Journal of Food Engineering 89 (2008) 204209

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Journal of Food Engineering

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a-Tocopherol nanodispersions: Preparation, characterization and

stability evaluation
Jean Ne Cheong a, Chin Ping Tan a,*, Yaakob B. Che Man a, Misni Misran b
a
Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia
b
Department of Chemistry, University of Malaya, 50603 Kuala Lumpur, Malaysia

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

Article history: A top down approach based on emulsicationevaporation technique was used to prepare nanodisper-
Received 21 November 2007 sion of a-tocopherol. Physicochemical properties of the prepared nanodispersions were investigated
Received in revised form 13 April 2008 under combination of the processing parameters (pressure and cycle) and ratio of aqueous:organic. Stor-
Accepted 19 April 2008
age study was performed for 3 months to evaluate the stability of all the prepared nanodispersions. The
Available online 28 April 2008
results showed that homogenization pressure have signicant (P < 0.05) inuence on the droplet diam-
eter and size distribution. On the contrary, the processing cycle had not signicant (P > 0.05) effect on
Keywords:
the droplet diameter and size distribution of the prepared nanodispersion. Droplet diameters in the range
a-Tocopherol
Nanodispersion
of 90120 nm were obtained for the prepared a-tocopherol nanodispersions. During storage duration,
High-pressure homogenization there were no signicant (P > 0.05) changes in mean diameters while the concentrations of a-tocopherol
Emulsicationevaporation were signicantly (P < 0.05) reduced for all prepared nanodispersions. In general, it is shown that emul-
Physicochemical properties sicationevaporation technique can be used as a suitable technique for the production of a-tocopherol
Storage stability nanodispersions with narrow size distribution.
2008 Elsevier Ltd. All rights reserved.

1. Introduction to the special feature of nanodispersions whereby it has been

Natural antioxidants such as a-, b-, c-, d-tocopherol and tocot-
rienols are widely used in vitamin supplementation and as antiox-
idants in the food, cosmetic and pharmaceutical industries
(Constantinides et al., 2006). The general trend of biological activ-
ity of some compounds of vitamin E has been reported as follows:
a-tocopherol > b-tocopherol > a-tocotrienol > c-tocopherol > b-
tocotrienol > d-tocopherol (Abidi, 2000). Vitamin E, a lipid-soluble
antioxidant, is often regarded to a-tocopherol.
Functional lipids are bioactive compounds with substantial
health benet. However, poor water solubility of these compounds
has made their use problematic in food formulation (Tan and Nak-
ajima, 2005). Most of the antioxidants are almost insoluble in
water or show a very low water solubility leading to insufcient
bioavailability especially after oral or parenteral/intravenous
administration and very often below the therapeutic level (Muller
and Peters, 1998). Hence, issue related to the solubility of this
ingredient has become one of the major considerations in the food
industry.
Great attentions have been paid for the research and develop-
ment to overcome this problem. Nanodispersion is an alternative
and promising approach to overcome bioavailability problems
(Krause and Muller, 2001). The increase in bioavailability is due
* Corresponding author. Tel.: +60 3 89468418; fax: +60 3 89423552.
E-mail address: tancp@putra.upm.edu.my (C.P. Tan).
shown that nanosized dispersion increases the surface areas and
dissolution velocity thus increasing the saturation solubility (Kra-
use and Muller, 2001; Jacobs et al., 2001). The emulsication
evaporation technique is one the techniques used for preparing
the nanosized dispersions (Mehnert and Mader, 2001). This tech-
nique has been used in incorporating an anti-helmintic drug: pra-
ziquantel into poly(lactic-co-glycolic acid) nanoparticles in the
treatment of schistosomiasis (Mainardes and Evangelista, 2005).
Lemos-Senna et al. (1998) prepared amphiphilic cyclodextrin nan-
ospheres using the emulsication-solvent evaporation method as a
promising carrier for hydrophobic drugs. Meanwhile, Jaiswal et al.
(2004) prepared biodegradable cyclosporine nanoparticles, a po-
tent immunosuppressive drug, by high-pressure emulsication-
solvent evaporation process to improve the oral bioavailability
and pharmacokinetics.
In general, the control in size reduction processes inuences the
properties of the produced materials. Size greatly relates to func-
tionality of food materials. A smaller size means bigger surface area
and is desirable to improve absorption and bioavailability of bioac-
tive compounds. High-pressure homogenization has been exten-
sively used to emulsify, disperse, mix and process the products in
various industrial sectors: chemical, pharmaceutical, specialty
foods, food science research and biotechnology (Floury et al.,
2004a; Roesch and Corredig, 2003; Jafari et al., 2007a). Homogeni-
zation processes are mostly possible in reducing the mean droplet
size, and simultaneously narrows the width of the size distribution

