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Innovative Food Science and Emerging Technologies 11 (2010) 169176

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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

Identication and antioxidant activity of anthocyanins extracted from the seed and
cob of purple corn (Zea mays L.)
Zhendong Yang , Weiwei Zhai
Food Engineering Department of Jiangsu Food Science College, Huaian, Jiangsu 223003, China

a r t i c l e

i n f o

Article history:
Received 7 March 2009
Accepted 13 August 2009
Editor Proof Receive Date 3 September 2009
Purple corn
Antioxidant activity

a b s t r a c t
The total anthocyanin content (TAC) and the antioxidant activity of the seed and cob from Chinese purple
corn (Zea mays L., cv Zihei) extracts were determined by pH-differential method, and DPPH, FRAP, and TEAC
methods, respectively. TAC in purple corn cob anthocyanins (PCCAs) extract was higher than TAC in purple
corn seed anthocyanins (PCSAs) extract. Compared to bulylated hydroxytoluene (BHT), PCCAs and PCSAs
possessed signicantly higher antioxidant activities, according to the DPPH, FRAP and TEAC assays. A satisfactory correlation between TAC and antioxidant activity was observed. Result indicated that cyanidin-3glucoside, pelargonidin-3-glucoside and peonidin-3-glucoside were components in PCSAs extracts, and
seven kinds of anthocyanin had been detected and six kinds of anthocyanin in PCCAs extracts were separated
and identied them as cyanidin-3-glucoside, pelargonidin-3-glucoside and peonidin-3-glucoside, and their
respective malonated counterparts as their anthocyanins using HPLCMS analysis.
Industrial relevance: In the last decades, in interest in anthocyanin pigments has increased because of their
possible utilization as natural food colorants and especially as antioxidant and anti-inammatory agents.
Purple corn is a pigmented variety of Z. mays L., originally cultivated in Latin America. Now, this corn variety
is mainly grown in China, especially in Shanxi and Anhui Province, could be new and interesting sources to
obtain extracts rich in anthocyanins for their use in food, pharmaceutical and cosmetic industries. Our results
indicated that the seed and cob of purple corn possessed excellent antioxidant activity, which could lead to
increased application of these natural food colorants by the food industry.
2009 Elsevier Ltd. All rights reserved.

1. Introduction
Anthocyanins, as a group of phenolic compounds widely existing
in the plant kingdom, present a spectrum from orange to blue in
colour in the natural world, satisfying consumers' desire for food
colours. They possess known pharmacological properties and are used
for therapeutic purpose (Smith, Marley, Seigler, Singletary, & Meline,
2000; Wang et al., 2000; Cooke, Steward, Gescher, & Marczylo, 2005;
Pergola, Rossi, Dugo, Cuzzocrea, & Sautebin, 2006). In this way,
anthocyanins have been noted not only as a food colorant but also as a
health food material.
There are various kinds of corn in the world, and the corn has
various colours such as white, yellow, red, purple, brown, green and
blue. Purple corn is a pigmented variety of Zea mays L., originally cultivated in Latin America and was introduced in China long time ago.
This kind of corn is mainly grown in China, especially in Shanxi and
Anhui province. Now, purple corn is an important source of antho-

Corresponding authors. Tel./fax: +86 0517 87088869.

E-mail address: (Z. Yang).
1466-8564/$ see front matter 2009 Elsevier Ltd. All rights reserved.

