Journal of Food Composition and Analysis 24 (2011) 615–619

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Journal of Food Composition and Analysis

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Short Communication

Effect of extraction solvents on the phenolic compounds and antioxidant activities

of bunga kantan (Etlingera elatior Jack.) inflorescence
M.M. Jeevani Osadee Wijekoon, Rajeev Bhat *, Alias A. Karim
Food Technology Division, School of Industrial Technology, Universiti Sains Malaysia, Penang 11 800, Malaysia

A R T I C L E I N F O A B S T R A C T

Article history: In the present study, we investigated the effect of methanol, acetone (50%, 90% and 100%, v/v) and
Received 11 December 2009 distilled water on the extractability of some of the antioxidant compounds (total phenols, tannins,
Received in revised form 9 September 2010 flavonoids, anthocyanins) of bunga kantan inflorescence (Etlingera elatior Jack.). The antioxidant activity
Accepted 9 September 2010
of each individual extract was also evaluated through DPPH (2,2-diphenyl-1-pic-rylhydrazyl) radical
Available online 7 December 2010
scavenging activity and ferric reducing antioxidant power (FRAP) assay. Of all the solvents employed,
50% acetone extract showed highest amount of total phenols (687.0 mg GAE/100 g) and total flavonoids
Keywords:
(1431 mg QE/100 g), while 50% methanol extract showed maximum (5.9 mg c-3-gE/100 g) recovery for
Antioxidants
Anthocyanins
anthocyanins. Tannin extractability was found to be highest with 100% methanol (467.8 mg CE/100 g).
Bioactive compounds The results obtained suggest the use of bunga kantan inflorescence as a potential source of natural
Bunga kantan antioxidants for food and nutraceutical applications.
Phenolic compounds ß 2010 Elsevier Inc. All rights reserved.
Flavonoids
Tannins
Torch ginger
Food analysis
Food composition

1. Introduction communities raw, as a condiment, or cooked. The inflorescences of

During the normal metabolic process in aerobic cells, free
radicals are generated that readily react with the lipids, nucleic acids,
proteins, sugars and sterols (Lee et al., 2004), which might lead to the
development of neurodegenerative diseases. To prevent the onset of
these diseases, high amount of antioxidants need to be present in the
body. The most applicable and healthy way to improve the
antioxidant levels in the body is by consuming various naturally
available food resources. Hence studies about commercial explora-
tion and utilization of plant source as antioxidants are of increasing
interest (Rice-Evans et al., 1997; Alothman et al., 2009).
Etlingera elatior (Jack.), sometimes called ‘‘Torch ginger’’, is a
plant that belongs to Zingiberaceae family in which the ginger
plant species are categorized. This plant is popularly known in
Malaysia as ‘‘bunga kantan’’ (Jaafar et al., 2007), and is commonly
found in the South Asian countries. This plant is considered to be a
native of Sumatra, Indonesia and is referred to as ‘‘kecombrang’’ or
as ‘‘kincung’’. Torch ginger plant has extensive traditional uses: the
young shoots, flower buds or fruits are consumed by indigenous
* Corresponding author at: Food Technology Division, Universiti Sains Malaysia,
Minden Campus, Penang 11 800, Malaysia. Tel.: +60 4653 5212; fax: +60 4657 3678.
E-mail addresses: rajeevbhat1304@gmail.com, rajeevbhat@usm.my (R. Bhat).
this plant are very popular as a spice for food flavouring; because of
them, the plant is also used as a garden ornamental. In some
traditional foods of Malaysia (like asam laksa, nasi kerabu and nasi
ulam) the inflorescence is a key ingredient (Chan et al., 2007).
Earlier studies have reported (Mohamad et al., 2005; Abas et al.,
2006; Chan et al., 2009) on the analysis of antioxidant activities of
leaves and rhizomes of this plant. However, to our knowledge no
detailed studies on the antioxidant activities of the inflorescence
have been reported.
Generally, for the extraction of polyphenols or other bioactive
compounds from plant materials, water and organic solvents
(ethanol, methanol, acetone, diethyl ether) are used. Additionally,
during the extraction process, the percent recovery depends
mainly on the type of solvent and the extraction methods being
adapted (Sun and Ho, 2005; Turkmen et al., 2006; Hayouni et al.,
2007). Depending on the plant materials, the nature of the
bioactive compound present also varies. Hence, in general, it is
very difficult to recommend a suitable extracting solvent for
individual plant materials. The main objective of the present study
was to evaluate the effect of various solvents (methanol, acetone
and water) on the extractability of antioxidant compounds such as
total phenols, total flavonoids, tannins, anthocyanin and antioxi-
dant activity (as percent DPPH inhibition activity and FRAP assay)
of the inflorescence of bunga kantan. We therefore hope to provide

