International Food Research Journal 24(2): 518-524 (April 2017)

Journal homepage: [Link]

Comparative study of organic solvents and extraction conditions on colour

and antioxidant capacity in red cabbage
1*
Junka, N., 2Rattanamechaiskul, C., 3,4Wongs-Aree, C. and 3,4Kanlayanarat, S.
1
Division of Crop production Technology, Faculty of Science and Technology, Nakhon Pathom
Rajabhat University, Nakhon Pathom 73000, Thailand
2
Department of Engineering, King Mongkut’s Institute of Technology Ladkrabang, Prince of
Chumphon Campus, Chumphon 86160
3
Postharvest Technology Program, School of Bioresources and Technology, King Mongkut’s
University of Technology Thonburi, Bangkhuntien Campus, Bangkok 10150, Thailand
4
Postharvest Technology Innovation Center, Commission on Higher Education, Bangkok 10400,
Thailand

Article history Abstract

Received: 11 January 2016 In industrially natural-dye production, there are many organic solvent with different
Received in revised form: efficiencies for extracting colour and antioxidants of red cabbage. The objective of this study
4 April 2016
Accepted: 20 April 2016
was to investigate appropriate organic solvent and extract condition on colour and antioxidant
activity. Two organic solvents were used for extraction: ethanol for hydrophilic antioxidants
and hexane for lipophilic antioxidants. Experimental results showed that red cabbage extracted
by ethanol was significantly higher in purple colour, anthocyanin content, total phenolic
Keywords content, antioxidant activity by 2,2-diphenyl-1-picrylhydrazyl radical assay (DPPH), and ferric
reducing/antioxidant power assay (FRAP) than extraction by hexane due to anthocyanins being
Red cabbage the main phenolic compound in extract. To find the appropriate concentration and extraction
Organic solvent extraction time, the results revealed that the red cabbage extracted by 70% ethanol for 18 h was suitable
Anthocyanins for extracting bioactive polar compounds, such as hydrophilic antioxidants.
Antioxidant activity
© All Rights Reserved

Introduction include: antioxidant activity, inhibition of DNA

damage in the cancer cell, prevention of coronary
Colour is an important sensory property in food heart disease, and preservation of eyesight. Tamura
products to attract consumer demand. At present, and Yamagami (1994) found that the pericarp of the
natural dyes from colourful plants for use in food Muscat Bailey grape is two times more effective than
products are increasingly in demand because they (+)-catechin and α-tocopherol. The anthocyanin can
are a good choice to avoid potentially mutagenic resist the mutation of cells because it inhibits DNA
and carcinogenic effects from synthetic dyes. In the damage in the cancer cells of internal organ of human
United States, for example, grape extract has been such as lung, gastric and colon (Chen et al., 2006a;
used as a colorant to mix with beverages or soft drinks Chen et al., 2006b; Wang and Stoner, 2008; Huang
(Francis, 1989; Bridle and Timberlake, 1997). Yang et al., 2011). Sumner et al. (2005) reported that in
et al. (2008) have reported that the purple maize cob, a patient who had ischaemic coronary heart disease
which was a by-product of the maize industry, could (CHD) daily consumption of pomegranate juice
be used as a source of purple colour in food. There could improve stress-induced myocardial ischaemia
are several colour plants, such as red radish, beetroot, within 3 months. The bilberry anthocyanins were
carrot, etc. whose extracts could replace synthetic used as medicine for the physiological renewal and
dyes (Sapers, 1994; Chethana et al. 2007). Red homeostasis of corneal epithelial cell because of the
cabbage is one of many plants that could address the ability of bilberry anthocyanins to improve nutrition
mentioned demand because its leaves are colourful of the retina and restore primary night vision (Song
and have a large quantity of phenolic compound, et al. 2010; Gottikh and Tashlitskii, 2010). As
especially anthocyanins. mentioned above, the advantages of anthocyanins are
Anthocyanins are the basis of the red or purple desirable for cancer patients or consumers who are
pigments that are a group of natural phenolic interested in the maintenance of good health.
compounds. The significant properties of anthocyanins Red cabbage is of interest in natural dye functions

