Accepted Manuscript

Title: PRODUCTION OF VIRGIN COCONUT OIL

MICROCAPSULES FROM OIL-IN-WATER EMULSION
WITH SUPERCRITICAL CARBON DIOXIDE SPRAY
DRYING

Authors: Y.Y. Hee, C.P. Tan, R.Abdul Rahman, M. Noranizan,

R.L. Smith Jr., G.H. Chong

PII: S0896-8446(17)30246-2
DOI: http://dx.doi.org/doi:10.1016/j.supflu.2017.07.037
Reference: SUPFLU 4004

To appear in: J. of Supercritical Fluids

Received date: 11-4-2017

Revised date: 28-7-2017
Accepted date: 29-7-2017

Please cite this article as: Y.Y.Hee, C.P.Tan, R.Abdul Rahman, M.Noranizan,
R.L.Smith, G.H.Chong, PRODUCTION OF VIRGIN COCONUT OIL
MICROCAPSULES FROM OIL-IN-WATER EMULSION WITH SUPERCRITICAL
CARBON DIOXIDE SPRAY DRYING, The Journal of Supercritical
Fluidshttp://dx.doi.org/10.1016/j.supflu.2017.07.037

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PRODUCTION OF VIRGIN COCONUT OIL MICROCAPSULES FROM OIL-IN-

WATER EMULSION WITH SUPERCRITICAL CARBON DIOXIDE SPRAY DRYING

Y. Y. Hee1, C. P. Tan1, R. Abdul Rahman1,2, Noranizan, M.1,2, R. L. Smith Jr3., G. H. Chong1,2*

1
Faculty of Food Science and Technology, University Putra Malaysia, 43400 UPM Serdang,

Malaysia.

2
Supercritical Fluid Centre, University Putra Malaysia, 43400 UPM Serdang, Malaysia.

3
Research Center of Supercritical Fluid Technology, Tohoku University, Aramaki Aza Aoba, 6-

6-11-403, Aoba-Ku, Sendai, Miyagi 980-8579, Japan.

*corresponding author

Graphical abstract

1
Highlights
 Virgin coconut oil encapsulated with maltodextrin, sodium caseinate, soy lecithin.
 Spherical particles, (27 – 72) μm, d90=36 μm
 High encapsulation efficiency, 80 % (16 MPa, 40 ºC).
 High antioxidant activity, (0.6 – 1.0 mmol BHT equivalent/g oil).

Abstract

Virgin coconut oil (VCO) has been shown to have health-promoting effects due to its high

medium-chain fatty acid (MCFA) content and antioxidant properties, however, its taste and

feeling of greasiness are not acceptable to many consumers. In this study, VCO was

microencapsulated to develop a new and healthy consumable food product. Spray drying VCO

with supercritical carbon dioxide (SC-CO2) at pressures from 12-16 MPa, temperatures from 40-

60 ºC was studied for an emulsion feed flow rate that was varied from 3-5 mL/min. The

encapsulation efficiencies ranged from 73 % to 80 %, the microcapsules were spherical with

diameters that ranged from 27 to 72 µm and the antioxidant activities of the retained

microencapsulated oil ranged from 0.6 to 1.0 mmol butylated hydroxytoluene (BHT)

equivalent/ml oil and 0.8 to 1.3 mmol trolox equivalent/ml oil for 2,2-diphenyl-1-picrylhydrazyl

(DPPH) tests and 2,2'-azino-bis (ABTS) tests, respectively. SC-CO2 spray drying is an effective

method to encapsulate VCO and other food oils.

Keywords: particle formation; emulsion; oil powder; encapsulation efficiency; particle size

1. Introduction

2
Virgin coconut oil (VCO) is obtained from the meat of coconut (Cocos nucifera L.) with

physical or thermal methods that do not alter its nature [1]. VCO that is not refined, bleached or

deodorized retains its nutritional value and distinctive aroma and taste. VCO has many potential

health benefits due to its rich medium-chain fatty acids (MCFAs), vitamins and antioxidants.