0889-1575/$ – see front matter ß 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2010.09.018
616 M.M.J.O. Wijekoon et al. / Journal of Food Composition and Analysis 24 (2011) 615–619
a suitable base for commercial exploitation as a source of natural
antioxidant for food and nutraceutical applications.
2. Materials and methods
2.1. Standards and reagents
Folin’s reagent (Folin-Ciocalteu reagent), gallic acid, quercetin,
catechin, vanillin and 2,2-diphenyl-1-picrylhydrazyl (DPPH) were
purchased from Sigma–Aldrich1, USA. Acetone, aluminium chlo-
ride, ferrous sulphate, ferric chloride, methanol, potassium chloride,
sodium acetate, sodium carbonate and sodium hydroxide were
purchased from R & M chemicals (Essex, UK). All the other chemicals
used in the present study were of analytical grade quality.
2.2. Sample preparation
Inflorescences of bunga kantan (Fig. 1) were purchased from the
local wet market in Pulau Penang, Malaysia. Fresh, unopened
inflorescences selected were of equal maturity and uniform colour
with no apparent physical defects. Soon after the collection, the
samples were brought to the laboratory and surfaced cleaned with
distilled water (to remove adherent dust particles). Further, the
samples were cut into uniformly sized small pieces (approximately
0.5 cm  0.5 cm) and were subjected to freeze drying for 48 h.
(Model 7754511, Labconco Corporation, Kansas City, Missouri,
USA). On completion of freeze drying, the samples were ground to
a fine powder using a commercial kitchen blender (Model BL 335,
Kenwood, Selangor, Malaysia) and stored at 4 8C in amber coloured
glass bottles, covered with aluminium foil (to prevent direct
exposure to light) until further analysis.
2.3. Solvent extraction
One gram of accurately weighed freeze dried sample powder
was mixed with 40 ml of the desired solvent and extracted using
the magnetic stirrer and hotplate at 1200 rpm (Model FSMQ
143551, Fisher Scientific, Kuala Lumpur, Malaysia) for one hour at
room temperature (25  1 8C). Extracts were then filtered under
suction by using Whatman No. 1 filter paper. The remaining residue
on the filter paper was transferred back into the same flask and re-
extracted for two more times following the same procedure until the
residue became colourless. Filtrates collected from all the three
successive extractions were pooled and collected in reagent bottle
covered with aluminium foil (to avoid light exposure). For the
extraction, distilled water and different concentrations of methanol
and acetone (50%, 90% and 100%, v/v) were used. Each of the
extraction was carried out in replicates (n = 3).
Fig. 1. Inflorescence of Etlingera elatior.
2.4. Determination of total phenolic content
Determination of total phenols in the extract was done using
Folin-Ciocalteu (FC) assay as described by Singleton and Rossi
(1965) with slight modifications. Properly diluted (until the
absorbance unit measured in the spectrophotometer was <1.2)
extract of 450 ml was added to 2.25 ml of FC reagent (ten fold