*Corresponding author.
Email: nittaya717@[Link]
519 Junka et al./IFRJ 24(2): 518-524

for industrial foods (Markakis, 1982; Malien-Aubert The aqueous red cabbage samples were filtered
et al., 2001). In Japan, anthocyanins from red through Whatman paper No. 1 for separation of the
cabbage were used for industrial extraction because it aqueous part from the residue. Then, the aqueous part
had a high yield, large quantities and sites compared was collected in plastic vials.
with berries (Bridle and Timberlake, 1997; Piccaglia
et al., 2002). In addition, red cabbage anthocyanins Determination of visual aqueous colours of extracted
were resistant to heat and light and are biologically samples
harmless. In the extraction process, there are many Tristimulus parameters as lightness (L*), redness
organic solvents with different efficiencies for (a ), yellowness (b*) and hue angle (h°) values were
*

extracting either hydrophilic antioxidants or lipophilic measured from aqueous samples in 3 replicates using
antioxidants or both (Anwar et al., 2013; Chavan and a colorimeter that was calibrated with a standard
Amarowicz, 2013; Benmeziane et al., 2014; Othman white plate (Minolta model CR 400, Japan). The
et al., 2014; Nur Syukriah et al., 2014). The times results are expressed as L*, a*, b* and h°.
and concentrations of solvent during extraction
significantly influence the antioxidant activities of Measurement of anthocyanin content
the plant extract (Sultana et al., 2009; Ahmad et al., The pH differential technique of Giusti and
2011; Chew et al., 2011; Woo et al., 2013; Thoo et Wrolstad (2005) was used for measuring anthocyanin
al., 2013; Bachir bey et al., 2014; Candrawinata et al., content in this experiment. Potassium chloride buffer
2014) To obtain the optimum extract, a comparative (0.025 M KCl, pH 1.0) and sodium acetate (0.4 M
study of the type of organic solvent and extraction CH3CO2Na•3H2O, pH 4.5) were used for examining
conditions on colour and antioxidant capacity in red anthocyanin contents. One hundred microliters of
cabbage is therefore needed. Until now, information extract from red cabbage were added to the test tube.
on this subject has not been reported. Two tubes of one extract sample were followed by the
The objective of this work was to investigate the two buffers. Nine hundred microliters of potassium
effects of polar and nonpolar, extracting duration time chloride buffer and sodium acetate buffer were added.
and appropriate concentrations of organic solvent on The test tubes were mixed and incubated for 15 min
key quality attributes of red cabbage extracted. The at room temperature (25°C). The anthocyanins were
quality parameters considered included the colour of measured by spectrum scanning at 320 to 700 nm
the extracts by tristimulus colorimetry as lightness using a UV-visible spectrophotometer (Shimadzu
(L*), redness (a*), yellowness (b*) and hue angle (h°) model 1601, Japan). The absorbance of the diluted
values, anthocyanin content, total phenolic content extracts was calculated as shown in equation (1)
and antioxidant activities by DPPH and FRAP assays.
A= (Aλvis-max - Aλ700)pH 1.0 - (A λvis-max - Aλ700)pH 4.5
Materials and Methods (1)

Plant materials Anthocyanin pigment concentration in the red

Fresh red cabbages obtained from the Royal cabbage extract was calculated using equation (2):
Project Foundation Center, Bangkok, Thailand were
used in this study. The plant material, extraction anthocyanin pigment (mg/L) = (A x MW x DF x
procedures and chemical analyses were conducted 1000)/(ε x l)
at the Postharvest Technology Laboratory, King (2)
Mongkut’s University of Technology Thonburi,
Bangkok. The samples were prepared for extraction where A is the absorbance of the mixed solution of
with different solvents, extraction times and the anthocyanin sample, MW is cyanidin-3-glucoside
concentrations. which has molecular weight as 449.2 (g/mol), DF is
the dilution factor, ε is the molar absorptive as 26,900
Sample extraction (L/mol*cm), and l is the path length of the cuvette
The method for extraction was that of Rodriguez- (cm). Then, the values of anthocyanin content in
Saona and Wrolstad (2005). The 99.8% hexane (H) units of mg/L were converted to mg/100 g fresh
and 50, 70, and 95% ethanol (E) solvents were added weight (FW).
to fresh red cabbage samples in plastic tubes and
homogenised. The extracted sample by hexane was Measurement of total phenolic content
stored for 18 h at 4°C in refrigerator while extracted Total phenolic content assay was determined by
samples by ethanol were stored for 2, 8, and 18 h. the method of Swain and Hillis (1959). One hundred
 Junka et al./IFRJ 24(2): 518-524 520