cyanins with peculiar features regarding coloring of foods. As natural

colorant, purple corn anthocyanins have been using for coloring beverages, jellies, candies and so on in China. The individual anthocyanins
of Andean purple corn, leaves, cobs, and seeds have been characterized, including cyanidin-3-dimalonylglucoside, cyanidin-3-glucoside,
pelargonidin-3-glucoside, peonidin-3-glucoside, and their respective
malonated counterparts as their main anthocyanins (Aoki, Kuze, &
Kato, 2002; Pascual-Teresa, Santos-Buelga, & Rivas-Gonzalo, 2002;
Harborne & Self, 1997). Likewise, a similar anthocyanin composition
was also found in corn owers (Fossen, Slimestad, & Andersen, 2001).
However, the literature is mainly focused on purple corn of Peruvian
varieties or derived products (Pascual-Teresa et al., 2002; CevallosCasals & Cisneros-Zevallos, 2003; Moreno, Snchez, Hernadez, &
Lobato, 2005; Pedreschi & Cisneros-Zevallos, 2007). Only a limited
number of works deal with Chinese varieties of purple corn (Zhang,
Mu, & Gong, 1998; Zhao et al., 2008), whereas the anthocyanin
composition and antioxidant activity of Chinese purple corn hybrids is
still not well documented. Hence, the anthocyanin content, antioxidant activity and individual anthocyanin composition of Chinese
purple corn hybrid (from Anhui province) were determined for the
rst time in this work.


Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176

To our knowledge, there has been no report on antioxidant activity between PCSAs extracts and PCCAs extract. Therefore, the main
purpose of the present study was to determine the total anthocyanins content (TAC) by pH-differential method and determine the
antioxidant activity of PCSAs and PCCAs from China, by use of the three
commonly used spectrophotometric methods: DPPH (2,2-diphenyl-1picrylhydrazyl) free radical-scavenging assay, TEAC (Trolox equivalent
antioxidant capacity) assay, and FRAP (ferric ion reducing antioxidant
power) assay. Furthermore, the main anthocyanins in PCSAs extracts
and PCCAs extract were analyzed high performance liquid chromatography (HPLC) and HPLCmass spectroscopy with electrospray ionization source.

2.3. Determination of total anthocyanin content (TAC)

The TAC was determined according the pH-differential method
(Giusti & Worsltad, 2001). An aliquot of the sample (1 mg) was placed
into 25 ml volumetric ask and made up to the nal volume with pH
1.0 buffer. Another 1 mg of the sample was also placed into a 25 ml
volumetric ask, made up to a nal volume with pH 4.5 buffer. Absorbance was measured by a spectrophotometer (UV-2802, UNICO) at
510 and 700 nm, respectively. Absorbance was calculated as Abs =
(A510 A700) pH 1.0 (A510 A700) pH 4.5 with a molar extinction
coefcient for cyanidin-3-glucoside of 26900. TAC was calculated
using the following equation and expressed as milligrams of cyanidin3-glucoside equivalents per 100 g dry seeds (Eq. (1)). All TCA shown
was made as cyanidin-3-glucoside equivalents.

2. Materials and methods


2.1. Material

TAC mg=100 g

The seed and cob of purple corn (Zea mays L., cv Zihei) (Fig. 1)
were generously supplied by Professor Zhong Zhang of Anhui
Technical Teachers College (Fengyang City, Anhui Province, China).
The seed and cob were dried in a heated air drier (50 C) (ZT-3,
Jiangdu City, Jiangsu Province, China), pulverized by the disintegrator
(FSD-100A, Taizhou City, Zhejiang Province, China) and sifted through
a 100 mesh sieve, respectively. The purple corn seed and purple corn
cob powder (water content b 12%) was sealed in a brown bottle and
kept at 4 C, respectively. ()-Epigallocatechin gallate (EGCG), ()epicatechin gallate (ECG), ()-epigallocatechin (EGC), and ()epicatechin (EC) were purchased from Funakoshi Co., Ltd. (Tokyo,
Japan). Caffeine, chlorogenic acid, DPPH, 2,2-azino-bis (3-ethylbenzothiazolin-6-sulfonate) diammonium salt (ABTS), theobromine,
theophylline, and bulylated hydroxytoluene (BHT) were obtained
from Sigma Chemical Co. (St. Louis, MO). FolinCiocalteu reagent,
2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) and 6-hydroxy- 2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox) were obtained from Fluka
(Buchs, Switzerland). All other chemicals were of analytical grade.

