Food Chemistry 76 (2002) 69–75

[Link]/locate/foodchem

An investigation of antioxidant capacity

of fruits in Singapore markets
L.P. Leong*, G. Shui
Food Science and Technology Programme, Department of Chemistry, National University of Singapore, Singapore 117543, Singapore

Received 27 February 2001; received in revised form 21 May 2001; accepted 21 May 2001

Abstract
The antioxidant capacity of a group of fruits obtained in the Singapore markets was investigated. A total of 27 fruit pulps were
tested for their general antioxidant capacity based on their ability to scavenge 2,20 -azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)
(ABTS) free radical. The contribution of l-ascorbic acid (AA) to the total antioxidant activity of fruits was investigated by using
RP-HPLC. The antioxidant capacity of the fruit pulp was measured by monitoring the change of absorbance of the free radical
solution at 414 nm in the test reaction mixture following addition of the fruit extract, as compared with AA. The results were
expressed as mg of AA equivalents per 100 g, i.e. the quantity of AA required to produce the same scavenging activity as the extract
in 100 g of sample (l-ascorbic acid equivalent antioxidant capacity, AEAC). Total antioxidant capacities of AA acid, trolox,
hydroquinone, pyrogallol and several fruits were also evaluated based on its ability to scavenge the 1,1-diphenyl-2-picryl-hydrazyl
(DPPH) radical. Results obtained were compared with those of ABTS assay. Every mol of AA, trolox or hydroquinone, was found
to reduce about 2 mol of ABTS + or DPPH . However, 4 mol of DPPH or 7 mol of ABTS + were scavenged by every mol of
pyrogallol. A good correlation of AEAC was observed between the two methods. Both methods have been recommended to be
useful tools to evaluate antioxidant capacities of fruits. According to the AEAC value of binary extract solution of fruits in the
ABTS model, ciku shows the highest antioxidant capacity, followed by strawberry, plum, star fruit, guava, seedless grape, salak,
mangosteen, avocado, orange, solo papaya, mango, kiwi fruit, cempedak, pomelo, lemon, pineapple, apple, foot long papaya,
rambutan, rambutan king, banana, coconut pulp, tomato, rockmelon, honeydew, watermelon and coconut water. The AA con-
tribution to AEAC of fruits varied greatly among species, from 0.06% in ciku to 70.2% in rambutan. # 2001 Elsevier Science Ltd.
All rights reserved.
Keywords: ABTS; DPPH; HPLC; Antioxidant capacity; l-Ascorbic acid; Fruits

1. Introduction delay the oxidation of an oxidisable substrate in a chain

reaction, would therefore seem to be very important in
High consumption of fruits and vegetables has been the prevention of these diseases (Ames et al., 1993;
associated with a lowered incidence of degenerative dis- Aruoma, 1998; Jacob & Burri, 1996; Steinberg et al.,
eases including cancer, heart disease, inflammation, 1991; Maxwell & Lip, 1997; Pratico & Delanty, 2000;
arthritis, immune system decline, brain dysfunction and Wang et al., 1996). However, knowledge of the potential
cataracts (Ames, 1983; Ames, Shiganaga, & Hagwn, antioxidant compounds present in a food does not
1993; Feskanich et al., 2000; Gordon, 1996; Haegele et necessarily indicate its antioxidant capacity. Further-
al., 2000; Halliwell et al., 1996; Michels et al., 2000). more, the synergistic effect which could exist between
These protective effects are considered, in large part, to different antioxidants (termed synergism) means that the
be related to the various antioxidants contained in total antioxidant effect may be greater than the sum of
them. There is overwhelming evidence to indicate that the individual antioxidant activities and the isolation of
free radicals cause oxidative damage to lipids, proteins, one compound will not exactly reflect the overall action
and nucleic acids. Antioxidants, which can inhibit or (Jia, Zhou, Yang, Wu, & Liu, 1998; Poeggeler, Reiter,
Hardeland, Sewerenek, Melchiorri, & Barlow-Warden,
* Corresponding author. Tel.: +65-874-2917; fax: +65-779-1691. 1995; Wu, Sugiyama, Zeng, Mickle, & Wu, 1998).
E-mail address: laipeng@[Link] (L.P. Leong). In addition, there are many different antioxidant
0308-8146/01/$ - see front matter # 2001 Elsevier Science Ltd. All rights reserved.
PII: S0308-8146(01)00251-5
70 L.P. Leong, G. Shui / Food Chemistry 76 (2002) 69–75