0260-8774/$ - see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2008.04.018
 J.N. Cheong et al. / Journal of Food Engineering 89 (2008) 204209
by reducing the number of microparticles and the polydispersity
index (Muller and Peters, 1998).
The main objective of this study was to prepare and
characterize the a-tocopherol nanodispersions based on the
emulsicationevaporation technique. Various processing parame-
ters were investigated during the production of nanodipsersions.
Nanodispersion containing a-tocopherol were obtained based on
the top-down approach by varying the power density of the
homogenizer and the number of homogenization cycles. In the
top-down approach, nano-objects are constructed from larger
entities (bulk system) without atomic-level control. In addition,
the prepared a-tocopherol nanodispersions were stored at 4 C to
determine their physicochemical stability.
2. Materials
a-Tocopherol (95%) was purchased from SigmaAldrich (M)
Sdn. Bhd. (Missouri, USA). Polyoxyethylene sorbitan mono-laurate
(Tween 20), analytical grade hexane and HPLC grade methanol
were purchased from Fisher Scientic (Leicestershire, UK).
3. Preparation of a-tocopherol nanodispersions
3.1. Pre-emulsication step
The organic phase containing a-tocopherol (1%, w/w) was rst
dissolved in hexane before dispersing into the aqueous phase con-
taining (1% w/w) of Tween 20 in deionized water using a conven-
tional homogenizer (Heidolph Diax-900, Schwabach, Germany) at
5000 rpm for 5 min to produce a coarse oil-in-water emulsion.
Three organic to aqueous phase ratios (in weight fraction) namely
1:9, 2:8 and 3:7 were prepared.
3.2. Preparation of nanodispersion
The resulting coarse pre-emulsion was immediately passed
through a high-pressure homogenizer (APV Lab 1000, Albertslund,
Denmark). Solvent was then removed from the ne emulsion by
using a rotary evaporator under reduced pressure (Eyela NE-
1001, Tokya Rikakikai Co. Ltd, Tokyo, Japan). Several batches were
prepared, using the mentioned steps, by varying the homogenizing
pressure (2080 MPa) and cycle (13 cycles). In addition, two dif-
ferent ratios of organic to aqueous phase (1:9 and 2:8) were used.
Each dispersion sample was then characterized for droplet size dis-
tribution, a-tocopherol concentration and for further storage eval-
uation (4 C with sampling at 4 weeks intervals for a period of 12
weeks).
4. Characterization of a-tocopherol nanodispersions
4.1. Particle size analysis
The dispersions were characterized in terms of particle size and
size distribution. Particle size analysis measurements were per-
formed using ZetaSizer Nano ZS (Malvern Instrument Ltd, Malvern,
UK.) Storage stability was assessed by monitoring the size as a
function of time (up to 3 months). The mean diameter measuring
the uctuation of the intensity of the scattered light which is
caused by the particle movement. The particle size of the prepared
a-tocopherol nanodispersions was described by the volume-
weighted mean diameter (D4,3). By denition, the volume-
weighted mean islet volume is the mean islet volume if droplets
are weighted proportional to their volume. This parameter can be
estimated without assumptions regarding the shape of the droplets
and provides unbiased information of three-dimensional size, in
205
contrast to the commonly used two-dimensional estimates of
mean droplet prole area. The polydispersity index (PI) is a mea-
sure for the width of the distribution. It is a measure for the width
of the distribution ranging from 0 (monodispersed) to 0.500 (rela-
tively broad distribution).
4.2. Sample preparation for a-tocopherol determination
The sample preparation procedures were modied from Iwases
work (2000). Bond Elut C18 cartridges (Varian, Harbor City, CA,
USA) were conditioned by washing with 5 ml of methanol and then
with 10 ml of deionized water prior to use. One-milliliter of sample
was applied to the conditioned Bond Elut C18 cartridge. The car-
tridge was then washed with deionized water (10 ml) and then
each with 5%, 25%, and 50% aqueous acetonitrile solution (5 ml)
sequentially, followed by elution with acetonitrile. The eluate
(10 ml) was further ltered with a membrane lter. An aliquot
(25 ll) of ltrate was injected into the HPLC.
4.3. Determination of a-tocopherol content
HPLC separation was performed with Shimadzu liquid chroma-
tography system (CT-10A VP, Shimadzu, Kyoto, Japan), equipped
with SPD-10AV UV-Vis detector, a LC-10AT pump system, and a
CT0-10A oven. Quantitative measurement of a-tocopherol content
was done at 295 nm. The a-tocopherol was separated on a
4.6  50 mm 3 lm silica gel column (Purospher STAR RP-18 en-
capped column; Merck, Darmstadt, Germany) with mobile phase
of methanol: water (99:1 v/v) at 1.0 ml/min. Oven temperature
was set at 40 C. The calibration of peak area versus a-tocopherol
concentration was linear in the concentration range of 0.2
1.0 mg/ml (R2 = 0.9931). Injections, in duplicate, were done at each
concentration for standards and samples. All results were ex-
pressed in mg/ml.
4.4. Storage stability
Prepared nanodispersions were placed in duplicate in amber
bottles and stored at 4 C for 90 days. The samples were used to
determine the droplet size, size distribution and retention of a-
tocopherol content at the interval of 4 weeks.
4.5. Statistical analysis
All experiments and measurements were duplicated. Data were
statistically analyzed using one-way analysis of variance procedure
(Minitab (Minitab 13) software package, State College, USA). The
signicant difference level was set a 0.05. Each reported value
was the mean of four analyses from two replications.
5. Results and discussions
5.1. General
The ability to alter the solubility of functional lipids is an attrac-
tive application, as the poor water solubility of lipids makes them
problematic in food formulations. Tan and Nakajima (2005) have
successfully prepared the preparation of b-carotene in nanodisper-
sions. They investigated the inuence of phase ratio and homoge-
nization conditions on droplet size and b-carotene content. As a
continuation from those studies, this paper focused on the
production of the a-tocopherol nanodispersions by high-pressure
homogenization (a top down approach) based on the emulsica-
tionevaporation technique. In top down approach, size reduction
is performed by application of force and the degree of control in
206 J.N. Cheong et al. / Journal of Food Engineering 89 (2008) 204209

