food and bioproducts processing 8 9 ( 2 0 1 1 ) 6772

Contents lists available at ScienceDirect

Food and Bioproducts Processing

journal homepage: www.elsevier.com/locate/fbp

Comparison of different extraction methods for the

extraction of major bioactive avonoid compounds from
spearmint (Mentha spicata L.) leaves
Mandana Bimakr a, , Russly Abdul Rahman a,b , Farah Saleena Taip a , Ali Ganjloo b ,
Liza Md Salleh a,d , Jinap Selamat c , Azizah Hamid c , I.S.M. Zaidul c
a

Department of Process and Food Engineering, Faculty of Engineering, University Putra Malaysia, 43400 Serdang, Selangor, Malaysia
Department of Food Technology, Faculty of Food Science and Technology, University Putra Malaysia, 43400 Serdang, Selangor, Malaysia
c Department of Food Science, Faculty of Food Science and Technology, University Putra Malaysia, 43400 Serdang, Selangor, Malaysia
d Faculty of Chemical and Natural Resources Engineering, Universiti Teknologi Malaysia, 81310 Skudai, Johor, Malaysia
b

a b s t r a c t
Different bioactive avonoid compounds including catechin, epicatechin, rutin, myricetin, luteolin, apigenin and
naringenin were obtained from spearmint (Mentha spicata L.) leaves by using conventional soxhlet extraction (CSE)
and supercritical carbon dioxide (SC-CO2 ) extraction at different extraction schemes and parameters. The effect of
different parameters such as temperature (40, 50 and 60 C), pressure (100, 200 and 300 bar) and dynamic extraction time (30, 60 and 90 min) on the supercritical carbon dioxide (SC-CO2 ) extraction of spearmint avonoids was
investigated using full factorial arrangement in a completely randomized design (CRD). The extracts of spearmint
leaves obtained by CSE and optimal SC-CO2 extraction conditions were further analyzed by high performance liquid
chromatography (HPLC) to identify and quantify major bioactive avonoid compounds prole. Comparable results
were obtained by optimum SC-CO2 extraction condition (60 C, 200 bar, 60 min) and 70% ethanol soxhlet extraction.
As revealed by the results, soxhlet extraction had a higher crude extract yield (257.67 mg/g) comparing to the SCCO2 extraction (60.57 mg/g). Supercritical carbon dioxide extract (optimum condition) was found to have more main
avonoid compounds (seven bioactive avonoids) with high concentration comparing to the 70% ethanol soxhlet
extraction (ve bioactive avonoids). Therefore, SC-CO2 extraction is considered as an alternative process compared
to the CSE for obtaining the bioactive avonoid compounds with high concentration from spearmint leaves.
2010 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
Keywords: Spearmint (Mentha spicata L.); Bioactive avonoid; Supercritical carbon dioxide (SC-CO2 ) extraction; Conventional soxhlet extraction (CSE); High performance liquid chromatography (HPLC)

1.

Introduction

Spearmint belongs to the genus Mentha in the family Labiateae (Lamiaceae) (Sweetie et al., 2007). A number of studies
have found that herbs of the Lamiaceae family are a potential
source of natural antioxidants (Choudhury et al., 2006). Rosemary (Rosmarinus ofcinalis L.), sage (Salvia ofcinalis L.), thyme
(Thymus vulgaris L.) and lavender (Lavendula angustifolia Mill.)
are native to the Mediterranean region; balm (Melissa ofcinalis L.) and spearmint (Mentha spicata L.) are common plants

in Britain and other European countries (Wang et al., 2004;

Paranjpe, 2001).
Different surveys have shown that herb extracts are useful as stabilizers of edible oils. Most studies on antioxidant
compounds in the Lamiaceae family are directed to phenolic
diterpenes, avonoids and phenolic acids (Kivilompolo and
Hyotylainen, 2007). Flavonoids which are widely distributed
in the leaves, seeds, bark and owers of plants are a broad
class of low molecular weight compounds. Flavonoids are a
kind of highly effective antioxidant and less toxic than syn-

Corresponding author. Tel.: +60 142679858; fax: +60 3 89423552.

