Food Hydrocolloids 43 (2015) 622e628

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Iron microencapsulation with blend of gum arabic, maltodextrin and

modified starch using modified solvent evaporation method e Milk
fortification
Chitra Gupta a, Prince Chawla a, Sumit Arora a, *, S.K. Tomar b, A.K. Singh c
a
Dairy Chemistry Division, National Dairy Research Institute, Karnal, Haryana, 132001, India
b
Dairy Microbiology Division, National Dairy Research Institute, Karnal, Haryana, 132001, India
c
Dairy Technology Division, National Dairy Research Institute, Karnal, Haryana, 132001, India

a r t i c l e i n f o a b s t r a c t

Article history: Iron microcapsules were prepared with blend of gum arabic, maltodextrin and modified starch using
Received 22 May 2014 modified solvent evaporation method. Process parameters were optimized for obtaining maximum
Accepted 19 July 2014 encapsulation efficiency and stability of microcapsules. Effect of different concentration of alcohol,
Available online 14 August 2014
different ratio of mixture to absolute alcohol, different composition of wall material and different amount
of iron salt on the encapsulation efficiency (EE) of iron microcapsules were evaluated. Microcapsules
Keywords:
prepared with gum arabic, maltodextrin and modified starch in the ratio of 4:1:1 and mixture to absolute
Microencapsulation
alcohol ratio 1:10 showed maximum encapsulation efficiency (91.58%) and stability. External
Gum arabic
Maltodextrin
morphology of iron microcapsules revealed slightly circular structure with minimum cracks and dents on
Modified starch (HiCap 100) the surface. Particle size as analysed by inverted light microscope was in the range of 6.84e33.42 mm.
Iron fortification Iron microcapsules were added to milk and evaluated for sensory characteristics and oxidative stability.
Milk Sensory scores of iron salt fortified milk were significantly lower (P < 0.05) as compared to iron mi-
crocapsules fortified milk during storage, however, this difference could be observed only upto the 5th
day of storage. Iron microcapsules fortified milk showed significantly higher (P < 0.05) in-vitro
bioavailability of iron as compared to control (unfortified) and iron salt fortified milk.
© 2014 Elsevier Ltd. All rights reserved.

1. Introduction fortification, not only due to world-wide consumption by all

Iron is an essential trace element, naturally present in the
structure of cytochrome, enzymes, haemoglobin and myoglobin
(Abbasi & Azari, 2011) and stored in the form of ferritin and
haemosiderin in liver (Allen, Benoist, Dary, & Hurrell, 2006). Its
deficiency causes lower growth rate, impaired cognitive scores in
children, poor pregnancy outcomes and lower working capacity in
adults (Derbyshire, Brennan, Li, & Bokhari, 2010). In India, inade-
quate dietary intake of iron and low bioavailability of iron from
foods are identified as major cause of anaemia. Among multiple
strategies to control iron deficiency, food fortification is an effec-
tive measure to increase the intake of iron without causing a
change in the existing dietary patterns (Tripathi & Platel, 2011).
Milk and milk products are always good candidates for iron
* Corresponding author. Tel.: þ91 (0) 184 2259156 (O), þ91 9896054444 (M).
E-mail addresses: chitragupta97@gmail.com (C. Gupta), princefoodtech@gmail.
com (P. Chawla), sumitak123@gmail.com, sumitak123@yahoo.com (S. Arora),
sudhirndri@gmail.com (S.K. Tomar), aksndri@gmail.com (A.K. Singh).
groups at risk of deficiency, but also because of their high nutri-
tional value, absorption processes and positive effect on growth,
cognition and morbidity (Boccio & Montiero, 2004). Iron fortifi-
cation of milk seems to be an effective nutritional strategy to
correct dietary iron deficiency (Bhawana et al., 2011). Adding iron
directly to milk results in reaction with milk components (lipids
and protein) and decreased bioavailability, along with develop-
ment of organoleptic problems such as colour, odour and taste
(Gaucheron, 2000). Microencapsulation technique has multitude
applications and has been widely used to protect iron from
oxidation by forming an impermeable membrane as barrier to
oxygen diffusion, to mask the unacceptable flavour caused by iron
salt and to increase bioavailability (De souza et al., 2013). Selection
of an appropriate wall material and physicochemical properties of
wall material are critical issues in governing the functionality of
microcapsule systems (Sarkar, Gupta, Variyar, Sharma, & Singhal,
2013; Wang, Wang, Li, Adhikari, & Shi, 2011).