size reduction processes inuences the properties of the produced Table 1

materials (Sanguansri and Augustin, 2006). In general, the mean
diameters of the nanodispersions were considerably small after
the homogenization process by a high-pressure homogenizer. In
a high-pressure homogenizer, the oil and water mixture is sub-
jected to intense shear forces, turbulence, cavitations, pressure gra-
dient and also recirculating regions, which lead to the break up of
the dispersed phase into smaller size (Floury et al., 2004b. In this
study, the dispersion was forced through a narrow homogenization
gap with a very high velocity, which lead to droplets size in the
nanometric range (typically having the range of 90120 nm) after
the emulsicationevaporation process. Bouchemal and co-work-
ers (2004) has successfully reported to produce a-tocopherol nano-
dipsersion of 171 2 nm using spontaneous emulsication
technique. However, high-pressure emulsication technique will
be more suitable and applicable in the modern emulsication tech-
nique because of the extreme emulsication conditions that is cre-
ation of very high density at a very short time on a small volume of
dispersion.
In the present study, a-tocopherol was rst dissolved in hexane
before dispersing to the continuous phase containing emulsier in
deionized water using a conventional homogenizer. The pre-coarse
dispersion in the micron range was further homogenized using a
high-pressure homogenizer. Hexane was then removed using an
evaporator under reduced pressure. The formation of a-tocopherol
nanoparticles happened as hexane diffuses into the aqueous phase
and evaporates at the water/air interface. Due to the high interfa-
cial tension between the organic and aqueous phases, emulsier
such as Tween 20 is used to allow the existence of interfacial ten-
sion gradients, which is crucial for formation of stable droplets.
Emulsiers play major roles in formation of nanoemulsion: by low-
ering the interfacial tension, Laplace pressure is reduced and hence
the stress needed to break up a drop is reduced (Tadros et al.,
2004). Marie and co-workers (2002) have shown that with emulsi-
ers concentration higher than 1% in the aqueous phase did not in-
duce a decrease in particle size. This may due to the low critical
micelle concentration value (60 mg/l) for Tween 20 (Helenius
et al., 1979). Stabilizer used has an effect on the long-term stability
(avoidance of aggregates), but had no effect on maximum disper-
sity and no effect on the shape of the produced nanodispersions
(Keck and Muller, 2005). Therefore, in this study we xed the type
and concentration of emulsier, while the inuence of organic to
aqueous phase ratio and homogenization conditions was investi-
gated. Tween 20, a non-ionic emulsier that absorbs very quickly
at the oilwater interface, has shown results in small particle for
various applications. For the use of suitable concentration of Tween
20, a preliminary study showed that 1% of Tween 20 in water was
adequate to produce a-tocopherol dispersion within the accept-
able nano range (data not shown).
Characteristic of droplet size (D4,3, nm) of a-tocopherol nanodispersions prepared
with different ratios of mixtures using two different homogenization pressuresA
Organic:aqueous phase ratio Pressure (MPa)B
20 80
1:9 106.6 2.7a 95.8 3.2a
2:8 119.6 1.5b 106.2 2.2b
3:7 146.0 5.0c 129.5 1.9c
A
Each value in the table represents the mean standard deviation of four mea-
surements from two replications.
B
Means within each column with different letters (a, b, or c) are signicantly
different from each other (p < 0.05).
portion would lead to a signicant (P < 0.05) increase in the parti-
cle size of the prepared nanodispersion regardless of the pressure
applied. The increase in the droplet sizes could be partly attributed
to an increase in the concentration of dispersed phase, which sub-
sequently leading to a higher probability of recoalescence of drop-
lets. Furthermore, as the organic volume increases, incomplete
covering of droplet interface by emulsier molecules, which favors
droplet aggregation and thus increases the mean droplet sizes
(Jafari et al., 2007b). In addition, this observation might be also
attributed to the reduction of the shear stress during homogeniz-
ing process, resulting from higher viscosity of the organic phase
and consequently a less favorable mixing efciency and larger
emulsion droplets (Jumaa and Muller, 1998). Due to this reason,
organic to aqueous ratio of 3:7 was not selected for subsequent
study.
5.3. Effect of homogenization parameters on the physicochemical
properties of a-tocopherol nanodispersions
To study the effect of different homogenization pressures in
preparing the a-tocopherol nanodispersions, two different
organic:aqueous ratios were xed (1:9 and 2:8). In general, a
high-pressure homogenization pressure ensures a good emulsica-
tion process and therefore leads to smaller particles. The efciency
of the emulsication process was recorded by measuring the mean
droplets diameter. The homogenization was performed using a
two-stage homogenizer with pressure ranging from 20 to
80 MPa. Table 2 shows the mean droplet diameters of nanodisper-
sions prepared in different operative conditions.
Generally, the results demonstrated that all of the pressures ap-
plied lead to a size reduction, although no consistent trend shown.
This shows that the intense turbulent and shear ow elds in the
gap generated at the applied homogenization pressure were
sufciently enough to break droplets, which were distinctly larger
than the gap width in the high pressure homogenizer. However,

