Innovative Food Science and Emerging Technologies 11 (2010) 485–490

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Innovative Food Science and Emerging Technologies

j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / i f s e t

Optimization of supercritical fluid extraction of bioactive compounds from grape

(Vitis labrusca B.) peel by using response surface methodology
Kashif Ghafoor a, Jiyong Park a, Yong-Hee Choi b,⁎
a
Biomaterial Process Engineering Lab, Department of Biotechnology, Yonsei University, 134-Sinchon Dong, Seodaemun Gu, Seoul 120-749, Republic of Korea
b
Food Engineering Laboratory, Department of Food Science & Technology, Kyungpook National University, 1370-Sankyuk Dong, Puk Gu, Daegu, 702-701, Republic of Korea

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

Article history: Supercritical fluid extraction (SFE) was applied for the extraction of valuable compounds from grape (Vitis
Received 6 August 2009 labrusca B.) peel. Extraction was carried out according to an orthogonal array design (OAD) and independent
Accepted 28 January 2010 variables selected were temperature, pressure and modifier concentration. SFE process was optimized by
Editor Proof Receive Date 9 February 2010 using response surface methodology (RSM) for the extract yield, total phenols, antioxidants and total
anthocyanins from grape peel. Effects of extraction temperature and pressure were found to be significant on
Keywords:
all responses. Optimal SFE conditions were identified as 45–46 °C temperature, 160–165 kg cm− 2 pressure
Supercritical fluid extraction and 6–7% ethanol as modifier for maximum extract yield (12.31%), total phenols (2.156 mg GAE/100 mL),
Optimization antioxidants (1.628 mg/mL) and total anthocyanins (1.176 mg/mL). Experimental values for response
Orthogonal array design variables at these optimal conditions match well with the predicted values. Grape peel extracts obtained by
Grape peel SFE showed more than 93% DPPH radical scavenging activities.
Bioactive compounds Industrial relevance: This study describes the response surface optimization of supercritical fluid extraction
Response surface methodology (SFE) process for the enhanced recovery of total phenols, antioxidant and anthocyanins from grape peel. SFE
uses CO2 as supercritical fluid which is environment friendly solvent; allows extraction at lower temperature
and the extracts obtained possess higher quality and safety. Industrially, it may be used as a promising
technique for the extraction of bioactive compounds from plant materials.
© 2010 Elsevier Ltd. All rights reserved.

1. Introduction of grape and grape derivatives are believed to be mainly due to the
antiradical and antioxidant properties of phenolic constituents
Grapes are among the most widely consumed fruits. They are rich (Lurton, 2003).
in polyphenols, with approximately 75% of grape polyphenols existing Extraction of functional components from plant materials is an
in the seeds and skin. Grape skin phenols may be classified as cell-wall important process and various techniques have been studied in this
phenols, which are bound to polysaccharides by hydrophobic regard (Spigno, Tramelli, & De-Faveri, 2008). Extraction is a major
interactions and hydrogen bonds; non-cell-wall phenols, encompass- step in the isolation, identification and use of phenolic compounds
ing phenols confined in the vacuoles of plant cells and phenols (Stevigny, Rolle, Valentini, & Zeppa, 2007). Extensive studies are being
associated with the cell nucleus (Pinelo, Arnous, & Meyer, 2006). carried out for the development of extraction processes which are
Phenolic compounds can be used in different therapeutic procedures novel and applicable to a variety of bioactive compounds. Supercrit-
with the purpose of free radical neutralization in biological systems ical fluid extraction (SFE) with supercritical CO2 has been used for the
(Heim, Tagliaferro, & Bobilya, 2002; Yilmaz & Toledo, 2004) and extraction from natural products and researchers have paid consid-
inhibition of oxidation of human low-density lipoproteins (Meyer, Yi, erable attention towards various aspects of this process (Lu et al.,
Pearson, Waterhouse, & Frankel, 1997). Proanthocyanidins are major 2007). Supercritical CO2 is an inert, non-toxic, environmentally safe
polyphenols in the grape skin and include condensed cyanidine-3- solvent and allows extraction at lower temperatures and relatively
glucosides, malvidin-3-glucosides, and peonidine-3-glucosides low pressures. The extracts obtained by SFE are of superior quality as
(Koeppen & Basson, 1966). Grapes anthocyanins possess strong compared to those obtained by conventional organic solvent
biological functions such as anti-inflammatory and antioxidant extraction methods (Friedrich & List, 1982; Gomez, Lopez, & La-
activities (Kong, Chia, Goh, Chia, & Brouillard, 2003; Vatai et al., Ossa, 1996). SFE extracts are also generally recognized as safe (GRAS)
2008) Positive physiological effects associated with the consumption to be used in food products, therefore it may serve as a very promising
technology in food processing (King, 2000).
The aim of our study was to optimize SFE variables such as
⁎ Corresponding author. Tel.: + 82 53 950 5777; fax +82 53 950 6772. temperature, pressure and concentration of ethanol as modifier for
E-mail address: [email protected] (Y.-H. Choi). the maximum extract yield, total phenolic compounds, antioxidants

