability (which was affected by the
polarity, density, porosity, homogeneity and thickness of shell wall materials) (Donhowe & Fennema, 1993). Alginate is pH sensitive (Whistler
& BeMiller, 1997) rendering it suitable for the intestinal delivery systems. Alginate solutions are chemically stable at pH 5.510 at room
temperature (McDowell, 1977). When pH N pKa, the carboxylate
groups are ionised in solution and alginate polymer acts like a polyelectrolyte. When pH b pKa, the carboxyl groups remain unionised and
alginate polymer behaves more like a neutral polymer depending on
the ratio of -D-mannuronic acid (M) and -L-guluronic (G) acid
(Bergfeldt, Piculell, & Tjerneld, 1995). When pH b 4, alginate is
converted to alginic acid forming a viscous acid gel (Rayment et al.,
2009), thus retarding the loss of encapsulated phenolics. Alginate concentration also affects the porosity of its gel matrix (Goh, Heng, &
Chan, 2012). Blandino, Macas, and Cantero (2001) found that capsules
prepared with 1% sodium alginate were more robust than those with
lower alginate concentrations, because a more densely cross-linked
gel structure could be formed. Thus, it is not surprising that a slightly
greater amount of phenolics were released from the 0.67%A beads
than the 1%A beads.
Mixing alginate with HM pectin modied the characteristics of bead
shells and the release of substances from bead cores (Madziva et al.,
2005; Sun-Waterhouse et al., 2012). The gelation of pure HM pectin involves various intermolecular interactions, with the junction zones
being stabilised by both hydrogen bonds and hydrophobic interactions
involving ester methyl groups (Oakenfull & Scott, 1984; Walkinshaw
& Arnott, 1981). In pure HM pectin, the proportion of the ionised
carboxyl groups present is smaller (b50%) than the proportion of methylated carboxylate groups. Thus, only a small number of hydrogen
bonds are formed with stronger interactions like ion-dipole and generation of electrostatic complexes between anionic alginate and pectin
being dominant. The addition of HM pectin naturally reduces the electrostatic repulsion and permits molecular association with alginate carboxyls in free acid form and facilitates subsequent gelling (Morris &
Chilvers, 1984). The interaction between pectin and alginate in mixed
gels is a heterogeneous association between the poly-G blocks of alginate and the methyl ester regions of pectin of low charge, packing together in rigid ribbons (Voo, Ravindra, Tey, & Chan, 2011). The pH is
critical to the gel formation for pectins especially HM pectins. Low pHs
increase the percentage of unionised carboxyl groups, thus reducing
electrostatic repulsion between adjacent pectin chains (Whistler &
BeMiller, 1997). The pH levels of the 1%A solution and 0.67%A solution
were ~7.2 and ~6.9, respectively. Pectin is a weak acid with a pKa
value of ~4 and 1% HM pectin solution has a pH ~3.5. When 1%A solution was mixed with 1% HM pectin solution at a volume ratio of 2:1
for encapsulation, the pH of the mixed shell solution was ~5.6. When
the AP beads were exposed to acidied water at pH 3, the HM pectin
component in the AP shell should be still stable (because HM pectin
generally has excellent stability at all temperatures at pH of 2.54.5)
(Oakenfull & Scott, 1984; Whistler & BeMiller, 1997). The alginate component in the AP shell, however, shrank and became more viscous
843
(Rayment et al., 2009). The different responses to pH of alginate and

