Powder Technology: Berta N. Estevinho, Ioana Carlan, Alexandra Blaga, Fernando Rocha
Powder Technology: Berta N. Estevinho, Ioana Carlan, Alexandra Blaga, Fernando Rocha
Powder Technology
a r t i c l e i n f o a b s t r a c t
Article history: Vitamins are important micronutritional compounds which are involved in many biochemical functions in the
Received 4 February 2015 human body but are not synthesized by it; so, they have to be supplied through diet. However, vitamins are
Received in revised form 24 September 2015 very sensitive which provoke a significant loss during the food processes and storage. So, microencapsulation
Accepted 9 November 2015
can be used to minimize the loss of vitamins, to minimize the factors that interfere with their stability, to allow
Available online 10 November 2015
a controlled release process and to mask its undesirable taste, increasing their applicability
Keywords:
In the present work, the microencapsulation of two vitamins, by a spray-drying process, was studied: vitamin
Encapsulating agent B12, considering that is the most chemically complex and the largest of all the vitamins and vitamin C which is
Microencapsulation the most popular vitamin in the food industry
Microcapsules The microparticles were prepared using a spray-dryer BÜCHI B-290 (Flawil, Switzerland) with a standard 0.5 mm
Spray drying nozzle, under the following conditions: solution and air flow rates, air pressure and inlet temperature were set at
Vitamin C 4 ml/min (15%), 32 m3/h (80%), 6.0 bar and 120 °C, respectively. The prepared microparticles were characterized
Vitamin B12 and their physicochemical structures were analyzed by scanning electron microscopy (SEM) and by Fourier
transform infrared spectroscopy (FTIR). The presence of vitamins in the microparticles was also evaluated by
UV-method, validated and optimized for this objective. The evaluation of the vitamin B12 was based on absor-
bance values read at 361.4 nm, and for the vitamin C the absorbance was read at 260.6 nm
A product yield ranging from 41.8 to 55.6% for the microparticles prepared with vitamin B12 and ranging from
43.6 to 45.4% for the microparticles formed with vitamin C was obtained and microparticles with a mean diam-
eter around 3 μm were observed, for all the biopolymers tested (chitosan, modified chitosan and sodium
alginate). The microparticles formed with chitosan presented a very rough surface; on the other hand, the parti-
cles formed with sodium alginate or modified chitosan presented a very smooth surface. The performed tests
yield significant results and prove the success of the vitamins microencapsulation
This work shows that it is possible to encapsulate vitamins using different biopolymers, through a spray-drying
process
© 2015 Elsevier B.V. All rights reserved.
1. Introduction Vitamin B12 is also called cobalamin because it has cobalt in its
structure and is the most chemically complex and largest of all the vita-
Vitamins are bioactive compounds; in food are physiologically active mins. Vitamin B12 belongs to the group “corrinoids,” which is a group of
components that provide health benefits beyond their nutritional role compounds having a corrin macrocycle. Vitamin B12 has a molecular
[1]. Vitamins are important micronutritional substances involved in weight of 1355.4, is stable in aqueous solution of pH 4–7 and can be
many biochemical functions in the human body, but they are not heated at 120 °C without significant losses [3,4]. Vitamin B12 is involved
synthesized by it; so, they have to be supplied through diet [2]. One in the cell metabolism (DNA synthesis and regulation), in the normal
diet poor in vitamins can lead to many deficiency diseases like perni- operation of the brain and nervous system, and in the formation of
cious anemia, scurvy, pellagra, ariboflavinosis, dermatitis and enteritis, blood. Vitamin B12 is produced by certain bacteria and is concentrated
among others. This research is focused on two main water-soluble in the bodies of higher predators in the food chain. Therefore, foods de-
vitamins: vitamin B12 and vitamin C. rived from animals are considered to be the major dietary sources of
B12 [3,4]. People with a limited intake of food with an animal source
have a high risk of suffering B12 deficiency. Vitamin B12 is related to
⁎ Corresponding author. some diseases, like pernicious anemia. One solution is to consume vita-
E-mail address: [email protected] (B.N. Estevinho). min B12-fortified foods or vitamin B12-containing dietary supplements
http://dx.doi.org/10.1016/j.powtec.2015.11.019
0032-5910/© 2015 Elsevier B.V. All rights reserved.