1466-8564/$ – see front matter © 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.ifset.2010.01.013
486 K. Ghafoor et al. / Innovative Food Science and Emerging Technologies 11 (2010) 485–490

and total anthocyanins from Campbell Early grape (Vitis labrusca B.) A 200 μL properly diluted extract or a standard solution of varying
peel by using orthogonal array design and response surface concentrations was mixed with 400 μL Folin Ciocalteu reagent. The
methodology. deionized water was used for dilution and control. The solution was
diluted to a total volume of 4.6 mL and thoroughly mixed. After
2. Materials and methods incubation for 10 min at room temperature, 1 mL of 20% Na2CO3
solution was added then immediately mixed and incubated for 2 h.
2.1. Materials The absorbance was read at 765 nm on a spectrophotometer (TU-
1800; Human Corporation, Seoul, Korea). Gallic acid of 1 mg/mL was
Freshly harvested ripened grapes were purchased from a local used as the standard and the total phenolic compounds of the samples
farm in Gyeongbuk province of Korea and the grape cultivar was were expressed in milligram gallic acid equivalent per 100 mL (mg
identified as ‘Campbell Early’. Grapes were excised from the stems and GAE/100 mL).
washed. Peels were manually removed from grape berries and oven
dried at 40 °C until the moisture level was constant (5.5% w/w). Dried 2.5. Determination of antioxidant activity
grape peels were ground to a powdered form using an electrical
grinder and passed through a 0.5 mm sieve. All the chemicals used The antioxidant activity of the grape peel extracts was evaluated
were of analytical grade and they were purchased from Sigma by the phosphomolybdenum complex method (Prieto, Pineda, &
Chemical Co. (St. Louis, MO) and Duksan Pure Chemical Co. (Ansan, Aguilar, 1999). The assay is based on the reduction of Mo(VI) to Mo
Korea). (V) by the extract and subsequent formation of a green phosphate/Mo
(V) complex at acid pH (Abdel-Hameed, 2009). In brief, 0.4 mL of
2.2. Supercritical fluid extraction sample solution (100 μL of grape peel extract dissolved in 1 mL of
methanol) was combined with 4 mL of reagent solution containing
Supercritical fluid extraction (SFE) system consisted of CO2 0.6 M sulphuric acid, 2 mM sodium phosphate and 4 mM ammonium
cylinder, cool water circulator (VTR-620, Jeio Tech., Seoul, Korea), molybdate. The blank solution contained 4 mL of reagent solution and
column thermostat (CO-1560, JASCO Corporation, Tokyo, Japan), the 1 mL of methanol. Test tubes were caped and placed in hot water
solvent (CO2 and modifier) pumps (PU-1580, JASCO), UV/VIS detector for 90 min at 95 °C. After samples were cooled to room temperature,
(UV-1575, JASCO) and back pressure regulator (880-81, JASCO). 3 g the absorbance was measured at 695 nm against blank. Antioxidant
powdered sample of grape peel was kept in the extraction vessel and activity was expressed relative to that of ascorbic acid.
placed in the column thermostat set at a specific temperature.
Pressure was adjusted at the back pressure regulator and solvent 2.6. Analysis for total anthocyanins
pumps. The flow rates for CO2 and modifier were fixed at 2 mL/min.
Once the set temperature and pressure (at solvent pumps and back Determination of total anthocyanins in grape peel extracts was
pressure regulator) were achieved after turning on the injection valve based on the method described by Iland, Cynkar, Francis, Williams, &
and the system was in equilibrium, the extraction was carried out for Coombe, (1996) with some modifications. In 1 mL of sample, 10 mL
30 min in each experimental run. Extract was collected in a flask 50% ethanol was added and sample was centrifuged at 1800 g for