Lauric acid is the dominant fatty acid in VCO and contributes to its strong antiviral and

antibacterial activity [2]. MCFAs of VCO can increase metabolism rates in the liver when

ingested and can help prevent obesity [3]. VCO has a cardioprotective effect that is beneficial in

lowering lipid components, increasing antioxidant enzymes and reducing lipid peroxide content

due to its polyphenolic content [4,5]. However, many consumers do not accept the texture of

VCO because it contains saturated fatty acids that are solid at room temperature. Therefore,

liquid VCO is typically converted into a powder form via spray drying to give consumers a new

perception of VCO as a food product [6].

The application of microencapsulation in the food industry includes providing a protective

barrier to sensitive functional compounds, masking unpleasant tastes and smells and stabilizing

and increasing the bioavailability of the bioactive compound [7]. Spray drying is the mostly

broadly technique applied in oil microencapsulation, however, it has several drawbacks,

including the destruction of heat-sensitive compounds, large particle sizes, wide particle size

distribution, low precipitation yields and difficulty in removing any solvents used [8–12].

There is lack of reported studies on the microencapsulation of VCO with SC-CO2 particle

formation methods. Spray drying with SC-CO2 has the following advantages including mild

operating temperatures (ca. 60 ºC), small particle size and narrow particle size distribution,

favorable yields for many products, it is safe and its removal is simple by depressurization [13].

In this work, the VCO was prepared as an oil-in-water emulsion and products were processed at

3
40 to 60 ºC. The influence of operating conditions, namely, pressure, temperature and emulsion

feed flow rate, on the physicochemical characteristics of the VCO microcapsules was

investigated. Properties measured include encapsulation efficiency, particle size, particle size

distribution, particle morphology, powder flowability, antioxidant activity and fatty acid

composition.

2. Materials and methods

2.1. Materials

VCO was provided by Adirondack (M) Sdn. Bhd. (Petaling Jaya, Malaysia). As the

encapsulating materials, maltodextrin DE10 was purchased from Porrima (M) Sdn. Bhd. (Batu

Caves, Malaysia), and sodium caseinate was purchased from V.I.S. Foodtech Ingredient Supplies

Sdn. Bhd. (Sri Damansara, Malaysia). Soy lecithin was purchased from Se Scientific Supplies

(Ampang, Malaysia). Carbon dioxide (CO2) with 99.9 % purity was purchased from Mox-Linde

Gases Sdn. Bhd. (Petaling Jaya, Malaysia). Reagent grade chemicals methanol (≥ 95 %), hexane

(≥ 95 %), ammonia solution (≥ 95 %), diethyl ether (≥ 95 %), isopropanol (≥ 95 %) and

petroleum ether (≥ 95 %), were purchased from Merck Sdn. Bhd. (Bandar Sunway, Malaysia)

and used in the analyses. Reagents related to bioassays, 2,2-diphenyl-1-picrylhydrazyl (≥ 98 %),

butylated hydroxytoluene (≥ 99 %), potassium persulfate (≥ 99 %), trolox (97 %), 2,2’-azino-

bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (≥ 98 %) and fatty acid methyl

ester (FAME) mix standards (C8-C22) were purchased from Sigma-Aldrich (M) Sdn. Bhd.

(Petaling Jaya, Malaysia).

2.2. Particle formation with SC-CO2 solvent

4
An oil-in-water emulsion was produced with the formulation described by Hee et al. [6]:

reagents maltodextrin (13.0 %), sodium caseinate (4.4 %) and soy lecithin (1.0 %) were

dissolved in water (70.0 %) and then blended with VCO (11.6 %) using a blender (MX-800S,

Panasonic, Petaling Jaya, Malaysia); the wall material and oil had a ratio of 60:40. The pre-

homogenized emulsion was subjected to a high-pressure homogenizer (APV 1000, SPX

Corporation, Albertslund, Denmark) at 18 MPa for seven cycles and was then used as feed in a

SC-CO2 spray unit.