diluted with distilled water) and 1.8 ml of sodium carbonate (7.5%,
w/v). Further, the contents were mixed and allowed to stand for
30 min at room temperature (25  1 8C). The absorbance was
measured at 765 nm using UV–vis spectrophotometer (Shimadzu UV-
160A PC, Shimadzu Corporation, Kyoto, Japan). Total phenol content
was expressed as mg gallic acid equivalent (GAE), per 100 g wet
weight.
2.5. Determination of total flavonoid content
Total flavonoid content was determined using the aluminium
trichloride method as described by Liu et al. (2008). Briefly, 500 ml
of the extract solution was added to a test tube with 2.5 ml of
distilled water. Sodium nitrite solution (5%, w/v, 150 ml) was
added to the mixture and maintained for 5 min. After that, 300 ml
of aluminium chloride (10%, w/v) was added. After 6 min of
incubation, 1 ml of 1 M sodium hydroxide (NaOH) was added.
Then, the mixture was diluted with 550 ml of distilled water, and
shaken vigorously. The absorbance of the mixture was measured at
510 nm (UV–vis spectrophotometer, Shimadzu UV-160A PC,
Shimadzu Corporation, Kyoto, Japan) immediately, and the total
flavonoid content was expressed as mg quercetin equivalent (QE)/
100 g sample.
2.6. Determination of total anthocyanin content
The spectrophotometric pH differential method as described by
Giusti and Wrolstad (2001) was used to determine the total
anthocyanin content in the extracts of the inflorescence. In brief,
0.5 ml of the extract was mixed thoroughly with 3.5 ml of 0.025 M
potassium chloride buffer (at pH 1). The mixture was allowed to
stand for 15 min and the absorbance was measured at 510 and
700 nm (UV–vis spectrophotometer, Shimadzu UV-160A PC,
Shimadzu Corporation, Kyoto, Japan) against a blank of distilled
water. Following the same procedure, extract was then added to
0.025 M sodium acetate buffer at pH 4.5, and the absorbance was
again measured at 510 nm and 700 nm after 15 min. The total
anthocyanin content was calculated using the following equation:
A  MW  DF  1; 000
Total anthocyanin content ðmg=lÞ ¼
eC
where A is absorbance of the extract calculated as:
A ¼ ðA515  A700 ÞpH 1:0  ðA515  A700 ÞpH 4:5
MW is the molecular weight for cyanidin-3-glucoside = 449.2; DF is
the dilution factor of the samples, e is the molar absorptivity of
cyanidin-3-glucoside = 26,900; and C is the concentration of the
buffer in mg/ml. Results were expressed as mg of cyanidin-3-
glucoside equivalents (c-3-gE) for 100 g of sample.
2.7. Determination of tannin content
For the determination of tannins in the sample extracts,
vanillin–HCl method described by Broadhurst and Jones (1978)
was adapted. Briefly, 0.5 ml of the extract was added to 3 ml of
vanillin reagent (4%, w/v, vanillin in methanol) and mixed
thoroughly. To this mixture 1.5 ml concentrated hydrochloric acid
was added and vortex mixed. Content was kept in the dark for
 M.M.J.O. Wijekoon et al. / Journal of Food Composition and Analysis 24 (2011) 615–619 617