and fifty microliters of red cabbage extract and 2400 Results and Discussion
µL of distilled water were added to the test tubes.
Then, 150 µL of 0.25 N of Folin-Ciocalteu reagents Colour analysis
was added in the tube. The samples were mixed using Colour parameter analysis of red cabbage
a vortex mixer and allowed to react for 3 min. To the extracts at various organic solvents, durations
mixtures was added 300 µL of 1N Na2CO3 solution. and concentrations shows in the Table 1. In the
The mixtures were vortexed for a few minutes and Tristimulus diagram, the extracted colour could be
kept away from light at 25°C for 2 h. Then, the predicted by the direction of L*, a*, b* and h° value. At
absorbance was analysed at 725 nm, and the data an extraction time of 2 h, the colour parameter results
were expressed in gallic acid equivalents (GAE; reveal that solution extracted using 70% ethanol had
mg/100 g fresh weight) by a gallic acid concentration the lowest L* values as 26.4 while 50 and 95% ethanol
of 0.0-0.1 mg/mL. had values of 28.2 and 28.0, respectively. The L*
values of 50 and 95% ethanol were not significantly
2,2-Diphenyl-1-picrylhydrazyl radical scavenging different. Increasing the duration of extraction from
activity (DPPH) 2 h to 8 and 18 h caused a significant decrease in
Free radical-scavenging activity of extracts L* values compared to the same concentration of
was reacted with the 1,1-diphenyl-2-picrylhydrazyl ethanol solvent.
radical (DPPH) which was modified from Brand- The other colour parameters, a*, b* and h°, are quite
Williams et al. (1995). The stock solution of DPPH complicated to describe when the various extraction
was prepared by twelve milligrams of a purple DPPH conditions are compared. At the same durations of 2
powder, which was dissolved in 50 mL of methanol. and 8 h, the increase in the concentration of ethanol
The working solution was made by mixing 20 mL did not significantly affect the a*, b* and h° value.
of stock solution and 90 mL of methanol. Before The results seem to indicate that the change in those
analysis, the working solution was measured by parameters is related to the increase in duration
spectrophotometer at 515 nm to obtain an absorbance more than the concentration of ethanol. At the same
of 1.1 ± 0.02 units. Two-thousand eight-hundred and concentration, the value of a* increased and the values
fifty microliters of working solution and 150 µL of of b* and h° decreased when the extraction time was
extracted anthocyanins were mixed in a plastic tube. 18 h.
The tubes were kept in the dark for 30 min and then With the different organic solvent for extraction,
recorded at 515 nm. Next, absorbance values of the the extracted colours using 95% ethanol and 99.8%
samples were calculated as shown in formula (3) hexane for 18 h were obviously dissimilar in
appearance. The solution extracted using ethanol
% inhibition = [(Abs control – Abs sample)/Abs control] x 100 contained smaller values for L*, b* and h° at 23.2, 0.1
(3) and 0.6, respectively, and larger in a* values at 14.0
than those of values (L*, b*, h° and a*) of solution
Ferric-reducing antioxidant power assay (FRAP) extracted using hexane at 75.3, 0.5, 54.4 and 0.1,
The FRAP assay was conducted according to respectively. The lightness in extracts of hexane was
Benzie and Strain (1996) with some modifications. mainly attributed to the greater luminosity. For the
Fresh FRAP reagent consisted of 0.3 M acetate buffer colour of red cabbage extracts with 95% ethanol,
(pH 3.6), 0.01 M TPTZ solution in 40 mM HCl, and there was less transmission of light represented by the
0.02 M FeCl3•6 H2O solution in a ratio of 100:10:10 colour magenta while the other extract scarcely had
(v/v/v). The reagent was warmed at 37°C before use. colour. This result corresponds to previous research
The 150 µL of aqueous extract was mixed with 2850 by Khandare et al. (2011) who studied extraction
µL of fresh FRAP reagent. Then, the mixture solutions processing on the colour of black carrot and reported
were kept in the dark for 30 min. The samples that the extract with small L* value and large a* value
were measured at 593 nm by spectrophotometer would show intense coloration in the sample.
(Shimadzu model 1601, Japan). The absorption result
was compared with a standard curve of Trolox at 25 Anthocyanin content
to 800 µM, which was expressed in µM TE / g fresh Table 2 shows the anthocyanin contents of red
weight. cabbage obtained by extraction at different organic
solvents, durations and concentrations. The various
condition of extract contains anthocyanins in the
range 0.0-6.7 mg/100 g fresh weight (FW) with
significant differences in each treatment. At the same
521 Junka et al./IFRJ 24(2): 518-524

