Food Chemistry 81 (2003) 321–326

Antioxidant capacity of phenolic phytochemicals from
various cultivars of plums
Dae-Ok Kima, Seung Weon Jeongb, Chang Y. Leea,*

Department of Food Science and Technology, Cornell University, Geneva, NY 14456, USA
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

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, flavonoids, 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 flavonoid concentrations ranged from 118 to 237 mg catechin equivalents (CE)/100 g fresh weight. The concentrations of total phenolics
and flavonoids 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 different 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 flavonoids
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. Significant
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, flavour, taste) of fresh fruits,
vegetables and their products. In addition, many phenolic phytochemicals have antioxidative, anticarcinogenic, antimicrobial, antiallergic, antimutagenic
and antiinflammatory 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: (C.Y. Lee).

Tomono, Katase, Ogawa, & Yano, 1999; Kim, Choi, &
Chung, 2000). Some phytochemicals, including flavonoids 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 significant inverse relationship between dietary intake
of fruits and vegetables and the risk of coronary heart
disease (Knekt, Ja¨rvinen, Reunanen, & Maatela, 1996).
The distribution and composition of phenolic phytochemicals are affected 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,
Plums contain copious amounts of natural phenolic
phytochemicals, such as flavonoids and phenolic acids,

0308-8146/03/$ - see front matter # 2003 Published by Elsevier Science Ltd.
PII: S0308-8146(02)00423-5

.20 -azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical. ABTS as diammonium salt. were recently analyzed by HPLC-DAD-ESIMS. Biomedical Products Department. Long John. MO. Fruits Six cultivars of plums (Beltsville Elite B70197. Cao. & Martı´nez-Tome´. Extraction of phenolics The phenolics in powdered freeze-dried plums and apples were extracted by the ultrasound-assisted method (Kim & Lee. 2001). 2001) and under-researched worldwide. using Trolox equivalents. antioxidant activity of chlorogenic acid and its isomers.3. using a Sorvall RC-5B refrigerated superspeed centrifuge (Du Pont Company. In another report. blue-green 2. / Food Chemistry 81 (2003) 321–326 which may function as effective natural antioxidants in our daily diet. (St.2. having very high concentrations of phenolic phytochemicals. The Gala apple cultivar. Various kinds of antioxidant components in plums may play important roles in the combinative or synergistic contribution to total antioxidant activity. Plums demonstrated very good scavenger activity against oxygen-derived free radicals such as hydroxyl and peroxyl radicals (Murcia. Louis. After incubation for 90 min at 23  C. the latter being one of the most commonly consumed fruits in our diet. Chemicals Gallic acid. an aliquot (1 ml) of appropriately diluted extracts or standard solutions of gallic acid (20. 10 ml of 7% Na2CO3 solution was added with mixing. and Prior (1996) demonstrated that plums had 4. 2. where Trolox is a vitamin E analogue but not a natural compound. 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. One mililiter of Folin & Ciocalteu’s phenol reagent was added to the mixture and shaken. Pitted plums and sliced apples were frozen and lyophilized. Purposes of this study were to determine the content of total phenolic phytochemicals and flavonoids and to evaluate total antioxidant activity in various cultivars of plums using vitamin C equivalent antioxidant capacity (VCEAC). All other chemicals used were of analytical grade.. Kim et al. The two filtrates were combined and transferred into a 1 l evaporating flask with an additional 50 ml of 80% aqueous methanol.4. Dubuque. (Richmond. The concentration of total phenolics was measured by the method described by Singleton and Rossi (1965) with some modification. Total phenolic contents of plums and apples were expressed as mg gallic acid equivalents .4 times higher total antioxidant capacities than apples. Su. USA). The mixture of freeze-dried powder and 80% aqueous methanol was sonicated for 20 min with continual nitrogen gas purging. was employed for determination of total antioxidant activity. Immediately upon arrival in the pilot plant after harvest. 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. Kent. Cacak Best. Yet plums. plums and apples were stored in a 2–5  C cold room. The mixture was centrifuged at refrigerated temperatures. Wilmington.20 Azobis(2-amidino-propane)dihydrochloride (AAPH) was obtained from Wako Chemicals USA. 40. A simple chromogen. Plums were carefully cut in half and the pits removed. The solution was then immediately diluted to volume (25 ml) with ddH2O and mixed thoroughly. remain underutilized in the average American diet (Vinson.1. USA) at 10 K with GSA rotor for 20 min and stored at 4  C until analyses were performed. 2. in five different varieties of yellow and red plums. England) using a chilled Bu¨chner funnel and rinsing with 50 ml 100% methanol. the absorbance versus prepared blank was read at 750 nm. was evaluated (Nakatani et al. Phenolics of the fruits were extracted from 10 g ground freeze-dried samples using 100 ml of 80% aqueous methanol. 2. Stanley. Iowa. Jime´nez. Extraction of the residue was repeated using the same conditions. Apple slices were prepared by hand. Zubik & Bose. USA). Materials and methods 2. They reported the total antioxidant capacity of various fruits including plums.322 D. USA). The remaining phenolic concentrate was first dissolved in 50 ml of 100% methanol and diluted to a final volume of 100 ml using distilled deionized water (ddH2O) obtained with a NANOpure water system (Barnstead. After 5 min. The mixture was filtered through Whatman #2 filter paper (Whatman International Limited. 2. Wang. DE. isolated from prune. (+)-catechin and Folin & Ciocalteu’s phenol reagent were obtained from Sigma Chemical Co. French Damson. Various phenolic compositions. VA. picked at commercial maturity during the 2001 harvest season at the same location. Freeze-dried samples were ground to powder and then stored at 20  C until analyzed. A reagent blank using ddH2O was prepared. 80 and 100 mg/l) was added to a 25 ml volumetric flask containing 9 ml of ddH2O. 2002). was used as a reference sample. 2001). Inc. 60. Determination of total phenolics 2. 2000). Briefly. but no antioxidant activity was reported (Toma´s-Barbera´n et al. The solvent was evaporated using a rotary evaporator at 40  C.-O.