components in animal and plant tissues, it is relatively solo (Carica papaya L.), mangosteen (Garcinia mangos-
difficult to measure each antioxidant component sepa- tana L.), salak (Salacca edulis), avocado (Persea Amer-
rately. Several analytical methods have been proposed icana), foot long papaya (Carica papaya L.), pomelo
for determining total antioxidant activity of biological (Citrus grandis), orange (Citrus aurantium), cempedak
extracts in order to evaluate the total antioxidant capa- (Artocarpus integer Merr.), rambutan king (Nephelium
city of biological samples (Cano, Hernandez-Ruiz, mutabile), rambutan (Nephelium lappaceum L.), apple
Garcia-Canovas, Acosta, & Arnao, 1998; Cao, Alessio, (Malus pumila), pineapple (Ananas comosus Merr.),
& Culter 1993; Miller, Rice-Evans, Davies, Gopinathan, tomato (Lycopersicon esculentum), banana (Musa par-
& Milner, 1993; Robert, Pellegrini, Poteggente, Pan- idasiaca), rockmelon (Cucumis melo var. cantalupensis),
nala, Yang, & Rice-Evans 1999; Wayner, Burton, honeydew (Cucumis melo var. inodorus), watermelon
Ingold, & Locke, 1985; Whitehead, Thorpe, & Maxwell, (Citrullus vulgaris) and coconut (Cocos nucifera) .
1992).
Fruits from the tropical and subtropical climates are 2.1.2. Chemicals
known to be associated with many medicinal properties ABTS, DPPH, AA, pyrogallol, hydroquinone, potas-
(Morton, 1987). Many of these fruits are taken as sium persulfate and glycine were purchased from Sigma
remedies for coughs, intestinal bleeding, diarrhoea and (MO, USA); trolox from Acros Organics (NJ, USA);
other medical conditions. Pineapple juice, for example, ethanol, acetic acid, methanol and hydrochloric acid
can be taken to alleviate sore throat and seasickness. were purchased from Merck (Darmstadt, Germany);
Thus, it is likely that these fruits will also have protec- acetonitrile from EM Science (NJ, USA).
tive effects towards degenerative disease.
l-Ascorbic acid (AA) has numerous biological func- 2.2. Sample preparation
tions, which include the synthesis of collagen, hormones
and neurotransmitters (Block, 1993). It is believed that Coconut water was weighed, diluted and filtered for
the role of AA acid in disease prevention is due to its analysis. All other fruits, except seedless red grapes,
ability to scavenge free radical in the biological systems. strawberries, and tomatoes, were peeled. Ciku was tes-
Cancer, which is due to uncontrolled cell proliferation, ted before natural ripening. The edible portion of the
may be initiated by oxidative and free radical damage to fruit was homogenized using a blender and weighed into
DNA and cell. Since AA may act as an effective anti- a 50 ml centrifuge tube, and 25 ml of 50% aqueous
oxidant, it is able to slow down or prevent such damage ethanol (HPLC grade of pure water, the solution was
(Block, 1993). AA is abundant in many fruits and is sonicated by a Bransonics cleaner) was added (1:10 w/v)
more prevalent in most fruits than is vitamin E. There- and mixed in a vortex mixer for 60 s. The extract was
fore, it is believed that the AA contributes largely to the centrifuged at 2000 g for 5 min at room temperature.
antioxidant activity in these extracts. The objective of The supernatant was filtered and used directly for the
this study was to measure the antioxidant properties of ABTS, DPPH and AA assay.
some common fruits available in the Singapore market
by using an improved 2,20 -azino-bis-(3-ethylbenzthiazo- 2.3. ABTS free radical decolorization assay
line-6-sulfonic acid) (ABTS) free radical decolourisation
assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radi- The total antioxidant capacity assay was carried out
cal-scavenging assay. RP- HPLC was used to determine on the Ultraspec 3000 UV/Visible Spectrophotometer
the AA (Vitamin C) content, thus the contribution of total (Pharmacia Biotech Ltd., Cambridge CB4 4FJ, Eng-
antioxidant capacity in these fruits can be calculated. land). The procedure was based on a method developed
by Robert et al. (1999) with some modification.
ABTS + was generated by reacting ABTS (7.4 mM)
2. Materials and methods with potassium persulphate (2.6 mM). The solution was
diluted to obtain an absorbance of 1.4 units at 414 nm
2.1. Materials (molar extinction coefficient E=3.6104 mol1 l cm1,
Forni, Morla-Arellano, Packer, & Willison 1986) with
2.1.1. Fruits 50 mM glycine-HCl buffer (pH 4.5) before use. Three
Twenty-seven varieties of fruits were purchased on millilitres of the solution were added to 20–80 ml of AA,
several separate occasions from local supermarkets and trolox, hydroquinone, pyrogallol and fruit extracts
the wholesale centre. The 27 fruits comprised ciku separately. The changes in absorbance at 414 nm were
(Manilkara zapota), strawberry (Fragaria virginiana), recorded at 1, 3, 6, 10, 20, 30, 40, 60 and 90 min after
‘Flame seedless’ grape (Vitis vinifera), guava (Psidium mixing and until the absorbance reached a plateau. The
guajava), plum (Prunus domestics), star fruit (Averrhoa antioxidant capacities, obtained by comparing the
carambola L.), kiwi fruit (Actinidia chinensis), mango absorbance change at 414 nm in a test reaction mixture
(Mangifera indica L.), lemon (Citrus limon), papaya var. containing extract of fruit with that containing AA,
 L.P. Leong, G. Shui / Food Chemistry 76 (2002) 69–75 71