E-mail addresses: mandanabimakr@yahoo.com (M. Bimakr), russly@food.upm.edu.my (R.A. Rahman).
Received 23 December 2008; Received in revised form 2 March 2010; Accepted 3 March 2010
0960-3085/$ see front matter 2010 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.fbp.2010.03.002

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food and bioproducts processing 8 9 ( 2 0 1 1 ) 6772

thetic antioxidants such as BHA and BHT. Therefore, these

secondary plant phenolics have received the greatest attention and have been studied extensively (Heim et al., 2002;
Naczk, 2004; Syed and Sharma, 2001). Effective separation of
antioxidants (high extraction yield and concentration of bioactive compounds) from a complex plant matrix is a difcult
procedure due to co-extraction of other various compounds,
which are undesirable in antioxidant extract. Different extraction techniques such as percolation, soxhlet and supercritical
uid extraction have been used to isolate antioxidants from
plants. Conventional soxhlet extraction (CSE) is a standard
technique which has been used for a long time. The main
disadvantages of conventional soxhlet extraction are long
extraction time and consumption of large amounts of used
solvents. This extraction method is also not suitable for the
extraction of thermo-sensitive compounds due to the possibility of thermal decomposition of target compounds as
extraction usually occurs at the boiling point of used solvent for a long time. However, conventional soxhlet extraction
comparing to supercritical uid extraction (SFE) is still widely
used due to its simplicity (Grigonis et al., 2005). Supercritical uid extraction with CO2 is an attractive method for food
industry applications due to its unique properties (Lin et al.,
1999).
The unique characteristic of this system is application of
gases above their critical points to extract selective soluble
components from a raw material (Cavero et al., 2006). However, CO2 is not a suitable solvent for the extraction of polar
compounds because it behaves as non-polar uid for certain
conditions of temperature and pressure. Therefore, this is
the main limiting step in its use for the separation of polar
phenolic compounds. But, in order to increase the polarity
of extraction solvent some food grade modiers like ethanol
can be used (Qingyong Lang, 2001). The objectives of the
present work were to investigate the effect of different parameters, such as pressure, temperature and dynamic extraction
time on the supercritical uid extraction of spearmint leaves
avonoids and to compare the extraction yield and concentration of avonoids in the extracts obtained under conventional
soxhlet and supercritical uid extraction. To the best of our
knowledge, no report has been yet appeared on the supercritical uid extraction (SFE) of Malaysian spearmint leaves
avonoid compounds.

2.

Materials and methods

2.1.

Materials

The leaves of spearmint (M. spicata L.) were obtained from

Cameron Highland in Pahang, Malaysia. After harvesting, the
leaves were separated and washed under tap water. Leaves
were dried at 40 C in a ventilated drying oven (1350FX, USA)
for 24 h and then stored at ambient temperature (22 C) in
the dark. The samples were ground in grinding mill (MX-335,
Panasonic, Malaysia) for 10 s to produce a powder with an
approximate size of 0.525 mm.

2.2.

Reagents

Carbon dioxide (CO2 , SFE grade), contained in a diptube

cylinder, was purchased from MOX Company in Malaysia.
Methanol (MeOH, analytical grade), ethanol (EtOH, 99.5%,
analytical grade) and petroleum ether (analytical grade)
were purchased from Scharlau chemical, European Union.
Methanol (MeOH, HPLC grade) was purchased from Fisher scientic chemical, USA. TFA (triuoroacetic acid, 98%) was
obtained from SigmaAldrich, Germany. All avonoid standards including (+)-catechin, ()-epicatechin, apigenin, rutin,
luteolin, myricetin and naringenin were purchased from
SigmaAldrich, Germany.

2.3.

Conventional soxhlet extraction (CSE)

Three grams (3 g) of dried and ground spearmint leaves were

placed in a soxhlet apparatus. Extraction was performed with
150 ml of an appropriate solvent for 6 h. After extraction, a
rotary vacuum evaporator (Eyela, A-1000S, Japan) at 40 C was
used in order to remove solvent. In this experiment four solvents were used: pure ethanol, methanol, petroleum ether and
70% ethanol. All extractions were performed in duplicate.

2.4.

Supercritical carbon dioxide (SC-CO2 ) extraction

Supercritical CO2 (SC-CO2 ) extraction was performed on a

supercritical uid extractor (ABRP200, Pittsburgh, PA, USA)
with the extractor volume 500 ml (Fig. 1). The ow rate of CO2 ,
the extraction temperature and pressure were adjusted by the

Fig. 1 Schematic diagram of supercritical uid extractor.