connected to the back pressure regulator and stored at −20 °C before 10 min. 200 µL of the centrifuged extract was mixed with 3.8 mL of
further analysis for the extract yield and bioactive components. The 1 M HCl and incubated at room temperature for 3 h. The absorbance
percentage extract yield was measured by drying the liquid extract at (A) of acidified diluted extract was measured at 520 nm using 1 M HCl
105 °C until constant weight of the dried grape peel extract was as the blank solution. Anthocyanins were calculated as mg/mL of
obtained. extract solution using the absorbance (B) of a 1% w/v solution of
malvidin-3-glucoside as follows:
2.3. Experimental design
Anthocyanins ðmg = mLÞ = A × Dilution factor × 1000 = B
Orthogonal array design (OAD) was used to arrange SFE experi-
ments for the response surface optimization. OAD is a type of
experimental design in which an orthogonal array is used to assign 2.7. Determination of antiradical activity
factors to a series of experimental combinations and results can be
analyzed using a mathematical procedure (Evangelaras, Kolaiti, & The free radical activity of the grape peel extract was determined
Koukouvinos, 2006; Moore, McKay, & Campbell, 2006). Effects of SFE by using 1, 1-diphenyl-2-picrylhydrazyl (DPPH) (Lee et al., 1998)
temperature, pressure and modifier concentration were investigated Briefly, 1 mL solution of the peel extract at a concentration of 100 μL/
on the recovery of bioactives from grape peel. An L16 (45) orthogonal mL methanol was mixed with 2 mL of 10 mg/L methanolic solution of
matrix with three factors, each factor containing four levels, was DPPH. The mixture was shaken vigorously and allowed to stand at
selected to arrange the experiments. Extraction temperatures were room temperature for 5 min and absorbance was recorded at 517 nm
37, 40, 43 and 46 °C; pressures were 140, 150, 160 and 170 kg cm− 2 by using spectrophotometer. Lower absorbance of the sample
and modifiers were 5, 6, 7 and 8% ethanol. The levels for each process indicated the higher free radical scavenging activity. The antiradical
variables were selected from a series of preliminary trials without activity of the grape peel extract was expressed in percentage.
using particular experimental designs. Regression analysis was
performed on the data obtained by triplicate analysis for each of the 2.8. Statistical analysis
dependent variables. Response surface analysis was also applied on
the data from orthogonal array design for modeling and prediction of All the analysis was carried out in triplicates and the experimental
optimum conditions of SFE for the extract yield, total phenols, results obtained were expressed as means ± SD. Statistical analysis
antioxidants and total anthocyanins from grape peel. was performed by using the Statistical Analysis System (SAS, version
9.1, SAS Institute, Cary, NC). Data were analyzed by the analysis of
2.4. Analysis for total phenolic compounds variance and the mean values were considered significantly different
when p b 0.05. The optimal extraction conditions were estimated
The total phenolic compounds were analyzed using the Folin through regression analysis and three dimensional response surface
Ciocalteu method with some modifications (Singleton & Rossi, 1965). plots of the independent variables and each dependent variable.
 K. Ghafoor et al. / Innovative Food Science and Emerging Technologies 11 (2010) 485–490 487

Table 1
Orthogonal array design for supercritical fluid extraction from grape peel and the yield, total phenol, antioxidants and anthocyanin contents of the extracts.

Test Supercritical fluid extraction conditions Analytical resultsa

set
Temperature Pressure Modifier Extract yield Total phenols Antioxidant activity Total anthocyanin
(°C) (kgcm− 2) (% Ethanol) (%) (mg GAE/100 mL) (mg/mL) (mg/mL)