were expressed as mg of AA equivalents per 100 g of An aliquot of 20–80 ml of an antioxidant/fruit extract

homogenate (AEAC). The AEAC was calculated using solution was added to 3 ml of this solution. The
the following equation: decrease in absorbance at 517 nm was measured at 0, 1,
5 and then every 10 min until the reaction reached a
A 100 plateau. The decreased absorbance of DPPH remaining
AEAC ¼  CAA  V 
AAA W at the steady-state was calculated and expressed as mg
of AA equivalents per 100 g of homogenate (AEAC).
where, A is the change of absorbance after addition of
fruit extract, CAA is the concentration of AA standard 2.5. HPLC assay
solution (mg/ml), AAA is the change of absorbance
obtained from a calibration curve when the same AA standard solution (1.36 g/ml) was prepared daily
volume of AA standard solution as that of fruit extract by accurately weighing 68 mg and dissolving it in 50 ml
was added, V is the volume of filtrate (ml) and W is the of 2% acetic acid. This was then diluted to give 0.068,
weight of homogenate used for extraction (g). 0.034, 0.017 and 0.0085 mg/ml working standard solu-
tions. The HPLC system consisted of a Shimadzu
2.4. DPPH free radical-scavenging assay HPLC (Model LC-10ATvp two Pumps and DGU-14A
Degasser) equipped with a photo-diode array detector
The free radical-scavenging activity of fruits was (Model SPD-M10AVP; Shimadzu, Kyoto, Japan) inter-
measured using the method described by Brand-Wil- faced with IBM Pentium-III personal computer. The
liams (Brand-Williams, Cuvelier, & Berset, 1995) with separation was performed on a Shim-Pack VP-ODS
some modification. A 0.1 mM solution of DPPH (1,1- column (2504.6 mm i.d.; Shimadzu, Kyoto, Japan)
diphenyl-2-picrylhydrazyl) in methanol was prepared. with a guard column (GVP-ODS, 104.6 mm i.d.;

Table 1
AEAC of 27 selected fruits using ABTS.+ decolorization assay and their l-ascorbic acid content

Variety AEACa AA content by HPLCa Percentage contribution of Source Classification

(mg/100 g) (mg/100 g) AEAC by AA (%)b by AEAC

Ciku 3396387.9 2.0 0.7 0.06 Malaysia Extremely high

Strawberry 47292.9 53.9 11.2 11.4 Australia
Plum 31223.2 8.2 2.3 2.6 USA
Star fruit 27822.3 5.9 1.8 2.1 Malaysia High
Guava 27018.8 131 18.2 48.3 Thailand
Grape seedless 26483.6 0.5 0.2 0.2 USA
Salak 26032.5 2.4 1.5 0.9 Malaysia