15 min at room temperature and the absorbance was read at
500 nm using UV–vis spectrophotometer (Shimadzu UV-160A PC,
Shimadzu Corporation, Kyoto, Japan). A blank sample was
prepared using the same procedure, but replacing the 0.5 ml of
extract with the solvent used in extraction. The content of tannins
in the sample was expressed as mg catechin equivalent (CE)/100 g
sample.
2.8. DPPH free radical scavenging activity
The capacity of the extracts to scavenge the 2,2-diphenyl-1-
picrylhydrazyl (DPPH) radical was assessed according to the
method described by Sanchez-Moreno et al. (1998). Briefly, 0.1 ml
of the extract was added to 3.9 ml of methanolic solution of DPPH
radical (25 mg/l). The mixture was shaken vigorously and left in
the dark for 30 min. The absorbance of the mixture was read at
515 nm against a blank of absolute methanol without DPPH. The
results were expressed as percentage inhibition of DPPH according
to the following formula:
Acontrol  Asample
Percent inhibition of DPPH ¼  100
Acontrol
where Acontrol is the absorbance of the DPPH solution without
extract and the Asample is the absorbance of the sample with DPPH
solution.
2.9. Ferric reducing/antioxidant power assay (FRAP assay)
The method described by Benzie and Strain (1996) was adapted
for the assay of FRAP. FRAP reagent was freshly prepared by mixing
100 ml of acetate buffer (0.3 M, pH 3.6), 10 ml of 2,4,6-tris(2-
pyridyl)-5-triazine (TPTZ) solution (10 mM in 40 mM HCl) and
10 ml of FeCl36H2O (20 mM) solution. A 60 ml aliquot of the
extract was mixed with 4.5 ml of FRAP reagent. Then the reaction
mixture was incubated in the water bath at 37 8C for 4 min. The
absorbance of the mixture was measured at 593 nm (UV–vis
spectrophotometer, Shimadzu UV-160A PC, Shimadzu Corpora-
tion, Kyoto, Japan) against a blank prepared by replacing the
amount of extract with water. FRAP activity was expressed as
millimoles of Fe(II) per 100 g of sample, based on the calibration
curve prepared with FeSO47H2O.
2.10. Statistical analysis
The results of the present study (samples run in triplicates,
n = 3) are represented as mean values  S.D. Analysis of variance
was performed and significant differences between mean values
were determined by Tukey’s pair-wise comparison test at a level of
significance of p < 0.05. Statistical analyses were conducted using
SPSS 12.01 (SPSS Inc., Wacker Drive, Chicago, USA).
3. Results and discussion
3.1. Total phenols
Phenolic compounds in plants are known to act as free radical
scavengers and it has been opined that the antioxidant activity of
most of the plant produce is mainly due to the presence of phenolic
compounds (Skerget et al., 2005). Basically, antioxidant mecha-
nism of polyphenolic compounds is based on their hydrogen
donating and metal ion chelating abilities (Lee et al., 2004; Jacobo-
Velazquez and Cisneros-Zevallos, 2009).
Table 1 shows the extraction yield of total phenols with
different solvents. According to the obtained results, it was
evident that both 50% acetone and methanol extracts had
highest levels of total phenols followed by 90% and absolute
solvents. Overall, considering all the solvent systems, 50%
acetone (687.0 mg GAE/100 g) was found to be the most
effective solvent and water (90.7 mg GAE/100 g) was the least
effective solvent for phenol extraction. Our results are on par
with the earlier observations (in plant products) of Zhou and Yu
(2004) in wheat bran extracts and Turkmen et al. (2006) in
mate tea extracts, wherein 50% acetone was reported to be
the most effective solvent. In the samples extracted with
100% methanol, the amount of total phenols recovered was
361.2 mg GAE/100 g, which was comparatively higher than the
total phenols recovered from the inflorescence (295 mg GAE/
100 g) and leaves (3550 mg GAE/100 g) of torch ginger as
reported earlier by Chan et al. (2007). This difference might
be attributed to the variations in the sample preparation,
extraction methods and extraction time. These results suggest
that the extractability of polyphenols is influenced by the
polarity and viscosity of the solvents used (Turkmen et al., 2006;
Hayouni et al., 2007).
3.2. Total flavonoid content
Flavonoids are the most common and widely distributed
important single group of phenols present in plants that are highly
effective as antioxidants (Yanishlieva-Maslarova, 2001). Flavo-
noids can inhibit metal-initiated lipid oxidation by forming
complexes with metal ions (Lee et al., 2004). The potential
antioxidant activity of flavonoids is associated with the chemical
structures with o-diphenolic group, a 2–3 double bond conjugated
with the 4-oxo function and hydroxyl groups in positions 3 and 5
(Bravo, 1998).