was determined at 510 nm versus prepared water blank. At zero time. Samples were analyzed in five replications. Results 3.3 ml 10% AlCl3 was added. Kim et al. Beltsville Elite B70197 and French Damson.0  6.4 ml of ddH2O and thoroughly mixed. The total 2.9 mg GAE) and Long John (199  2. Determination of total flavonoids Total flavonoid contents of six fresh cultivars of plums and Gala apples are shown in Fig. All samples were analyzed in five replications.5 mM ABTS in phosphate-buffered saline (pH 7. as determined by scavenging blue-green ABTS radical anions. pink in colour.3. 323 3. 2 ml 1 M NaOH was added to the mixture. In brief. The mixture was incubated in a 37  C water bath under restricted light for 10 min.001 and P< 0. At 6 min. ST. Total phenolic and flavonoid contents were significantly different at P< 0. 1.4 mg GAE. 0. LJ. 1. The content of total flavonoids in 100 g fresh Gala apples was at the level of 62.D. Significant difference was statistically considered at the level of P < 0. Vitamin C equivalent antioxidant capacity (VCEAC) assay using ABTS radical ABTS radical anions were used. was expressed on a fresh weight basis as mg/100 g vitamin C equivalents (VCEAC). 60. 2. 80 and 100 mg/l) was added to a 10 ml volumetric flask containing 4 ml ddH2O. After 5 min. The decrease of absorbance at 734 nm was measured at an endpoint of 10 min. Yugoslavian Elite T101 (146  6. 3. Determination of total phenolics 2.0 mM AAPH. Total flavonoids of fruits were expressed on a fresh weight basis as mg/100 g catechin equivalents (CE).05.4 mg CE) and Stanley (118  2. YE. Twenty microliters of sample was added to 980 ml of the ABTS radical solution. GA. The total phenolic contents of the fresh plums per 100 g ranged from 174 1. Total phenolics of six plum cultivars and Gala apples are shown in Fig. Yugoslavian Elite T101.6. The data are displayed with meanstandard deviation (bars) of five replications. was mixed with 2. according to the method of Kim.8 mg GAE in French Damson.5 mg CE). A control (20 ml 50% methanol and 980 ml of ABTS radical solution) was run with each series of samples.6 mg CE). The resulting blue-green ABTS solution was adjusted to the absorbance of 0.7. 40. Stanley. The radical stock solution was freshly prepared.5 mg GAE in Stanley to 375  3. Statistical analysis Results are presented as mean value  standard deviation.1.0 mg CE). 1. 0.4. All tested samples were replicated six times.020 at 734 nm with additional phosphate-buffered saline.5.9 mg CE. Determination of total flavonoids Total flavonoids were measured by a colorimetric assay developed by Zhishen. / Food Chemistry 81 (2003) 321–326 (GAE)/100 g fresh sample. 1.3 3.4 mg GAE). are presented in Fig.5 mg GAE). 100 mM potassium phosphate buffer containing 150 mM NaCl). as determined by scavenging ABTS radical anions. and Lee (2002). Cacak Best (319  1. Cacak Best. Yugoslavian Elite T101 (217  4.05. Beltsville Elite B70197 exhibited the highest flavonoids content of 237  6. Fig. The total phenolic content of 100 g of fresh Gala apples was 118  1. CB. A 1 ml aliquot of appropriately diluted sample or standard solutions of catechin (20.1 mg GAE).-O.650  0.3 ml 5% NaNO2 was added to the flask. BE and FD stand for Gala apple. Lee.3 mg CE/100 g fresh sample. daily. Immediately. Cacak Best (200  2. Mengcheng. . respectively. 2. Absorbance of the mixture. The mixed solution was heated in a water bath at 68  C for 13 min. This was followed by French Damson (215  9. Long John (126. the reaction flask was diluted to volume with the addition of 2. Phenolic concentrations of the other plum varieties were as follows: Beltsville Elite B70197 (332  3. and Jianming (1999).001 or P < 0. a radical initiator. Total antioxidant capacity of plums and apples. 3. Statistical analysis between experimental results was based on Student’s t test.2.7 mg CE). Lee. VCEAC assay using ABTS radical The total antioxidant capacities of the fresh plum cultivars and Gala apples. Contents of total phenolics and flavonoids in various cultivars of plum and Gala apple. Long John. respectively.