HM pectin are expected to inuence shell porosity and density, and be
responsible for different degrees of loss of phenolics from the encapsulated beads.
3.4. Stability of core oil and total extracted phenolic content (TEPC) in beads
after storages
The degree of oxidation (evaluated based on PV, p-AV and Totox
values), extent of hydrolytic rancidity (evaluated based on FFA contents), change in unsaturation (evaluated based on IV values), and retention of phenolics (evaluated based on TEPC values) of the encapsulated
canola oil beads over 60 day storage trials were summarised in Table 3
(at 20 C) and Table 4 (at 38 C).
The PVs of each encapsulated oil increased with storage time, and the
increase was greater at 38 C (Tables 3 and 4). Such PV increases were
due to a higher rate for hydroperoxide formation rather than decomposition (Lee, Lee, & Choe, 2007). For each type of beads, addition of an antioxidant (i.e. quercetin, vitamin E or BHT) mostly reduced primary oil
oxidation but exceptions were found especially in the presence of vitamin
E The effectiveness decreased in the order quercetin N BHT N vitamin E
at either 20 or 38 C. After 60 days at 20 C, the quercetin-containing encapsulated oils had a similar PV (9 meq/kg oil) which was lower than
that of control oil (unencapsulated oil, 13.4 meq/kg oil) (Table 3). After
60 days at 38 C (Table 4), the PV of control oil was the highest
(16.5 meq/kg oil), while the quercetin-containing oil encapsulated by
1%A had a slightly lower PV (10.1 meq/kg oil) than that encapsulated
by 0.67%A or AP (~12.5 meq/kg oil).
The p-AVs of the control and encapsulated oils increased after
60 days at 20 C or 38 C, indicating an increased concentration of secondary oil oxidation products. At either storage temperature, little difference was detected in p-AV among the 3 types of antioxidant for
either the 0.67%A beads or the 1%A beads. But for the AP beads, vitamin
E was slightly more effective than BHT and quercetin in suppressing secondary oil oxidation, which may be associated with polarity differences
between these antioxidants and oil primary oxidation products. After
60 days and in the presence of quercetin, the 1%A beads had the lowest
p-AVs (2.20 and 2.98 at 20 C or 38 C, respectively).
Totox values estimate the overall oil oxidation status. After 60 days at
20 C, the control oil had a higher Totox value (29.4, close to the legally
acceptable value 30, O'Connor et al., 2007) than the quercetin-containing
encapsulated oils (2022). After 60 days at 38 C, the Totox value
decreased in the order control oil (36.9) N quercetin-containing AP
(30.1) N quercetin-containing 0.67%A (27.4) N quercetin-containing
1%A (23.2). After 30 days at either temperature and in the presence of
quercetin, the AP beads had lower Totox values compared to the
other 2 types of beads. Thus, in the presence of quercetin, AP may
form the best shell against oil oxidation for a short storage period (i.e.
30 days), but 1%A appeared to be the best shell for 60 day storage
periods.
The FFA of canola oil was low on Day 0 (0.08%) but increased after
storage, suggesting moisture capture and moisture permeability. In
the absence of an added antioxidant, the encapsulated oils had comparable FFA contents after 60 days storage. No discernable differences in
suppressing oil hydrolytic rancidity were observed among the 3 types
of antioxidants i.e. quercetin, vitamin E and BHT. The FFA contents
showed uctuation due to the nature of the lipid and its surrounding
environment (Hung & Slinger, 1981). After storage at 20 C and 38 C,
the differences among the FFA readings of different encapsulated oils
under the same storage conditions were insignicant.
Iodine value (IV) indicates unsaturation status of lipids thus providing an estimate of oil nutritional value (i.e. unsaturated oil is generally
considered good for health) and oil stability (i.e. unsaturated lipids are
more susceptible to rancidity) (Toscano, Riva, Pedretti, & Duca, 2012).
IV of oil typically decreases with storage time before levelling off, because oxidation decreases oil unsaturation via abstraction of hydrogen
844
Table 3
Peroxide value, p-anisidine value, free fatty acid content, iodine value and total extracted phenolic content of control and encapsulated oils after storage at room temperature.
Oil sample
Storage (day)
PV (meq/kg oil)
Control
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
5.0
9.7
13.4
4.9
9.3
13.1
4.8
6.9
9.0
4.9
8.4
12.7
4.8
7.6
11.0
4.8
7.9
12.8
4.9
7.0
9.0
4.8
7.9
12.2
4.9
7.6
11.2
4.9
8.0
14.3
4.8
5.5
9.0
4.9
8.1
13.3
4.7
6.8
11.0
A + O
A + O + Q
A + O + VE
A + O + BHT
A+O
A+O+Q
A + O + VE
A + O + BHT
A+P+O
A+P+O+Q
A + P + O + VE
A + P + O + BHT
0.1C,a
0.3B,a
0.2A,b
0.1C,a
0.6B,a
0.3A,b
0.1C,a
0.1B,c
0.7A,e
0.1C,a
0.6B,b
0.5A,b
0.1C,a
0.3B,bc
0.2A,c
0.1C,a
0.1B,b
0.1A,b
0.1C,a
0.3B,c
0.2A,d
0.1C,a
0.5B,b
0.3A,bc
0.3C,a
0.4B,bc
0.3A,c
0.1C,a
0.7B,b
0.6A,a
0.1C,a
0.3B,d
0.3A,d
0.1C,a
0.4B,b
0.7A,b
0.3C,a
0.5B,c
0.7A,c
p-AV
1.6
1.9
2.7
2.3
2.2
2.9
2.6
2.8
4.0
1.7
2.3
3.5
1.9
2.2
3.4
1.3
2.0
2.2
1.2
1.8
2.2
1.5
2.1
2.5
1.5
1.7
1.9
1.7
1.9
2.3
2.5
2.5
3.1
1.5
1.6
2.8
1.7
1.8
3.6
Totox
0.1C,de
0.1B,d
0.1A,de
0.1B,b
0.1B,c
0.1A,cd
0.1B,a
0.1B,a
0.2A,a
0.1C,d
0.1B,c
0.1A,b
0.1C,c
0.1B,c
0.1A,b
0.1B,f
0.1A,cd
0.1A,ef
0.1C,g
0.1B,de
0.1A,ef
0.1C,e
0.1B,cd
0.1A,e
0.1B,e
0.1A,de
0.1A,f
0.1B,d
0.1B,d
0.1A,e
0.1B,a
0.1B,b
0.1A,c
0.1B,e
0.1B,e
0.1A,d
0.1B,d
0.1B,de
0.1A,b
11.6
21.2
29.4
12.1
20.9
29.1
12.2
16.6
22.0
11.5
19.9
28.9
11.4
17.4
25.2
10.9
17.8
27.8
12.0
15.9
20.2
11.1
17.9
26.9
11.3
16.9
24.3
11.5
17.8
30.9
12.0
13.4
21.1
11.3
17.8
29.4
11.1
15.4
25.6
FFA (g/100 g oil)

