Food Chemistry 81 (2003) 321326

www.elsevier.com/locate/foodchem

Antioxidant capacity of phenolic phytochemicals from

various cultivars of plums
Dae-Ok Kima, Seung Weon Jeongb, Chang Y. Leea,*
a

Department of Food Science and Technology, Cornell University, Geneva, NY 14456, USA
b
Korea Food Research Institute, Bundang, Seongnam, Kyounggi 463-746, South Korea

Received 27 May 2002; received in revised form 9 September 2002; accepted 9 September 2002

Abstract
Polyphenolic phytochemical extractions of six cultivars of plums (Beltsville Elite B70197, Cacak Best, French Damson, Long
John, Stanley, Yugoslavian Elite T101) and Gala apples were performed using 80% aqueous methanol with ultrasound assistance
and extracts were analyzed for total phenolics, avonoids, and antioxidant capacity. The total phenolic contents of various cultivars
of plums were in a range of 174 to 375 mg/100 g, expressed as gallic acid equivalents (GAE), on a fresh weight basis. Total avonoid concentrations ranged from 118 to 237 mg catechin equivalents (CE)/100 g fresh weight. The concentrations of total phenolics
and avonoids in Gala apples were 118
1.4 mg GAE and 62.0
6.9 mg CE per 100 g fresh sample weight, respectively. The stable
radical chromogen, ABTS, commonly employed for the antioxidant activity measurement, was used to evaluate antioxidant
capacity of plums and apples. The total antioxidant capacities, expressed as vitamin C equivalent antioxidant capacity (VCEAC), of
fresh plums ranged from 266 to 559 mg/100 g. The order of total antioxidant capacity among dierent plum cultivars was as follows: Beltsville Elite B70197 >Cacak Best5French Damson> Yugoslavian Elite T101> Long John >Stanley. The total antioxidant
capacity of fresh Gala apple was 205
5.6 mg VCEAC/100 g. There was a good correlation between total phenolics or avonoids
contents and VCEAC at the high level of P< 0.001. Dietary polyphenolics from plums may supply substantial antioxidants, which
may provide health-promoting advantages to the consumer.
# 2003 Published by Elsevier Science Ltd.
Keywords: Antioxidant capacity; Free radical; Phenolic phytochemicals; Plums; Vitamin C equivalent antioxidant capacity (VCEAC)

1. Introduction
Phenolic phytochemicals are important aromatic secondary metabolites in plants, many of which are commonly substituted by sugar moieties such as glucose,
arabinose, xylose, rhamnose and galactose. Signicant
amounts of phenolic compounds frequently occur in
foods such as fruits and vegetables and are routinely
consumed in our diet. They importantly attribute to the
sensory qualities (colour, avour, taste) of fresh fruits,
vegetables and their products. In addition, many phenolic phytochemicals have antioxidative, anticarcinogenic, antimicrobial, antiallergic, antimutagenic
and antiinammatory activities (Cao & Cao, 1999;
Eberhardt, Lee, & Liu, 2000; Ito et al., 1998; Kawaii,
* Corresponding author. Tel.: +1-315-787-2271; fax: +1-315-7872284.
E-mail address: cyl1@cornell.edu (C.Y. Lee).

Tomono, Katase, Ogawa, & Yano, 1999; Kim, Choi, &

Chung, 2000). Some phytochemicals, including avonoids in fruits and vegetables, consumed as part of our
daily diet, may reduce the risk of cardiovascular disease
(Cook & Samman, 1996). Epidemiological studies show
a signicant inverse relationship between dietary intake
of fruits and vegetables and the risk of coronary heart
disease (Knekt, Jarvinen, Reunanen, & Maatela, 1996).
The distribution and composition of phenolic phytochemicals are aected by maturity, cultivars, horticultural practices, geographic origin, growing season,
postharvest storage conditions and processing procedures (Burda, Oleszek, & Lee, 1990; De Freitas &
Glories, 1999; Donovan, Meyer, & Waterhouse, 1998;
Kalt, Forney, Martin, & Prior, 1999; Kim, Koh, &
Koh, 2001; Lee & Jaworski, 1987; Spanos & Wrolstad,
1990).
Plums contain copious amounts of natural phenolic
phytochemicals, such as avonoids and phenolic acids,

0308-8146/03/$ - see front matter # 2003 Published by Elsevier Science Ltd.