Mangosteen 15023.3 4.1 1.2 2.7 Malaysia

Avocado 14316.5 9.0 2.1 6.3 Thailand
Orange 14222.6 36.1 15.9 25.5 Australia
Solo papaya 14126.7 67.8 12.6 48.0 Malaysia
Mango 13921.5 19.7 9.1 14.2 Philippines
Kiwi fruit 13618.2 52.8 22.5 38.7 New Zealand
Cempedak 12619.1 6.2 0.9 4.83 Malaysia Medium
Pomelo 10434.7 36.0 7.5 34.7 Malaysia
Lemon 93.39.8 49.6 6.8 53.2 Australia
Pineapple 85.621.3 54.0 7.9 63.0 Malaysia
Apple 78.92.7 2.1 0.9 2.7 China
Foot long papaya 72.52.6 45.2 10.3 62.3 Malaysia
Rambutan 71.57.6 50.2 6.5 70.2 Malaysia
Rambutan king 70.68.2 49.5 8.8 70.0 Malaysia

Banana 48.31.2 2.1 0.8 4.4 Philippines

Coconut pulp 45.86.5 0.9 0.3 19.7 Malaysia
Tomato 38.01.7 11.0 2.6 29.1 Malaysia
Rockmelon 26.23.5 2.7 0.6 10.3 Australia Low
Honeydew 19.60.8 3.9 0.4 19.9 Malaysia
Watermelon 11.90.1 3.7 0.2 31.0 Malaysia
Coconut water 11.52.2 0.7 0.3 6.1 Malaysia
a
Mean of three determinationsS.D. (standard deviation).
72 L.P. Leong, G. Shui / Food Chemistry 76 (2002) 69–75

Shimadzu, Kyoto, Japan), using 2% (v/v) acetic acid/ papaya, rambutan, rambutan king, banana, coconut
acetonitrile (95:5, v/v) as the mobile phase at a flow-rate pulp, tomato, rockmelon, honeydew, watermelon and
of 0.8 ml/min at 40  C oven temperature. Ten micro- coconut water. The total antioxidant capacity of ciku
litres of the extract, prepared earlier, were injected into was found to be extremely high (compared with the
the HPLC. The measured AA content was expressed as other fruits examined). We are currently looking into
mg AA/100 g edible portion. the compounds in ciku contributing such a high anti-
oxidant capacity. Several tropical fruits from Southeast
Asia, such as guava, star fruit and salak, are very good
3. Results and discussion sources of antioxidants. Other fruits, such as avocado,
papaya, mangosteen, pomelo, pineapple, cempedak,
3.1. Total antioxidant capacity of fruits by ABTS + lemon and rambutan, also contain considerable amount
decolorization assay of antioxidants. Wang et al. (1996) had previously
measured the total antioxidant activity of 12 fruits using
This method measures the relative antioxidant ability automated oxygen radical absorbance capacity (ORAC)
of fruits to scavenge the radical ABTS + in the aqueous assay. Of the eight fruits that were mutually tested, the
phase, as compared with a standard amount of AA. The ranking orders of total antioxidant capacity were as
ABTS +, generated by potassium persulfate, is pre- follows: strawberry>plum>orange>kiwi>banana>
sented as an excellent tool for determining the anti- apple>tomato>honeydew melon. The ranking order
oxidant activity of hydrogen-donating antioxidants was found to be the same here, except that apple was
(scavengers of aqueous phase radicals) and of chain- found to have a higher AEAC than banana. The differ-
breaking antioxidants (scavengers of lipid peroxyl radi- ence could be related to seasonal variations or variety.
cals). Rice-Evans and co-workers (Rice-Evans, Miller,
Bolwell, Bramley, & Pridham, 1995; Rice-Evans, Miller, 3.2. Content of AA in fruits
& Paganga, 1996) and Roberta Re (Roberta et al., 1999)
have demonstrated that ABTS + assay can be used to AA is one of the most effective antioxidants in fruits
measure the antioxidant activity of a broad diversity of and vegetables. People with high intakes of dietary AA
substances. acid or citrus fruits have repeatedly been associated with
The antioxidant defence system of the body is com- lowered risk of developing cancer. Because of different
posed of a mixture of antioxidants. Fruits are good criteria of adequacy and different interpretations of
sources of antioxidants that may be more effective and experimental evidence, the recommended dietary allow-
economical than supplements in protecting the body ance (RDA) for AA varies between 30 and 80 mg/day.
against oxidative damage under different conditions. The RDA of AA for adult men and women is 30, 60 and
Fruit antioxidants, which include AA, tocopherol, car- 80 mg/day in Singapore, United States and Nether-
otenoids and phenolics, vary greatly in their contents lands, respectively (Bender, 1993; Department of Nutri-
and profile among various fruits. As a result, the anti- tion, 1999). However, many scientific reports indicate
oxidant capacity of one fruit differs considerably from that an increased intake of AA is associated with a
another. By using the ABTS decolourisation assay, we reduced risk of chronic diseases such as cancer, cardio-
have measured, for the first time, the antioxidant capa- vascular disease, and cataract. One possible mode of
city of a variety of fruits available in the Singapore action of AA is through its antioxidant capability. It is
markets. likely that the current RDA for AA to prevent defi-
As shown in Table 1, total antioxidant capacity, in ciency disease, is not sufficient for optimum protection
AEAC of fruits tested, was found to vary over 300-fold against the chronic diseases mentioned above, therefore,
from that at the lowest value. Based on these values, the Carr and Frei (1999) suggested a new RDA of 120 mg
activity of the fruit extract to scavenge free radicals is AA/day.
classified into four categories. The fruit with AEAC of RP-HPLC was used to measure the content of AA
over 600 mg AAeq/100 g is classified as containing due to its high sensitivity, high reproducibility, easy
extremely high antioxidant capacity. On the other hand, operation and short analysis time. Table 1 shows AA
fruits with AEAC from 200 to 600 mg AAeq/100 g, 70 to contents of the 27 selected fruits analysed. The AA
200 AAeq/100 g and less than 70 mg AAeq/100 g are contribution to scavenge ABTS + varies extensively up
classified as containing high, medium and low anti- to 200 times the lowest value. AA accounts for a high
oxidant capacities respectively. On the basis of the wet percentage contribution to ABTS + scavenging activity
weight of fruit (edible portion), ciku shows the highest in rambutan and rambutan king (70%), pineapple
antioxidant capacity, followed by strawberry, plum, star (63%), guava (48.3%), lemon (53.2%) and solo papaya
fruit, guava, seedless grape, salak, mangosteen, avo- (48%), foot long papaya (62.3%), kiwi fruit (38.7%),
cado, orange, solo papaya, mango, kiwi fruit, cempe- pomelo (34.7%), watermelon (31.7%), tomato (29.1%)
dak, pomelo, lemon, pineapple, apple, foot long and orange (25.5%). Wang et al. (1996) suggested that
 L.P. Leong, G. Shui / Food Chemistry 76 (2002) 69–75 73