0.2C,b
0.5B,a
0.3A,b
0.1C,a
0.8B,a
0.5A,b
0.2C,a
0.1B,b
0.9A,cd
0.1C,b
0.9B,a
0.7A,b
0.2C,b
0.5B,ab
0.3A,c
0.2C,b
0.2B,ab
0.1A,bc
0.1C,a
0.4B,bc
0.3A,cd
0.2C,b
0.7B,ab
0.5A,bc
0.4C,b
0.5B,b
0.4A,c
0.2C,b
1.1B,ab
0.9A,a
0.2C,a
0.4B,c
0.4A,cd
0.2C,b
0.6B,ab
1.0A,b
0.4C,b
0.7B,bc
0.9A,c
0.1
0.2
0.4
0.1
0.2
0.4
0.1
0.2
0.2
0.1
0.2
0.3
0.1
0.2
0.3
0.1
0.2
0.4
0.1
0.2
0.2
0.1
0.2
0.2
0.1
0.2
0.3
0.1
0.2
0.4
0.1
0.2
0.2
0.1
0.1
0.3
0.1
0.1
0.3
0.1B,a
0.1B,a
0.1A,a
0.1B,a
0.1B,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1B,a
0.1B,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1B,a
0.1B,a
0.1A,a
0.1B,a
0.1B,a
0.1A,a
IV (g I2/100 g oil)
185
180
176
140
179
138
168
180
135
178
169
112
170
158
110
177
176
172
176
171
159
187
162
125
130
127
121
131
187
155
126
178
171
174
159
148
172
133
115
0.5A,ab
0.5A,b
0.4B,a
0.3B,d
0.3A,b
0.3B,d
0.1B,c
0.5A,b
0.9C,d
0.8A,b
0.4B,bc
0.5C,f
0.9A,bc
0.7B,c
0.9C,f
0.5A,b
0.4A,bc
0.3B,ab
0.3A,b
0.3A,bc
0.5C,b
0.1A,a
0.4B,c
0.7C,e
0.1A,de
0.1AB,d
0.9B,e
0.5C,de
0.1A,a
0.4B,bc
0.5B,e
0.7A,b
0.5A,ab
0.9A,b
0.1B,c
0.1C,c
0.1A,bc
0.3B,cd
0.9C,f
TEPC (mg GalE/kg oil)

21.2
23.0
26.9
24.2
18.8
29.5
36.1
23.5
32.6
30.8
26.9
46.5
31.4
30.8
40.3
25.2
26.2
22.5
48.7
32.3
37.8
28.2
26.0
43.2
32.3
26.8
35.1
23.2
23.5
25.9
35.0
23.0
31.8
28.9
26.3
29.8
28.1
32.3
38.4
0.1B,e
0.8B,c
0.4A,d
0.2B,d
0.6C,d
0.6A,cd
0.5A,b
0.3B,c
0.1A,cd
0.1B,c
0.3C,b
0.8A,a
0.1B,c
0.6B,ab
0.4A,ab
0.1A,d
0.6A,b
0.5B,e
0.1A,a
0.3C,a
0.4B,b
0.2B,cd
0.2C,b
1.4A,a
0.6B,bc
0.8C,b
1.8A,bc
0.5B,de
0.2B,c
0.4A,de
0.2A,b
0.2C,c
0.5B,c
0.3A,cd
0.3B,b
0.4A,cd
0.1C,cd
0.5B,a
0.3A,b
Note: PV = peroxide value, p-AV = p-anisidine value, FFA free = fatty acid, IV = iodine value, TEPC = total extracted phenolic content, GalE = gallic acid equivalents, A = 1% alginate,
A = 0.67% alginate, O = canola oil, Q = quercetin, vitamin E = VE, and P = High methoxyl pectin. ACvalues of the same analysis for the same oil sample but different storage periods
(Days 0, 30 and 60) with different superscripts are signicantly different (p b 0.05). afValues of the same analysis and on the same storage day but for different oil samples with different
superscripts are signicantly different (p b 0.05).
atoms adjacent to a double bond and leads to formation of free radicals