1 37 140 5 6.78 ± 0.74 0.887 ± 0.082 0.476 ± 0.028 0.520 ± 0.028

2 37 150 6 6.89 ± 0.25 0.952 ± 0.091 0.518 ± 0.072 0.580 ± 0.083
3 37 160 7 7.56 ± 0.12 1.265 ± 0.028 0.576 ± 0.009 0.840 ± 0.058
4 37 170 8 8.58 ± 0.45 1.347 ± 0.123 0.725 ± 0.028 0.840 ± 0.054
5 40 140 6 7.29 ± 1.11 1.059 ± 0.072 0.619 ± 0.042 0.540 ± 0.071
6 40 150 5 7.89 ± 0.24 1.286 ± 0.082 0.805 ± 0.071 0.660 ± 0.093
7 40 160 8 8.27 ± 0.33 1.285 ± 0.127 0.825 ± 0.101 0.840 ± 0.073
8 40 170 7 10.78 ± 0.45 1.545 ± 0.214 0.893 ± 0.055 0.925 ± 0.039
9 43 140 7 9.56 ± 0.47 1.193 ± 0.141 0.824 ± 0.081 0.780 ± 0.085
10 43 150 8 10.11 ± 0.39 1.364 ± 0.082 1.115 ± 0.068 0.820 ± 0.077
11 43 160 5 11.55 ± 0.85 1.698 ± 0.082 1.318 ± 0.047 1.080 ± 0.028
12 43 170 6 12.33 ± 0.74 1.838 ± 0.026 1.520 ± 0.172 1.240 ± 0.062
13 46 140 8 10.56 ± 0.69 1.457 ± 0.117 1.245 ± 0.075 1.040 ± 0.033
14 46 150 7 11.33 ± 0.19 1.880 ± 0.092 1.565 ± 0.117 1.125 ± 0.118
15 46 160 6 11.89 ± 0.44 2.225 ± 0.076 1.569 ± 0.072 1.140 ± 0.027
16 46 170 5 13.22 ± 0.59 2.584 ± 0.051 1.878 ± 0.063 1.240 ± 0.076
a
Analytical results are means ± SD (n = 3).

3. Results and discussions pressure, while the second variable remains constant, results in
enhancement of the extract yield. The relationship between process
3.1. Modeling of the extraction process from grape peel variables and the extract yield is presented in Eq. (1).

The orthogonal array design (OAD), to optimize the supercritical

Y1 = 10:85033−0:93624X1 −0:43694X2 −1:44027X3
fluid extraction (SFE) of total phenols, antioxidants and total
2
anthocyanins from grape peel, is represented in Table 1. The + 0:00243X1 X2 + 0:07413X1 X3 −0:00826X2 X3 −0:00674X1 ð1Þ
experimental values of extract yields, total phenols, antioxidant 2
+ 0:00226X2 −0:04188X3
2

activities and total anthocyanins of grape peel extracts at various

experimental conditions are also presented in Table 1. The results of
analysis of variance, goodness of fit and the adequacy of the models where Y1 is the extract yield (%) in grape peel extract, X1 is the
are summarized in Table 2. The data showed a good fit with p b 0.05 extraction temperature (°C), X2 the extraction pressure (kg cm− 2)
and adequate with satisfactory R2 values. The model was used for the and X3 the ethanol concentration (%). The equation was based on the
construction of three dimensional response surface plots to predict data of regression coefficients as presented in Table 2.
the relationships between independent variables and the dependent
variables. 3.3. Effects of process variables on total phenolic compounds

Total phenolic contents of grape peel extracts obtained by SFE are

3.2. Effects of process variables on percent extract yield
shown in Table 1. Regression analysis was performed on the experimental
data and the coefficients of model were evaluated for significance. The
The percent yields of grape peel extracts obtained by using SFE are
effect of extraction temperature was highly significant (pb 0.001) on the
presented in Table 1. The regression analysis of the data showed that the
extraction of phenolic compounds. The effect of CO2 pressure was also
extract yield was significantly (pb 0.05) affected by the extraction
significant (pb 0.05) while that of ethanol was insignificant. This is
temperature and pressure. The relationship of the extract yield and that
because high pressure and temperature increase the solvating power of
of extraction temperature and pressure is depicted in Fig. 1 and it was
the CO2 (Liu et al., 2009). The relationship between total phenols of grape
linear with R2 value of 0.969. An increase in either of temperature and
peel extract and main variables is depicted in Fig. 2. Response surface
analysis of data in Table 1 demonstrates that the relationship between
total phenols and the operating parameters is both linear and quadratic
with a good regression coefficient (R2 =0.988). Eq. (2) shows the
Table 2 relationship between temperature, pressure and ethanol concentration
Regression coefficients and analysis of the model for four response variables. for the extraction of total phenolic compounds.
Coefficient Coefficients estimated