Table 1. Colour parameters of red cabbage extracts at Table 2. Anthocyanin content and total phenolic content of
various organic solvents, durations and concentrations red cabbage extracts at various organic solvents, durations
and concentrations

Different superscripts in the same column denote that the mean

values are significantly different at p ≤ 0.01
Different superscripts in the same column denote that the mean
concentration of ethanol, the anthocyanin content values are significantly different at p ≤ 0.01
increased in extracts with longer extraction times. Remark *1= Anthocyanin content (mg/100 g FW)
*
2= Total phenolic content (mg gallic acid equivalents/100 g FW)
The duration of 18 h of extraction using 70% ethanol
yielded the greatest content of anthocyanin in the
FW. At the same concentration i.e., 70% ethanol,
extract. Lapornik et al. (2005) have reported that the
total phenolic content of the extract increased to
yield of total anthocyanins of black currant extracted
larger values: 177.8, 185.1 and 189.1 mg gallic acid
using 70% ethanol was enhanced 48% by increasing
equivalents/100 g FW with durations 2, 8, and 18 h,
the extraction time.
respectively (Lapornik et al., 2005; Turkmen et al.,
When the result of 95% ethanol and 99.8%
2007). From the result, the extraction obtained using
hexane for 18 h was comparison, the ethanol extract
70% ethanol for 18 h showed the largest total phenolic
contained an anthocyanin content of 6.5 mg/100 g
content, corresponding to the results of anthocyanins.
FW while the hexane extract had no anthocyanin
The total phenolic content was greater with greater
content. The better extraction using ethanol was
anthocyanin content as mentioned earlier.
most likely due to the major polar compounds in red
The large amounts of phenolics in the 70% ethanol-
cabbage leaves. Anthocyanins are polar molecules
extracted solution were due to the optimal combination
with aromatic rings that are conjugated with multi-
of water and organic solvent. This combination for
hydroxyl, methoxy and glycosyl groups. Harborne
extraction contributed to a greater polarity index
and Grayer (1988) reported that the polarity of
compared to 95% ethanol. Chandrasekhar et al.
anthocyanin molecules affected the extraction
(2012) reported that the increased proportion of the
and separation of anthocyanins from the samples.
water in the ethanol enhanced the polarity index
Commonly, polar organic solvents exhibit a better
of the solvent, subsequently increasing the degrees
ability to elute anthocyanins nonpolar solvents.
of extraction. However, the higher proportion of
Thus, anthocyanins would be better extracted by a
water in the solvent might affect identification and
polar solvent, 95% ethanol, than a non-polar organic
quantification of phenolics and interfere with organic
solvent, 99.8% hexane. This result with regard to
acids, sugars, and soluble proteins (Robards et al.,
the colours of the extracts as shown in the Table 1 is
1999; Lapornik et al., 2005; Liyana-Pathirana and
apparently related to the anthocyanin content, which
Shahidi, 2005).
increased with decreases in h° and an increase in the
As expected result of total phenolic contents of
a* value (Kim et al., 2007).
red cabbage extracts obtained using 95% ethanol and
99.8% hexane, the solution extracted using ethanol
Total phenolic content
contained a large total phenolic content. This result
Total phenolic contents of red cabbage at various
is due to the major polar compounds in red cabbage
extract conditions are shown in Table 2. The ethanol
leaves such as anthocyanins that are a one subgroup
extract showed the smallest value in 50% ethanol
of phenolic compounds (Arnnok et al. 2012). Thus,
for 2 h, 168.3 mg gallic acid equivalents/100 g
 Junka et al./IFRJ 24(2): 518-524 522