until complete oxidation of the accessible double bonds in FAs (Naz,
Sheikh, Siddiqi, & Sayeed, 2004). In this study, the IV readings of the
control and encapsulated oils generally decreased after storage. After
60 days at 20 C, the quercetin-containing AP beads had the highest
IV (171 0.5 g I2/100 g oil). The IV of 1%A and 0.67%A beads were
159 0.5 and 135 0.9 g I2/100 g oil, respectively. After 60 days at
38 C, the control oil had lower IV (126 0.1 g I2/100 g oil) compared
to the 3 types of quercetin-containing encapsulated oils. The IV of 1%A
beads (150 0.1 g I2/100 g oil) was higher than those of the AP and
0.67%A beads (143 0.4 and 140 0.3 g I2/100 g oil, respectively).
Quercetin generally exerted better preservation of unsaturated FAs in
the encapsulated oils compared to vitamin E or BHT. Some of the Day
30 IV readings of encapsulated oils were relatively high, which might result from interferents such as aldehydes and ketones newly formed during oil oxidation.
As expected, the TEPC values were higher for the beads with an
added antioxidant. Different TEPC patterns were found for the oil beads
encapsulated with different types of shell. In presence of quercetin, the
1%A beads had a slightly higher TEPC than the 0.67%A and AP beads
after 30 or 60 days at 20 C. However, at 38 C, the 1%A beads exhibited
a much lower TPEC (32.6 0.2 mg GalE/kg oil) compared with the
other 2 types of beads (53.7 0.4 and 54.5 0.4 mg GalE/kg oil)
after 60 days, whilst the AP beads had a slightly higher TEPC (40.8
0.7 mg GalE/kg oil) than the 0.67%A and 1%A beads (33.0 0.4 and
38.5 0.3 mg GalE/kg oil, respectively) after 30 days. The TEPC
detected by the FolinCiocalteu assay after extraction represents the extractable phenolic compounds present in the oil beads. Factors such as
bead matrix, storage temperature and time, and extraction method all
inuence TEPC values (Sun-Waterhouse et al., 2012). The amounts of
extracted canola oil from the 0.67%A, 1%A and AP beads were 86.6,
78.3 and 52.5 g/100 dried bead, respectively. The TEPC values were the
net results of self-degradation of antioxidants (especially at elevated
storage temperatures or prolonged storage times or both), the consumption of added antioxidants for protecting oil against deterioration, and
the different extractability of antioxidant from core canola oils encapsulated by different bead shell matrices.
Interplay exists between shell formulations and storage conditions
which ultimately inuence the detected PV, p-AV, FFA, IV and TEPC
readings. There was a close correlation between the TEPC values and
the status of oil deterioration, indicating that a signicant portion of
added antioxidants were used to suppress oil rancidity. Oil stability depends on chemical and physical factors such as light, temperature, pH,
FA composition and available oxygen (Frankel, Satu-Gracia, Meyer, &
German, 2002). BHT and tocopherols both contain phenolic components and are lipophilic organic compounds. Quercetin is quite polar
but has low water solubility (Huber, Rupasinghe, & Shahidi, 2009),
and in this study, was distributed in oil with the aid of PEG.
845
Table 4
Peroxide value, p-anisidine value, free fatty acid content, iodine value and total extracted phenolic content of control and encapsulated oils after storage at 38 C.
Oil sample
Storage (day)
PV (meq/kg oil)
Control
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
0
30
60
5.0
11.3
16.5
4.9
10.1
15.7
4.8
8.9
12.1
4.9
9.7
15.5
4.8
8.3
13.5
4.8
8.1
13.8
4.9
8.7
10.1
4.8
9.6
13.7
4.9
8.2
12.6
5.0
7.9
16.0
4.8
7.2
13.1
5.0
9.1
17.3
4.7
8.9
14.1
A + O
A + O + Q
A + O + VE
A + O + BHT
A+O
A+O+Q
A + O + VE
A + O + BHT
A+P+O
A+P+O+Q
A + P + O + VE
A + P + O + BHT
0.1C,a
0.2B,a
0.1A,b
0.1C,a
0.7B,b
0.3A,bc
0.1C,a
0.1B,c
0.1A,e
0.1C,a
0.1B,bc
0.1A,bc
0.1C,a
0.1B,cd
0.6A,cd
0.1C,a
0.2B,d
0.3A,cd
0.1C,a
0.3B,cd
0.2A,f
0.1C,a
0.7B,bc
0.9A,cd
0.3C,a
0.6B,d
0.4A,de
0.1C,a
0.4B,de
0.5A,b
0.1C,a
0.5B,e
0.7A,d
0.1C,a
0.6B,c
0.1A,a
0.3C,a
0.4B,c
0.2A,c
p-AV
1.6
2.1
3.9
2.3
2.7
3.3
2.6
2.4
3.2
1.7
2.6
3.5
1.9
2.3
3.1
1.3
2.8
3.3
1.2
2.4
3.0
1.5
2.5
3.7
1.5
2.2
3.4
1.7
3.4
4.5
2.5
3.0
3.9
1.5
2.2
4.3
1.7
2.7
4.9
Totox
0.1C,bc
0.1B,e
0.1A,bc
0.1C,ab
0.1B,c
0.1A,cd
0.1B,a
0.1B,d
0.1A,d
0.1C,b
0.1B,cd
0.1A,cd
0.1C,b
0.1B,de
0.2A,cd
0.1C,c
0.1B,c
0.1A,cd
0.1C,c
0.1B,d
0.1A,d
0.1C,bc
0.1B,cd
0.1A,c
0.1C,bc
0.1B,e
0.1A,cd
0.1C,b
0.1B,a
0.1A,ab
0.1C,a
0.1B,b
0.1A,bc
0.1C,bc
0.1B,e
0.1A,b
0.1C,b
0.1B,c
0.1A,a
11.6
24.8
36.9
12.1
22.9
34.7
12.2
20.3
27.4
11.5
21.9
34.5
11.4
18.8
30.1
10.9
18.9
30.9
12.0
19.7
23.2
12.1
21.7
31.1
11.3
18.7
28.6
11.5
19.1
36.5
12.0
17.4
30.1
11.3
20.3
38.9
11.1
20.4
33.1
FFA (g/100 g oil)

0.2C,b
0.3B,a
0.1A,a
0.1C,a
0.9B,ab
0.4A,ab
0.2C,a
0.1B,b
0.2A,bc
0.1C,b
0.2B,b
0.2A,ab
0.2C,b
0.2B,c
0.9A,b
0.2C,b
0.2B,c
0.4A,b
0.1C,a
0.4B,bc
0.2A,c
0.2C,a
0.9B,b
1.2A,b
0.4C,b
0.8B,c
0.5A,bc
0.2C,b
0.6B,c
0.7A,a
0.2C,a
0.6B,d
0.9A,b
0.2C,b
0.9B,b
0.1A,a
0.4C,b
0.5B,b
0.3A,ab
0.1
0.3
0.2
0.1
0.3
0.2
0.1
0.2
0.2
0.1
0.2
0.2
0.1
0.3
0.3
0.1
0.3
0.3
0.1
0.2
0.3
0.1
0.2
0.2
0.1
0.2
0.3
0.1
0.3
0.2
0.1
0.2
0.3
0.1
0.2
0.3
0.1
0.2
0.3
0.1B,a
0.1A,a
0.1AB,a
0.1B,a
0.1A,a
0.1AB,a
0.1A,a
0.2A,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1B,a
0.1A,a
0.1A,a
0.1B,a
0.1A,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1A,a
0.1A,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1B,a
0.1A,a
0.1AB,a
0.1B,a
0.1AB,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
0.1B,a
0.1AB,a
0.1A,a
IV (g I2/100 g oil)
185
170
126
139
162
133
167
166
140
169
140
112
168
123
118
177
175
142
176
151
150
182
126
118
130
118
113
132
178
145
126
149
143
155
128
121
165
130
110
0.7A,a
0.5B,ab
0.1C,bc
0.5C,de
0.5A,bc
0.5B,b
0.1A,c
0.5A,b
0.3B,ab
0.3A,c
0.4B,e
0.5C,cd
0.3A,c
0.4B,de
0.4C,cd
0.2A,b
0.2A,a
0.2B,ab
0.3A,b
0.2B,c
0.1B,a
0.1A,a
0.7B,de
0.7C,cd
0.1A,de
0.7B,e
0.6C,cd
0.4C,de
0.7A,a
0.2B,ab
0.4C,e
0.4A,c
0.4B,ab
0.9A,d
0.6B,de
0.7C,c
0.1A,c
0.3B,d
0.4C,d
TEPC (mg GalE/kg oil)