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food and bioproducts processing 8 9 ( 2 0 1 1 ) 6772

ICE software, and the extraction time was measured by the

stopwatch. Liquid CO2 was supplied from a gas cylinder. Before
liquid CO2 passed into the extraction vessel, lled with the
samples, by the means of a pump (P-50, Thar designs, Inc.
Pittsburg, PA, USA), it was pressurized to the desired pressure
and heated to the specied temperature in order to reach the
supercritical state. Absolute ethanol (EtOH) acted as the cosolvent and the ow rate was maintained at 3 g/min. In this
study, extractions were performed at three different temperatures (40, 50 and 60 C), three different pressure levels (100,
200 and 300 bar) and three different dynamic extraction time
(30, 60 and 90 min). The supercritical CO2 ow rate was maintained at 15 g/min and the duration of static extraction time
was xed to 30 min. The powdered plant material (30 g) was
mixed with 90 g glass beads (2.0 mm in diameter), placed into
the extractor vessel. The extractions were performed in duplicate. In this study, the SC-CO2 extraction of spearmint leaves
was planned according to the full factorial with complete randomized design (CRD) for the highest crude extract yield.

2.4.1.

Further processes

After extraction, the extract was collected and the co-solvent

(ethanol) was removed with vacuum rotary evaporator (Eyela,
A-1000 S, Japan) under reduced pressure at 40 C (water-bath
temperature). Then a gravimetric measurement was used
to obtain the amount of total crude extract weight. Afterward, to destruct glycosides bounds each sample was acid
hydrolysed as follows: 0.05 g sample + 16 ml H2 O + 24 ml pure
methanol + 5 ml HCl (6 M) (Perva-Uzunalic et al., 2006).
After 2 min manual homogenizing in order to complete the
effect of acid hydrolysis the sample was reuxed at 90 C for
2 h and nally cooled at room temperature. Then, the sample was ltered through lter paper 541, 70 mm. By using lter
syringe set, sample was ltered through 0.45-m nylon membrane lter. At the end, 20 l of extract obtained using SFE was
quantitatively analyzed by HPLC for the avonoid compounds
detection. The extractions were performed in duplicate.

2.5.
High performance liquid chromatography (HPLC)
analysis
The extracts obtained from optimum supercritical CO2 (SCCO2 ) extraction condition and conventional soxhlet extraction
(CSE) were analyzed using a HPLC which was composed of
a Water 600 pump Controller, 9486 tuneable absorbance UV
detector and equipped with an Eclipes XDR-C18 reversedphase column (25 cm 4.6 mm 5 m, Supelco, USA). The
volume of the injection loop was 20 l. Classic Millenium 2010
software was used for the manipulation of data processing.
The mobile phase used for analysis was solvent A: TFA (triuoroacetic acid) 2.5 pH in deionized water and solvent B
consists of methanol 100% (HPLC grade). MeOH 70% was also
required for washing the system. The avonoids were detected
at 280 nm. The temperature was set to room temperature and
ow rate was set at 1.0 ml/min. All of the main avonoid
compounds were identied by matching their retention time
against those of standard compounds. Quantity calculations
were made according to the linear calibration curves of standards.

2.6.

Statistical analysis

The optimization of the method can be carried out step by step

or by using an experimental design. In the present study the

process of extraction was optimized with complete randomized design (CRD) full factorial for a higher extraction yield
from spearmint (M. spicata L.) leaves. Independent variables for
SC-CO2 extraction were temperature (40, 50 and 60 C), pressure (100, 200 and 300 bar) and dynamic extraction time (30,
60 and 90 min). Data were subjected to analyses of variance
(ANOVA) and multiple comparison tests were performed using
LSDs test at the 95% of condence level. All the analyses were
carried out using the statistical software, Minitab v.14. A probability value of p < 0.05 was considered signicant. The factors
and levels investigated are reported in Table 1.

3.

Results and discussion

3.1.

Optimization of the experimental conditions

Optimization of the experimental conditions is a critical step

in the development of SC-CO2 extraction method due to the
effect of various parameters on the extraction yield. Generally,
extraction pressure, temperature and dynamic time are considered as important factors. The extraction yields obtained
under studied conditions were 30.1260.57 mg/g. In general,
a full evaluation of the effect of three selected factors from
three levels and duplicate experiments on the extraction yield
needed 54 (33 2) experiments.
The mean values of the extraction yields for the corresponding parameters at each level were calculated according
to the assignment of the experiment. The mean values of
the three levels of each parameter show how the extraction
yield changes when the level of that parameter is changed.
All the three studied parameters had signicant effect on the
extraction yield (p < 0.05). In Table 2 the average response of
each level about extraction yield was presented. Also, R-value
which is mentioned in Table 2 means range between three
average responses of each level about extraction yield. From
the R-value it can be concluded that temperature (R = 10.58)
had a dominant effect on the extraction yield followed by the
pressure (R = 10.38) and dynamic time (R = 8.12).