Table 1
Total phenol, flavonoid, anthocyanin and tannin contents of Etlingera elatior inflorescence extracted with various solvents (n = 3  S.D.).

Solvent (mg GAE/100 g)A Total phenols (mg QE/100 g)B Flavonoids (mg QE/100 g)B Anthocyanins (mg c-3-gE/100 g)C Tannins (mg CE/100 g)D
a a a
Water 90.7  1.7 182.5  3.2 0.90  0.5 37.2  1.5a
Methanol
100% 361.2  17.1b 762.8  44.5b 5.10  2.9b 467.8  36.4b
90% 431.4  25.8c 632.6  51.2c 4.50  2.1c 438.4  18.7c
50% 615.0  14.6d 717.6  41.0d 5.90  0.4d 293.0  18.9d
Acetone
100% 142.8  14.5a 301.6  57.0e 1.50  1.1a 255.5  9.6e
90% 502.8  15.9e 1025  97.1f 2.30  1.3a 151.4  8.8f
50% 687.0  43.5f 1431  34.0g 3.60  0.4a 360.7  8.2g

Letters followed by the same letter in the same column are not statistically significant from each other at p < 0.05.
A
Gallic acid equivalents.
B
Quercetin equivalents.
C
cyanidin-3-glucoside equivalents.
D
Catechin equivalents.
618 M.M.J.O. Wijekoon et al. / Journal of Food Composition and Analysis 24 (2011) 615–619
Table 1 shows the flavonoid content of the samples extracted in
different solvent systems. Samples extracted in 50% acetone
showed highest amount (1431 mg QE/100 g) of total flavonoids.
With the increasing concentration of organic solvent, a corre-
sponding significant decrease in the total flavonoids was observed.
These results highlight that the yield of extraction of phenolic
compounds is dependent on the polarity of the solvent being used.
3.3. Anthocyanins
Determination of anthocyanin content with pH differential
method gives precise results even in the presence of interfering
compounds, as this method is based on the formation of reversible
structural transformations of anthocyanin pigments (at pH 1 and
pH 4.5). At pH 1 anthocyanin exists as coloured (pink-orange)
oxonium form and at pH 4.5 they exist as colourless hemiketal
form (Giusti and Wrolstad, 2001). Based on the results in Table 1,
50% methanol gave the highest yield of anthocyanins and water
gave the lowest amount of anthocyanins. Earlier, methanol has
been reported to be the most effective solvent for extraction of
anthocyanins and has been shown to be 73% more effective than
water (Metivier et al., 1980).
3.4. Tannins
In case of tannins, the recovery was significantly increased with
increasing the concentration of methanol only. The tannin levels
recorded was 467.8 and 438.4 mg CE/100 g sample weight in 100
and 90% methanol, respectively. On comparing the yield of tannins
extracted with acetone, a significantly high extraction yield was
observed at 50% compared to other concentrations. Overall, the
levels of tannins were low in water compared to other solvents
under study (37.2 mg CE/100 g sample weight). It is a well known
fact that in addition to the numerous hydroxyl groups, tannins do
have some hydrophobic character due to the presence of the
benzene rings in the molecule. Therefore, it could be hypothesized
that an appropriate extraction solvent for tannins could have
hydrophilic character as well as a degree of hydrophobicity. Hence,
this might be the possible reason that methanol and acetone gave
much higher yields of tannins compared to aqueous/water extract.
3.5. Antioxidant activity (percent inhibition of DPPH and FRAP assay)
DPPH assay is widely used in the determination of free radical
scavenging activity of natural antioxidants, mainly due to its
simplicity and high sensitivity. In this assay, an antioxidant
donates hydrogen or electron which is then accepted by the DPPH
radicals (Moon and Shibamoto, 2009). Upon the reduction of
DPPH radical to stable diamagnetic molecule, the colour changes
from purple to yellow. DPPH radical itself shows a strong
absorption maximum at 515 nm which measured by using