21.2
19.6
26.1
24.2
23.1
27.1
31.5
33.0
53.7
26.8
50.3
60.2
27.0
66.8
67.9
25.2
24.1
28.3
48.7
38.5
32.6
26.6
50.0
74.9
30.2
49.7
73.1
23.2
22.4
24.6
38.8
40.8
54.5
32.1
33.4
55.0
30.6
27.8
55.4
0.1B,e
0.2C,f
0.3A,ef
0.3B,de
0.3B,e
0.1A,ef
0.3B,c
0.4B,d
0.4A,c
0.1C,cd
0.4B,b
0.6A,bc
0.2B,cd
0.3A,a
0.4A,b
0.1B,d
0.3B,e
0.1A,e
0.1A,a
0.3B,cd
0.2C,d
0.2C,cd
0.4B,b
1.4A,a
0.6C,c
0.1B,b
0.4A,a
0.5B,de
0.5B,ef
0.2A,f
0.2C,b
0.7B,c
0.4A,c
0.3B,bc
0.4B,d
0.9A,c
0.6B,c
0.2C,de
0.3A,c
Note: PV = peroxide value, p-AV = p-anisidine value, FFA free = fatty acid, IV = iodine value, TEPC = total extracted phenolic content, GalE = gallic acid equivalents, A = 1% alginate,
A = 0.67% alginate, O = canola oil, Q = quercetin, vitamin E = VE, and P = High methoxyl pectin. ACValues of the same analysis for the same oil sample but different storage periods
(Days 0, 30 and 60) with different superscripts are signicantly different (p b 0.05). afValues of the same analysis and on the same storage day but for different oil samples with different
superscripts are signicantly different (p b 0.05).
The difference in the chemical structure, solubility, lipophillicity/

hydrophicility and emulsifying properties in oils of BHT, vitamin E and
quercetin accounted for the detected differences in their corresponding
PV, p-AV, FFA and IV results. With hydrophilic groups (e.g. hydroxyl
groups), quercetin may migrate and concentrate at the wateroil interphase (e.g. the interface between the polysaccharide gel shell and the
core oil), which enables to more effectively combat with the penetrating
oxygen. During the early stages of storage trials (i.e. Days 030), less oil
rancidity or degradation of antioxidants that were distributed in hydrophobic oil occurred. With extended storage (i.e. from Days 30 to 60), the
breakdown of vitamin E and BHT resulted in the exposure of their phenolic units. As a result of quercetin degration, more potent intermediate
products were possibly generated via ring cleavage. Thus, relatively
high readings were detected by FolinCiocalteu assay in some cases
on Day 60, and in particular, an elevated storage temperature such as
38 C also facilitated increased extractability of phenolics (SunWaterhouse et al., 2012). Under harsher storage conditions (38 C for
60 days), more antioxidants were consumed to preserve oil and/or
underwent self-degradation, causing a simultaneous decrease in TEPC.
In this study, oil primary oxidation was much more dominant than secondary oxidation and hydrolytic rancidity. The 1%A and AP shell formulations are acceptable and comparable for preserving quercetincontaining canola oil, with 1%A being slightly better for 60 day storages
and the AP shell for short storage periods up to 30 days.
3.5. HPLC analysis of phenolics in the encapsulated quercetin-containing

canola oil beads
HPLC analyses indicate that the shell formulations and storage conditions determined the amount of quercetin, avonoids, non-avonoid
phenolic compounds and some non-phenolic substances (Table 5).
The absorption spectra of avonoids generally consist of two characteristic bands with maxima at 280 and 370 nm (Zvezdanovi, Stanojevi,
Markovi, & Cvetkovi, 2012). Quercetin can be quantied at the
370 nm wavelength using rutin as an internal standard, which appeared
in the HPLC chromatograms at ~7.2 min and ~3.5 min, respectively.
On Day 0 (Fig. 3A), quercetin (2.163.12 g/g dried bead) and two
avonoids at 3.3 and 4.1 min were detected in all the 3 types of encapsulated oil samples. After 30 days at 20 C (HPLC proles not shown),
no quercetin was identied but instead two avonoids (at 3.7 and
4.1 min) were detected in the 3 types of encapsulated beads. Flavonoid
III (at 4.1 min) was also present in all the Day 0 beads. After 30 days at
38 C (HPLC proles not shown), neither quercetin nor any other avonoid was detected, instead, a non-avonoid phenolic compound at
3.2 min was identied at 280 nm in the 3 types of beads. This phenolic
compound might be a degradation product derived from quercetin.
Moreover, non-phenolic substances were found at 4.2, 4.7, 6.8, 8.4
and/or 10.2 min in the 3 types of beads. They might originate from the
shell materials. The 0.67%A beads had two extra substances at 6.8 and
846
Table 5
Amount of identied compounds in encapsulated quercetin-containing oil beads.
Storage
Day 0
Bead
280 nm
No detected phenolic and non-phenolic compounds.
0.67% alginate
7. 2 min, quercetin, 2.17 0.01 g/g dried bead,