Gum arabic, commercial exudate gum obtained from Acacia
Senegal, is one of the most common wall material used in micro-
encapsulation due to its low viscosity, good emulsifying, high

http://dx.doi.org/10.1016/j.foodhyd.2014.07.021
0268-005X/© 2014 Elsevier Ltd. All rights reserved.
 C. Gupta et al. / Food Hydrocolloids 43 (2015) 622e628
stabilizing and film forming properties (Ali, Zaida, & Blunden,
2009; Sarkar et al., 2013). It has the ability of better retention of
volatile substances and effective protection against oxidation
(Gabas, Telis, Sobral, & Telis, 2007; Righetto & Netto, 2005). Gum
arabic has a highly branched structure which results in compact
spheroidal conformation (Tombs & Harding, 1998). Other wall
materials used are modified starch (HiCap 100) and maltodextrin.
Maltodextrins are acid or enzyme hydrolysed starch used for
encapsulation of oils and flavouring compounds (Carneiro, Tonon,
Grosso, & Hubinger, 2013). Maltodextrins have numerous proper-
ties that allow them to be used for diverse purposes in both the
food and pharmaceutical industries (Toure, Zhang, Jia, & Dong,
2007). Their ability to retain water and form gels explains their
choice as efficient food stabilizers (Chronakis, 1998). It has also
reported that maltodextrins have been used as stabilizers in the
microencapsulation of vitamins, minerals, colourants as well as fat
and oils (Toure et al., 2007). However, the poor film-forming ability,
hygroscopicity and turbidity of maltodextrins account for their
inability to protect volatile compounds during spray drying (Raja,
Sankarikutty, Sreekumar, Jayalekshmy, & Narayanan, 1989).
Modified starch (HiCap 100) has good film forming properties.
It has been used for microencapsulation of ascorbic acid and
showed high retention of ascorbic acid during storage (Wijaya,
Small, & Bui, 2011). Gums, starches, polysaccharides and proteins
show good compatibility with gum arabic (Krishnan, Bhosale, &
Singhal, 2005). Therefore, maltodextrin and modified starch
(HiCap100) are often used as microencapsulating material with
gum arabic to improve the encapsulation efficiency and binding
property (Carneiro et al., 2013; Kanakdande, Bhosale, & Singhal,
2007; Krishnan et al., 2005; Shaikh, Bhosale, & Singhal, 2006;
Vaidya, Bhosale, & Singhal, 2006). Zilberboim, Kopelman, and
Talmon (1986) reported cold dehydration as an alternative to
spray drying for encapsulation of highly volatile materials and
microencapsulate paprika oleoresin and aromatic esters using
alcohol as dehydrating liquid.
Iron was microencapsulated using modified solvent evaporation
method which involved alcohol as dehydrating medium. As soon as
coat material comes in contact with the alcohol, gets dehydrated
and resulted in the formation of microcapsules. Microcapsules
were separated from the alcohol or dehydrating medium and re-
sidual alcohol was evaporated at low temperature (4e7  C). Higher
temperature resulted in the disruption of coat material and leaking
of the encapsulated material (Richmond & Moss, 1983).
Limited attempts have been made so far to access the stability
and encapsulation efficiency of iron microcapsules and in-vitro
bioavailability of iron from these microcapsules prepared by
modified solvent evaporation method using plant based exudate
gums such as gum arabic, maltodextrin and modified starch as a
coating. Therefore, present work was designed to evaluate the
ability of gum arabic in combination with maltodextrin and
modified starch (HiCap 100) as alternative materials for iron
(ferrous sulphate) microencapsulation by modified solvent evapo-
ration method and to minimize the iron induced oxidation and safe
delivery of iron to consumers via milk fortification. Iron content,
ratio of wall materials, concentration of dehydrating medium and
ratio of mixture to dehydrating medium were also standardized for
obtaining stable microcapsules.