5.2. Effect of organic/aqueous phase ratio on the size distribution of a-

tocopherol nanodispersions Table 2
Characteristic of droplet size of a-tocopherol nanodispersions prepared using
Table 1 shows the average droplet size parameters of a-tocoph- different homogenization pressureA
erol nanodispersions determined by the photon correlation Pressure Organic:aqueous phase ratioB
spectroscopy (PCS) technique for three different ratios (MPa)
1:9 2:8
(organic:aqueous), namely 1:9, 2:8 and 3:7. For three of these
Droplet size, Polydispersity Droplet size, Polydispersity
organic/aqueous ratios, the average droplets sizes signicantly
D4,3 (nm) index (PI) D4,3 (nm) index (PI)
(P < 0.05) increased with an increase in the organic phase portion
20 106.6 2.7a 0.32 0.09a 119.7 1.5a 0.21 0.01a
for both applied pressures; 20 MPa and 80 MPa.
40 104.1 2.9a 0.40 0.19b 120.7 0.4b 0.24 0.01b
In general, for the pressure applied at 20 MPa, the droplets size 60 102.8 3.2a 0.38 0.12c 110.8 0.6a,b 0.24 0.02c
distribution extended from 100 to 120 nm with an increase in the 80 95.8 3.2b 0.43 0.14d 106.2 2.2c 0.29 0.03c,d
portion of organic phase. Similar particle size distribution trend A
Each value in the table represents the mean SD of four measurements from
can be observed when pressure was applied at 80 MPa, with the two replications.
droplet size distribution obtained extended from 90 to 110 nm. B
Means within each column with different letters (a, b, or c) are signicantly
These results clearly showed that an increase of the organic phase different from each other (p < 0.05).
 J.N. Cheong et al. / Journal of Food Engineering 89 (2008) 204209 207

increasing pressure from 20 MPa to 60 MPa had no signicant the homogenization pressures lead to the increase of temperature
(P > 0.05) effect on the droplet size. Possible explanation would in the homogenizers chamber during emulsication, and thus in-
be the fact that this pressure range may not be able to reach the crease the droplet size.
energy threshold necessary to break the particle apart forming In addition to the droplet size analysis, the concentration of pre-
the desired nanodispersions (Floury et al., 2000). In addition, at pared nanodispersions of a-tocopherol was investigated. Tables 4
low shear rates, the forces are not large enough to disrupt the and 5 illustrated the results on the loss of a-tocopherol during
bonds holding the particle together (interfacial forces < disruptive preparation steps for ratios of 1:9 and 2:8, respectively. The con-
forces). According to Keck and Muller (2005), there is no linear tent of a-tocopherol from a freshly prepared coarse emulsion
relationship between the decrease in size and increase in pressure. was approximately 755 mg/l and 1800 mg/l for ratios of 1:9 and ra-
They observed that increasing the pressure stepwise by 50 MPa has tio 2:8, respectively.
not lead to stepwise decrease in a linear relationship. Jafari et al. For both organic/aqueous phase ratios, a-tocopherol contents
(2007c) also observed the same phenomenon for the preparation were signicantly (P < 0.05) decreased with an increase in the
of nanoemulsion of maltodextrin combined with a surface-active pressures and number of homogenizing cycles. After the prepara-
biopolymer. tion steps, the contents of a-tocopherol decreases for both the
In spite of the observation mentioned earlier, it was observed 1:9 and 2:8 ratios at 80 MPa for 3 cycles. It is well known that a-
that by increasing pressure to 80 MPa lead to a signicant tocopherol is sensitive to light, oxygen and heat. In a dynamic high