Extract yield Total phenols Antioxidants Anthocyanins

Y2 = 15:67942−0:75458X1 + 0:04375X2 −0:00002X3

b0 10.85033 15.67945 8.96062 0.10865 + 0:00123X1 X2 −0:01866X1 X3 + 0:00262X2 X3 ð2Þ

b1 0.93624 −0.75458b − 0.44713 − 0.20203 2 2 2
+ 0:40256X1 + 0:00955X2 −0:00002X3
b2 − 0.43694 0.043749 − 0.00376 0.03523
b3 − 1.44027 −0.00002 − 0.60867 0.04470 where (Y2) represents total phenols in grape peel extract. The
b11 − 0.00674 0.40256a 0.00439 0.00343 equation was based on the data of regression coefficients presented in
b22 0.00226 0.00955 − 0.00007 0.00007 Table 2. Although an increase in temperature favors extraction of
b33 − 0.04188 −0.00002 0.01894 0.01149
b12 0.00243 0.00123 0.00097 − 0.00068
phenols by enhancing both the solubility of solute and the diffusion
b13 0.07413 −0.01866 0.00711 0.01049 coefficient, it cannot be increased indefinitely; since the denaturation
b23 − 0.00826 0.00262 0.00027 − 0.00218 of phenolic compounds may take place at temperatures above 50 °C
Probability of F value b 0.001 b 0.0001 b0.001 b0.01 (Pinelo et al., 2006). SFE allows the extraction of phenolic compounds
a
p b 0.01, bp b 0.05. at lower temperatures hence preserves the quality of the extract.
488 K. Ghafoor et al. / Innovative Food Science and Emerging Technologies 11 (2010) 485–490

Fig. 1. Response surface analysis for the extract yields during supercritical fluid Fig. 3. Response surface analysis for the supercritical fluid extraction of antioxidants
extraction from grape peel with respect to extraction temperatures and pressures. from grape peel with respect to extraction temperatures and pressures.

3.4. Effects of process variables on antioxidant compounds various researchers (Cai, Luo, Sun, & Corke, 2004; Kumaran &
Karunakaran, 2006).
The analytical results of antioxidant compounds in grape peel extracts
obtained by SFE are shown in Table 1. The results of regression analysis 3.5. Effects of process variables on anthocyanin contents
reflect that the main extraction parameters for antioxidant compounds
from grape peel were extraction temperature and pressure, former being Total anthocyanin contents of the grape peel extracts are shown in
highly significant (pb 0.001). The effect of ethanol concentration was not Table 1. Extraction temperature and pressure had significant (p b 0.05)
significant. The relationship between the extraction of antioxidant effects on total anthocyanins and that of ethanol as modifier was not
components, temperature and pressure is depicted in Fig. 3. Antioxidant significant. Fig. 4 represents the relationship between total anthocy-
activities of the grape peel extracts were affected by the linear and anin compounds and the extraction temperature and pressure in SFE
quadratic terms of extraction process variables and R2 value was 0.981. from grape peel. Unlike total phenols and antioxidants, the effect of
The relationship between extraction variable and antioxidant activities is process variables on the total anthocyanins was only linear with R2
represented in Eq. (3). value of 0.954. Based on the values of regression coefficients (Table 2),
Eq. (4) was developed to present the relationship between total
Y3 = 8:96062−0:44713X1 −0:00376X2 −0:60867X3 anthocyanins (Y4) and extraction variables.

+ 0:00097X1 X2 + 0:00711X1 X3 + 0:00027X2 X3 ð3Þ Y4 = 0:10865−0:20203X1 + 0:03523X2 + 0:04470X3 −0:00068X1 X2

+ 0:00440X12 −0:00007X22 + 0:01894X32 + 0:010489X1 X3 −0:00218X2 X3 + 0:00343X12 ð4Þ
+ 0:00007X22 + 0:01149X32
Y3 represents antioxidant activity of the grape peel extract. It can
be observed that there is a correlation between total phenols and It has been reported that the extraction of anthocyanins from
antioxidant activities of extracts. Such linear correlations between grapes is enhanced by increasing temperature (Vatai, Skerget, & Knez,
phenolic compounds and antioxidant activities has been observed by 2009). The application of high pressure processing such as SFE not

Fig. 2. Response surface analysis for the supercritical fluid extraction of total phenols Fig. 4. Response surface analysis for the supercritical fluid extraction of total
from grape peel with respect to extraction temperatures and pressures. anthocyanins from grape peel with respect to extraction temperatures and pressures.
 K. Ghafoor et al. / Innovative Food Science and Emerging Technologies 11 (2010) 485–490 489

Table 3
Estimated optimum conditions, predicted and experimental values of responses under these conditions.