72 B.N. Estevinho et al. / Powder Technology 289 (2016) 71–78
to prevent B12 deficiency [3,4]. In addition low values of vitamin B12 (w/V), prepared in a solution of acetic acid (1% (V/V)) has a viscosity
were reported in animals, and one of the solutions is the incorporation of 200 mPa·s (25 °C). Water soluble chitosan (pharmaceutical grade)
of vitamin B12 in additives [5–7]. was obtained from China Eastar Group (Dong Chen) Co., Ltd. (Batch
Vitamin C is a well-known bioactive compound and is a represen- no. SH20091010). Water soluble chitosan was produced by carboxyla-
tative water-soluble vitamin. Vitamin C has a variety of biological, tion and had a deacetylation degree of 96.5%. The solution of modified
pharmaceutical and dermatological functions. [8] Vitamin C helps chitosan at 1% (w/v), prepared in deionized water has a viscosity of
in fighting common colds by strengthening the immune system 5 mPa·s (25 °C). Sodium alginate (alginic acid, sodium salt) (Cat. No.
and is important for its potential role in minimizing the risk of seri- 180947) was from Aldrich (USA).
ous diseases such as cancer, heart disease, cataracts, and high lead All the three solutions were prepared at room temperature. The
levels. Deficiency in vitamin C is associated with the disease known chitosan solution was prepared with a concentration of 1% (w/V) in an
as scurvy [9]. Unfortunately, the human body is unable to synthesize acetic acid solution 1% (v/v) and with 2 h agitation at 1200 rpm
vitamin C and cannot store it. So, appropriate amounts must be sup- (magnetic agitator — MS-H-Pro, Scansci). The other two solutions, of
plied regularly through the diet to restock this valuable compound water soluble chitosan 1% (w/V) and sodium alginate 1% (w/V) were
[9]. Vitamin C is widely used in various types of foods as a vitamin prepared with deionized water and with 2 h agitation at 1200 rpm.
supplement. However, vitamin C is very unstable to air, moisture, To obtain the vitamin microparticles it was necessary to prepare so-
light, heat, oxygen and alkaline pH and easily decomposes into biologi- lutions containing the vitamins and the encapsulating agents. These so-
cally inactive compounds [8]. Furthermore, due to its acidic nature, it lutions were then fed to the spray dryer. Thus, the solution containing
can interact with other food components and thus negatively affect the vitamin was added and mixed with each one of the biopolymers
the sensory properties and shelf life of vitamin C-fortified foods [10]. aqueous solutions (encapsulating agents) at constant agitation speed
Therefore, microencapsulation could be used as an alternative to of 1200 rpm, during 10 min at room temperature. The concentration
minimize the factors that interfere with the stability of the vitamins, of the vitamin in the fed solution to the spray-dryer was 2.0% (w/w).
allow for the controlled release and mask its undesirable taste, which Also, microparticles without vitamin were prepared, in order to study
can be unpleasant [8,11]. It is important to microencapsulate these the effect of the vitamin on the microparticles produced, under the
vitamins in order to increase their applicability in food processes. same conditions.
For example, some authors studied the possibility of microencapsu-
lating vitamins with the purpose of increasing their resistance to the 2.2. Experimental conditions —spray-drying process
cooking process or storage [9]. Different studies concluded that,
vitamin C, vitamin B9 and vitamin B6 are less stable during high- Spray-drying was performed using a spray-dryer BÜCHI B-290
temperature processing as compared to retinol, thiamine, riboflavin (Flawil, Switzerland) with a standard 0.5 mm nozzle. The same proce-
and niacin. Almost all of these studies were made with encapsulating dure was followed for all the microparticles prepared with vitamin
agents or processes that request the presence of organic solvents that B12 and vitamin C and also for microparticles prepared without vitamin.
can increase the toxicity of the particles produced. New solutions and The solutions were spray-dried, under the following conditions: solu-
techniques are requested. Borrmann et al. [12] microencapsulated pas- tion and air flow rates, air pressure and inlet temperature were set at
sion fruit juice (vitamin C) with n-octenyl succinate-derivatised starch 4 ml/min (15%), 32 m3/h (80%), 6.0 bar and 120 °C, respectively. The
using a spray-dryer and stored at two different temperatures. Bastos outlet temperature, a consequence of the other experimental conditions
et al. [13] microencapsulatated cashew apple (Anacardium occidentale, and of the solution properties, was around 65 °C. The operating condi-
L.) juice (vitamin C) using also a spray-drying process. tions have been selected considering preliminary studies. All the exper-
The spray-drying process is flexible and produces microparticles of iments were made in duplicate, with a coefficient of variation smaller
good quality and is also, a relatively low cost technology, rapid, repro- than 10%.
ducible, allowing easy scale-up, when compared with other microen-
capsulation techniques, justifying the preference in industrial terms [2, 2.3. SEM characterization
14–20].