2. Materials and methods
2.1. Materials
Gum arabic (spray dried) (GA), maltodextrin (DE 16.5e19.5)
(MD), iron salt (Ferrous sulphate hepta hydrate), cyclohexanone,
thiobarbituric acid (TBA), ammonium sulphate, citric acid, sodium
623
hydroxide, phenolphthalein, ethanol and L-ascorbic acid were
procured from Sigma Aldrich, St. Louis, USA. Chemicals used were
of AR grade. Modified Starch (HiCap 100) (MS) was obtained from
National Starch Chemicals Corporation, Mumbai, India. Fresh cow
and buffalo milk were collected from cattle yard of National Dairy
Research Institute, Karnal, India. a-amylase (EC 3.2.1.1), human
pancreatic lipase (EC 3.1.1.3), colipase, cholesterol esterase (EC
3.1.1.13), phospholipase A2 (EC 3.1.1.4), mucin, bovine serum albu-
min, pepsin (2080 units mg1 of protein), pancreatin and taur-
ocholate salts were purchased from Sigma Chemical Co (Madrid,
Spain). Cellulose dialysis membranes (flat width, 25 mm; internal
diameter, 16 mm; molecular weight cut-off, 12,000e14,000 Da)
were procured from Himedia Laboratories Pvt., Ltd., Mumbai, India.
Milli Q water (10 mS) and acid washed glassware were used
throughout the experiments.
2.2. Methods
2.2.1. Preparation of microcapsules
Microcapsules were prepared by dissolving a blend of GA, MD
and MS in the ratio of 4:1:1 (6 g) in 10 mL deionized water at 60  C
and kept it for rehydration under refrigerated condition (4e7  C)
for 12 h. Iron salt (ferrous sulphate) and ascorbic acid (used as
antioxidant) in the ratio of 15:1 were dissolved in 10 mL deionized
water and added to rehydrated blend solution. Ferrous sulphate
hepta hydrate has been used as core material for preparation of iron
microcapsules as it is the cheapest iron source with high iron
bioavailability. Solution of core and coat materials were mixed well
and kept in water bath maintained at 5  C, followed by sonication
using a probe sonicator (Model VCx750, Sonics and Materials Inc.,
New Town, USA) at 5  C with 5.0 s pulse rate for 15 min. During
sonication, proper mixing of core and coat materials occur. Iron
may interact with the carboxylic group of gum arabic and forms the
ionic/electrostatic bond. This mixture was then sprayed in chilled
alcohol. It was kept on a magnetic stirrer and stirred at 500 rpm.
Airless paint sprayer (Wagema Professional Quality, Pretoria, South
Africa) was used for spraying of the mixture in chilled alcohol
which was operated at 1.5 kg/cm2. After spraying, it was left un-
disturbed for 5 min. Finally, filtration was carried out with filter
paper (Whatman No. 1) using vacuum filtration assembly. Micro-
capsules retained on the filter paper had residual amount of alcohol
which was easy to evaporate at low temperature. The retentate was
spread in a petridish and stored at 4e7  C for 12e14 h for complete
removal of alcohol. Powdered microcapsules were stored in air
tight glass containers at room temperature. This method has an
advantage over other methods, since the solvent can be reused by
distillation of filtered alcohol.
2.2.1.1. Optimization of process parameters. Effect of different con-
centration of alcohol, different ratio of mixture to absolute alcohol,
different composition of wall material and different amount of iron
salt on the encapsulation efficiency (EE) of iron microcapsules were
evaluated.
2.2.1.1.1. Concentration of alcohol. Different concentrations of
chilled alcohol (80%, 90% and absolute alcohol) were used for the
spraying of mixture and its effect on EE of iron microcapsules was
evaluated.
2.2.1.1.2. Ratio of mixture to absolute alcohol. Mixture prepared
after sonication was sprayed in different amount of alcohol.
Amount of chilled alcohol was varied according to the ratio of
mixture to absolute alcohol. Different ratios of mixture to absolute
alcohol (1:5, 1:7.5 and 1:10) were used for preparation of iron mi-
crocapsules and its effect on the EE of iron microcapsules was
evaluated.
624 C. Gupta et al. / Food Hydrocolloids 43 (2015) 622e628
2.2.1.1.3. Composition of wall material. Various compositions of
wall materials were used for the preparation of microcapsules. GA,
MD and MS in the ratio of 4:1:1, 1:4:1, 1:1:4, 2:2:2, 3:2:1, 3:1:2,
6:0:0 were used and evaluated for its effect on EE.