The particle formation process was performed with an aqueous spraying unit (FeyeCon,

Development & Implementation BV, Weesp, the Netherlands). The precipitation chamber (25 L)

was set to the desired temperature; a high-pressure pump (NP16/14-210C, Speck-

Kolbenpumpenfabrik, Geretsried, Germany) pumped the SC-CO2 into the precipitation chamber

through a full cone spray nozzle with an internal mix setup (PNR, Voghera, Italy) with a

regulated volumetric flow rate. When the required conditions were achieved, the feed was

delivered into the precipitation chamber by a syringe pump (260 D, Teledyne ISCO, Lincoln,

USA) through the spray nozzle. SC-CO2 was continuously fed into the vessel to sustain stable

operation. Once the emulsion was depleted, the liquid pump was discontinued and CO2 was

allowed to flow through the chamber for 30 min to eliminate the remaining liquid, which was

solubilized in SC-CO2. The gas within the precipitation chamber was slowly depressurized until

it returned to atmospheric pressure. The powder was collected from a filter bag located in the

precipitation vessel and immediately vacuum packed and stored at -2 ºC until further

characterization tests were performed.

During the particle formation process, the CO2 flow rate was maintained at 150 L/h (at the

operating conditions). The mass flow rate of CO2 at different operating conditions was listed in

5
Table S1. In this study, three main factors, namely, precipitation pressure (12, 14, and 16 MPa),

precipitation temperature (40, 50, and 60 ºC) and emulsion flow rate (3, 4, and 5 mL/min) were

evaluated. All experiments were performed in duplicate. The range for each variable was

determined via preliminary experiments.

2.3. Powder characterization

2.3.1. Encapsulation efficiency

The encapsulation efficiency was defined as the percentage of encapsulated oil divided by the

total amount of oil, as given by eq. (1):

total oil (g) - surface oil (g)

Encapsulation efficiency (%)= ×100 (1)
total oil (g)

where the total oil is the mass of oil in the microparticles and the surface oil is the mass of non-

encapsulated oil on the surface of the microcapsules.

The surface oil on the microcapsules was quantified by a method reported by Calvo et al. [14].

Five grams of microcapsules was placed in a beaker and mixed with 50 mL of hexane. The

mixture was stirred for 15 s to remove the surface oil and then filtered. Next, the collected

solvent was removed by a rotary evaporator (Laborota 4000eco, Heidolph Instruments GmbH &

Co. KG, Schwabach, Germany). The amount of surface oil was obtained after evaporating the

solvent [15].

The total oil was quantified by the Rose-Gottlieb method as in Manirakiza et al. [16]. Six

milliliters of boiling water were added to a 50 mL falcon tube that contained 1 g of sample. The

mix was vortexed for 60 s. One milliliter of 25 % ammonia solution and 7.5 mL of methanol

6
were added to the mixture when it was cooled. The solution was vortexed for 2 min after each

addition of the ammonia/methanol solution. Subsequently, 17 mL of diethyl ether and 17 mL of

petroleum ether were added, and the mixture was agitated by hand for extraction after each

addition. The resulting mixture was centrifuged to form two layers; the upper layer was pipetted

into a pre-weighed flask. The solvents were removed with a rotary evaporator, and the extract

was dried at 105 ºC for 1 h with an oven.

2.3.2. Moisture content

One gram of powder was evenly spread on the sample pan of the infrared moisture analyzer (XM

120; Precisa Instruments Ltd, Dietikon, Switzerland), the chamber was heated to 105 ºC and

drying was stopped when the sample achieved constant weight.

2.3.3. Flowability

The flowability was evaluated based on the compressibility index (CI) [17], defined by eq. (2):

CI = 100∙(TD – AD)/TD (2)

where TD is the tapped density and AD is the aerated bulk density. AD is the mass per unit

volume of microcapsules when the microcapsules are not subjected to any compression. TD was

obtained after hand-tapping the 25 mL graduated cylinder that contained 2.5 g of powder until no

noticeable change in volume was observed.