4.1 min avonoid III, 0.82 0.01 g/g dried bead,
3.3 min, avonoid I, a big shoulder peak.
7.2 min quercetin, 3.12 0.02 g/g dried bead,
7.2 min, quercetin, 2.16 0.02 g/g dried bead,
No quercetin detected,
3.7 min avonoid II, 0.79 0.01 g/g dried bead,
4.1 min avonoid III, 0.09 0.01 g/g dried bead.
3.7 min avonoid II, 0.71 0.01 g/g dried bead,
No quercetin detected, 3.7 min avonoid II, 0.99 0.02 g/g dried bead,
No quercetin and other avonoid detected.
1% Alginate
Alginatepectin
0.67% Alginate
3.3 min, avonoid I, a shoulder peak, smaller than Day 0.
4.1 min, avonoid III, 0.61 0.03 g/g dried bead,
3.3 min, avonoid I, a very small shoulder peak.
0.67% Alginate
1% Alginate
Alginatepectin
Day 30, RT
0.67% Alginate
1% Alginate
Alginatepectin
Day 30, 38 C
Day 60, RT
1% Alginate
Alginatepectin
Day 60, 38 C
Identied compounds at different wavelengths

370 nm
0.67% Alginate
1% Alginate
Alginate-pectin
4.5 min, a very small phenolic peak;

no detected non-phenolic compounds.
No detected phenolics;
an unknown non-phenolic compound at 10.2 min.

3.2 min a phenolic compound, 2.89 0.03 g/g dried bead;
unknown non-phenolic compounds at 4.2, 4.7, 6.8, 8.4 and 10.2 min.
unknown non-phenolic compounds at 4.2, 4.7 and 10.2 min.
unknown non-phenolic compounds at 4.2, 4.7 and 10.2 min.
3.2 min, a phenolic compound, 1.74 0.01 g/g dried bead;
unknown non-phenolic compounds at 4.7 and 10.2 min.
Unknown non-phenolic compounds at 4.2, 4.7, 5.5, 5.8, 6.9, 8.4, 8.9,
10.2, 12.5 and 13.9 min.
Unknown non-phenolic compounds at 4.2, 4.7, 8.4, 10.2 min.
Unknown non-phenolic compounds at 4.2, 4.7, 5.8, 6.8, 8.4, 10.2, 12.5
and 13.9 min.
Note: RT = room temperature.
8.4 min compared with the 1%A and AP beads. After 60 days at 20 C,
no quercetin was detected, but a avonoid at 3.3 min and a nonavonoid phenolic compound at 3.2 min were found in the 3 types of
beads. Furthermore, only one non-phenolic substance (at 10.2 min)
was detected in the 1%A and AP beads, and two non-phenolic substances (at 4.7 and 10.2 min) were found in the 0.67%A beads. After
60 days at 38 C (Fig. 3B), neither quercetin nor non-avonoid phenolics were found, but two avonoids (at 3.3 and 4.1 min) were seen in
the 3 types of beads. Ten and eight non-phenolic substances were detected in the 0.67%A and AP beads, respectively, but only four nonphenolic species in the 1%A beads. This result agrees with the storage
trial nding about the better stability of the 1%A beads. The nonphenolic compounds detected at 280 nm might originate from alginate
and/or pectin.
The HPLC results suggest the interplay between storage conditions and shell wall formulations (which determined the pathways
and rates of quercetin decomposition). The hydroxyl group at C-3
on the C-ring, adjacent to the 2,3-double bond and the 4-carbonyl,
is easily oxidised due to the electron donation of the ketone structure
at C-4 (Lin, Wang, Qin, & Bergensthl, 2007; Rice-Evans, Miller, &
Paganga, 1996). Previous studies published in the literature have
suggested the possible phenolic products derived from quercetin
decomposition such as 3,4-dihydroxybenzoic (protocatechuic)
acid, 1,3,5-trihydroxybenzene (phloroglucinol), 1,3,8-Trihydroxy-9aH,
11H-benzofuro[3,2-b]-[1]benzopyran-7,11-dione cyclic ether, 2,5,7,
3,4-pentahydroxy-3,4-avandione, 2-(3,4-Dihydroxybenzoyl)-2,4,6-
trihydroxy-3(2H)-benzofuranone, 2-(3,4-dihydroxyphenyl)-2,3-dihydroxyprop-2-en-1-al (Buchner, Krumbein, Rohn, & Kroh, 2006; Lei