where Abs is absorbance, e is cyanidin-3-glucoside molar absorbance

(26900), L is the cell path length (1 cm), MW is the molecular weight
of anthocyanin (449.2 Da), D is the dilution factor, V is the nal volume
(ml), and M is the dry weight (mg).
2.4. Colour coordinates
Tristimulus parameters (L*, C* and h) were calculated using CRS4w software (CR400, Konica Minolta, Japan), based on CIELAB colour
space, and with reference to standard observation angle of 10 visual
eld and a standard illuminator D65 at of 5 nm (Fan, Han, Gu, & Gu,
2.5. Antioxidant activity determination
Three methods, DPPH, TEAC, and FRAP, based on reaction with
electron-donating or hydrogen radicals (H) producing compounds/
antioxidants according to the reaction R + Aox-H RH + Aox, were

2.2. Preparation of PCSAs and PCCAs extracts

Seed (100 g) and cob (100 g) were macerated with 400 ml methanol for 24 h in the dark at 4 C, respectively. Both crude extracts
obtained were ltered through lter paper and the remaining residues were washed with 200 ml of methanol, and then collected the
crude extract ltrate. The methanol in the crude extract was evaporated by the rotary evaporator (0.1 MPa, 35 C). The extracts were
then subjected to the measurement of TAC and antioxidant activity,
and HPLCMS analysis.

2.5.1. DPPH assay

The DPPH free radical-scavenging activity of each sample was determined (Leong & Shui, 2002; Hu, Lu, Huang, & Ming, 2004). Briey, a
0.1 mM solution of ethanolic DPPH solution was prepared. The initial
absorbance of the DPPH in ethanol was measured at 517 nm and did
not change throughout the period of assay. An aliquot (0.1 ml) of each
sample (with appropriate dilution if necessary) was added to 3.0 ml of
ethanolic DPPH solution. Discolorations were measured at 517 nm
after incubation for 30 min at 30 C in the dark. Measurements were

Fig. 1. Picture of purple corn seed and purple corn cob.

Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176


performed at least in triplicate. The percentage of DPPH(%DPPHsc)

was calculated as:
% DPPHsc = AcAs 100 = Ac

where Ac is the absorbance of the control, and As is the absorbance

of the sample. IC50 values calculated denote the concentration of a
sample required to decrease the absorbance at 517 nm by 50%.
2.5.2. TEAC assay
The ABTS free radical-scavenging activity of each sample was
determined according to the method described by Pavel and Vlastimil
(2006). The radical cation ABTS+ was generated by persulfate oxidation of ABTS. A mixture (1:1, v/v) of ABTS (7.0 mM) and potassium
persulfate (4.95 mM) was allowed to stand overnight at room temperature in the dark to form radical cation ABTS+. A working solution
was diluted with phosphate buffer solution to reach absorbance values
between l.0 and 1.5 at 731 nm (constant initial absorbance values
must be used for standard and samples). An aliquot (0.1 ml) of each
sample (with appropriate dilution if necessary) was mixed with the
working solution (3.9 ml), and the decrease of absorbance was measured at 731 nm after 10 min at 37 C in the dark. Aqueous phosphate
buffer solution (3.9 ml, without ABTS+ solution) was used as a control.
The ABTS+ scavenging rate was calculated to express the antioxidant
ability of the sample. IC50 values calculated denote the concentration of
sample required to decrease the absorbance at 731 nm by 50%.
2.5.3. FRAP assay
The ability to reduce ferric ions was measured using a modied
version of the method described by Benzie and Strain (1996). An aliquot (0.2 ml) of each sample (with appropriate dilution if necessary)
was added to 3.8 ml of FRAP reagent (10 parts of 300 mM sodium
acetate buffer at pH 3.6, 1 part of 10.0 mM TPTZ solution, and 1 part of
20.0 mM FeCl36H2O solution), and the reaction mixture was incubated at 37 C for 30 min. The increase in absorbance at 593 nm was
measured. Fresh working solutions of FeSO4 were used for calibration.
The antioxidant capacity based on the ability to reduce ferric ions of
the samples were calculated from the linear calibration curve and
expressed as mmol FeSO4 equivalents per gram of sample (DW).
2.6. HPLCDADMS analysis of anthocyanins
A high performance liquid chromatography (HPLC) Waters 2690
and a Waters 2996 photodiode array detector were used. Data analysis
was performed with Waters HPLC chem-station software. Solvents
and samples were ltered through a 0.45 m lter. Chromatographic
analysis was carried out using a Lichrospher C-18 (5 m 2.1 mm
250 mm i.d.) prodigy. Simultaneous monitoring was performed at
520 nm at a ow rate 0.3 mL/min. The running temperature was
at 35 C and the injection volume was 10 l. Mobile phase A was a
mixture of 0.05% (v/v) triuoroacetic acid (TFA) in distilled water,
whereas Mobile B consisted of 100% HPLC-grade acetonitrile. Separation of anthocyanins was carried out for 40 min. The elution prole
was a linear gradient elution with 9580% solvent A from 0 to 20 min,
8060% from 20 to 50 min, at a ow rate of 0.8 ml/min. The identity of
anthocyanins were checked using electrospray mass spectrometry