(P < 0.05) reduction of the droplet size distribution for both the or- pressure system, such as the high pressure homogenizer, tempera-
ganic/aqueous phase ratios. With the increased of pressure to ture rise in the chamber is expected, triggering a loss in the pre-
80 MPa, the energy required to deform the droplets are sufcient pared a-tocopherol in the prepared nanodispersion. In addition,
to overcome the Laplace pressure. These observations showed that the presence of heat, light and oxygen during evaporation process
the intensity of the high shear forces, and the turbulent and/or cav- also contributed to the losses a-tocopherol content. Therefore, a
itations produced during the homogenization process determining loss in the content of a-tocopherol in the sample, after high pres-
the droplet size. These results are in broad agreement with those sure homogenization, is predicted.
reported by Tan and Nakajima (2005). This is a good indication that
droplet size distributions can be controlled by adjusting the 5.4. Stability evaluation of prepared a-tocopherol nanodispersions
homogenization pressure. during storage
Considering both the two ratio of the aqueous:organic phases,
ratio 1:9 showed the smaller droplet size but wider distribution All prepared nanodispersions showed a good stability in term
than ratio 2:8. The main reason for this observation may not be ex- of droplet size distribution. Figs. 1 and 2 show the inuence of
actly claried. Tentatively, the differences might be due to the
organic:aqueous phase variations. Table 4
Becher (1967) showed that the number of times a product Changes in a-tocopherol concentration after the preparation steps (for the
passed through a homogenizer affected the mean droplet size organic:aqueous ratio of 1:9)A
and the droplet size distribution of the nal product. Increasing Pressure Number of After high-pressure After evaporation (mg/l)
of cycle numbers resulted in a decrease in average droplet size (MPa) cycles homogenization (mg/l)
and narrowing the droplet size distribution. However, the observa- ConcentrationB Loss ConcentrationB Loss
tion is not in agreement with our ndings (Table 3). In general, (%) (%)
there is no further decrease in mean droplet size after three 20 1 690 0a 8.6 645 21a 14.6
homogenizing cycles. There was a slight increase in the mean 40 1 650 14b 13.9 625 7ab 17.2
droplet size indicating the maximum dispersity at the given power 60 1 640 14b 15.2 610 28b 19.2
density has been reached. One of the possible explanations is that 80 1 615 7c 18.5 570 14c 24.5
80 2 605 21d 19.9 565 21c 25.2
the droplet size has getting more and more perfect and has reach
80 3 545 21e 27.8 475 7d 37.1
its maximum dispersivity (over-processing), which means that
A
the force and energy required to break the droplets seems to in- Concentrtaion of a-tocopherol after 1st stage homogenization was
755 7.1 mg/l.
crease rather exponentially. Furthermore, temperature can be an B
Each value in the column represents the mean standard deviation of four
important parameter in emulsication affecting both viscosity of analyses from two replications. Means within the column with different super-
the dispersed and continuous phase and also the nature of the scripts are signicantly (P < 0.05) different.
emulsier as a consequence of phase inversion temperature and
its solubility, which can lead to deterioration and would favors
aggregation and consequently affects its stability (Joscelyne and Table 5
Tragardh, 2000; Marie et al., 2002; Floury et al., 2000). Increasing Changes in a-tocopherol concentration after the preparation steps (for the
organic:aqueous ratio of 2:8)A