The present work shows the recent developments and the new Structural analysis of the surface of the particles was performed
applications of the spray drying technology for microencapsula- by scanning electron microscopy (Fei Quanta 400 FEG ESEM/EDAX
tion of two different vitamins (vitamin B12 and vitamin C) with Pegasus X4M). The surface structure of the particles was observed
different biopolymers: chitosan, modified chitosan and sodium by SEM after sample preparation by pulverization of gold in a Jeol
alginate, considering all the advantages of these biopolymers. On JFC 100 apparatus at Centro de Materiais da Universidade do Porto
the other hand, the two selected vitamins were chosen considering (CEMUP).
their complexity (vitamin B12) and their high applicability (vita-
min C). 2.4. FTIR analysis
concentration of released vitamin was estimated by mass balance, For vitamin C, the calibration was developed and validated in the
considering the amount of reagents used, proportions vitamin/bio- concentration range of 0.0005 g/L to 0.022 g/L with 10 standards
polymer and specifications of the spray drying process, and was (prepared in deionized water), with a correlation coefficient of 0.985
also determined experimentally considering the amount of vitamin and a detection limit of 0.001 g/L.
released from the microparticles. The released vitamin was deter-
mined in continuous absorbance measurements (intervals of 30 s) 3. Results and discussion
until the maximum value of the released vitamin was obtained and
stabilized. The spray drying process was performed with previously fixed oper-
The determination of the presence of the vitamin B12 and vitamin C ating conditions, in order to compare the microcapsules formed with
in the microparticles was based on absorbance values, read at room tem- different encapsulating agents (chitosan, modified chitosan and sodium
perature in an UV–Visible spectrophotometer (SCANSPEC SP110070 alginate).
from SCANSCI) at 361.4 nm and 260.6 nm respectively. The product yield (quantity of powder recovered reported to the
Two calibration curves were developed to evaluate the concentra- quantity of raw materials used) for the microparticles with vitamin
tion of vitamin released to the solution. The calibrations were made in B12 was 41.8%, 55.6% and 42.4% when prepared with sodium alginate,
duplicated with coefficients of variation smaller than 10% for all the chitosan and modified chitosan, respectively. In the case of the micro-
standards. particles with vitamin C a product yield of 43.6%, 44.5% and 45.4%,
For vitamin B12, standard solutions were prepared in deionized respectively, was obtained. The product yields obtained for these micro-
water. The determination of vitamin B12 was validated in the concen- particles are very similar with the ones obtained by Estevinho et al. [21]
tration range of 0.0025 g/L to 0.1 g/L with 12 standards and with a to prepare β-galactosidase microparticles with the same encapsulating
correlation coefficient of the method of 0.985. The detection limit deter- agents. Estevinho et al. [22] also discussed the existence of small prod-
mined for vitamin B12 was 0.006 g/L. uct yields, around 30–50%, for the microencapsulation by a spray drying
Fig. 1. SEM images of the microparticles with vitamins and without vitamins with different biopolymers: sodium alginate, chitosan and modified chitosan. Amplified 30,000 times, beam
intensity (HV) 1000 kV, distance between the sample and the lens (WD) less than 12 mm.
74 B.N. Estevinho et al. / Powder Technology 289 (2016) 71–78
technique. When the inlet temperature is lower, as in the present case polysaccharides with similar functional groups, that will provoke a
(120 °C), the probability of obtaining low product yields increases. At high similarity between the spectra.