Response variables R2 R2- F- p- Optimum SFE conditions Maximum values

adjusted value value
Temp Pressure Ethanol Estimate Actuala
value
(°C) (kgcm− 2) (%)

Extract yield (%) 0.969 0.907 20.68 0.0008 44.31 166.70 6.44 12.31 12.25 ± 1.68
Total phenols (mg GAE/100 mL) 0.988 0.961 53.54 b.0001 45.66 160.1 6.25 2.156 2.215 ± 0.12
Antioxidant activities (mg/mL) 0.981 0.940 33.96 0.0002 45.64 160.8 6.52 1.628 1.652 ± 0.28
Total anthocyanins (mg/mL) 0.954 0.862 13.83 0.0023 45.23 163.3 6.58 1.176 1.172 ± 0.06
a
Analytical results are means ± SD (n = 3).

only increases the quantity of anthocyanins in the extract but also the purple DPPH to a yellow colored diphenyl picrylhydrazine and the
color stability upon storage (Vatai et al., 2008). remaining DPPH. The DPPH radical scavenging activities of 16 grape
peel extracts, according to OAD as listed in Table 1, are shown in Fig. 5.
3.6. Optimization of the extraction process All the extracts showed higher DPPH radical scavenging activities and
the maximum activity (98%) was observed for the extract 8 obtained
The optimum SFE conditions for the extract yields, total phenols, using SFE temperature of 40 °C and CO2 pressure of 180 kg cm− 2. It
antioxidant activities and total anthocyanins from grape peel as has been reported that food materials rich in antioxidants with higher
obtained by using OAD and response surface methodology (RSM) are free radical scavenging abilities are protective against certain types of
presented in the Table 3. SFE temperatures of 45–46 °C, extraction cancer and may also reduce the risk of cardiovascular and cerebro-
pressures in the range of 160–165 kgcm− 2 and lower concentrations vascular disorders (Miraliakbari & Shahidi, 2008).
(6–7%) of ethanol as modifier, can result in optimal extract yield Research has been carried out for the optimal extraction of
(12.31%) total phenols (2.156 mg GAE/100 mL), antioxidant phenolic compounds using SFE and the modifier was 30% ethyl acetate
(1.628 mg/mL) and total anthocyanins (1.176 mg/mL) from grape or methanol (Palmal & Taylor, 1999). We used ethanol as modifier in
peel. R2 and R2-adjusted values are also presented in Table 3. The low concentrations and observed that it has non-significant effects on
predicted results matched well with the experimental results SFE from grape peel. Extraction recovery in SFE is similar or better
obtained using optimum extraction conditions which validated the than solvent extraction methods and it depends on sample matrix,
RSM model with a good correlation. The prediction of one set of temperature, pressure, and modifier. Using an organic modifier is
optimal conditions for four response variables was also done by using essential to obtain good recovery, but 10% modifier gives essentially
desirability function approach. A total desirability value of 0.87 was the same recovery as 30% modifier (Seo, Burri, Quan, & Neidlinger,
obtained on a scale of 0 to 1 where 0 represents a completely 2005). Modifiers increase the solubility of sample material and the
undesirable response and 1 represents the most desirable response. At extracted bioactives can be easily recovered from the extraction
this desirability, SFE from grape peel by using 44.83 °C temperature, system. Ethanol is being used as modifier for SFE from plant materials
162.1 kg cm− 2 of pressure and 6.03% ethanol as modifier can results and it is regarded as a safe alcohol for food applications (Ovando,
in 12.37% extract yield, 2.161 mg GAE/100 mL total phenols, 1.66 mg/ Hernandez, Hernandez, Rodriguez, & Vidal, 2009). Studies have also
mL antioxidants and 1.27 mg/mL total anthocyanins. been carried out on the extraction of oil from grape seed using SFE and
parameters such as CO2 pressure and enzyme treatment of grape seed
3.7. Antiradical activities of the grape peel extracts were studied. The use of cell-wall degrading enzymes and higher
pressures of up to 200 bar can significantly improve the extraction
The antiradical activities of the grape peel extracts obtained by yield (Passos, Silva, Silva, Coimbra, & Silva, 2009). The internal mass
using SFE were also assessed using DPPH (1, 1-diphenyl-2-picrylhy- transport parameters during SFE were also studied using solubility
drazyl) radical scavenging assay. It is quick, reliable and reproducible data dealing with extractor vessel with non-negligible void volume
method to search in vitro general antiradical activities of pure (Fiori, 2007). During the extraction of resveratrol from Vitis inifera
compounds as well as plant extracts (Koleva, Van-Beek, Linssen, De- grape peel using SFE, it was observed that the extraction yield was
Groot, & Evstatieva, 2002). This method depends on the reduction of higher at 40 °C temperature, 150 bar pressure, 7.5% ethanol as
modifier and 15 min extraction time (Pascual-Martí, Salvador, Chafer,
& Berna, 2009). SFE extracts are safer as compared to those prepared
by conventional extraction techniques due to the use of carbon
dioxide as extraction solvent which is non-toxic and environmental
friendly (Friedrich & List, 1982; King, 2000). We can infer that the
main process variables of SFE are the extraction temperature and
pressure. Grape and grape products have been extensively documen-
ted as healthy foods. Extracts obtained from grape peel and seed have
been tested to possess anticancer and cancer chemopreventive
properties during in vivo and in vitro studies (Kaur, Agarwal, &
Agarwal, 2009). Phenolic compounds contained in these extracts are
reported to have beneficial effects on other chronic diseases such as
coronary heart disease (Forester & Waterhouse, 2009). These health
effects are reported to be due to antiradical and antioxidant properties
of phenolics in grapes and grape derivatives (Lurton, 2003).