Table 1
TAC and CIELAB value of PCSAs and PCCAs extracts.

TAC (mg/100 g)




55.8 1.5 a
92.3 2.1 b

30.69 0.39a
25.70 0.19b

12.65 0.25b
21.27 0.14a

2.25 0.13b
12.12 0.18a

All the trials were performed in triplicate (n = 3) and all the data were reported as
means SD; TAC expressed as mg of cyanidin-3-glucoside per 100 g of dry seeds. Data
in the same column with different letters are signicantly different (p b 0.05).

Fig 2. Antioxidant activities of PCCAs, PCSAs and BHT determined by DPPH free radicalscavenging assay (A), TEAC assay (B), and FRAP assays (C).

(MS) with a Waters Platform ZMD 4000 mass spectrometer equipped

with an ion spray interface (ISV = 4400) operated in the positive ion
mode. Spectra were recorded in positive ion mode between m/z 200
and 1200.
Table 2
Antioxidant activities of anthocyanins with different extracted solvents and BHT using
the DPPH assay, TEAC assay, and FRAP assay.



FRAP value
(mmol FeSO4/g DW)


40.1 0.8b
48.5 0.5c
175.7 0.4a

38.6 0.3c
43.1 0.5b
55.8 0.6a

18.7 0.4a
16.2 0.5b
8.6 0.4c

Data are present as the mean standard deviation (n N 3).

The antioxidant activity was evaluated as the concentration of the test sample
required to decrease the absorbance at 517 nm by 50%.
The antioxidant activity was evaluated as the concentration of the test sample
needed to decrease the absorbance at 731 nm by 50%.


Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176

Table 3
Correlation coefcients for relations between total anthocyanin contents and the
results from DPPH assay, FRAP assay, and TEAC assay.

FRAP value
Total anthocyanin content



FRAP value




the quantitative component of chromaticity, which is a bidimensional parameter that correlates with the visual sensation attribute colorfulness. The main colour differences among the PCSAs and PCCAs
extracts can be measured in terms of C* values, resulting in 12.65
0.25 and 21.27 0.14 CIELAB units for PCCSA extract and PCSAs
extract, respectively. In addition, a qualitative indicator of Hue angle
(h), was also calculated in the present study, which were 2.25 0.13
and 12.12 0.18 for PCSAs extract and PCCAs extract, respectively.

2.7. Statistical analysis

3.2. Determination of antioxidant activity of extracts

Values shown in tables and graphs were means standard deviations (n = 3). The IC50 values were calculated from linear regression
analysis. The correlation coefcients between TAC and the three
methods of radical-scavenging activity were measured by the SAS
software (Version 6.0).