low temperatures, the deposition of particles on the cylinder or/and Alginate is a natural, linear, unbranched polysaccharide containing
on the cyclone wall of spray dryer was observed, leading to a lower 1,4′-linked beta-D-mannuronic and alpha-L-guluronic acid residues
product yield. On the other hand, the particles formed by this method [23]. For sodium alginate microparticles the more important absorption
are very small (around 3 μm), and the efficiency of the cyclone to sepa- bands at frequency values that justify the existence of the corre-
rate small particles decreases, some of them being aspirated with the air sponding functional groups (bonds), are for example: O–H (3700–
leaving the spray dryer. Also, the sample volume influences the product 3000 cm − 1 ; stretch vibration), C–H (3000–2850 cm − 1 ; stretch
yield; small volumes implying higher relative losses [22]. vibration), CO 2 – (1600 cm − 1 antisymmetric CO 2 -stretch), CO 2 –
The prepared microparticles were characterized and their physico- (1400 cm− 1 symmetric CO2–stretch), at 1300 cm− 1 skeletal vibra-
chemical structures were analyzed by SEM and by FTIR. tion and at 1100–1000 cm − 1 antisymmetric stretch C–O–C. These
bands are consistent with the results of, for example, Lawrie et al.
3.1. Microparticles characterization [24].
Chitosan is an attractive biopolymer, although a water-insoluble ma-
3.1.1. Scanning electron microscopy (SEM) analysis terial; however it is possible to modify the structure in order to produce
Spherical microparticles were produced in all the cases (Fig. 1). The an easily soluble chitosan in neutral aqueous solutions [25–27]. Water
surface of the microparticles presented different textural characteristics. soluble chitosan can be useful for drug carriers and for food industrial
In the case of the particles formed with chitosan the surface was very applications [19,28].
rough. The particles formed with sodium alginate had a smooth surface For chitosan microparticles the more important absorption bands
and the microparticles formed with modified chitosan presented a very are at frequency values: 3700–3000 cm−1 (O–H and N–H, stretch vibra-
regular shape and a smooth surface. Microparticles with a mean diame- tion), 3000–2850 cm−1 (C–H, stretch vibration), 1645 cm−1 (Amide I),
ter around 3 μm were observed, for all the biopolymers tested (chitosan, 1584 cm−1 (N–H bending from amine and amine II), 1410 cm−1 (–CH2
modified chitosan and sodium alginate). In SEM images, the size of the bending), 1375 cm−1 (CH3 symmetrical deformation), 1150 cm−1 anti-
microparticles containing vitamins appears to be similar to the size of symmetric stretch C–O–C and C–N stretch and at 1030 skeletal vibration
the microparticles produced without vitamins. Estevinho et al. [21] of C–O stretching. The band at 1560 cm−1 has a larger intensity than at
found similar results, surface, textural characteristics and size, for the 1655 cm−1, which suggests effective deacetylation of chitosan. The
microencapsulation of β-galactosidase with the same encapsulating band at 1656 cm−1 corresponds to the amide I stretching of C_O, as de-
agents. Also the microparticles size without enzyme appears to be scribed by Lawrie et al. [19,24]. The spectrum of the microparticles with
similar to the size of the microparticles with enzyme. modified chitosan is similar to the spectrum of the chitosan; only the
size of some absorption bands is different.
3.1.2. FTIR analysis In Fig. 3, the spectra of the microparticles with vitamin B12 and vita-
FTIR studies give information about the molecular structure of min C are presented. These microparticles have only 2% of vitamin and
chemical compounds and are useful for the characterization of 98% of encapsulating agent, which makes it difficult to identify the pres-
biopolymers. ence of the vitamins in the spectra, when compared with the spectra of
In this work, 3 different biopolymers (sodium alginate, chitosan and the microparticles made only with the encapsulating agent. Some im-
modified chitosan) were tested, taking into account their high bio- portant absorption bands of the vitamins are overlapped by the absorp-
compatibility. In Fig. 2, it is possible to evaluate the FTIR spectrum tion bands of the encapsulating agent making it difficult to distinguish
of the microparticles made with these biopolymers. All of them are them.
Fig. 2. FTIR spectra for samples with microparticles made with sodium alginate, chitosan and modified chitosan. Spectra obtained with KBr at 99%, at 21 scans/min, with a resolution of
4 cm−1 and expressed in transmittance in the 4000–650 cm−1 range.
B.N. Estevinho et al. / Powder Technology 289 (2016) 71–78 75
Fig. 3. FTIR spectra for samples with microparticles with vitamin B12 and vitamin C made with: A — sodium alginate, B — chitosan and C — modified chitosan. Spectra obtained with KBr at
99%, at 21 scans/min, with a resolution of 4 cm − 1 and expressed in transmittance in the 4000–650 cm − 1 range.