Table 3 Pressure Number of After high-pressure After evaporation (mg/l)

Characteristics of droplet size distribution of a-tocopherol nanodispersions prepared (MPa) cycles homogenization (mg/l)
with different homogenization cycles and two different ratios of mixture (at 80 MPa)A
ConcentrationB Loss ConcentrationB Loss
Ratio Number of Droplet size, D4,3 Polydispersity index (%) (%)
(organic:aqueous) cycle (nm)B (PI)B
20 1 1750 14a 2.8 1650 36a 8.3
1:9 1 95.8 3.2a 0.43 0.14 40 1 1770 7a 1.7 1620 34a 10.0
2 108.8 0.8b 0.32 0.10 60 1 1690 7b 6.1 1500 72a 16.7
3 109.7 0.6b 0.32 0.05 80 1 1560 14b 13.3 1350 67b 25.0
2:8 1 106.2 2.2a 0.29 0.03 80 2 1460 35b 18.9 1420 67b 21.1
2 107.3 1.5a 0.29 0.01 80 3 1460 21c 18.9 1430 64b 20.6
3 114.9 2.1b 0.24 0.01 A
Concentration of a-tocopherol after 1st stage homogenization was
A
Each value in the table represents the mean SD of four measurements from 1800 2.12 mg/l.
B
two replications. Each value in the column represents the mean standard deviation of four
B
Means within each column with different letters (a, b, or c) are signicantly analyses from two replications. Means within the column with different super-
different from each other (P < 0.05). scripts are signicantly (P < 0.05) different.
208 J.N. Cheong et al. / Journal of Food Engineering 89 (2008) 204209

120 700

Alpha tocopherol concentration (mg/l)

115 600

110 500

105 400
Droplet Size (nm)

100 300
20MPa, 1 cycle
40Mpa, 1 cycle
200
95 60MPa, 1 cycle
20MPa, 1 cycle 80MPa, 1 cycle
40MPa, 1 cycle 100 80MPa, 2 cycles
90
60MPa, 1 cycle 80MPa, 3 cycles
80MPa, 1 cycle 0
85 0 4 8 12
80MPa, 2 cycles
Storage Duration (week)
80MPa, 3 cycles
80
Fig. 3. Changes in a-tocopherol content for a-tocopherol nanodispersions prepared
0 4 8 12
using various homogenization conditions during storage at 4 C (for organic:aque-
Storage Duration (Week) ous phase ratio of 1:9).