Figure 1. Antioxidant activity of red cabbage extracts Figure 2. Antioxidant activity of red cabbage extracts
obtained using various ethanol concentrations after an obtained using 70% ethanol and various durations
extraction time of 2 h

the total phenolic content of flavonoids including at 2 h were significantly different. The extracts were
anthocyanins would be better extracted by 95% in a range of 280.6-308.0 µM TE/g FW. The FRAP
ethanol, a polar organic solvent, than hexane. value of 70% ethanol had the largest value as 308.0
µM TE/g FW. By increasing the extraction times as
2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical shown in Figure 2, extracts had greater antioxidant
scavenging activity activity. The results illustrated that a long duration of
Figure 1 shows the antioxidant activity of red 18 h with 70% ethanol-extracted solution yielded the
cabbage extracts using various ethanol concentrations greatest the activity as determined in the FRAP assay.
at a duration of 2 h. That per cent inhibition activity An increase in antioxidant capacity in red cabbage
of all extracts illustrated a range of 78.1%-83.2%. extracts was correlated with anthocyanins and total
The 70% ethanol extract had the largest value of phenolic contents. As a result, solution extracted
83.2% while that of 50% ethanol extract had the using 70% ethanol for 18 h was the best condition for
smallest value. When increasing the duration to 8 extracting the antioxidant properties of red cabbage.
and 18 h, the only antioxidant activity obtained from On the other hand, hexane solvent had no
70% ethanol was compared as shown in Figure 2. antioxidant activity analysed by FRAP assay.
The percent inhibition significantly increased with Although solution extracted by using hexane had a low
increasing the duration time, and it increased up to total phenolic content, it had no anthocyanins. One
90.4% at a duration of 18 h. could conclude from this result that the anthocyanins
The antioxidant activity at the duration time play important role in antioxidant activity as tested
for 18 h of the sample extracted using 95% ethanol by FRAP assay.
solvent was greater than that obtained using 99.8%
hexane (data not shown). The results seem that Conclusions
the percentage of inhibition activity is related to
total phenolic content; the inhibition activity was Ethanol was suitable for extracting natural colorant
greater with greater total phenolic content. In from red cabbage leaves with significant amounts of
fact, the phenolic compounds belong to a group of antioxidant compounds, such as anthocyanins, and
phytochemical substances whose molecules are total phenolic contents because of its high degree of
easy to oxidise such that they can donate hydrogen polarity compared to the non-polar solvent hexane.
atoms to reactive compounds and resist free radicals The adjustment of polarity properties when adding
(Castañda-Ovando et al. 2009). Fleschhut et al. water showed a moderate portion of 70% ethanol
2006 have been reported that cyanidin, which is was better than 50 and 95% ethanol. A greater quality
a natural organic compound in a particular type of of extracts came with an increase in extraction time.
anthocyanins, possesses o-dihydroxy substitution, The solution extracted using 70% ethanol for 18 h
which would increase the antioxidant activity of the was appropriate for obtaining colorant extracts with
extract. large quantities of anthocyanins and hydrophilic
antioxidants from red cabbage.
Ferric-reducing antioxidant power value (FRAP)
From Figure 1, the FRAP values of ethanol of
the various concentrations of 50%, 70%, and 95%
523 Junka et al./IFRJ 24(2): 518-524