2.2.1.1.4. Amounts of iron salt. Different amounts of iron salt
(300, 500, 800 and 1000 mg) were used for the preparation of iron
microcapsules and its effect on EE was evaluated.
2.2.2. Estimation of iron content
Iron content of microcapsules was estimated by atomic ab-
sorption spectrophotometer (AAS) (AA-7000, Shimadzu, Tokyo,
Japan) using the method of AOAC (2005). Microcapsules were
subjected to ashing (550  C for 8 h), solubilized in tri acid mixture
(HNO3:HClO4:H2SO4 in 3:2:1 ratio) and heated for complete
dissolution. Samples were diluted suitably for analysis by AAS at
lmax 248.3 nm.
2.2.3. Encapsulation efficiency (EE)
During preparation, iron microcapsules were separated from
alcohol by filtration. It was assumed that iron in the retentate was
in encapsulated/bound form. Non encapsulated iron was present in
the filtrate. Retentate was dried under refrigerated conditions
(4e7  C) for 12e14 h. Encapsulated iron in retentate was estimated
using AAS. Iron content which was added initially for the prepa-
ration of iron microcapsules was considered as total iron content
and bound iron was estimated in the retentate. Filtrate contained
majorly alcohol which was volatile and difficult to measure volu-
metrically and gravimetrically, therefore, bound iron was measured
in the retentate. EE was calculated as follows:
Bound iron
Encapsulation efficiency ¼  100
Total iron
2.2.4. Scanning electron microscopy
The external structure of powdered microcapsules was exam-
ined by scanning electron microscopy (Carl Zeiss EV018, 18th edi-
tion, Cambridge, UK). Microcapsules were attached on scanning
electron microscopy stub by double coated adhesive tape. Mounted
samples were coated with gold (20 nm thickness) on ion coater
(Eiko IB3, Tokyo, Japan) at 0.05e0.07 torr for 4 min maintaining the
ion current at 6 mA. Samples were finally examined by scanning
electron microscopy at an acceleration voltage of 15 KV under high
vacuum (9.0*105 torr) and micrographs were recorded.
2.2.5. Size analysis of iron microcapsules
Microcapsules were kept on the glass slides and dispersed in
one or two drops of distilled water. Distilled water is used for
proper spreading of the microcapsules on the glass slide. These
were observed (magnification 400-fold) under an inverted light
microscope (Nikon Eclipse Ti, Tokyo, Japan) and photographed
using a fitted digital camera (Nikon Digital Sight, Tokyo, Japan). Size
of the microcapsules was measured with the help of inbuilt soft-
ware (Nikon Basic Research Imaging Software (v 3.1)) with
microscope.
2.2.6. Fortification in milk
Fresh cow and buffalo milk were mixed in the ratio of 1:1 and
standardized to the fat and solid not fat (SNF) percent of toned milk.
Toned milk (3% fat and 8.5% SNF) was fortified with iron salt
(ferrous sulphate at 25 ppm iron) and iron microcapsules (@25 ppm
iron). Addition of iron salt and iron microcapsules was accompa-
nied by mixing for 10 min with the help of magnetic stirrer for
complete dissolution of fortificants. After mixing, the milk samples
were pasteurized at 63  C for 30 min followed by cooling to 4  C.
2.2.7. Sensory evaluation of fortified milk
Prepared milk samples were evaluated by the sensory panelists.
Sensory panel of ten trained judges from the institute were asked to
grade fortified milk samples for changes in colour and appearance,
odour, taste and mouthfeel. Composite score card for sensory
analysis of milk as approved by BIS (IS: 7768, 1975) was used. In
taste characteristics, the main focus was on metallic, rancid and
oxidized taste. Other mentioned taste characteristics e.g. cowy,
acidic, astringent, bitter, cooked, flat, foreign, malty, salty, barny etc.
were excluded. The sensory booth environment was held at a
constant temperature (20  C), red lighting was used to obscure any
colour differences between the samples and a positive airflow
removed any odour from the testing area. Saline water (0.89% so-
dium chloride solution) (at room temperature) was provided as
palate cleanser for rinsing mouth and cleaning the tongue before
sensory evaluation of each sample.
2.2.8. Thiobarbituric acid (TBA) value of fortified milk
TBA value of milk samples were evaluated by the method
described by Hegenauer, Saltman, Ludwig, Ripley, and Bajo (1979).