76 B.N. Estevinho et al. / Powder Technology 289 (2016) 71–78
The major band for the vitamin B12 occurs at frequency values of 3.2. Evaluation of the presence of vitamin B12 and vitamin C in the
1664 cm− 1 and is due to the amide I C_O stretching mode of the microparticles
propionamide side chains of the corrin ring [29]. These authors also de-
scribe a medium intensity band at 1572 cm−1, attributed to a breathing For both of the vitamins the release was total. So, the presence of vi-
mode of the corrin ring of vitamin B12 and another one at a frequency tamin B12 and vitamin C in the microparticles was confirmed and also
2135 cm−1 (cyanide stretching), proving the cobalt-carbon distance in obtained the different release profiles, for the different encapsulating
cyanocorrinoids of vitamin B12 [29]. agents. The total amount of the vitamin was recovered in different
The strongest absorption bands for the vitamin C occur at frequency times depending on the encapsulating agents (Figs. 4 and 5). For exam-
values: 1764 cm−1 (C_O stretching), 1675 cm−1 (C_C ring stretching) ple, for vitamin B12 the total amount of vitamin was released in 120 min
and 3216–3626 cm−1 (OH stretching). Various vibrational bands can be for microparticles made with chitosan, in 15 min for microparticles
observed in the region 1200–1500 cm−1 which are connected with the made with alginate and 10 min for the microparticles made with mod-
CH2 scissoring, twisting and wagging and the C–H deformation modes. ified chitosan. Similar results have been obtained for the microparticles
The band at 1277 cm−1 is originated by C–O–C stretching. Other C–O– with vitamin C.
C stretches can be seen at 1142, 1121, 1113, 1077 and 1046 cm−1 in Comparing the SEM images with the release profiles, a slow release
the vitamin C spectrum [30]. was associated to a rougher surface (microparticles with chitosan).
In FTIR spectra of the microparticles made with alginate (Fig. 3A) the So, depending on the type of application intended for the vitamins,
presence of the vitamins was very difficult to recognize. There are only different encapsulating agents need to be selected to allow the more ad-
small differences in the size or in the proportion of the bands. equate controlled release of the vitamins. For instance if a slow release
In the case of chitosan (Fig. 3B), for the microparticles with vita- of the vitamin in one aqueous solution is wanted the best option for
min C the bands at frequency values of 1764 cm− 1, 1675 cm− 1 and the encapsulating agent will be the chitosan. On the other hand if it
1277 cm− 1 are different and appear to be bigger than in the case of is intended a fast release of the vitamin the best option can be the micro-
the microparticles made only with chitosan. For the microparticles encapsulation of vitamins with modified chitosan or alginate. These two
with vitamin B12 the band at the frequency 2135 cm− 1 (cyanide encapsulating agents can be used, for example, for microencapsulated
stretching) increases, proving the presence of vitamin B12. Finally, vitamins used in drinks prepared instantaneously from powder formu-
for the case of the microparticles with modified chitosan (Fig. 3C), lations. Thus the vitamins will be protected from oxidation, light, mois-
and for the case of vitamin B12, the bands at frequency values of ture and other factors during the storage time.
1664 cm − 1 (C_O stretching) and 1572 cm− 1 (attributed to a As referred by Murugesan and Orsat microencapsulation and
breathing mode of the corrin ring) have other size relation than in nanoencapsulation are the best ways to preserve vitamins [2]. Some au-
the spectrum of microparticles made only with modified chitosan. thors used with success the spray drying technique to microencapsulate
It is also possible to see a small band at a frequency of 2135 cm − 1 vitamins such as vitamin A [31,32], vitamin E [33] and vitamin C [8]. Vi-
(cyanide stretching). For vitamin C, the bands at frequency values tamin C was successfully encapsulated in tripolyphosphate (TPP) cross-
of 1764 cm− 1 (C_O stretching), 1675 cm− 1 (C_C ring stretching) linked chitosan (TPP-chitosan) microspheres by the spray-drying meth-
and at 1277 cm− 1 (C–O–C stretching) increased. For both of the od. The effect of adding a crosslinking agent and how this crosslinking
cases of the microparticles made with vitamins, the size of the increases the stability of the microparticles was studied [8,34]. The
band 3600–3000 cm− 1 (OH stretching) increased, when compared sphericity of chitosan microspheres was lost at higher volume of
with the spectrum of the microparticles made only with modified crosslinking agent. The TPP-chitosan microspheres loaded with vitamin
chitosan. C were spherical and had smooth surface and the release of vitamin C
The differences between the spectra with and without vitamins are from these microspheres was sustained and affected by the volume of
very small, but they give support to the idea that microparticles have vi- crosslinking agent added. [8,34]. In general the crosslinking agents pro-
tamins in their composition. To confirm this, analytical methods have voke changes in the structure of the microparticles and delay the release
been developed to quantify the presence of vitamins in the microparticles. of the compounds from the microparticles, improving the controlled
Fig. 4. Release of vitamin B12 from microparticles made with different encapsulating agents.