Fig. 1. Changes in mean droplet diameter for a-tocopherol nanodispersions during

the duration of storage for organic:aqueous phase ratio 1:9 (12 weeks).
1800

1600
Alpha tocopherol concentration (mg/l)

140
1400

130 1200

1000
120
Droplet size (nm)

800
20MPa, 1 cycle
600 40MPa, 1 cycle
110
60Mpa, 1 cycle
400
80MPa, 1 cycle
100 20MPa, 1 cycle 80MPa, 2 cycles
200
40MPa, 1 cycle 80MPa, 3 cycles
60MPa, 1 cycle 0
90 80MPa, 1 cycle 0 4 8 12
80MPa, 2 cycles Storage Duration (week)
80MPa, 3 cycles
80 Fig. 4. Changes in a-tocopherol content for a-tocopherol nanodispersions prepared
0 4 8 12 using various homogenization conditions during storage at 4 C (for organic:aque-
Storage Duration (Week) ous phase ratio of 2:8).

Fig. 2. Changes in mean droplet diameter for a-tocopherol nanodispersions during

the duration of storage for organic:aqueous phase ratio of 2:8 (12 weeks).
storage on the droplet size distribution of a-tocopherol nanodi-
spersions for 1:9 and 2:8 ratios, respectively. The droplet diameter
and size distribution are important parameters affecting emulsion
stability (Saito et al., 2006; Heurtault et al., 2003). Results indicated
that the emulsicationevaporation technique proved to be an ef-
cient way to stabilize the droplet size of the prepared nanodisper-
sions for both ratios. a-Tocopherol nanodispersions prepared from
both organic/aqueous ratios were physically stable throughout the
12 weeks of storage period at 4 C.
However, during storage at 4 C, there was a decrease in the a-
tocopherol concentration. Figs. 3 and 4 show the degradation pro-
le of a-tocopherol as a function of storage time. The degree of
degradation of a-tocopherol content was found to increase with
a decrease in the droplet diameter, with the higher degradation
for the ratio 1:9. Numerous factors had an inuence on the degra-
dation, which include process factor (temperature, homogeniza-
tion equipment) and storage condition. The process of breaking
up droplets during emulsication caused the increase in the cham-
bers temperature to rise. The exposure of temperature, the inten-
sity of light, and oxidation might have promotes degradation of
content during storage. Another possible explanation would be
the large surface area of the prepared nanodispersions due to the
nanometer range. This may signicantly reduce the stability of
the prepared a-tocopherol content.
6. Summary
The observations presented in this study conrmed that the
nanosized droplets formed using emulsicationevaporation is rel-
atively a simple and effective technique for producing a-tocoph-
erol nanoemulsions in a range of 100120 nm. Droplet size of
a-tocopherol can be produced in a controlled way by adjusting the
processing parameters pressures and cycle numbers accordingly.
In study, smaller droplet size was obtained for nanodispersion pre-
pared using 80 MPa during the emulsication step. This holds true
as long as the a-tocopherol/interfacial area ratio is large enough to
cover the newly formed interface in such a way that recoalescence
are minimized. In the case of homogenizing cycle, recoalescence or
the so-called over-processing occurs with increasing number of
homogenizing cycles. The a-tocopherol nanoemulsions showed
 J.N. Cheong et al. / Journal of Food Engineering 89 (2008) 204209
good physical stability. However, the tocopherol content of the
prepared nanodispersions was signicantly (P < 0.05) reduced dur-
ing storage. The number of poorly soluble functional lipids are
steadily increasing, especially lipids which simultaneously poorly
soluble in water and in non-aqueous media. Therefore, there will
be a high demand for formulations overcoming the problem re-
lated to these solubility problems. Further study will be required
to develop suitable emulsier system to protect the prepared func-
tional lipid of nanodispersion from degradation during prolonged
storage.
Acknowledgement
Financial support of this work by Ministry of Science, Technol-
ogy and Innovation of Malaysia through research Grant IRPA 09-
02-04-10059 EAR is gratefully acknowledged.
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