In this method, TBA reagent was prepared immediately before use
by mixing equal volumes of freshly prepared 0.025 M TBA
(neutralized with NaOH) and 2 M H3PO4/2 M citric acid. Reaction
was terminated by mixing 5.0 mL of milk sample with 2.5 mL TBA
reagent into a glass centrifuge tube. The mixture was heated
immediately in boiling water bath for exactly 10 min and then
cooled on ice. 10 mL cyclohexanone and 1 mL of 4 M ammonium
sulphate were then added and centrifuged (Kubota Corporation,
Gyeonggi-do, South Korea) at 5000 g for 5 min at room temper-
ature. The orange-red cyclohexanone supernatant was decanted
and its absorbance was measured at 532 nm using spectropho-
tometer (SPECORD 200 Analytik Jena, Jena, Germany) in a 1 cm
light path.
2.2.9. In-vitro bioavailability of iron
In-vitro bioavailability of iron was determined by simulated
gastro-intestinal model as described by Herrero-Barbudo,
Granado-Lorencio, Blanco-Navarro, and Perez-Sacristan (2009)
with slight modification. Compositions and concentrations of
inorganic and organic solutions, saliva, gastric juice, duodenal
juices and bile constituents were carefully duplicated as described
by (Granado-Lorencio et al., 2007). Control (unfortified), iron salt
and iron microcapsules fortified milk were assessed for bioavail-
ability of iron.
2.2.9.1. Preparation of membrane. Cellulose dialysis membrane
were poured in deionized water and boiled for 5 min in deionized
water before use.
2.2.9.2. In-vitro digestion model. Approximately, 15 ml milk sam-
ples (control, iron salt and iron microcapsules fortified) were
transferred to a flask and saliva solution (9 ml, pH 6.5) con-
taining organic and inorganic components and a-amylase
(700 mg/L of saliva solution) was added after which the samples
were incubated in a shaking water bath at 37  C, 95 rpm for
5 min. Gastric juice (13.5 ml) with organic and inorganic solu-
tions, mucin (6 g/L of gastric juice), bovine serum albumin (2 g/L
of gastric juice), and pepsin (2 g/L of gastric juice) from porcine
stomach was added. pH was adjusted to 1.1 with HCl and the
solution was incubated for 1 h at 37  C. Duodenal juice (25 ml,
organic plus inorganic solutions containing porcine pancreatin
6 g/L of duodenal juice) and bile solutions (9 ml, containing bile
 C. Gupta et al. / Food Hydrocolloids 43 (2015) 622e628
salt 12 g/L of bile solution), prepared on the day of assay, were
introduced after neutralization of the pH (7.8) and the
human pancreatic lipase (5 units), colipase (25 mg), cholesterol
esterase (10 units), phospholipase A2 (50 ml) and taurocholate
salts (39.8 mg) were added. Final volume was approximately
70 ml, the mixture was then incubated up to 3 h at 37  C.
Dialysis sac was formed by closing the one end of dialysis
tube with membrane closure, the digested contents thus ob-
tained were poured in this dialysis sac and sac was closed from
opposite side also. The contents along with this dialysis sac were
dipped in distilled water kept in a 1000 ml beaker for 16 h at
37  C. Iron content in the retentate was determined by AAS
to estimate the digestibility of the added nutrient under simu-
lated gastro-intestinal conditions. Bioavailability of nutrient
(iron) was calculated from the amount of the nutrient (iron) that
had passed the dialysis membrane proportional to the total
nutrient (iron) content of the sample. Bioavailability was calcu-
lated as:
Bioavailability ð%Þ ¼ D=C  100
where,
D ¼ Iron content in the dialysate and
C ¼ Iron content of sample
2.2.10. Statistical analysis
Means and standard error mean (SEM) were calculated using
Microsoft Excel, 2007 (Microsoft Corp., Redmond, WA). Significant
Fig. 1. Encapsulation efficiency of microcapsules as affected by (A) Different concentration of alcohol, (B) Different ratio of mixture to alcohol ratio, (C) Composition of gum arabic
(GA), maltodextrin (MD) and modified starch (MS) & (D) Iron concentration. aebSamples represented with different letters are significantly different (P < 0.05) from each other. Error
bars show the variations of three determinations in terms of standard error of mean.