2.3.4. Particle size distribution

The particle size distribution of the powders was determined using a Malvern Mastersizer

Scirocco 2000 laser-based static light scattering particle size analyzer (Malvern Instruments Ltd.,

7
Worcestershire, UK). A small sample was placed on the dry powder feeder tray and the sample

was dispersed by a flow of clean dry air. The refractive index of the sample was 1.5, the

absorption was zero, the vibration feed rate was 30 % and the dispersive air pressure was 0.2

MPa. The particle size was determined as a mean volumetric size (D43; De Brouckere mean

diameter).

2.3.5. Particle morphology

A thin layer of powder was adhered onto a scanning electron micrography (SEM) stub. Next, the

SEM stub was coated with gold/palladium under vacuum from 3-5 mA. The powder was

examined with a scanning electron microscope (JSM 5800, JEOL, Tokyo, Japan) at 15 kV with

3,000× magnification.

To view the inner structure of the microparticles, the microparticles were mechanically broken

with a blade, attached to SEM stub and coated with gold. The morphology of the broken

microparticles was viewed with a Hitachi VP-SEM SU1510 (Hitachi, Tokyo, Japan) at 15 kV

with 3,000x magnification.

2.3.6. Antioxidant activity

2.3.6.1. Radical scavenging activity

The 2,2-diphenyl-1-picrylhydrazyl (DPPH) measurement was performed as described by Lee et

al. [18]. Fifty-six microliters of oil samples were dissolved in 5 mL of 0.10 mM DPPH in a 15

mL centrifuge tube. The sample was placed in the dark for 30 min, and the absorbance was read

with a UV/VIS-spectrophotometer (UVmini – 1240, Shimadzu, Kyoto, Japan) at 517 nm. The

8
radical scavenging capacity of the oils was expressed in mmol butylated hydroxytoluene (BHT)

equivalent/ml oil.

2.3.6.2 Radical cation ABTS•+ scavenging capacity

The 2,2'-azino-bis (ABTS) test was based on the method of Uluata and Özdemir [19]. An equal

amount of 7.0 mM ABTS stock solution and 2.45 mM potassium persulfate was reacted in the

dark for 16 h to produce the ABTS radical cation stock solution. The mixture was topped up with

isopropanol until the absorbance ranged from 0.700 ± 0.020 at 765 nm. One hundred microliters

of the isopropanol-diluted oil sample and 2.9 mL of isopropanol-diluted ABTS•+ solution were

mixed and vortexed for 20 s. After being placed in the dark for 6 min, the absorbance was read at

765 nm. The radical cation ABTS•+ scavenging activity of the samples was expressed in mmol

Trolox equivalent/ml oil.

2.3.9. Fatty acid composition

The fatty acid composition (FAC) of the oils was analyzed using AOCS method Ce 1f-96 and

AOCS procedure Ce 2-66, respectively [20]. The FAC was analyzed with an Agilent 6890N gas

chromatography system that was equipped with a flame ionization detector and an Agilent

7683B automatic liquid sampler (Agilent Technologies, Santa Clara, United States). The FAMEs

were separated with a SGE BPX70 GC capillary column (30 m × 0.32 mm, 0.25 µm film). One

microliter of sample was employed. First, the oven was heated to 100 ºC and maintained for 2

min. Next, the oven temperature was increased to 230 ºC at 6 ºC/min and maintained at that

temperature for 34 min. The injector and detector temperatures were fixed at 250 ºC. The FAC

9
was determined by matching the peak retention time with FAME standards and quantified based

on peak area of the FAMEs.

2.4. Statistical analysis

Results are expressed as the mean ± standard deviation and were statistically calculated using a

one-way analysis of variance (ANOVA). Significant differences in the mean values were

determined by Tukey’s test at a significance level of p<0.05 using the Minitab 16 software

(Minitab Inc., Pennsylvania, USA).

3. Results and discussion

A total of seven runs (Table 1) made from fourteen trials were carried out for which the

encapsulation efficiency of microcapsules ranged from 73 % to 80 % and particle sizes ranged

from 27 to 72 µm. Microcapsules produced had low moisture content (≤ 4 %) and a

compressibility index of less than 20 %.