PII: S0308-8146(02)00423-5

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D.-O. Kim et al. / Food Chemistry 81 (2003) 321326

which may function as eective natural antioxidants in

our daily diet. Wang, Cao, and Prior (1996) demonstrated that plums had 4.4 times higher total antioxidant capacities than apples, the latter being one of
the most commonly consumed fruits in our diet. They
reported the total antioxidant capacity of various fruits
including plums, using Trolox equivalents, where Trolox is a vitamin E analogue but not a natural compound. Plums demonstrated very good scavenger
activity against oxygen-derived free radicals such as
hydroxyl and peroxyl radicals (Murcia, Jimenez, &
Martnez-Tome, 2001). Various phenolic compositions,
in ve dierent varieties of yellow and red plums, were
recently analyzed by HPLC-DAD-ESIMS, but no antioxidant activity was reported (Tomas-Barberan et al.,
2001). In another report, antioxidant activity of chlorogenic acid and its isomers, isolated from prune, was
evaluated (Nakatani et al., 2000).
The measurement of antioxidant activity of individual
compounds may lead to a misleading conclusion due to
frequently observed antagonistic or synergistic interactions of various components of foods. Various kinds of
antioxidant components in plums may play important
roles in the combinative or synergistic contribution to
total antioxidant activity. Yet plums, having very high
concentrations of phenolic phytochemicals, remain
underutilized in the average American diet (Vinson, Su,
Zubik & Bose, 2001) and under-researched worldwide.
Purposes of this study were to determine the content of
total phenolic phytochemicals and avonoids and to
evaluate total antioxidant activity in various cultivars of
plums using vitamin C equivalent antioxidant capacity
(VCEAC). A simple chromogen, blue-green 2,20 -azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)
radical, was employed for determination of total antioxidant activity.

2.2. Chemicals
Gallic acid, ABTS as diammonium salt, (+)-catechin
and Folin & Ciocalteus phenol reagent were obtained
from Sigma Chemical Co. (St. Louis, MO, USA). 2,20 Azobis(2-amidino-propane)dihydrochloride
(AAPH)
was obtained from Wako Chemicals USA, Inc. (Richmond, VA, USA). All other chemicals used were of
analytical grade.
2.3. Extraction of phenolics
The phenolics in powdered freeze-dried plums and
apples were extracted by the ultrasound-assisted
method (Kim & Lee, 2002). Phenolics of the fruits were
extracted from 10 g ground freeze-dried samples using
100 ml of 80% aqueous methanol. The mixture of
freeze-dried powder and 80% aqueous methanol was
sonicated for 20 min with continual nitrogen gas purging. The mixture was ltered through Whatman #2
lter paper (Whatman International Limited, Kent,
England) using a chilled Buchner funnel and rinsing
with 50 ml 100% methanol. Extraction of the residue
was repeated using the same conditions. The two ltrates were combined and transferred into a 1 l evaporating ask with an additional 50 ml of 80% aqueous
methanol. The solvent was evaporated using a rotary
evaporator at 40  C. The remaining phenolic concentrate was rst dissolved in 50 ml of 100% methanol
and diluted to a nal volume of 100 ml using distilled
deionized water (ddH2O) obtained with a NANOpure
water system (Barnstead, Dubuque, Iowa, USA). The
mixture was centrifuged at refrigerated temperatures,
using a Sorvall RC-5B refrigerated superspeed centrifuge (Du Pont Company, Biomedical Products
Department, Wilmington, DE, USA) at 10 K with GSA
rotor for 20 min and stored at 4  C until analyses were
performed.