B.N. Estevinho et al. / Powder Technology 289 (2016) 71–78 77
Fig. 5. Release of vitamin C from microparticles made with different encapsulating agents.
release systems [18]. In the present work, 3 encapsulating agents have References
been studied and compared, without crosslinking agents. The use of
crosslinking agents will be subject of future works. [1] A. Teleki, A. Hitzfeld, M. Eggersdorfer, 100 years of vitamins: the science of formula-
tion is the key to functionality, KONA Powder Part. J. 30 (2013) 144–163, http://dx.
doi.org/10.14356/kona.2013015.
[2] R. Murugesan, V. Orsat, Spray drying for the production of nutraceutical
4. Conclusion ingredients—a review, Food Bioprocess Technol. 5 (2011) 3–14, http://dx.doi.org/
10.1007/s11947-011-0638-z.
[3] F. Watanabe, Y. Yabuta, Y. Tanioka, T. Bito, Biologically active vitamin B12
In the present work two different vitamins (vitamin B12 and vitamin compounds in foods for preventing deficiency among vegetarians and elderly sub-
C) were microencapsulated by a spray drying process using three differ- jects, J. Agric. Food Chem. 61 (2013) 6769–6775, http://dx.doi.org/10.1021/
ent encapsulating agents: chitosan, modified chitosan and sodium jf401545z.
[4] F. Watanabe, Y. Yabuta, T. Bito, F. Teng, Vitamin B12-containing plant food
alginate. sources for vegetarians, Nutrients 6 (2014) 1861–1873, http://dx.doi.org/10.
A product yield around 45% was obtained for both of vitamins in all 3390/nu6051861.
the assays. Microparticles with a mean diameter around 3 μm were ob- [5] N.D. Grace, S.O. Knowles, Trace element supplementation of livestock in New
Zealand: meeting the challenges of free-range grazing systems, Vet. Med. Int.
served, for all the biopolymers tested. The microparticles formed with 2012 (2012) 639472, http://dx.doi.org/10.1155/2012/639472.
chitosan presented a very rough surface but the particles formed with [6] N.D. Grace, D.H. Lewis, An evaluation of the efficacy of injectable microencapsulated
sodium alginate or modified chitosan presented a very smooth surface. vitamin B12 in increasing and maintaining the serum and liver vitamin B12 concen-
trations of lambs, N. Z. Vet. J. 47 (1999) 3–7, http://dx.doi.org/10.1080/00480169.
Finally, the presence of vitamins in the microparticles was confirmed
1999.36099.
and evaluated by UV-method, validated and optimized for this objec- [7] N.D. Grace, The effect of increasing the vitamin B12 status of Romney ewes on foetal
tive. Different release profiles were obtained for both of the vitamins liver vitamin B12, milk vitamin B12 and liver vitamin B12 concentrations in suckling
with the different encapsulating agents (chitosan, modified chitosan lambs, N. Z. Vet. J. 47 (1999) 97–100, http://dx.doi.org/10.1080/00480169.1999.36121.
[8] K.G.H. Desai, H.J. Park, Encapsulation of vitamin C in tripolyphosphate cross-linked
and sodium alginate). In general, the release time of the total amount chitosan microspheres by spray drying, J. Microencapsul. 22 (2005) 179–192,
of vitamins was around 120 min for microparticles made with chitosan, http://dx.doi.org/10.1080/02652040400026533.