625
difference between values was verified by one way or two way
analysis of variance and comparison between means was made by
critical difference value (Snedecor & Cochran, 1994).
3. Results and discussion
3.1. Optimization of preparation process
Process was optimized by varying one parameter and keeping
the others constant. The ratio of GA, MD, MS as 4:1:1, iron salt
(300 mg) and ascorbic acid (20 mg) in the ratio of 15:1 were dis-
solved in 20 mL deionized water and absolute alcohol was used as a
dehydrating medium in the ratio of mixture to absolute alcohol
1:10 for optimization of process parameters.
Different concentrations of alcohol, 80%, 90% and absolute
alcohol were used as dehydrating medium (Fig. 1A). Absolute
alcohol showed maximum EE as compared to 80% and 90% alcohol
and hence was the most suitable dehydrating medium required for
dehydration of microcapsules. Our results were in agreement with
Zilberboim et al. (1986) who observed that the reduction in alcohol
concentration reduced the retention capacity of the microcapsules,
as lower concentration resulted in slow release of water from the
microcapsules and longer dehydration time. Lower concentration
of alcohol also resulted in particle agglomeration into sticky mass
which was difficult to filter.
The ratio of mixture to absolute alcohol was varied as 1:5, 1:7.5
and 1:10 for the preparation of microcapsules with maximum EE
(Fig. 1B). 1:10 resulted in highest EE as compared to 1:5 and 1:7.5.
626 C. Gupta et al. / Food Hydrocolloids 43 (2015) 622e628

Fig. 2. (A) Scanning electron microscopy (500) and (B) light microscopic (400) observation of 300 mg iron salt containing microcapsules.

EE significantly decreased (P < 0.05) with decreasing concentration microscopy and particle size analysis and also used for fortification
of alcohol. 1:10 mixture to alcohol ratio was the most suitable for of milk.
microencapsulation of iron. Our results were in agreement with
Zilberboim et al. (1986) who reported that alcohol to emulsion
3.2. Scanning electron microscopy and particle size analysis
ratio below 10:1 reduced the retention efficiency of the micro-
capsules. However, at higher concentration it reduced the water
Scanning electron microscopy was used to evaluate morpho-
content without affecting the retention efficiency as higher ratio of
logical structures of microcapsules obtained from GA/MD/MS
alcohol to emulsion resulted in rapid drying of microcapsules and
(4:1:1) and mixture to absolute alcohol ratio (1:10) showed slightly
formed a protective crust that prevented the extensive leaching of
circular, uniform and minimum cracks and dents on the surface of
core material. Cho and Park (2003) also evaluated the process
microcapsules (Fig. 2A). Spherical shape results in maximum sta-
parameters for oil in water in oil (O/W/O) multiple emulsion
bility due to minimum surface area to volume ratio. Microcapsules
method for flavour encapsulation and reported that increase in the
were not as spherical as we obtain in the spray drying technique. In
GA content resulted in more stable emulsion and highest flavour
spray drying, immediate dehydration of microcapsules occur
retention (71%) was observed with ethanol to mixture ratio of 9:1
which resulted in proper spherical shape of microcapsules. How-
as a dehydrating agent.
ever, in modified solvent evaporation method slow dehydration of
Absolute alcohol and ratio of mixture to absolute alcohol 1:10
microcapsules occur which resulted in the slight disruption of the
showed highest EE. Therefore, keeping these two conditions
spherical structure. Kanakdande et al. (2007), Krishnan et al.
constant, iron microcapsules were prepared with different ratios
of GA, MD and MS and evaluated for EE (Fig. 1C). Ratio of 4:1:1
showed highest EE as compared to all other microcapsules and a Table 1
ratio of 6:0:0 showed significantly lower (P < 0.05) EE as Effect of iron fortification (@25 ppm of iron) on sensory scores of milk during storage
at 4e7  C.