the contribution of AA to ORAC activity of a fruit was studies are underway to measure the total phenolic
usually less than 15%, except for kiwi fruit and honey- content in these fruit extracts.
dew melon. However, in this study, it was found that
almost half of the fruits tested showed high contribu- 3.3. Scavenging DPPH free radicals
tions of AA to their total antioxidant capacity. Since
Wang et al. (1996) did not test the antioxidant capacity The model of scavenging the stable DPPH radical is a
of fruits having a high AA contribution in this study, widely used method to evaluate antioxidant activities in
the real L-ascorbic acid contribution to ORAC activity a relatively short time compared with other methods.
among these fruits needs to be confirmed. The con- The effect of antioxidants on DPPH radical scavenging
tribution of AA to AEAC among other fruits was low, was thought to be due to their hydrogen donating abil-
especially for ciku, plum, star fruit, salak, seedless ity (Baumann, Wurn, & Bruchlausen 1979). AEAC of
grape, mangosteen, apple and cempedak. It seems fruits some fruits by DPPH and ABTS assay and their AA
with a high AEAC value are more likely to have a low content are shown in Fig. 1.
percentage contribution from AA to AEAC. Most fruits tested with a high AEAC in the ABTS
As discussed above, this set of data shows that the model also have a high AEAC in the DPPH model. This
contribution of AA to the AEAC of fruits can differ high correlation may partly result from a similar
extensively from one fruit to another. Therefore, con- mechanism and, also, both the antioxidants are soluble
tributions of other compounds to AEAC of these fruits in aqueous/ethanol systems. Fig. 2 shows that a good
must not be neglected. Among the most important correlation exits between two assay methods:
compounds that contribute to the high AEAC value of AEACDPPH=0.9203AEACABTS and R2=0.9045 (not
these fruits are the polyphenolic compounds. Future including ciku, R2=0.9984 if ciku was included). This

Fig. 1. AEAC of 12 fruits by ABTS- and DPPH-scavenging assay.