spectrophotometer.
In FRAP assay, the reducing ability of an antioxidant is
measured. In the presence of antioxidants, Fe3+–TPTZ (2,4,6-
tris(2-pyridyl)-5-triazine) complex is reduced to Fe2+–TPTZ in the
acidic medium. The reduced form of TPTZ has an intense blue
colour which has absorption at 593 nm (Moon and Shibamoto,
2009).
In the present study, results of the DPPH and FRAP assay
showed identical trends to that of total phenol content. Of all the
solvent systems studied, 50% acetone had the highest percentage
inhibition (94.9%) with DPPH. Similarly, the FRAP value for the 50%
acetone was the highest (4.2 mM Fe(II)/100 g). Based on the results
shown in Table 2, it was evident that the percentage inhibition of
DPPH and FRAP values increased with the corresponding reduction
in the concentration of methanol and acetone.
Table 2
DPPH and FRAP values of bunga kantan inflorescence extracted with various
solvents (n = 3  S.D.).
Solvent Inhibition of DPPH (%) FRAP (mM Fe(II)/100 g)
Water 39.6  4.8b 0.9  0.1a
Methanol
100% 89.1  2.7c 2.6  0.1b
90% 93.9  1.3d 2.8  0.4b
50% 94.7  0.1d 3.6  0.3c
Acetone
100% 25.2  12.2a 0.5  0.1a
90% 87.0  2.5c 3.8  0.2c
50% 94.9  0.4d 4.2  0.1d
Letters followed by the same letter in the same column are not statistically
significant from each other at p < 0.05.
Earlier, it has been opined that with the change of solvent
polarity, vapour pressure and viscosity, the type of antioxidant
compound being dissolved in the solvent also varies. Solvents with
low viscosity have low density and high diffusivity that allows
them to easily diffuse into the pores of the plant materials to leach
out the bioactive constituents (Naczk and Shahidi, 2006). As a
result of this, the antioxidant activity of the extract observed also
varies (Zhou and Yu, 2004; Turkmen et al., 2006; Alothman et al.,
2009).
4. Conclusion
This study shows that, depending on the type of solvent, the
type of antioxidant compounds being extracted also vary.
Additionally, further research is warranted to explore the
individual or major polyphenolic groups and other bioactive
compounds in the inflorescence of bunga kantan and their
contribution to health. It is also envisaged that the inflorescence
might possess high antimicrobial activities with promising
potential to be used in food industries as a natural preservative
as well.
Acknowledgements
One of the authors (Ms. Jeevani Osadee Wijekoon) thanks
Universiti Sains Malaysia, Malaysia for the Postgraduate fellowship
awarded. The authors gratefully acknowledge the anonymous
referees for comments and constructive suggestions provided for
improving the manuscript.
References
Abas, F., Lajis, N.H., Israf, D.A., Khozirah, S., Kalsom, Y.U., 2006. Antioxidant and nitric
oxide activities of Malay traditional vegetables. Food Chemistry 95, 566–573.
Alothman, M., Bhat, R., Karim, A.A., 2009. Antioxidant capacity and phenolic content
of selected tropical fruits from Malaysia, extracted with different solvents. Food
Chemistry 115, 785–788.
Benzie, I.F.F., Strain, J.J., 1996. The ferric reducing ability of plasma (FRAP) as a
measure of antioxidant power: the FRAP assay. Analytical Biochemistry 239,
70–76.
Bravo, L., 1998. Polyphenols: chemistry, dietary sources, metabolism and nutrition-
al significance. Nutrition Reviews 56, 317–333.
Broadhurst, R.B., Jones, W.T., 1978. Analysis of condensed tannins using acidified
vanillin. Journal of the Science of Food and Agriculture 29, 788–794.
Chan, E.W.C., Lim, Y.Y., Omar, M., 2007. Antioxidant and antibacterial activity of
leaves of Etlingera species (Zingiberaceae) in Peninsular Malaysia. Food Chem-
istry 104, 1586–1593.