3.1. Encapsulation efficiency

As shown in Table 1, the total oil content of the powder for different conditions ranged from 34 %

to 37 %. The total oil content in the microparticles was less than the incorporated amount (40 %),

which indicated a loss of oil in the production of the powder. The total oil content increased with

an increase in pressure and decreased with temperature, but was not significantly influenced by

the emulsion feed flow rate, whereas the encapsulation efficiency of the microcapsules was

significantly affected by the pressure, temperature and emulsion feed flow rate. The

encapsulation efficiency was lower (p<0.05) at the emulsion feed flow rate of 5 mL/min than the

10
encapsulation efficiency at 3 and 4 mL/min (Table S2). When the pressure increased from 12 to

16 MPa (Table 1) at emulsion feed flow rate of 4 mL/min, the percentage of the surface oil

decreased and the encapsulation efficiency of the powders increased. Increasing the temperature

from 40 to 60 ºC increased the surface oil percentage and decreased the encapsulation efficiency

of the powder.

The increase in total oil content and encapsulation efficiency with increasing pressure and

decreasing temperature may be related to the variation in the density of the SC-CO2. After the

emulsion was atomized at supercritical conditions, competing mutual diffusion occurs between

the absorption of SC-CO2 into the droplet and water evaporation. In other words, CO2 probably

caused the oil in the emulsion droplet to swell and thus to increase in surface area which

promoted removal of the water that was followed by material collapse and encapsulation of the

oil within the wall matrix [21]. The feed is an oil-in-water emulsion, and the encapsulation

agents, namely maltodextrin and sodium caseinate, were dissolved in the water, so that the oil is

surrounded by an encapsulation agent in the emulsion. The precipitation of the encapsulation

agents (water being removed by SC-CO2) entrap VCO, so it can be expected that the oil is

dispersed in the matrix of encapsulation agents. As the SC-CO2 exits the droplets, it may carry

some oil because VCO is relatively soluble in SC-CO2. VCO has a solubility of 0.0109 g/g CO2

at 40 ºC and 16 MPa as calculated with the Chrastil model from data of Zuknik et al. [22].

CO2 has a high density and a high solvent power at high pressures (ca. 16 MPa) and moderate

temperatures (ca. 40 ºC). Thus, the particle formation process occurs faster, and oil is trapped

inside the particles more efficiently for runs at high pressure and low temperature. As a result,

the dry particles retain more oil and have a lower amount of surface oil, which produces high

encapsulation efficiency. The highest encapsulation efficiency (80.2 %) was achieved at 16 MPa,

11
40 ºC and 3 mL/min emulsion feed flow rate, which was comparable to that obtained by spray

drying (80.5 %) [6].

3.2. Characterization of the powder

Table 1 shows that the mean particle size ranged from 40 to 60 µm as the pressure increased

from 12 to 16 MPa. Conversely, as the temperature increased from 40 to 60 ºC, the mean

diameters of the particles decreased. The mean particle size was the smallest (27 µm) at the

highest temperature of 60 ºC. The diffusivity of CO2 decreases with an increase in pressure (at

fixed temperature) or a decrease in temperature (at fixed pressure). This change reduces the mass

transfer among the droplets and the CO2 around them, which slows the particle formation process

and produces large particles, thus, particle sizes increased with an increase in pressure or a

decrease in temperature [23]. The effect of the emulsion feed flow rate was evaluated over a

range from 3-5 mL/min. The mean particle size decreased from 67 to 48 µm when the emulsion

feed flow rate increased from 3 to 4 mL/min; however, further increase in the emulsion feed flow

rate to 5 mL/min formed larger particles with a diameter of 72 µm. The largest particle sizes

were obtained at a feed flow rate of 5 mL/min. At a constant CO2 flow rate, the SC-CO2/water

ratio became smaller with increasing emulsion feed flow rate, which probably slowed down the

mass transfer between the droplets and the surrounding CO2 such that larger particles were

formed [24].