compared to 4:1:1. Hence, 4:1:1 ratio of GA, MD, and MS was
most suitable for microencapsulation of iron. EE significantly Characteristics Storage Control Iron salt Iron
decreased (P < 0.05) with decreasing content of GA. Our results time (unfortified) fortified microcapsules
(days) fortified
were in accordance with Kanakdande et al. (2007) and Krishnan
et al. (2005) who observed that a blend of GA, MD and MS in the Colour and 0 9.18 ± 0.111dB 8.96 ± 0.040cA 9.10 ± 0.100dB
ratio of 4:1:1 gave better results as compared to 100% GA. Mi- appearance (10) 3 8.86 ± 0.098cB 8.60 ± 0.100bA 8.66 ± 0.093cA
5 8.56 ± 0.040bC 8.22 ± 0.136aA 8.42 ± 0.107bB
crocapsules prepared with GA, MD and MS in the ratio of 1:4:1 7 8.40 ± 0.100aB 8.28 ± 0.116aA 8.30 ± 0.122aA
and 1:1:4 form a sticky mass after spraying in alcohol, therefore, Odour (20) 0 18.24 ± 0.112cC 15.70 ± 0.255cA 17.50 ± 0.158dB
which was difficult to separate from alcohol as they blocked the 3 17.30 ± 0.122bC 15.30 ± 0.122bA 16.60 ± 0.187cB
pores of filter paper (Whatman No. 1). This might be due to the 5 16.30 ± 0.374aC 15.40 ± 0.292bA 15.90 ± 0.292bB
7 16.50 ± 0.224aB 15.10 ± 0.332aA 15.20 ± 0.255aA
high water holding capacity of these two wall materials. The
Taste (40) 0 36.00 ± 0.632cA 35.80 ± 0.663dA 36.20 ± 0.735dA
resulting microcapsules showed lower EE as compared to others, 3 36.00 ± 0.447cC 31.80 ± 0.374cA 34.20 ± 0.374cB
therefore, 0:6:0 and 0:0:6 ratio of GA, MD and MS were not tried 5 34.20 ± 0.374bC 30.20 ± 0.374bA 31.20 ± 0.374bB
for preparation of iron microcapsules. It was also evident from 7 32.80 ± 0.374aC 27.40 ± 0.678aA 27.80 ± 0.663aA
the above results that MS enhanced the EE more as compared to Mouthfeel (30) 0 27.50 ± 0.224cA 27.20 ± 0.123dA 27.20 ± 0.123cA
3 27.20 ± 0.374cB 26.40 ± 0.400cA 26.90 ± 0.100cB
the MD. 5 26.60 ± 0.245bB 25.20 ± 0.374bA 25.00 ± 0.548bA
Finally, iron content was optimized by adding 300 (60 mg iron), 7 25.90 ± 0.245aB 24.10 ± 0.400aA 24.40 ± 0.224aA
500 (100 mg iron), 800 (160 mg iron) and 1000 (200 mg iron) mg Total (100) 0 90.92 ± 0.637dC 87.66 ± 0.430dA 90.00 ± 0.837dB
iron salt to the selected blend of GA/MD/MS (4:1:1) and mixture to 3 89.36 ± 0.806cC 83.10 ± 0.400cA 86.36 ± 0.427cB
5 85.66 ± 0.398bC 79.02 ± 0.510bA 80.52 ± 0.648bB
absolute alcohol ratio (1:10) for stable microencapsulation (Fig. 1D).
7 83.60 ± 0.660aC 74.88 ± 0.441aA 75.80 ± 0.490aB
Iron microcapsules containing 300 mg showed highest EE as
compared to all other microcapsules. EE significantly decreased Data are presented as means ± SEM (n ¼ 10).
aeb
Means within columns with different lowercase superscript are significantly
(P < 0.05) with increasing iron concentration. different (P < 0.05) from each other.
Iron microcapsules prepared by optimized process containing AeB
Means within rows with different uppercase superscript are significantly
300 mg iron was further subjected for scanning electron different (P < 0.05) from each other.
 C. Gupta et al. / Food Hydrocolloids 43 (2015) 622e628
Table 2
Effect of iron fortification on TBA value of milk during storage at 4e7  C.
Milk samples 0th day
Control (unfortified) 0.016 ± 0.0006aA
Iron salt fortified 0.017 ± 0.0007aA
Iron microcapsules fortified 0.017 ± 0.0003aA
Data are presented as means ± SEM (n ¼ 3).
aeb
Means within columns with different lowercase superscript are significantly different (P < 0.05) from each other.
AeB
Means within rows with different uppercase superscript are significantly different (P < 0.05) from each other.