3.2.1. Determination by DPPH assay

DPPH radical-scavenging activity has been extensively used for
screening antioxidants, such as polyphenols and anthocyanins, from
fruits. DPPHis scavenged by polyphenols and anthocyanins through
the donation of hydrogen, forming the reduced DPPH-H*. The colour
changes from purple to yellow after reduction can be quantied by
its decrease of absorbance at wavelength 517 nm (Bondet, BrandWilliams, & Berset, 1997; Chang et al., 2007). DPPH free radicalscavenging activities of PCSAs, PCCAs and BHT are shown in Fig. 2. For
each sample, ve concentrations (mg/ml) were tested. PCSAs and
PCCAs exhibited considerably higher DPPH free radical-scavenging
activities than BHT. The free radical-scavenging activity of PCSAs was
less than that of PCCAs, which may result from the TAC in purple corn
cob, was lower than that of purple corn seed. Among them (Fig. 2A),
the PCSAs and PCCAs exerted strong activities, showing 98.3 and
95.7% of DPPH radical-scavenging activities at the concentration of
1.0 mg/ml, respectively. On the other hand, BHT showed relatively
low DPPH radical-scavenging activity (70.2%). In order to quantify the
antioxidant activity further, the IC50 was calculated (Table 2). The
lower the IC50 value is, the greater the free radical-scavenging activity
is. The IC50 values of the DPPH radical-scavenging activities were
40.1 0.8, 48.5 0.5 and 175.7 0.4 for PCCAs, PCSAs and BHT, respectively. These results revealed that PCCAs and PCSAs had significantly (p b 0.05) higher scavenge power than BHT.

3. Results and discussion

3.1. The content and CIELAB value of total anthocyanins in PCSAs and
PCCAs extracts
As it can be observed in Table 1, the TAC measured in PCSAs and
PCCAs extracts were calculated to be 55.8 1.5 and 92.3 2.1 mg/
100 g, expressed in cyanidin-3-glucoside, respectively. The TAC in
PCCAs extract showed a signicant (p b 0.05) higher than that in PCSAs
extract. The Chinese purple corn contained higher amounts of anthocyanins than those reported for pigmented corns from the United
States (30.7 mg/100 g), Mexico (72.1 mg/100 g), and Canada
(127.7 mg/100 g) (Dep Pozo-Insfran, Brenes, Serna Saldivar, & Talcott,
2006). It was thus conrmed that cultivars and the growth conditions
of the purple corn can also inuence its anthocyanin content (Ferreyra,
Vina, Mygridge, & Chaves, 2007). To prove this, Moreno et al. (2005)
found that TAC of four Mexican blue corn varieties ranged from 54 to
115 mg/100 g without consistence.
In relation to the colour properties of the PCSAs and PCCAs extracts,
lightness (L*) is an attribute related with the transmission of light
observed. It was 30.44 0.39 and 25.70 0.19 for PCSAs extract
and PCCAs extract in the present study, respectively. Chroma (C*) is

3.2.2. Determination by TEAC assay

The ABTS+ radical formed from the reaction ABTSe ABTS+
reacts quickly with electron/hydrogen donors to form colourless ABST. The reaction is pH-independent. A decrease of the

Fig. 3. Chromatogram of anthocyanin extracts from PCSAs (A) and PCCAs (B) recorded at 520 nm.

Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176

ABTS+ concentration is linearly dependent on the antioxidant

concentration, including Trolox as the calibration (van den Berg,
Haenen, van den Berg, van der Vijgh, & Bast, 2000). The results
correlate very well with biological redox properties of phenolic
substrates. The antioxidant abilities of the MA, EA and BHT by TEAC
method were shown in Fig. 2B. For each sample, ve concentrations
(mg/ml) were tested. The highest ABTS+ scavenging rate was
found for PCCAs, while the lowest was found for BHT. The antioxidant
activities values decreased in the order of PCCAs N PCSAs N BHT. This
rank order was similar to that of the DPPH assay. The IC50 was further
calculated and the values were 30.6 0.3, 43.1 0.5 and 55.8 0.6 for
PCCAs, PCSAs and BHT, respectively.