Chan, E.W.C., Lim, Y.Y., Wong, S.K., Lim, K.K., Tan, S.P., Lianto, F.S., Young, M.Y., 2009.
Effects of different drying methods on the antioxidant properties of leaves and
tea of ginger species. Food Chemistry 113, 166–172.
Giusti, M.M., Wrolstad, R.E., 2001. Characterization and measurement of antho-
cyanins by UV–visible spectroscopy. In: Wrolstad, R.E. (Ed.), Current Protocols
in Food Analytical Chemistry. John Wiley and Sons, New York, Unit F1.2.1-13.
Hayouni, A., Abedrabba, M., Bouix, M., Hamdi, M., 2007. The effects of solvents and
extraction method on the phenolic contents and biological activities in vitro of
Tunisian Quercus coccifera L. and Juniperus phoenicea L. fruit extracts. Food
Chemistry 105, 1126–1134.
 M.M.J.O. Wijekoon et al. / Journal of Food Composition and Analysis 24 (2011) 615–619
Jaafar, F.M., Osman, C.P., Ismail, H., Awang, K., 2007. Analysis of essential oils of
leaves, stems, flowers and rhizomes of Etlingera elatior (Jack) R.M. Smith. The
Malaysian Journal Analytical Sciences 11, 269–273.
Jacobo-Velazquez, D.A., Cisneros-Zevallos, L., 2009. Correlations of antioxidant
activity against phenolic content revisited: a new approach in data analysis
for food and medicinal plants. Journal of Food Science 74, 107–113.
Lee, J., Koo, N., Min, D.B., 2004. Reactive oxygen species, aging and antioxidative
nutraceuticals. Comprehensive Reviews in Food Science and Food Safety 3,
21–33.
Liu, X., Zhao, M., Wang, J., Yang, B., Jiang, Y., 2008. Antioxidant activity of methanolic
extract of emblica fruit (Phyllanthus emblica L.) from six regions in China. Journal
of Food Composition and Analysis 21, 219–228.
Metivier, R.P., Francis, F.J., Clydesdale, F.M., 1980. Solvent extraction of anthocya-
nins from wine pomace. Journal of Food Science 45, 1099–1100.
Mohamad, H., Lajis, N.H., Abas, F., Ali, A.M., Sukari, M.A., Kikuzaki, H., Nakatani, N.,
2005. Antioxidative constituents from Etlingera elatior. Journal of Natural
Products 68, 285–288.
Moon, J., Shibamoto, T., 2009. Antioxidant assays for plants and food components.
Journal of Agricultural and Food Chemistry 57, 1655–1666.
Naczk, M., Shahidi, F., 2006. Phenolics in cereals, fruits and vegetables: occurrence,
extraction and analysis. Journal of Pharmaceutical and Biomedical Analysis 41,
1523–1542.
619
Rice-Evans, C.A., Miller, N.J., Paganga, G., 1997. Antioxidant properties of phenolic
compounds. Trends in Plant Science 2, 152–159.
Sanchez-Moreno, C., Larrauri, J.A., Saura-Calixto, F., 1998. A procedure to measure
the antiradical efficiency of polyphenols. Journal of the Science of Food and
Agriculture 76, 270–276.
Singleton, V.L., Rossi, J.A., 1965. Colorimetry of total phenolics with phosphomo-
lybdic–phosphotungstic acid reagents. American Journal of Enology and Viti-
culture 16, 144–158.
Skerget, M., Kotnik, P., Hadolin, M., Hras, A.R., Simonic, M., Knez, Z., 2005. Phenols,
proanthocyanidins, flavones and flavonols in some plant materials and their
antioxidant activities. Food Chemistry 89, 191–198.
Sun, T., Ho, H., 2005. Antioxidant activities of buckwheat extracts. Food Chemistry
90, 743–749.
Turkmen, N., Sari, F., Velioglu, Y.S., 2006. Effects of extraction solvents on concen-
tration and antioxidant activity of black and black mate tea polyphenols
determined by ferrous tartrate and Folin-Ciocalteu methods. Food Chemistry
99, 835–841.
Yanishlieva-Maslarova, N.V., 2001. Inhibiting oxidation. In: Pokorny, J., Yanishlieva,
N., Gordon, M.H. (Eds.), Antioxidants in Food: Practical Applications. Woodhead
Publishing Limited, Cambridge, pp. 22–70.
Zhou, K., Yu, L., 2004. Effects of extraction solvent on wheat bran antioxidant
activity estimation. Lebensmittel-Wissenschaft und-Technologie 37, 717–721.