Acknowledgements Vidal, C. A. 2009. Chemical studies of anthocyanins:

A review. Food Chemistry 113: 859-871.
The authors express their appreciation to the Chandrasekhar, J., Madhusudhan, M. C. and Raghavarao,
Development Institute Nakhon Pathom Rajabhat K. S. M. S. 2012. Extraction of anthocyanins from red
cabbage and purification using adsorption. Food and
Universiy, Postharvest Technology Program, School
bioproducts processing 90: 615-623.
of Bioresources and Technology, King Mongkut’s Chavan, U. D. and Amarowicz, R. 2013. Effect of various
University of Technology Thonburi, and Postharvest solvent systems on extraction of phenolics, tannins
Technology Innovation Center, Commission on and sugars from beach pea (Lathyrus maritimus L.).
Higher Education for their equipment supporting. International Food Research Journal 20(3): 1139-
1144.
Reference Chen, P. N., Chu, S. C., Chiou, H. L., Kuo, W. H., Chiang,
C. L. and Hsien, Y. S. 2006a. Mulberry anthocyanins,
Ahmad, N., Anwar, F., Hameed, S. and Boyce, M.C. 2011. cyanidin 3-rutinoside and cyanidin 3-glucoside,
Antioxidant and antimicrobial attributes of different exhibited an inhibitory effect on the migration and
solvent extracts from leaves and flowers of akk invasion of a human lung cancer cell line. Cancer
[Calotripis procera (Ait.) Ait. F]. Journal of Medicinal Letters 235: 248-259.
Plants Research 5(19): 4879-4887. Chen, P. N., Kuo, W. H., Chiang, C. L., Chiou, H. L., Hsien,
Anwar, F., Kalsoom, U., Sultana, B., Mushtaq, M., Y. S. and Chu, S. C. 2006b. Black rice anthocyanins
Mehmood, T. and Arshad, H.A. 2013. Effect of drying inhibit cancer cells invasion via repressions of MMPs
method and extraction solvent on the total phenolics and u-PA expression. Chemico-Biological Interactions
and antioxidant activity of cauliflower (Brassica 163: 218-229.
oleracea L.) extracts. International Food Research Chethana, S., Chetan, A. N. and Raghavarao, K. S. M. S.
Journal 20(2): 653-659. 2007. Aqueous two phase extraction for purification
Arnnok, P., Ruangviriyachai, C., Mahachai, R., and concentration of betalains. Journal of Food
Techawongstien, S. and Chanthai, S. 2012. Engineering 81: 679-687.
Determination of total phenolics and anthocyanin Chew, K. K., Khoo, M. Z., Ng, S. Y., Thoo, Y. Y., Wan
contents in the pericarp of hot chilli pepper (Capsicum Aida, W. M. and Ho, C. W. 2011. Effect of ethanol
annuum L.). International Food Research Journal concentration, extraction time and extraction
19(1): 235-243. temperature on the recovery of phenolic compounds
Bachir bey, M., Meziant, L., Benchikh, Y. and Louaileche, and antioxidant capacity of Orthosiphon stamineus