2. Materials and methods

2.4. Determination of total phenolics
2.1. Fruits
Six cultivars of plums (Beltsville Elite B70197, Cacak
Best, French Damson, Long John, Stanley, Yugoslavian
Elite T101) were picked at commercial maturity during
the 2000 harvest season at the New York State Agricultural Experiment Station orchard in Geneva, New
York. The Gala apple cultivar, picked at commercial
maturity during the 2001 harvest season at the same location, was used as a reference sample. Immediately upon
arrival in the pilot plant after harvest, plums and apples
were stored in a 25  C cold room. Plums were carefully
cut in half and the pits removed. Apple slices were prepared by hand. Pitted plums and sliced apples were frozen
and lyophilized. Freeze-dried samples were ground to
powder and then stored at 20  C until analyzed.

The concentration of total phenolics was measured by

the method described by Singleton and Rossi (1965)
with some modication. Briey, an aliquot (1 ml) of
appropriately diluted extracts or standard solutions of
gallic acid (20, 40, 60, 80 and 100 mg/l) was added to a
25 ml volumetric ask containing 9 ml of ddH2O. A
reagent blank using ddH2O was prepared. One mililiter
of Folin & Ciocalteus phenol reagent was added to the
mixture and shaken. After 5 min, 10 ml of 7% Na2CO3
solution was added with mixing. The solution was then
immediately diluted to volume (25 ml) with ddH2O and
mixed thoroughly. After incubation for 90 min at
23  C, the absorbance versus prepared blank was read
at 750 nm. Total phenolic contents of plums and
apples were expressed as mg gallic acid equivalents

D.-O. Kim et al. / Food Chemistry 81 (2003) 321326

(GAE)/100 g fresh sample. All samples were analyzed

in ve replications.

323

3. Results
3.1. Determination of total phenolics

2.5. Determination of total avonoids

Total avonoids were measured by a colorimetric
assay developed by Zhishen, Mengcheng, and Jianming
(1999). A 1 ml aliquot of appropriately diluted sample
or standard solutions of catechin (20, 40, 60, 80 and 100
mg/l) was added to a 10 ml volumetric ask containing
4 ml ddH2O. At zero time, 0.3 ml 5% NaNO2 was
added to the ask. After 5 min, 0.3 ml 10% AlCl3
was added. At 6 min, 2 ml 1 M NaOH was added to
the mixture. Immediately, the reaction ask was diluted to volume with the addition of 2.4 ml of ddH2O
and thoroughly mixed. Absorbance of the mixture,
pink in colour, was determined at 510 nm versus prepared water blank. Total avonoids of fruits were
expressed on a fresh weight basis as mg/100 g catechin equivalents (CE). Samples were analyzed in ve
replications.
2.6. Vitamin C equivalent antioxidant capacity (VCEAC)
assay using ABTS radical
ABTS radical anions were used, according to the
method of Kim, Lee, Lee, and Lee (2002). In brief, 1.0
mM AAPH, a radical initiator, was mixed with 2.5 mM
ABTS in phosphate-buered saline (pH 7.4; 100 mM
potassium phosphate buer containing 150 mM NaCl).
The mixed solution was heated in a water bath at 68  C
for 13 min. The resulting blue-green ABTS solution
was adjusted to the absorbance of 0.650
0.020 at 734
nm with additional phosphate-buered saline. Twenty
microliters of sample was added to 980 ml of the ABTS
radical solution. The mixture was incubated in a 37  C
water bath under restricted light for 10 min. A control
(20 ml 50% methanol and 980 ml of ABTS radical solution) was run with each series of samples. The decrease
of absorbance at 734 nm was measured at an endpoint
of 10 min. Total antioxidant capacity of plums and
apples, as determined by scavenging blue-green ABTS
radical anions, was expressed on a fresh weight basis as
mg/100 g vitamin C equivalents (VCEAC). The radical
stock solution was freshly prepared, daily. All tested
samples were replicated six times.