3.1.1.

Effect of temperature

Fig. 2 shows the effect of temperature on the extraction yield

of spearmint (M. spicata L.) leaves in SC-CO2 extraction at three
temperature levels of 40, 50 and 60 C. The density of CO2 at
constant pressure is reduced with increasing temperature and
leading to reduce the solvent power of supercritical CO2 . The
effect of temperature on solute solubility is different at pressures in the critical range. Near the system critical pressure,
the uid density is very sensitive to temperature. This might
be the reason that avonoid yield was changed signicantly
when temperature was changed over the range of 4060 C. A
moderate increase in temperature can lead to a large decrease
in uid density, with a consequent reduction in solute solubility (Roop et al., 1989). However, the increase in temperature
will also accelerate mass transfer and improve the extracTable 1 Experimental levels of the factors used in
complete randomized design (CRD) full factorial.
Factors

Pressure (bar)
Temperature ( C)
Dynamic time (min)

Levels
1

100
40
30

200
50
60

300
60
90

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food and bioproducts processing 8 9 ( 2 0 1 1 ) 6772

Table 2 Results obtained at the experimental condition using complete randomized design (CRD) full factorial.
Parameter

Yielda (mg/g)
L1c

Yielda (mg/g)
L2c

Yielda (mg/g)
L3c

Pressure
Temperature
Time

40.69 4.43
40.48 5.32
40.28 3.45

51.07 3.21
45.36 5.61
48.21 4.43

45.13 4.72
51.06 5.43
48.40 6.3

a
b
c

Rb
10.38
10.58
8.12

Values are mean SD of duplicate runs.

R-value means range between three average responses of each level about extraction yield.
Average responses of each level about extraction yield.

Fig. 2 Effect of temperature on the extraction yield of

crude extract at the constant pressure.
tion yield (Wang et al., 2008). The increase of temperature
can increase the vapour pressure of the extractable compounds. Thus, the tendency of the compounds to be extracted
is increased to pass in the supercritical uid phase (Reverchon
and De Marco, 2006). For a volatile solute, there is competition between its solubility in supercritical carbon dioxide and
its volatility (Pourmortazavi and Hajimirsadeghi, 2007). Therefore, it is difcult to predict the effect of temperature. In the
present study, the extraction yield increased with temperature
and the highest extraction yield (60.57 mg/g) was obtained at
60 C. In this manner, the solute vapour pressure played a key
role leading to increase in the extraction yield.

3.1.2.

Effect of pressure

Fig. 3 presents the effect of pressure on the extraction yield

of spearmint (M. spicata L.) leaves in SC-CO2 using three pressure levels of 100, 200 and 300 bar. According to the obtained

Fig. 4 Effect of dynamic extraction time on the extraction

yield of crude extract at the constant pressure.
results, the extraction yield increased with pressure from 100
to 200 bar, which was due to increase of SC-CO2 density at
higher pressures. However, an increase in the pressure level
above 200 bar led to an unexpected reduction in the extraction yield. This unexpected result can probably be related to
the reduced diffusion rates of the extracted compounds from
the plant matrix to the supercritical uid medium (Rezaei and
Temelli, 2000).
An increase of pressure can result in an increase in the
uid density, which alters solute solubility. Gomes et al. (2007)
have indicated that a higher recovery of volatile fractions and
a lower recovery of non-volatile fractions are obtained at high
pressure. Therefore, it is interesting to control the composition of the extract using pressure. In this study, the avonoid
yield increased with increasing pressure to a certain value.
Over this range of pressure, increasing uid density is presumably the main mechanism leading to a higher avonoid yield.
Above this range of pressure, a decreasing avonoid yield with
increasing pressure was observed. The volatility and polarity of extracted analytes might be responsible for the result
(Gomes et al., 2007; Wang et al., 2008).

3.1.3.

Fig. 3 Effect of pressure on the extraction yield of crude

extract at constant temperature.