The operating conditions had no considerable effect on the range of the particle size distribution

(PSD) when varying the operating parameters (Figure S1- S3). However, bimodal PSDs were

noted except at 60 ºC The particle size distribution of VCO microcapsules produced at 16 MPa,

60 ºC and emulsion feed flow rate of 4 mL/min gave the smallest particle sizes as shown in

12
Figure 1. The distribution curve (Figure 1) demonstrated unimodal PSD with narrow particle size

(d10 = 11 µm and d90 = 36 µm). Formation of fine particles with homogenous distribution of size

was most likely due to the high mass transfer between the sample droplets and CO2 that enhances

the diffusion of CO2 into the droplet and evaporation of solvent from droplets which leads to

precipitation of small particles with narrow distribution [25]. It was found that the distribution

curve of the VCO particles was close to a Gaussian distribution pattern which is observed for

many particle types [26–29].

The moisture content of the powders ranged from 2.9 % to 4.1 % as shown in Table 1, with no

significant differences due to process conditions. The moisture content of VCO microcapsules

obtained in this study was slightly higher than VCO microcapsules obtained from spray drying

(2.4 % – 2.9 %) [6] which can be explained by the water having low solubility in SC-CO2 which

results in the water evaporation rate not being as efficient as that in spray drying due to the low

temperature used in the supercritical process.

The operating parameters did not have a significant effect (p>0.05) on the bulk density (0.3

g/cm3) of the powder (Table 1) and were comparable to the bulk density values of other essential

oil microcapsules (0.24 to 0.33 g/cm3) [30,31]. High bulk density is favorable for oil powders as

this may signify that less gas is occluded in the spaces between the particles [32].

Powders obtained at 16 MPa, 60 ºC, 4 mL/min had the lowest compressibility index (9.9)

whereas powders produced at 14 MPa, 50 ºC, 4mL/min had the highest compressibility index

(19.1) as shown in Table 1. The compressibility index of the powders produced was not

significantly influenced by the pressure, temperature and emulsion feed flow rate (Table S2).

The compressibility index was inversely proportional to the flowability, which indicates that the

13
materials are easier to compress than they are flowable. In general, a powder with a

compressibility index of less than 20 % indicates superior flowability [17].

3.3. Morphology

SEM images of particles with the highest (Figure 2a) and lowest (Figure 2b) encapsulation

efficiency are presented in Figure 2. The microcapsules obtained at both conditions were

spherically shaped and well-dispersed. Microcapsules produced at 16 MPa, 40 ºC, 3 mL/min

(Figure 2a) had smooth surfaces with no apparent cracks or fissures. On other hand, the

microcapsules produced at 12 MPa, 50 ºC, 4 mL/min showed wrinkled surfaces with pores on

some particles (Figure 2b) which could be the result of released encapsulated oil. It can be

deduced that the encapsulation conditions had no strong effect on the morphology of the

microcapsules for the conditions studied. Similar spherically shaped particles have been reported

for other oil microcapsules [33–37]. Morphology of particles in this study was very different

from those that are typically obtained with spray drying. Spray dried powders often have

concave, dented, shriveled, rough or distorted surfaces or deflated spherical shapes due to

inconsistent shrinkage of particle walls during high-temperature drying [15,31,38–43].

The inner structure of VCO microparticle is shown in Figure 3. The micrograph shows that VCO

was dispered in the wall matrix as small droplets and had a homogeneous core distribution. This

result is similar to the inner structure of other oil microcapsules reported in the literature [44–46].

3.4. Antioxidant activity

Table 2 shows the antioxidant activities (AA) of the raw VCO and the encapsulated VCO oil.