15 min for microparticles made with alginate and 10 min for the micro- [9] S. Abbas, C. Da Wei, K. Hayat, Z. Xiaoming, Ascorbic acid: microencapsulation tech-
niques and trends—a review, Food Rev. Int. 28 (2012) 343–374, http://dx.doi.org/10.
particles made with modified chitosan 1080/87559129.2011.635390.
Comparing the SEM images with the release profiles, a slow release [10] T.A. Comunian, A. Abbaspourrad, C.S. Favaro-Trindade, D.A. Weitz, Fabrication of solid
was associated to a rougher surface (microparticles with chitosan). lipid microcapsules containing ascorbic acid using a microfluidic technique, Food
Chem. 152 (2014) 271–275, http://dx.doi.org/10.1016/j.foodchem.2013.11.149.
This work shows that it is possible to encapsulate vitamins using [11] T.A. Comunian, M. Thomazini, A.J.G. Alves, F.E. de Matos Junior, J.C. de Carvalho
different biopolymers through a spray-drying process, and depending Balieiro, C.S. Favaro-Trindade, Microencapsulation of ascorbic acid by complex coac-
on the type of application pretended for the vitamins, different encapsu- ervation: Protection and controlled release, Food Res. Int. 52 (2013) 373–379,
http://dx.doi.org/10.1016/j.foodres.2013.03.028.
lating agents need to be selected to allow more adequate controlled [12] D. Borrmann, A.P.T.R. Pierucci, S.G.F. Leite, M.H.M.D.R. Leão, Microencapsulation of
release of the vitamins passion fruit (Passiflora) juice with n-octenylsuccinate-derivatised starch using
spray-drying, Food Bioprod. Process. 91 (2013) 23–27, http://dx.doi.org/10.1016/j.
fbp.2012.08.001.
[13] D.D.S. Bastos, M.D.P. Gonçalves, C.T. De Andrade, K.G.D.L. Araújo, M.H.M. Da Rocha
Acknowledgments Leão, Microencapsulation of cashew apple (Anacardium occidentale, L.) juice using
a new chitosan-commercial bovine whey protein isolate system in spray drying,
The authors thank Fundação para a Ciência e a Tecnologia (FCT) for Food Bioprod. Process. (2015)http://dx.doi.org/10.1016/j.fbp.2012.04.005 (in
press).
the postdoctoral grant SFRH/BPD/73865/2010 of Berta Estevinho and [14] J. Pu, J.D. Bankston, S. Sathivel, Developing microencapsulated flaxseed oil contain-
to the Erasmus program for the scholarship of Ioana Carlan. ing shrimp (Litopenaeus setiferus) astaxanthin using a pilot scale spray dryer,
78 B.N. Estevinho et al. / Powder Technology 289 (2016) 71–78
Biosyst. Eng. 108 (2011) 121–132, http://dx.doi.org/10.1016/j.biosystemseng.2010. [25] H. Sashiwa, N. Kawasaki, A. Nakayama, Chemical modification of chitosan. 14:1
11.005. synthesis of water-soluble chitosan derivatives by simple acetylation.
[15] A.L.R. Rattes, W.P. Oliveira, Spray drying conditions and encapsulating composition Biomacromolecules 3 (2002) 1126–1128 (http://www.ncbi.nlm.nih.gov/
effects on formation and properties of sodium diclofenac microparticles, Powder pubmed/12217063, accessed April 18, 2012).
Technol. 171 (2007) 7–14, http://dx.doi.org/10.1016/j.powtec.2006.09.007. [26] H. Zhang, S. Wu, Y. Tao, L. Zang, Z. Su, Preparation and characterization of water-sol-
[16] N. Schafroth, C. Arpagaus, U.Y. Jadhav, S. Makne, D. Douroumis, Nano and micropar- uble chitosan nanoparticles as protein delivery system, J. Nanomater. 2010 (2010)
ticle engineering of water insoluble drugs using a novel spray-drying process, 1–5, http://dx.doi.org/10.1155/2010/898910.