et al., 2008; Zenkevich et al., 2007).
3.6. FT-IR analysis of encapsulated canola oil beads
FT-IR was used to examine the composition of macromolecules in
the complex bead systems. Fig. 4A shows the FT-IR-spectra of the ingredients alginate, HM pectin, quercetin, canola oil and water that were
used to prepare the 3 types of quercetin-containing canola oil beads.
All spectra were consistent with literature reports for these ingredients
(Che Man & Rohman, 2012; Ismail, Ramli, Hani, & Meon, 2012;
Krishnakumar, Prabu, & Sulkkarali, 2012; Manrique & Lajolo, 2002).
FT-IR spectra (Fig. 4B, C and D) of the freeze dried quercetincontaining oil beads encapsulated by 0.67%A, 1%A and AP, respectively,
were similar and dominated by signals from the core substance (canola
oil) and shell material (alginate and HM pectin). The canola oil signals
were as follows; 3009 cm1 (CH stretching vibration of the cisdouble bond CH), 2925 cm1 and 2854 cm1 (symmetric and asymmetric stretching vibration of the aliphatic CH2 group), 1746 cm1
(ester carbonyl functional group of the triglycerides), 1465 cm1 (bending vibrations of the CH2 and CH3 aliphatic groups), 1377 cm1 (bending
vibrations of CH2 groups), 1238 cm1 and 1163 cm1 (stretching vibration of the CO ester groups) and 723 cm1 (overlapping of the CH2
rocking vibration and the out-of-plane vibration of cis-disubstituted olens) (Che Man & Rohman, 2012; Muik, Lendl, Molina-Daz, & Ayora-
Caada, 2005; Vlachos et al., 2006). The dominant oil signals reect the
high mass fraction of canola oil in the encapsulated beads especially in
the freeze dried form. As seen in Fig. 4A, the assignment of the absorption
bands for HM pectin in the beads was difcult, as most of these bands
overlapped with those of alginate and canola oil that were at higher
concentration than HM pectin. Unambiguous assignment of FT-IR absorbance signals for quercetin was not possible because of its low concentration in these freeze dried oil beads. Reactions between quercetin or
its degradation products, and the core oil encapsulated by alginate
alone or alginate-pectin matrix during storages are evident for data in
Tables 35 (Sun-Waterhouse et al., 2013). Storage at 20 C or 38 C for
847
30 and 60 days did not cause major changes in the FT-IR spectra of all
the 0.67%A, 1%A and AP beads, reecting the chemical stability of the
encapsulated beads. Where the intensity of oil-related signals decreased,
there seemed to be an increase of water-related signals i.e. the bands at
3300 and 1635 cm1 due to asymmetric OH stretching and HOH
bending modes, respectively (Thygesen, Lkke, Micklander, & Engelsen,
2003). The 3009 cm1 signal is associated with the unsaturation
degree of canola oil (Muik et al., 2005), slightly decreased in some
cases (e.g. the 0.67%A and AP bead after 60 days at 38 C). The intensity
of the 723 cm1 band (overlapping of the CH2 rocking vibration and the
out-of-plane vibration of cis-disubstituted olens) generally remained
A
A+O+Q, Day 0,280 nm
A+O+Q, Day 0,370 nm
A+O+Q, Day 0,280 nm
A+O+Q, Day 0,370 nm
A+P+O+Q, Day 0, 280 nm
A+P+O+Q, Day 0, 370 nm
Fig. 3. High performance liquid chromatograms (at 280 and 370 nm) for 0.67% alginate (A + O + Q), 1% alginate (A + O + Q) and alginate + high methoxyl (HM) pectin
(A + P + O + Q) encapsulated beads on A) Day 0, B) Day 60 at 38 C.
848
B
A'+O+Q, Day 60, 38C, 280 nm
A'+O+Q, Day 60, 38C, 370 nm
A+O+Q, Day 60, 38C, 280 nm
A+O+Q, Day 60, 38C, 370 nm
A+P+O+Q, Day 60, 38C, 280 nm
A+P+O+Q, Day 60, 38C, 370 nm
Fig. 3 (continued).
strong for the 1%A and AP beads (Fig. 3C and D), but decrease for the
0.67%A beads (Fig. 4B) after storage. These results agree with the ndings
in storage trials that the 1%A and AP beads offered better oil protection.
In general, polysaccharide polymer gels create a cross-linked hydrophilic network matrix with a porous structure, formed via chemical
interactions (e.g. covalent bonds) or physical interactions (e.g. noncovalent hydrogen bonding, hydrophobic, and ionic interactions)
(Whistler & BeMiller, 1997). In the network structures, interactions between molecules of the same type are favoured over those between
molecules of different types forming permanent junction zones or
temporary entanglements in the network (Choi & Yoo, 2008). HM

pectins form rapid-set gels mainly by hydrophobic interactions and hydrogen bonds (Oakenfull & Scott, 1984), which may facilitate a rapid
separation (also a reduced contact) between canola oil and oxygen in
air, causing decreased oil deterioration. With increasing temperature,
the increase in entropy reduces the hydration of pectin chains, and hydrophobic interactions are strengthened and become the most important contribution to the macromolecular interactions (Oakenfull &
Fenwick, 1977). Thus, the addition of HM pectin to alginate shell
might enhance the stability of the shell and core oil during prolonged
A, ingredients
B, 0.67% alginate
A'+O+Q day 60 38C
Absorbance (arbitrary units)
Sodium alginate
849
HM pectin
Canola oil
A'+O+Q day 60 RT
A'+O+Q day 30 38C
A'+O+Q day 30 RT
Quercetin dihydrate
A'+O+Q day 0
Water
4000
3500
3000
2500
2000
1500
1000
4000
3500
3000
2500
2000
1500
1000
1500
1000
Wavenumber (cm-1)
C, 1% alginate
D, alginate-pectin
A+O+Q day 60 38C
A+P+O+Q day 60 38C
Wavenumber (cm-1)
A+O+Q day 60 RT
A+O+Q day 30 38C
A+P+O+Q day 60 RT
A+P+O+Q day 30 38C
A+P+O+Q day 30 RT
A+O+Q day 30 RT
A+P+O+Q day 0
A+O+Q day 0
4000
3500
3000
2500
2000
1500
1000
Wavenumber (cm-1)
4000
3500
3000
2500
2000
Wavenumber (cm-1)
Fig. 4. Normalised FT-IR absorbance spectra for A) ingredients, and freeze dried encapsulated quercetincanola oil beads before and after a storage at room temperature (RT) or 38 C:
B) 0.67% alginate alone (A + O + Q), C) 1% alginate alone (A + O + Q), D) alginate + high methoxyl (HM) pectin (A + P + O + Q). The spectra have been normalised and offset
vertically for clarity.
storage especially at higher temperature (e.g. 38 C) (Endress &