Fig. 3. Correlation of [ABTS.+] and [DPPH.] reduced by AA

Fig. 2. Comparison of AEAC of fruits in ABTS and DPPH models. [ABTS.+]= 1.0088[DPPH.].
74 L.P. Leong, G. Shui / Food Chemistry 76 (2002) 69–75

shows that the two methods are compatible when used orange, solo papaya, mango, kiwi fruit, cempedak,
to assess free radical-scavenging activity. pomelo, lemon, pineapple, apple, foot long papaya,
rambutan, rambutan king, banana, coconut pulp,
3.4. Correlation of ABTS assay and DPPH assay tomato, rockmelon, honeydew, watermelon and coco-
nut water. The contribution of AA to AEAC varies
Fig. 3 shows a linear correlation between reduced markedly from one fruit to another. A good correlation
ABTS + and DPPH after different concentrations of of AEAC, between ABTS model and DPPH model, was
AA were added. Similar results were obtained when observed among 11 fruits. Both models are recom-
trolox, hydroquinone and pyrogallol were added, and mended as useful tools for evaluating the total anti-
their linear correlation could be described as oxidant capacity of fruits.
[ABTS +]=0.963[DPPH ] (R2=0.9993), [ABTS +]=
0.9987 [DPPH] (R2=0.9989) and [ABTS +]= 1.688
[DPPH ] (R2=0.9989) respectively.
In this study, it is found that 1 mol of AA reacts with Acknowledgements
approximately 2 mol of ABTS radicals or DPPH radi-
cals. The result is consistent with a previous report that Thanks are given to Professor Fereidoon Shahidi and
1 mol of AA reduces 2 mol of ABTS radicals (Cano et Associate Professor Philip J Barlow for helpful sugges-
al., 1998). Similarly, 2 mols of ABTS or DPPH radicals tions and corrections.
are scavenged by 1 mol of trolox or hydroquinone. The
stoichiometry of the reaction between pyrogallol and
ABTS or DPPH radical is found to be 1:7 and 1:4, References
respectively.
For the antioxidants tested, the stoichiometry of Ames, B. N. (1983). Dietary carcinogens and anticarcinogens: oxygen
radicals and degenerative diseases. Science, 221, 1256–1263.
reactions between antioxidant and ABTS.+ or DPPH. Ames, B. N., Shigenaga, M. K., & Hagwn, T. M. (1993). Oxidants,
is very close. However, when pyrogallol was used, the antioxidants, and the degenerative diseases of aging. Proc. Natl.
stoichiometry differs. Lissi, Modak, Torres, Escobar, Acad. Sci. USA, 90, 7915–7922.
and Urzua (1999) determined that total antioxidant Aruoma, O. I. (1998). Free radicals, oxidative stress, and antioxidants
in human health and disease. Journal of the American Oil Chemists’
potential of resinous exudates from Heliotropium spe-
Society, 75, 199–212.
cies showed significant differences by employing ABTS Baumann, J., Wurn, G., & Bruchlausen, F. V. (1979). Prostaglandin
and DPPH scavenging assay. Wang et al. (1998) found synthetase inhibiting O 2 radical scavenging properties of some fla-
that some compounds, which have ABTS + scavenging vonoids and related phenolic compounds. Naunyn-Schmiedebergs
activity may not show DPPH scavenging activity. Arch. Pharmacol., 308, R27.
Therefore, a linear correlation between two models may Bender, D. A. (1993). Micronutrients: the vitamins and minerals. In An
introduction to nutrition and metabolism (2nd ed.), UCL Press, London.
not be obvious among some other biological samples Block, G. (1993). Vitamin-C, Cancer and aging. Age, 16, 55–58.
that may contain a variety of antioxidants. Brand-Williams, W., Cuvelier, M. E., & Berset, C. (1995). Use of a
However, the results obtained in this study, from 11 free radical method to evaluate antioxidant activity. Lebensmittel-
fruits show a good correlation between the two models. Wissenschaft und Technologie, 26, 25–30.
Among the eleven fruits tested in both models, six fruits Cano, A., Hernandez-Ruiz, J., Garcia-Canovas, F., Acosta, M., &
Arnao, M. B. (1998). An end-point method for estimation of the
have a relative AEAC difference of less than 20%, only total antioxidant activity in plant material. Phytochemical Analysis,
two fruits more than 30% and none less than 50%. 9, 196–202.
Since no significant difference of AEAC was observed Carr, A. C., & Frei, B. (1999). Toward a new recommended dietary
among fruits by employing the two models, any one of allowance for vitamin C based on antioxidant and health effects in
humans. American Journal of Clinical Nutrition, 69, 1086–1107.
the two models may be a useful tool for evaluating the
Cao, G., Alessio, H. M., & Culter, R. G. (1993). Oxygen-radical
total antioxidant capacity of fruits. Further research on absorbance capacity assay for antioxidants. Free Radical Biology &
the stoichiometry of reactions between pure fruit anti- Medicine, 14, 303–311.
oxidants and ABTS + or DPPH is currently under Department of Nutrition, Ministry of Health, Singapore. (1999). Sin-
investigation. gapore food facts.
Feskanich, D., Ziegler, R. G., Michaud, D. S., Giovannucci, E. L.,
Speizer, F. E., Willett, W. C., & Colditz, G. A. (2000). Prospective
study of fruit and vegetable consumption and risk of lung cancer
4. Conclusion among men and women. Journal of the National Cancer Institute,
92, 1812–1823.
The AEAC of 27 fruits in the ABTS model and the Forni, L. G., Morla-Arellano, V. O., Packer, J. E., & Willison, R. L.
contributions of AA to the AEAC of fruits are mea- (1986). Nitrogen dioxide and related free radicals: electron-transfer
reactions with organic compounds in solutions containing nitrite or
sured. Among the 27 fruits tested, ciku had the highest nitrate. J. Chem. Soc. Perkin Trans., 2, 1–6.
AEAC value, followed by strawberry, plum, star fruit, Gordon, M. H. (1996). Dietary antioxidants in disease prevention.
guava, seedless grape, salak, mangosteen, avocado, Natural Product Reports, 265–273.
 L.P. Leong, G. Shui / Food Chemistry 76 (2002) 69–75 75