4. Conclusions

Fig. 5. Antiradical activities of grape peel extracts (1–16 according to OAD) obtained by SFE is an effective technology in the recovery of biologically
supercritical fluid extraction. Bars represent standard error of the means (n = 3). valuable components from grape peel. Extraction process was
490 K. Ghafoor et al. / Innovative Food Science and Emerging Technologies 11 (2010) 485–490

significantly affected by extraction temperature and pressure and the Kumaran, A., & Karunakaran, J. (2006). In vitro antioxidant activities of methanol
extracts of five Phyllanthus species from India. LWT-Food Science and Technology, 40,
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can be used as an alternative to conventional organic solvent Lee, S. K., Mbwambo, Z. H., Chung, H. S., Luyengi, L., Games, E. J. C., & Mehta, R. G. (1998).
extraction methods. SFE involves the application of high pressures; Evaluation of the antioxidant potential of natural products. Combinational
Chemistry & High Throughput Screening, 1, 35−46.
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preserve the quality of extracts. By optimizing SFE process variables, dioxide extraction of seed oil from Opuntia dillenii Haw. and its antioxidant activity.
yields of total phenols, antioxidants and total anthocyanins can be Food Chemistry, 114(1), 334−339.
Lu, T. J., Gaspar, F., Marriott, R., Mellor, S., Watkinson, C., & Al-Duri, B. (2007). Extraction
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peel is a good source of phenolic compounds, antioxidants and total modeling. Journal of Supercritical Fluids, 41, 68−73.
anthocyanins. The SFE extracts of grape peel also possess strong Lurton, L. (2003). Grape polyphenols: New powerful health ingredients. Innovations in
Food Technology, 18, 28−30.
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Meyer, A. S., Yi, O. S., Pearson, D. A., Waterhouse, A. L., & Frankel, E. N. (1997). Inhibition
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Miraliakbari, H., & Shahidi, F. (2008). Antioxidant activity of minor components of tree
The authors are grateful to Korean Research Foundation, Republic nut oils. Food Chemistry, 111, 421−427.
of Korea for the financial support to conduct this research work. Moore, L. M., McKay, M. D., & Campbell, K. S. (2006). Combined array experiment
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Ovando, A. C., Hernandez, M. L. P., Hernandez, M. E. P., Rodriguez, J. A., & Vidal, C. A. G.
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