H. 2014. Deployment of response surface methodology extracts. International Food Research Journal 18(4):
to optimize recovery of dried dark fig (Ficus carica 1427-1435.
L., var. Azenjar) total phenolic compounds and Fleschhut, J., Kratzer, F., Rechkemmer, G., & Kulling, S.
antioxidant activity. International Food Research E. 2006. Stability and biotransformation of various
Journal 21(4): 1477-1482. dietary anthocyanins in vitro. European Journal of
Benmeziane, F., Djamai, R., Cadot, Y. and Seridi, R. 2014. Nutrition 45(1): 7-18.
Optimization of extraction parameters of phenolic Francis, F.J. 1989. Food colorants: anthocyanins. Critical
compounds from Algerian fresh table grapes (Vitis Reviews in Food Science and Nutrition 28: 273-314.
Vinifera). International Food Research Journal 21(3): Giusti, M. M. and Wrolstad, R. E. 2005. Characterization
1061-1065. and measurement of anthocyanins by UV-visible
Benzie, I. F. F. and Strain, J. J. 1996. The ferric reducing spectroscopy. In: Wrolstad, R. E., Acree, T. E.,
ability of plasma (FRAP) as a measure of antioxidant Decker, E. A., Penner, M. H., Reid, D. S., Schwartz, S.
power: The FRAP Assay. Analytical Chemistry 239: J., Shoemaker, C. F., Smith, D. and Sporns, P. (Eds.).
70-76. Handbook of Food Analyical Chemistry (Vol 2):
Brand-Williams, W., Cuvelier, M. E. and Berset, C. 1995. Pigments, Colorants, Flavors, Texture, and Bioactive
Use of a free radical method to evaluate antioxidant Food Components. p.19-31. New Jersey: John Wiley
activity. Lebensmittel Wissenschaft und Technologie and Sons, Inc.
Food Science and Technology 28: 25-30. Gottikh, M. B. and Tashlitskii, V. N. 2010. Determination
Bridle, P. and Timberlake, C. F. 1997. Anthocyanins as of the qualitative and quantitative composition
natural food colour-selected aspects. Food Chemistry of antocyan pigments as components of dietary
58: 103-109. supplements and drugs for vision. Vestnik oftalmologii
Candrawinata, V. I., Golding, J. B., Roach, P. D. and 126(5): 34-37.
Stathopoulos, C. E. 2014. Total phenolic content and Harborne, J. B. and Grayer, R. J. 1988. The anthocyanins.
antioxidant activity of apple pomace aqueous extract: In: Harborne, J. B. (ed.). The Flavonoids. p. 1-20.
effect of time, temperature and water to pomace ratio. New York. Chapman and Hall.
International Food Research Journal 21(6): 2337- Huang, H. P., Chang, Y. C., Wu, C. H., Hung, C. N. and
2344. Wang, C. J. 2011. Anthocyanin-rich mulberry extract
Castañda-Ovando, A., Pacheco-Hernández, M. L. M., inhibit the gastric cancer cell growth in vitro and
Páez-Hernández, M. E., Rogríguez, J. A., Galán- xenograft mice by inducing signals of p38/p53 and
 Junka et al./IFRJ 24(2): 518-524 524