The AA of encapsulated VCO was negatively influenced by the encapsulation process in which

the AA is probably related to the thickness of the encapsulating material. The AA of the

14
encapsulated oil was not statistically different (p>0.05) for the DPPH assays from 40 ºC to 60 ºC

(Table S3), which indicates that the moderate temperatures employed in the particle formation

with the SC-CO2 environment process could prevent the AA degradation of VCO. The AA of the

encapsulated oil was not significantly different (p>0.05) for the ABTS assay at emulsion feed

flow rates from 3 to 5 mL/min. In ABTS assays, the AA of encapsulated VCO of this study had a

maximum retention of 84 % whereas spray dried VCO microcapsules with the same wall

materials had 79 % of AA retention [6]. Several researchers have reported that the

microencapsulation process reduces the antioxidant properties of encapsulated oils for which the

AA of crude palm oil and extra virgin olive oil have been reported to be degraded [14,47]. The

present encapsulated VCO showed a 20 to 30 % degradation in AA.

3.5. Fatty acid composition (FAC)

Chromatographic analyses of the encapsulated products revealed that the VCO contained a high

percentage of saturated MCFAs. The FAC of the encapsulated oils (Table 3) was similar to the

FAC of the raw oil, where the predominant fatty acid was lauric acid (C12:0). The process

conditions did not significantly affect the majority of the fatty acids (Table S4). The FAC for

both the encapsulated oil and the raw oil satisfy the specification of the VCO standard in

accordance with the Codex standard (Codex Alimentarius Commision, 2003) [48] and the

Malaysian Standard (2007) [49]. Many authors have reported that the FAC of encapsulated oil is

not affected or was only slightly affected by microencapsulation [14,50–52].

4. Conclusions

VCO was successfully microencapsulated with SC-CO2 spray drying of an oil-in-water emulsion.

Encapsulation efficiency and particle size was positively associated with pressure but negatively

15
correlated with temperature. The moisture content, bulk density, compressibility index and

morphology of the powder were not significantly affected by the operating parameters. The

process variables were shown to have minor effects on the antioxidant activity and fatty acid

composition of the encapsulated oil. It can be concluded that SC-CO2 spray drying is an efficient

method for encapsulating VCO and has excellent potential for producing functional foodstuffs.

Acknowledgments

Financial support of this study was provided by Geran Putra Universiti Putra Malaysia,

vot number 9419700. We gratefully acknowledge Adirondack (M) Sdn. Bhd. for supplying the

virgin coconut oil. Research was performed in cooperation of the MoU between Research Center

of Supercritical Fluids (Tohoku University) and Supercritical Fluid Centre (Universiti Putra

Malaysia).

16
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24
Figure captions:

Figure 1. Particle size distribution curve of virgin coconut oil microcapsules encapsulated by
maltodextrin and sodium caseinate precipitated with supercritical carbon dioxide spray drying
with supercritical CO2 conditions at 16 MPa, 60 ºC and 4 mL/min of emulsion feed flow rate at
27 ºC and 16 MPa.

25
(a)

(b)

10 µm
JEOL 15KV x3,000 17mm

10 µm
JEOL 15KV x3,000 19mm

Figure 2. Morphology of virgin coconut oil microparticles precipitated with supercritical CO2
conditions at (a) 16 MPa, 40 ºC and 3 mL/min of emulsion feed flow rate (27 ºC and 16 MPa) (b)
12 MPa, 50 ºC and 4 mL/min of emulsion feed flow rate (27 ºC and 12 MPa).

26
 QUASI-S 15.0 kV 5.9mm x3.00k SE 10.0 µm

Figure 3. Micrograph of broken virgin coconut oil microparticles precipitated with supercritical CO2
conditions at 16 MPa, 40 ºC and 4 mL/min of emulsion feed flow rate (27 ºC and 16 MPa).

27
Table 1. Effect of operating variables on the encapsulation efficiency of virgin coconut oil
microcapsules using supercritical carbon dioxide spray drying and characteristics of the resulting
powdera.