3.2.3. Determination by FRAP assay

FRAP assay is based on the ability of the antioxidant to reduce Fe3+
to Fe2+ in the presence of TPTZ, forming an intense blue Fe2+TPTZ
complex with an absorption maximum at 593 nm. This reaction is
pH-dependent. The absorbance decrease is proportional to the antioxidant content (Benzie & Strain, 1996). In the present study, the trend
for ferric ion reducing activities of PCCAs, PCSAs and BHT are shown
in Fig. 2C. For, PCCAs and PCSAs, the absorbance clearly increased,
due to the formation of the Fe2+TPTZ complex with increasing
Being expressed in FeSO4 equivalents, the FRAP value was applied
to determine the antioxidant ability of purple corn anthocyanins

Fig. 4. Positive ion mass spectra of purple corn cob anthocyanins (PCCAs) and purple corn seed anthocyanins (PCSAs): (peak 1) cyanidin-3-glucoside; (peak 2) pelargonidin-3glucoside; (peak 3) penodin-3-glucoside; (peak 4) cyanidin-3-(6-malonylglucoside); (peak 5) pelargonidin-3-(6-malonylglucoside); (peak 6) penodin-3-(6-malonylglucoside).


Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176

Fig. 4. (continued).

(Table 2). The FRAP values were 20.7 0.4, 16.2 0.5 and 8.6 0.4
for PCCAs, PCSAs and BHT, respectively. The signicantly (p b 0.05)
higher reducing activity was found for PCCAs and PCSAs, compared
to that of BHT. Similar to the results obtained from the DPPH assay
and TEAC assay. Both PCCAs and PCSAs showed signicantly stronger
ferric ion-reducing activities than that of BHT.
3.3. Correlations of DPPH, TEAC, and FRAP with the total anthocyanin
Anthocyanins have been reported to be responsible for the antioxidant activities of botanical extracts. The DPPH, TEAC, and FRAP

assays have been used to measure the antioxidant activity and their
results should correlate with those of the total anthocyanin contents.
A direct correlation between radical-scavenging activity and anthocyanin content or the extracts was analyzed by the linear regression
analysis. With reference to Table 3, the correlations of the total anthocyanin content against the antioxidant activity based on their DPPH
assay, TEAC assay, and FRAP assay were satisfactory (r N 0.828). The
results indicate that anthocyanins in purple corn extract are largely
responsible for the antioxidant activities. A strong correlation between the mean values of the total anthocyanin content and FRAP
deserves further detailed attention, as it implies that anthocyanins
in purple corn are capable to reduce ferric ions. Some works have

Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176

reported similar correlations between anthocyanins and antioxidant activity measured by various methods (Rivero-Prez, Muiz, &
Gonzlez-Sanjos, 2008; Lachman et al., 2009).