Effect of dynamic extraction time

Fig. 4 shows the effect of dynamic time on the extraction

yield of spearmint (M. spicata L.) leaves in SC-CO2 by applying
three different levels of dynamic time including: 30, 60 and
90 min. At 100 bar pressure the extraction yield was increased
with dynamic time until 90 min. However, at higher pressures
(200 and 300 bar) the extraction yield increased with dynamic
extraction time until 60 min. It can be concluded that the solvent power of supercritical CO2 density is reduced at 100 bar
pressure due to the lower CO2 density and maximum yield
was obtained at 90 min. However, at higher pressures (200 and
300 bar) the extraction rate is higher and as a consequence the

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food and bioproducts processing 8 9 ( 2 0 1 1 ) 6772

Table 3 Identication and quantication of bioactive avonoid compounds extracted by conventional soxhlet extraction
(CSE) and supercritical carbon dioxide (SC-CO2 ) extraction.
Extraction mode

Extraction yield
(mg/g)a

Flavonoid content (mg/g)

Catechin Epicatechin

Conventional soxhlet extraction (CSE)

Methanol
267.33
Ethanol (99.5%)
218.0
Ethanol:water (70:30)
257.66
Petroleum ether
30.47
SC-CO2 extraction
(60 C, 200 bar and 60 min)

Rutin

Myricetin Luteolin Apigenin Naringenin

3.12e
4.24c
3.47d
2.34a

0.144
0.081
0.117

0.163
0.114
0.149

0.161
0.109
0.140

0.041
0.090

0.093
0.154
0.146

0.392
0.246
0.305

0.054

60.566 3.14b

0.140

0.156

0.148

0.117

0.657

0.270

0.249

Values in the yield column followed by letters are signicantly different (p-value < 0.05).
a

Values are mean SD of duplicate runs.

extraction yield kept increasing but after 60 min of extraction

time, the extraction yield dropped (Reverchon and De Marco,
2006). Therefore, as shown in Fig. 4, the highest extraction
yield was achieved at 60 min dynamic extraction time.

3.2.

Conventional soxhlet extraction (CSE) yields

Different solvents with different polarities were used to determine which one gives the highest recoveries of bioactive
avonoid compounds. Four solvents were used: (1) methanol,
(2) pure ethanol, (3) ethanol (70%) and (4) petroleum ether.
The extraction yield obtained by using each solvent is presented in Table 3. Based on the obtained results, the highest
extraction yield (267.3 mg/g) was found with methanol extraction and then with a little difference followed by ethanol
70% extraction (257.6 mg/g). The lowest extraction yield was
obtained by using petroleumether (30.4 mg/g), suggesting that
polar compounds in the plant matrix would be easier to
extract with a more polar solvent while lower polarity solvents enable to obtain the extracts with higher concentration
of bioactive compounds. The extracts which were obtained
from methanol, pure ethanol, ethanol 70% and petroleum
ether were then analyzed by HPLC to identify and quantify the
major bioactive avonoid compounds prole. With different
solvent, different avonoid compounds were extracted. The
avonoid compounds concentration in the extract obtained
with petroleum ether was undetectable. Using pure ethanol
ve avonoid compounds including catechin, epicatechin,
rutin, luteolin and apigenin were extracted from spearmint
(M. spicata L.) leaves. Apigenin had the highest concentration
(0.246 mg/g) among the other avonoids, which are obtained
with pure ethanol conventional soxhlet extraction.
The highest extraction yield (267.33 mg/g) was obtained
with methanol solvent, which extracted seven avonoid
compounds including catechin, epicatechin, rutin, myricetin,
luteolin, apigenin and naringenin. However, the concentrations of myricetin and naringenin were low (0.041 and
0.054 mg/g, respectively) and apigenin had the highest concentration (0.392 mg/g). The obtained extraction yield from
ethanol 70% conventional soxhlet extraction was near to the
obtained extraction yield from methanol soxhlet extraction
but extracted avonoids were same as compounds which are
extracted with pure ethanol soxhlet extraction. Five avonoid
compounds catechin, epicatechin, rutin, luteolin and apigenin
were extracted from spearmint leaves with pure ethanol and
ethanol 70%. But higher concentrations of more bioactive
avonoid compounds were detected with ethanol 70% due to

its higher polarity than pure ethanol. For example, in ethanol

soxhlet extraction apigenin with 0.246 mg/g had the highest
concentration in contrast to ethanol 70% the concentration
of dominant detected avonoid, apigenin, was reached to
0.305 mg/g. By adding water to the pure ethanol up to 30% for
preparing ethanol 70% the polarity of solvent was increased.
Therefore, polar compounds, avonoids, could be isolated better from herb matrix.