Rentschler, 1999). However, a remarkable difference in the behaviour
of HM pectin as a function of storage temperature is not expected in
this study due to the small temperature difference between 20 C and
38 C. In comparison, the alginate alone gels where hydrogen bonding
or electrostatic interactions are the only signicant interactions for

stabilising the polymer network (Andresen & Smidsrod, 1977), a monotonous decrease of the elastic modulus with increasing temperature
is expected. Hence, the alginate alone oil beads may be less robust
than AP beads as a function of temperature.
850
4. Conclusions
Co-extrusion encapsulation using alginate and alginate-HM pectin
shell materials is a feasible approach for encapsulating unsaturated canola oil. Encapsulation conditions inuenced the characteristics of the
oil beads, with the core and shell ow rates being the critical to realising
stable spherical oil beads. Bead size, core oil stability and retained phenolic content vary with the shell wall formulations. Core oil fortied
with quercetin is more effective than fortication with vitamin E or
BHT for suppressing oil oxidation at 38 C. FT-IR studies conrmed
the chemical stability of the three types of quercetin-containing encapsulated beads during storage. HPLC analyses revealed that the degradation pathway of quercetin depended on the shell formulation and
storage conditions.
Both 1% alginate and alginateHM pectin shell formulations are acceptable and comparable for preserving quercetin-fortifying canola oil.
Considering the nutritional value introduced by the pectin addition,
the alginatepectin shell represents a good option for encapsulating unsaturated oil for food ingredient application. The quercetin-containing
canola oil beads encapsulated with alginate-based polysaccharide
shell, which can be consumed directly or used as bioactive ingredients,
simultaneously deliver the goodness of unsaturated oil, quercetin antioxidant and bre polysaccharides to consumers.
Acknowledgments
We acknowledge research funding from the University of Auckland
and Plant & Food Research.
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Food Research International 54 (2013) 837851

Contents lists available at ScienceDirect

Food Research International

Co-extrusion encapsulation of canola oil with alginate: Effect of quercetin

Corresponding author. Tel.: +64 9 3737599x89024; fax: +64 9 3737422.

including quercetin can impart a bitter taste to foods (Jaeger, Axten,

W. Wang et al. / Food Research International 54 (2013) 837851

2.2. Preparation of encapsulated canola oil beads

W. Wang et al. / Food Research International 54 (2013) 837851

Bead parameters and oil loading efciency

Oil loading efciency

After freeze drying

2.2.4. Measurement of oil loading efciency and bead breakage

(away from light) for 30 or 60 days at room temperature (in cooled

W. Wang et al. / Food Research International 54 (2013) 837851

2.8. Oil stability evaluation

2.9. High performance liquid chromatography (HPLC) phenolic proling

An analytical HPLC (HP Agilent 1100 series, Agilent Technologies,

2.8.2. Peroxide value (PV) determination

2.10. Statistical analysis

W. Wang et al. / Food Research International 54 (2013) 837851

the increased shell ow rate led to the presence of small individual

W. Wang et al. / Food Research International 54 (2013) 837851

1%A (magnication 400) and AP (magnication 300) oil beads had

are both polyuronates, and the Ca2+ binding mechanism occurred in

W. Wang et al. / Food Research International 54 (2013) 837851

Total phenolic content

Water solution for pH treatment

0.67%A N 1%A N AP beads. The release characteristics of bead shells are

(Rayment et al., 2009). The different responses to pH of alginate and

W. Wang et al. / Food Research International 54 (2013) 837851

FFA (g/100 g oil)

TEPC (mg GalE/kg oil)

atoms adjacent to a double bond and leads to formation of free radicals

W. Wang et al. / Food Research International 54 (2013) 837851

FFA (g/100 g oil)

TEPC (mg GalE/kg oil)

The difference in the chemical structure, solubility, lipophillicity/

3.5. HPLC analysis of phenolics in the encapsulated quercetin-containing

W. Wang et al. / Food Research International 54 (2013) 837851

7. 2 min, quercetin, 2.17 0.01 g/g dried bead,

No quercetin and other avonoid detected.

No quercetin and other avonoid detected.

Identied compounds at different wavelengths

No detected phenolic and non-phenolic compounds.

4.5 min, a very small phenolic peak;

No detected phenolic and non-phenolic compounds.

Note: RT = room temperature.

trihydroxy-3(2H)-benzofuranone, 2-(3,4-dihydroxyphenyl)-2,3-dihydroxyprop-2-en-1-al (Buchner, Krumbein, Rohn, & Kroh, 2006; Lei

W. Wang et al. / Food Research International 54 (2013) 837851

A+O+Q, Day 0,370 nm

A+O+Q, Day 0,280 nm

A+O+Q, Day 0,370 nm

A+P+O+Q, Day 0, 280 nm

A+P+O+Q, Day 0, 370 nm

W. Wang et al. / Food Research International 54 (2013) 837851

A'+O+Q, Day 60, 38C, 370 nm

A+O+Q, Day 60, 38C, 280 nm

A+O+Q, Day 60, 38C, 370 nm

A+P+O+Q, Day 60, 38C, 280 nm

A+P+O+Q, Day 60, 38C, 370 nm

temporary entanglements in the network (Choi & Yoo, 2008). HM

W. Wang et al. / Food Research International 54 (2013) 837851