(2005) and Vaidya et al. (2006) observed the microcapsules pre-
pared by spray drying method using scanning electron microscopy
and found that GA, MD and MS in the ratio of 4:1:1 gave spherical
microcapsules with smooth surface. Microcapsules from GA alone
were found to be nearly spherical but had many dents on the
surface, whereas the microcapsules obtained from MD and MS
were partially disrupted. Average particle size of microcapsules
was observed as 15.54 mm (range 6.84e33.42 mm) by light
microscope (Fig. 2B).
3.3. Milk fortification, sensory evaluation and oxidative changes
Three type of milk samples i.e. control (unfortified), iron salt and
iron microcapsules fortified (@25 ppm of iron) milk were prepared.
Colour and appearance, odour and taste scores of iron salt and iron
microcapsules fortified milk samples were significantly different
(P < 0.05) from each other upto the 5th day of storage whereas
mouthfeel scores were significantly different (P < 0.05) upto the 3rd
day of storage (Table 1). Total sensory scores of iron salt and iron
microcapsules fortified milk samples were slightly but significantly
different (P < 0.05) from each other upto the 7th day of storage. TBA
value of iron microcapsules fortified milk was significantly lower
(P < 0.05) than iron salt fortified milk upto the 5th day of storage
(Table 2). Iron salt fortified milk showed highest TBA value due to
the presence of free iron, however, iron microcapsules fortified milk
showed lower value due to presence of bound iron. These micro-
capsules slowly release the iron in the milk, therefore, TBA value of
milk increases.
3.4. In-vitro bioavailability of iron
In-vitro bioavailability of iron was determined for three types of
milk i.e. control (unfortified), iron salt and iron microcapsules for-
tified (@25 ppm of iron) milk under simulated gastro-intestinal
conditions. All the three samples showed significant difference
(P < 0.05) in iron bioavailability (Table 3). Iron microcapsules for-
tified milk showed significantly higher (P < 0.05) in-vitro
bioavailability of iron as compared to control (unfortified) and iron
salt fortified milk. Control (unfortified) milk showed lowest iron
bioavailability (%), might be due to the strong binding of iron with
casein which makes it insoluble in the gastro-intestinal conditions
(Jackson & Lee, 1992). Iron salt fortified milk also showed signifi-
cantly lower (P < 0.05) iron bioavailability as compare to iron
Table 3
In-vitro bioavailability of iron from control (unfortified), Iron salt and iron
microcapsules fortified milk.
Milk samples % Bioavailability
Control (unfortified) 19.86 ± 0.23a
Iron salt fortified 54.31 ± 0.36b
Iron microcapsules fortified 63.78 ± 0.23c
Data are presented as means ± SEM (n ¼ 3).
aeb
Means within columns with different lowercase superscript are signifi-
cantly different (P < 0.05) from each other.
627
3rd day 5th day 7th day
0.029 ± 0.0009aB 0.049 ± 0.0009aC 0.062 ± 0.0009aD
0.061 ± 0.0012cB 0.083 ± 0.0009cC 0.103 ± 0.0009bD
0.043 ± 0.0012bB 0.076 ± 0.0012bC 0.101 ± 0.0009bD
microcapsules fortified milk. Iron microcapsules are less affected by
the presence of inhibitors, therefore resulting in high iron
bioavailability (Olivares, 2002).
4. Conclusion
Stable iron microcapsules were successfully prepared with a
blend of GA/MD/MS using modified solvent evaporation method.
GA/MD/MS (4:1:1) and mixture to absolute alcohol ratio (1:10)
proved to be a superior microcapsules composition for iron
microencapsulation with EE of 91.58%. External morphological
characteristics revealed circular and uniform structure with
minimum cracks and dents on the surface of iron microcapsules.
The average particle size of the microcapsules was 15.54 mm.
These iron microcapsules were added to milk and showed sig-
nificant difference in total sensory scores from iron salt fortified
milk during storage. TBA value of iron microcapsules fortified milk
was significantly lower (P < 0.05) than milk fortified with iron salt
upto the fifth day of storage. Iron microcapsules fortified milk
showed significantly higher (P < 0.05) in-vitro bioavailability of
iron as compared to control (unfortified) and iron salt fortified
milk. The results of this study revealed the potential of GA, MD
and MS for use as a superior and convenient wall material for iron
microencapsulation.
Acknowledgement
The author would like to acknowledge National Starch Chem-
icals Corporation, Mumbai, India for providing modified starch
(HiCap 100) used during the study.
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