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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
Keywords:
Purple corn
Anthocyanins
Antioxidant activity
DPPH
FRAP
TEAC
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-
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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.
2.1. 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.
Z. Yang, W. Zhai / Innovative Food Science and Emerging Technologies 11 (2010) 169176
171
Table 1
TAC and CIELAB value of PCSAs and PCCAs extracts.
Solvent
TAC (mg/100 g)
L*
C*
PCSAs
PCCAs
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).
IC50/DPPH
(ug/ml)a
IC50/TEAC
(ug/ml)b
FRAP value
(mmol FeSO4/g DW)
PCCAs
PCSAs
BHT
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
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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.
IC50/TEAC
FRAP value
Total anthocyanin content
IC50/DPPH
IC50/TEAC
FRAP value
0.977
0.828
0.842
0.913
0.851
0.936
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.
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).
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
173
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).
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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
content
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).
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