Total phenolics of six plum cultivars and Gala apples

are shown in Fig. 1. The total phenolic contents of the
fresh plums per 100 g ranged from 174
1.5 mg GAE in
Stanley to 375
3.8 mg GAE in French Damson. Phenolic concentrations of the other plum varieties were as
follows: Beltsville Elite B70197 (332
3.1 mg GAE),
Cacak Best (319
1.4 mg GAE), Yugoslavian Elite
T101 (217
4.9 mg GAE) and Long John (199
2.5 mg
GAE). The total phenolic content of 100 g of fresh Gala
apples was 118
1.4 mg GAE.
3.2. Determination of total avonoids
Total avonoid contents of six fresh cultivars of
plums and Gala apples are shown in Fig. 1. Beltsville
Elite B70197 exhibited the highest avonoids content of
237
6.3 mg CE/100 g fresh sample. This was followed
by French Damson (215
9.7 mg CE), Cacak Best
(200
2.5 mg CE), Yugoslavian Elite T101 (146
6.0
mg CE), Long John (126.3
3.4 mg CE) and Stanley
(118
2.6 mg CE). The content of total avonoids in
100 g fresh Gala apples was at the level of 62.0
6.9 mg
CE.
3.3. VCEAC assay using ABTS radical
The total antioxidant capacities of the fresh plum
cultivars and Gala apples, as determined by scavenging
ABTS radical anions, are presented in Fig. 2. The total

2.7. Statistical analysis

Results are presented as mean value
standard
deviation. Statistical analysis between experimental
results was based on Students t test. Signicant dierence was statistically considered at the level of P < 0.001
or P < 0.05.

Fig. 1. Contents of total phenolics and avonoids in various cultivars

of plum and Gala apple. The data are displayed with mean
standard
deviation (bars) of ve replications. Total phenolic and avonoid
contents were signicantly dierent at P< 0.001 and P< 0.05, respectively. GA, ST, LJ, YE, CB, BE and FD stand for Gala apple, Stanley,
Long John, Yugoslavian Elite T101, Cacak Best, Beltsville Elite
B70197 and French Damson, respectively.

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D.-O. Kim et al. / Food Chemistry 81 (2003) 321326

Fig. 2. Vitamin C equivalent antioxidant capacity (VCEAC) of various cultivars of plum and Gala apple using free blue-green ABTS
radical anions. The data are displayed with mean
standard deviation
(bars) of six replications. VCEACs showed signicant dierence
except between CB and FD at the level of P<0.05. GA, ST, LJ, YE,
FD, CB and BE stand for Gala apple, Stanley, Long John, Yugoslavian Elite T101, French Damson, Cacak Best and Beltsville Elite
B70197, respectively.

antioxidant capacity of Gala apples was 205
5.6 mg

VCEAC/100 g. Beltsville Elite B70197, Cacak Best,
French Damson, Long John, Stanley and Yugoslavian
Elite T101 showed total antioxidant capacities of
559
12.9, 535
9.2, 524
25.5, 288
16.2, 266
14.5
and 317
21.7 mg VCEAC/100 g, respectively.

4. Discussion
The total phenolic content of 100 g fresh plums ranged from 174 to 375 mg GAE and for Gala apples,
118
1.4 mg GAE (Fig. 1). Based on the total phenolics, the six plum cultivars studied may be classied
into two groups, one exhibiting relatively high levels of
polyphenolic phytochemicals and the other, low levels.
The cultivars having relatively high concentrations of
phenolic phytochemicals are French Damson, Beltsville
Elite B70197 and Cacak Best, whereas the cultivars with
lower concentrations are Stanley, Long John and
Yugoslavian Elite T101. The total phenolic content of
French Damson was about 2.2-fold higher than that of
Stanley. French Damson and Stanley plums showed
about 3.2 and 1.5 times higher total phenolic level than
Gala apples, respectively. Plums were previously shown
to have a higher total phenolic content than apples
(Proteggente et al., 2002). The averages of total phenolic
content of plums and apples were signicantly dierent at
a level of P< 0.001. Total phenolic content of a mixture of
red plums (Black Star, Ciruela Santa Rosa, Raviota, June
Black), cultivars of which were dierent from ours, was
reported to be at the level of 320 mg GAE/100 g (Proteggente et al., 2002). Another study reported that total
phenols of black plums was 144 mg/100 g, expressed as