c-jun. Food Chemistry 129: 1703-1709. extracts on cultured human corneal limbal epithelial
Khandare, V., Walia, S., Singh, M. and Kaur, C. 2011. cells (HCLEC). Phytotherapy Research 24(4): 520-
Black carrot (Daucus carota ssp. sativus) juice: 524.
Processing effects on antioxidant composition and Sultana, B., Anwar, F. and Ashraf, M. 2009. Effect of
color. Food and Bioproducts Processing 89: 482-486. extraction solvent/ technique on the antioxidant
Kim, M. J., Hyun, J. N., Kim, J. A., Park, J. C., Kim, M. activity of selected medicinal plant extracts. Molecules
Y., Kim, J. G., Lee, S. J., Chun, S. C., and Chung, I. 14: 2167–2180.
M. 2007. Relationship between phenolic compounds, Sumner, M. D., Elliott-Eller, M., Weidner, G., Daubenmier,
anthocyanins content and antioxidant activity in J. J., Chew, M. H., Marlin, R. Raisin, C. J., Ornish,
colored barley germplasm. Journal of Agricultural and D. 2005. Effects of pomegranate juice consumption
Food Chemistry 55: 4802-4809. on myocardial perfusion in patients with coronary
Lapornik, B., Prošek, M. and Wonder, A. G. 2005. heart disease. The American Journal of Cardiology
Comparison of extract prepared from plant by products 90(6):810-814.
using different solvents and extraction time. Journal of
Food Engineering 71: 214-222. Swain, T. and Hillis, W. E. 1959. The phenolic constituents
Liyana-Pathirana, C. and Shahidi, F. 2005. Optimization of Prunus domestica I- the quantitative analysis of
of extraction of phenolic compounds from wheat phenolic constituents. Journal of the Science of Food
using response surface methodology. Food Chemistry and Agriculture 10: 63-68.
93: 47-56. Tamura, H. and Yamagami, A. 1994. Antioxidative activity
Malien-Aubert, C., Dangles, O. and Amiot, M. J. 2001. of mono-acylated anthocyanins isolated from Muscat
Color stability of commercial anthocyanin-based Bailey A grape. Journal of Agriculture and Food
extracts in relation to the phenolic composition. Chemistry 42(8): 1612-1615.
Protective effects by intra- and intermolecular Thoo, Y. Y., Ng, S. Y., Khoo, M. Z., Wan Aida, W. M. and
copigmentation. Journal of Agricultural and Food Ho, C. W. 2013. A binary solvent extraction system
Chemistry 49: 170-176. for phenolic antioxidants and its application to the
Markakis, P. 1982. Stability of anthocyanins in foods. In: estimation of antioxidant capacity in Andrographis
Markakis, P. (ed.). Anthocyanins as Food Colors. p. paniculata extracts. International Food Research
163-180. New York: Academic Press. Journal 20(3): 1103-1111.
Othman, A., Mukhtar, N. J., Ismail, N. S. and Chang, S. K., Turkmen, N., Velioglu, Y. S., Sari, F., and Polat, G. 2007.
2014. Phenolics, flavonoids content and antioxidant Effect of extraction conditions on measured total
activities of 4 Malaysian herbal plants. International polyphenol contents and antioxidant and antibacterial
Food Research Journal 21(2): 759-766. activities of black tea. Molecules 12: 484-496.
Nur Syukriah, A. R., Liza, M. S., Harisun, Y. and Fadzillah, Wang, L.S. and Stoner, G.D. 2008. Anthocyanins and their
A. A. M. 2014. Effect of solvent extraction on role in cancer prevention. Cancer Letters 269: 281-
antioxidant and antibacterial activities from Quercus 290.
infectoria (Manjakani). International Food Research Woo, P. F., Yim, H. S., Khoo, H. E., Sia, C. M. and
Journal 21(3): 1067-1073. Ang, Y. K. 2013. Effects of extraction conditions on
Piccaglia R., Marotti M. and Baldoni G., 2002. Factors antioxidant properties of sapodilla fruit (Manilkara
influencing anthocyanin content in red cabbage zapota). International Food Research Journal 20(5):
(Brassica olaracea var capitata L f. rubra (L) Thell). 2065-2072.
Journal of the Science of Food and Agriculture 82: Yang, Z., Fan, G., Gu, Z., Han, Y. and Chen, Z. 2008.
1504-1509. Optimization extraction of anthocyanins from purple
Robards, K., Prenzler, P. D., Ticker, G., Swatsitang, P. corn (Zea mays L.) cob using tristimulus colorimetry.
and Glover, W. 1999. Phenolic compounds and their European Journal of Lipid Science and Technology
role in oxidative process in fruits. Food Chemistry 66: 227: 409-415.
401-436.
Rodriguez-Saona, L.E. and Wrolstad, R.E. 2005.
Extraction, isolation, and purification of anthocyanins.
In: Wrolstad, R. E., Acree, T. E., Decker, E. A., Penner,
M. H., Reid, D. S., Schwartz, S. J., Shoemaker, C. F.,
Smith, D. and Sporns, P. (Eds.). Handbook of Food
Analyical Chemistry (Vol 2): Pigments, Colorants,
Flavors, Texture, and Bioactive Food Components.
p.7 - 17. New Jersey: John Wiley and Sons, Inc.
Sapers, G. M., 1994. Colour characteristics and stability of
nonbleeding cherries dyed with carotenoid pigments.
Journal of Food Science 59: 135-138.
Song, J., Li, Y., Ge, J., Duan, Y., Sze, S. C., Tong, Y., Shaw,
P.C., Ng, T. B., Tsui, K.C., Zhuo, Y., Zhang, K. Y. 2010.
Protective effect of bilberry (Vaccinium myrtillus L.)