3.3.
Comparison of SC-CO2 extraction and CSE
methods
Different methods of natural matter extraction have different extraction yield and efciencies. Higher concentration
of natural bioactive compounds in the extracts is an important factor in the production of natural products while a
primary task in the industries is lower economic cost which
can be achieved by better extraction yield (Grigonis et al.,
2005). Based on the results obtained with methanol conventional soxhlet extraction the extraction yield and recovery
of avonoid compounds were better but due to its toxicity and less consumption in food industry comparison was
performed between the obtained results of optimum SC-CO2
extraction condition and ethanol 70% conventional soxhlet
extraction (Table 3). The composition of the extracts obtained
from SC-CO2 and conventional soxhlet extraction was strongly
different. According to the extraction yield results, ethanol
70% conventional soxhlet extraction (257.6 mg/g) had a higher
yield comparing to the SC-CO2 extraction (60.57 mg/g). But,
supercritical carbon dioxide extract (optimum condition) was
found to have better quality and more main avonoid compounds (seven avonoids) comparing to the ethanol 70%
conventional soxhlet extraction (ve avonoids). Flavonoid
compounds of plants have usually been extracted by the other
conventional extraction methods such as solvent extraction,
steam distillation. Some of the main disadvantages of all these
methods include: long extraction time, losses of volatile compounds and degradation of unsaturated compounds, resulting
unfavourable off-avour compounds due to heat (Grigonis et
al., 2005). Supercritical CO2 (SC-CO2 ) extraction has different
advantages over conventional soxhlet extraction (CSE) method
such as low operating temperature, thus no thermal degradation of most of the labile compounds, shorter extraction
duration and high selectivity in the extraction of target compounds. SC-CO2 extraction also, seems to be a cost-effective
process at laboratory scale, but a precise economic evaluation
will need additional experiments for establishing large-scale

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food and bioproducts processing 8 9 ( 2 0 1 1 ) 6772

units (Qingyong Lang, 2001). Therefore, it can be recommended as a suitable extraction method to isolate bioactive
avonoid compounds from spearmint (M. spicata L.) leaves.
However, for the complete extraction of the other avonoid
compounds may be higher pressure and extraction times
are needed. Further studies are in progress to quantitatively
assess the extract antioxidant power and the enrichment of
antioxidants at different extraction conditions.

4.

Conclusion

Generally, it may be possible to concentrate the avonoid compounds in spearmint (M. spicata L.) extracts by manipulating
extraction condition of SC-CO2 extraction. In this study the
effect of the three tested parameters including temperature,
pressure and dynamic extraction time were investigated and
it is revealed they have signicant effect on the extraction
yield. The highest extraction yield achieved at 60 C, 200 bar
and 60 min. The product of best condition (60 C, 200 bar
and 60 min) was analyzed by HPLC to identify and quantify major bioactive avonoid compounds. Seven avonoid
compounds including catechin, epicatechin, rutin, myrecitin,
luteolin, apigenin and naringenin with different concentration were identied and quantied in mentioned extract.
Based on the obtained conventional soxhlet extraction results,
polar solvents show better recoveries of avonoids and solvents with lower polarity enable to extract high concentration
of avonoids. Pure ethanol and ethanol 70% were safe solvents with lower toxicity than methanol. Also, good yield and
high concentration of bioactive avonoid compounds could be
isolated with these safe solvents from plant matrix. Despite
good results obtained with the conventional soxhlet extraction, supercritical CO2 extraction was tested to search for
a faster and better extraction method consuming less solvent, especially those that are undesirable in food industry.
Soxhlet extraction comparing to supercritical uid extraction
(SFE) possessing some disadvantages but it is still widely used
due to its simplicity. Conventional soxhlet extraction (CSE) is
not always acceptable for industrial applications due to long
extraction time, large consumption of hazardous solvents and
some other disadvantages. Therefore, supercritical CO2 (SCCO2 ) extraction could be an alternative extraction method.

Acknowledgment
The authors are grateful for the nancial support received
from the RMC, the Universiti Putra Malaysia for this project.

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