Colloids Surf. B: Biointerfaces 90 (2011) 8–15, http://dx.doi.org/10.1016/j.colsurfb. [27] M.N.V.R. Kumar, A review of chitin and chitosan applications, React. Funct. Polym. 46
2011.09.038. (2000) 1–27 (http://www.sciencedirect.com/science/article/pii/S1381514800000389,
[17] P. de Vos, M.M. Faas, M. Spasojevic, J. Sikkema, Encapsulation for preservation of accessed April 18, 2012).
functionality and targeted delivery of bioactive food components, Int. Dairy J. 20 [28] B.N. Estevinho, A.M. Damas, P. Martins, F. Rocha, Study of the inhibition effect on the
(2010) 292–302, http://dx.doi.org/10.1016/j.idairyj.2009.11.008. microencapsulated enzyme β-galactosidase, Environ. Eng. Manag. J. 11 (2012)
[18] B.N. Estevinho, F. Rocha, L. Santos, A. Alves, Microencapsulation with chitosan by 1923–1930.
spray drying for industry applications — a review, Trends Food Sci. Technol. 31 [29] L. Jin, P. Lu, H. You, Q. Chen, J. Dong, Vitamin B12 diffusion and binding in
(2013) 138–155, http://dx.doi.org/10.1016/j.tifs.2013.04.001. crosslinked poly(acrylic acid)s and poly(acrylic acid-co-N-vinyl pyrrolidinone)s,
[19] B.N. Estevinho, F. Rocha, L. Santos, A. Alves, Using water soluble chitosan for flavour Int. J. Pharm. 371 (2009) 82–88, http://dx.doi.org/10.1016/j.ijpharm.2008.12.022.
microencapsulation in food industry, J. Microencapsul. 30 (2013) 571–579, http:// [30] C. Yohannan Panicker, H. Tresa Varghese, D. Philip, FT-IR, FT-Raman and SERS spec-
dx.doi.org/10.3109/02652048.2013.764939. tra of vitamin C, Spectrochim. Acta A Mol. Biomol. Spectrosc. 65 (2006) 802–804,
[20] B.N. Estevinho, A.M. Damas, P. Martins, F. Rocha, The influence of microencapsula- http://dx.doi.org/10.1016/j.saa.2005.12.044.
tion with a modified chitosan (water soluble) on β-galactosidase activity, Dry. [31] Y. Xie, H. Zhou, X. Liang, B. He, X. Han, Study on the morphology, particle size and
Technol. 32 (2014) 1575–1586, http://dx.doi.org/10.1080/07373937.2014.909843. thermal properties of vitamin A microencapsulated by starch octenylsucciniate,
[21] B.N. Estevinho, A.M. Damas, P. Martins, F. Rocha, Microencapsulation of β- Agric. Sci. China 9 (2010) 1058–1064, http://dx.doi.org/10.1016/S1671-
galactosidase with different biopolymers by a spray-drying process, Food Res. Int. 2927(09)60190-5.
64 (2014) 134–140, http://dx.doi.org/10.1016/j.foodres.2014.05.057. [32] Y.-L. Xie, H.-M. Zhou, Z.-R. Zhang, Effect of relative humidity on retention and stabil-
[22] B.N. Estevinho, I. Ramos, F. Rocha, Effect of the pH in the formation of β- ity of vitamin A microencapsulated by spray drying, J. Food Biochem. 31 (2007)
galactosidase microparticles produced by a spray-drying process, Int. J. Biol. 68–80, http://dx.doi.org/10.1111/j.1745-4514.2007.00099.x.
Macromol. 78 (2015) 238–242, http://dx.doi.org/10.1016/j.ijbiomac.2015.03.049. [33] J. Hategekimana, K.G. Masamba, J. Ma, F. Zhong, Encapsulation of vitamin E: effect of
[23] K. Möbus, J. Siepmann, R. Bodmeier, Zinc-alginate microparticles for controlled pul- physicochemical properties of wall material on retention and stability, Carbohydr.
monary delivery of proteins prepared by spray-drying, Eur. J. Pharm. Biopharm. 81 Polym. 124 (2015) 172–179, http://dx.doi.org/10.1016/j.carbpol.2015.01.060.
(2012) 121–130, http://dx.doi.org/10.1016/j.ejpb.2012.01.018. [34] K.G. DESAI, H.J. PARK, Effect of manufacturing parameters on the characteristics of
[24] G. Lawrie, I. Keen, B. Drew, A. Chandler-Temple, L. Rintoul, P. Fredericks, et al., Inter- vitamin C encapsulated tripolyphosphate-chitosan microspheres prepared by
actions between alginate and chitosan biopolymers characterized using FTIR and spray-drying, J. Microencapsul. 23 (2006) 91–103.
XPS, Biomacromolecules 8 (2007) 2533–2541.