catechin equivalents (Karakaya, El, & Tas , 2001). Total

phenolic concentrations of various apple cultivars were
reported to be in the range of 50.9 to 140 mg GAE/100 g
(Lee & Smith, 2000). These variations are due to dierences among cultivars, growing seasons, other agricultural practices and also variations in assays protocols.
Beltsville Elite B70197 ranked second highest in total
phenolics and had the highest concentration of total
avonoids among the six cultivars of plums tested, followed by French Damson, Cacak Best, Yugoslavian
Elite T101, Long John and Stanley. Beltsville Elite
B70197, having the highest content of total avonoids
and Stanley, the lowest content, displayed levels 3.8-fold
and 1.9-fold higher than Gala apples, at 62.0 mg CE/
100 g, respectively. The total avonoid contents of the
plum cultivars and Gala apples were signicantly different at a signicance level of P < 0.05. Total phenolic
content and total avonoid content of plums and apples
showed a linear relationship with a positive correlation
coecient of r2=0.934 (data not shown).
Reactive free radicals, such as superoxide anion (O
2 ),
hydroxyl radical (OH), and peroxy radical (ROO), are
extremely reactive and are known to be a biological
product in reducing molecular oxygen (Williams & Jeffrey, 2000). Damage mediated by free radicals results in
the disruption of membrane uidity, protein denaturation, lipid peroxidation, oxidative DNA and alteration
of platelet functions (Fridovich, 1978; Kinsella, Frankel, German, & Kanner, 1993), which has been generally considered to be linked with many chronic health
problems such as cancers, inammation, aging and
atherosclerosis. An antioxidant, which can quench
reactive free radicals, can prevent the oxidation of other
molecules and may, therefore, have health-promoting
eects in the prevention of degenerative diseases. The
VCEAC procedure, a decolorization assay using free
blue-green ABTS radicals, was shown to be a very useful tool for expeditiously measuring the antioxidant
activity in individual chemical compounds or complex
fruits extracts (Kim et al., 2002). This method expresses
antioxidant capacity on the basis of vitamin C equivalent. Vitamin C is ubiquitously present in fruits and has
been shown to have preventive eects against carcinogenesis (Lee, Lee, Kang, & Lee, 2002).
The total antioxidant activity of plums and apples, in
terms of VCEAC, ranked as follows: Beltsville Elite
B70197> Cacak Best5French Damson > Yugoslavian
Elite T101> Long John > Stanley > Gala apple (Fig. 2).
Beltsville Elite B70197 showed 1.8 times higher VCEAC
than Stanley with the lowest VCEAC among plum cultivars. However, the VCEAC of Stanley plums was
1.3-fold higher than that of Gala apples. All plum varieties in this study had higher VCEAC than Gala apple.
At the signicance level of P < 0.05, VCEACs, among
various cultivars of plums and Gala apples, showed
signicant dierence except between Cacak Best and

D.-O. Kim et al. / Food Chemistry 81 (2003) 321326

French Damson. Good correlation (r2=0.938) between

total phenolic content and VCEAC was observed with a
high signicance level (P < 0.001) (Fig. 3A). A similar
relationship (r2=0.942) with high signicance
(P < 0.001) was also obtained between total avonoid
content and VCEAC (Fig. 3B). This positive and signicant relationship between total phenolic content and
total antioxidant activity, observed in this study was
previously reported using other assays (Kalt et al., 1999;
Proteggente et al., 2002; Velioglu, Mazza, Gao, &
Oomah, 1998).
The results of this study imply that dietary polyphenolic phytochemicals from plums may supply
substantial antioxidants, which, in turn, may provide
health-promoting eects to consumers. Since it has
been suggested that a higher consumption of fruits
and vegetables with high phytochemicals can inhibit,
prevent or retard chronic diseases (Birt, Hendrich, &
Wang, 2001; Cook & Samman, 1996; Singletary,
2000; Yang, Landau, Huang, & Newmark, 2001) and
the plums in our study showed substantially more

Fig. 3. Relationship between total phenolics and VCEAC (A) and

between total avonoids and VCEAC (B).

325

antioxidant activity than apples, an increased consumption of this fruit is therefore recommended in
our diet.

Acknowledgements
The authors thank Ms. Nancy Smith, research support specialist, for revising the text and providing technical support and Mr. Jay Freer for selecting and
providing the cultivars of plums.

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