French Damson. cultivars of which were different from ours.8 times higher VCEAC than Stanley with the lowest VCEAC among plum cultivars.7 mg VCEAC/100 g. whereas the cultivars with lower concentrations are Stanley. the lowest content. Yugoslavian Elite T101. This method expresses antioxidant capacity on the basis of vitamin C equivalent. Beltsville Elite B70197.2-fold higher than that of Stanley. 266 14.9-fold higher than Gala apples. French Damson. & Kanner. The averages of total phenolic content of plums and apples were significantly different at a level of P< 0. Beltsville Elite B70197 ranked second highest in total phenolics and had the highest concentration of total flavonoids among the six cultivars of plums tested.4 mg GAE (Fig. which can quench reactive free radicals. The total phenolic content of French Damson was about 2. German. VCEACs showed significant difference except between CB and FD at the level of P<0. respectively. have health-promoting effects in the prevention of degenerative diseases. An antioxidant. At the significance level of P < 0. Total phenolic content and total flavonoid content of plums and apples showed a linear relationship with a positive correlation coefficient of r2=0. respectively. Total phenolic concentrations of various apple cultivars were reported to be in the range of 50.3-fold higher than that of Gala apples. Long John and Stanley. Lee. The VCEAC procedure. El. respectively. oxidative DNA and alteration of platelet functions (Fridovich. 4. Cacak Best and Beltsville Elite B70197. ranked as follows: Beltsville Elite B70197> Cacak Best5French Damson > Yugoslavian Elite T101> Long John > Stanley > Gala apple (Fig. Beltsville Elite B70197 and Cacak Best. low levels. Vitamin C equivalent antioxidant capacity (VCEAC) of various cultivars of plum and Gala apple using free blue-green ABTS radical anions.05. other agricultural practices and also variations in assays’ protocols. aging and atherosclerosis. followed by French Damson. inflammation.9. a decolorization assay using free blue-green ABTS radicals.2.324 D. Raviota.0 mg CE/ 100 g. 2). Total phenolic content of a mixture of red plums (Black Star.934 (data not shown). and peroxy radical (ROO). 2002). growing seasons. French Damson and Stanley plums showed about 3. CB and BE stand for Gala apple. one exhibiting relatively high levels of polyphenolic phytochemicals and the other. Based on the total phenolics.5 times higher total phenolic level than Gala apples.. ST. Cacak Best. / Food Chemistry 81 (2003) 321–326 Fig. 1). showed significant difference except between Cacak Best and . 2002). Kang. Discussion The total phenolic content of 100 g fresh plums ranged from 174 to 375 mg GAE and for Gala apples.8-fold and 1. in terms of VCEAC. 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. 288  16.. having the highest content of total flavonoids and Stanley. Kim et al. protein denaturation.2.05.-O. & Tas¸ . hydroxyl radical (OH).6 mg VCEAC/100 g. 118 1. The data are displayed with meanstandard deviation (bars) of six replications. displayed levels 3. Vitamin C is ubiquitously present in fruits and has been shown to have preventive effects against carcinogenesis (Lee. 1978. Kinsella. Yugoslavian Elite T101. Beltsville Elite B70197 showed 1. which has been generally considered to be linked with many chronic health problems such as cancers. YE. & Lee. FD. June Black). Reactive free radicals. are extremely reactive and are known to be a biological product in reducing molecular oxygen (Williams & Jeffrey. Plums were previously shown to have a higher total phenolic content than apples (Proteggente et al. Beltsville Elite B70197.001. the six plum cultivars studied may be classified into two groups.9 to 140 mg GAE/100 g (Lee & Smith.05. 524  25. 2000). 535 9. 2002). can prevent the oxidation of other molecules and may. Damage mediated by free radicals results in the disruption of membrane fluidity. GA. 2000). expressed as catechin equivalents (Karakaya. such as superoxide anion (O 2 ). These variations are due to differences among cultivars. 2002). The cultivars having relatively high concentrations of phenolic phytochemicals are French Damson. at 62. Frankel. Stanley. LJ. was reported to be at the level of 320 mg GAE/100 g (Proteggente et al. Cacak Best. lipid peroxidation.. antioxidant capacity of Gala apples was 205  5. 2. However. Ciruela Santa Rosa.5 and 317  21. Long John and Yugoslavian Elite T101. 2001). 1993).2 and 1. therefore. respectively. VCEACs. Another study reported that total phenols of black plums was 144 mg/100 g. All plum varieties in this study had higher VCEAC than Gala apple. among various cultivars of plums and Gala apples.5. Long John. The total flavonoid contents of the plum cultivars and Gala apples were significantly different at a significance level of P < 0. The total antioxidant activity of plums and apples. the VCEAC of Stanley plums was 1. Stanley and Yugoslavian Elite T101 showed total antioxidant capacities of 559 12. Long John.

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