Press Tempera Emuls Total Surfa Encapsul Particl Moist Bulk Compressi
ure ture (ºC) ion oil ce oil ation e size, ure density bility index
(MPa feed (%) (%) efficiency D(4,3) conte (g/cm3
) flow (%) (µm) nt (%) )
rate*
(mL/m
in)
36.1± 9.8± 39.5±1 4.1±1. 0.32±0
12 50 4 72.7±0.5 6 .04 14.5±5.0
0.4 0.2 3.8

35.8± 9.4± 59.9±1 3.9±0. 0.32±0

14 50 4 73.8±0.5 6 .03 19.1±3.3
0.6 0.1 4.7

37.5± 8.6± 51.5±1 2.9±0. 0.30±0

16 50 4 77.0±0.4 5 .03 14.6±4.7
0.4 0.2 6.2

37.2± 7.5± 48.3±1 3.4±0. 0.31±0

16 40 4 79.8±0.8 2 .02 14.9±2.0
0.3 0.3 0.9

34.9± 9.5± 26.7±1 4.0±0. 0.30±0

16 60 4 72.9±0.6 3 .01 9.9±2.1
0.5 0.2 0.8

37.0± 7.3± 66.9±1 3.9±0. 0.31±0

16 40 3 80.2±0.6 4 .04 11.6±1.7
0.4 0.2 4.4

37.0± 8.2± 72.1±1 3.7±0. 0.29±0

16 40 5 77.8±0.7 5 .02 12.7±5.2
0.7 0.2 3.8
Spray dried VCO powder
38.6 7.47 80.5 229.7 2.35 0.29 -
[6]
a) Ratio of VCO to encapsulating agent was 2:3 where VCO loaded was 40 % in dry weight.
Data are indicated as means ± standard deviation

* Flow rates at 27 oC and at the respective pressure.

28
Table 2. Antioxidant activity of the encapsulated virgin coconut oil (VCO) for the range of
operating parameters studied for spray-drying of VCO emulsions with supercritical CO2.

Pressure (MPa) Temperature (ºC) Emulsion feed DPPH value, ABTS value,
flow rate* mmol BHT mmol trolox
(mL/min) equivalent/ml oil equivalent/ml oil

12 50 4 0.86±0.04 0.90±0.06

14 50 4 0.64±0.10 0.80±0.04

16 50 4 1.00±0.08 0.90±0.06

16 40 4 0.99±0.08c 1.25±0.03

16 60 4 1.00±0.04 1.18±0.06

16 40 3 0.83±0.04 1.20±0.02

16 40 5 1.05±0.10 1.21±0.02

Raw VCO 1.13±0.08 1.48±0.13

Notes: Data are indicated as averaged values ± standard deviation.

* Flow rates at 27 oC and at the respective pressure.

29
Table 3. Fatty acid composition of the encapsulated virgin coconut oil (VCO) for the range of
operating parameters studied for spray-drying of VCO emulsions with supercritical CO2.

Press Tempera Emulsion feed C8: C1 C1 C14: C1 C1 C18: C18:

ure ture (ºC) flow rate* 0 0:0 2:0 0 6:0 8:0 1 2
(MPa (mL/min)
)

7.7 6.4 50. 17.6 7.8 3.2

12 50 4 5.43 0.95
7 0 77 5 3 1

7.1 6.1 50. 18.3 8.2 3.2

14 50 4 5.49 0.97
1 9 54 0 0 1

7.2 6.1 49. 18.6 8.5 3.2

16 50 4 5.41 0.96
1 4 85 9 4 2

7.7 6.3 50. 18.1 8.0 3.1

16 40 4 5.26 0.92
3 1 37 9 9 3

7.0 6.1 50. 18.2 8.3 3.3

16 60 4 5.72 1.01
1 5 13 1 5 6

7.2 6.1 49. 18.5 8.4 3.2

16 40 3 5.46 0.97
3 6 88 8 8 5

7.8 6.2 49. 18.5 8.3 3.1

16 40 5 5.26 0.93
2 5 68 5 8 4

Raw VCO 8.0 6.2 48. 18.9 8.5 3.1 5.15 0.89
3 8 99 8 8 0

Notes: Data are indicated as mean values.

* Flow rates at 27 oC and at the respective pressure.

30