ing conditions (Fossen, Slimestad, & Andersen, 2001; Ferreyra et al.,

4. Conclusions

3.4. Identication of anthocyanin from PCSAs and PCCAs extracts by

HPLCMS analysis
Reverse phase HPLC and MS analysis were used to rapidly identify
the anthocyanin from purple corn extracts. The identication was
carried out by comparing their retention time and conrming the
molecular weight with ESIMS (Aoki et al, 2002; Pascual-Teresa et al.,
As it can be observed in Fig. 3A, a total of three peaks appeared
in the chromatograms, indicating the existence of three kinds of
anthocyanins, accounting for 75.7%, 8.3% and 16.0% respectively of the
total amount of all the anthocyanins, and their retention times were
13.5 min, 15.7 min and 16.6 min, respectively. These pigments were
identied by their HPLC retention times, elution order, spectroscopic
characteristics and fragmentation pattern by comparison with the
previous reported literature (Aoki et al., 2002), who reported that six
kinds of anthocyanins from Peruvian purple corn seed. Fig. 3B shows
the chromatograms of anthocyanins from PCCAs. Seven peaks appears
in the chromatograms, indicating the existence of seven anthocyanins.
Four of these anthocyanins were majors ones, accounting for 58.5, 7.1,
12.4 and 10.9%, respectively of total amount of all the anthocyanins,
and their retention times were 13.5, 15.7, 16.6 and 18.1 min. Pedreschi
and Cisneros-Zevallos (2007) reported that six different anthocyanins
were isolated and identied in Peruvian purple corn cob by HPLCMS,
The structures of PCSAs and PCCAs estimated on the basis of
spectral means are shown in Fig. 4.
Peak 1 (PCSAs and PCCAs), was cyanidin-3-glucoside with molecular ion [M+H]+ at m/z 449 and a fragment ion [M+H-162]+ at m/z
287 was corresponded to cyanidin, which loss of glucose from cyanidin3-glucoside.
Peak 2 (PCSAs and PCCAs), with [M+H]+ at m/z 433, was identied as pelargonidin-3-glucoside. Fragment ion [M+H-162]+ at m/z
271 was corresponded to pelargonidin, loss of glucose from pelargonidin-3-glucoside.
Peak 3 (PCSAs and PCCAs), with [M+H]+ at m/z 463, was identied as peonidin-3-glucoside. The fragment ion at m/z 301, loss of
162 from peonidin-3-glucoside, was characterized as peonidin.
Peak 4 (PCCAs), with [M+H]+ at m/z 535, was identied as
cyanidin-3-(6-malon)-glucoside. The fragment ion [M+H-162-86]+
at m/z 287 was characterized as cyanidin, which cyanidin-3(6-malon)glucoside lost a glucose (162) and a malonic acid (86).
Peak 5 (PCCAs), with [M+H]+ at m/z 519, was identied as
pelargonidin-3-(6-malon)-glucoside. The fragment ion at m/z
433, loss of 162 from pelargonidin-3-glucoside, was characterized
as pelarginidin.
Peak 6 (PCCAs), with [M+H]+ at m/z 549, was identied as
peonidin-3-(6-malon)-glucoside. The fragment ions at m/z 463and
301 referred to peonidin-3-glucoside and peonidin, respectively.
Peak 7 (PCCAs), with [M+H]+ at m/z 575 (picture not shown). The
fragment ions at m/z 449 may be cyanidin-3-glucoside. However, we
cannot supply further information or make suggestions about the
possible identity of the remaining residue of 126 amu.
In this study, the individual anthocyanins in PCCAs extract showed
similar with those reported by Pascual-Teresa et al. (2002) and
Pedreschi and Cisneros-Zevallos (2007). However, the individual anthocyanins in PCSAs extract showed remarkable differences with those
reported by Aoki et al. (2002), cyanidin-3-glucoside, pelargonidin-3glucoside and peonidin-3-glucoside were only anthocyanin in PCSAs
extract, whereas cyanidin-3-(6-malonylglucoside), pelargonidin-3-(6malonylglucoside) and peonidin-3-(6-malonylglucoside) were not
found. The difference may be due to the different variants and grow-

In conclusion, the total anthocyanin content from purple corn seed

and purple corn cob were determined by pH-differential methods
and its (PCSAs and PCCAs) antioxidant activities were determined by
DPPH assay, TEAC assay, and FRAP assay. Form the data obtained in
these three methods, the antioxidant activities decreased in the order
or PCCAs N PCSAs N BHT. The antioxidant activities of purple corn may
be related to its anthocyanin substrates. The extracts of purple corn
contained a large amount of anthocyanins. The results provide useful
information on pharmacological activities associated with free radicals
of this crop, being useful for their application as colorants or antioxidants in foods.
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