Haegele, A. D., Gillette, C., O’Neill, C., Wolfe, P., Heimendinger, J., breaking antioxidants ascorbate, trolox and glutathione. Neu-
Sedlacek, S., & Thompson, H. J. (2000). Plasma xanthophyll car- roendocrinological letters, 17, 87–92.
otenoids correlate inversely with indices of oxidative DNA damage Pratico, D., & Delanty, N. (2000). Oxidative injury in diseases of the
and lipid peroxidation. Cancer Epidemiology Biomarkers & Preven- central nervous system: focus on Alzheimer’s disease. American
tion, 9, 421–425. Journal of Medicine, 109, 577–585.
Halliwell, B. (1996). Antioxidants in human health and disease. Ann. Rice-Evans, C., Miller, N. J., Bolwell, P. G., Bramley, P. M., & Prid-
Rev. Nutr., 16, 33–50. ham, J. B. (1995). The relative antioxidant activities of plant-derived
Jacob, R. A., & Burri, B. J. (1996). Oxidative damage and defense. polyphenolic flavonoids. Free Radical Research, 22, 375–383.
American Journal of Clinical Nutrition, 63, S985–S990. Rice-Evans, C., Miller, N. J., & Paganga, G. (1996). Structure-anti-
Jia, Z. S., Zhou, B., Yang, L., Wu, L. M., & Liu, Z. L. (1998). Anti- oxidant activity relationships of flavonoids and phenolic acids. Free
oxidant synergism of tea polypenols and a-tocopherol against free Radical Biology & Medicine, 20, 933–956.
radical induced peroxidation of linoleic acid in solution. J. Chem. Robert, R. E., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M.,
Soc., Perkin Trans., 2, 911–915. & Rice-Evans, C. (1999). Antioxidant activity applying an improved
Lissi, E. A., Modak, B., Torres, R., Escobar, J., & Urzua, A. (1999). ABTS radical cation decolorization assay. Free Radical Biology and
Total antioxidant potential of resinous exudates from Heliotropium Medicine, 26, 1231–1237.
species, and a comparison of the ABTS and DPPH methods. Free Steinberg, D. (1991). Antioxidants and atherosclerosis: a current
Radical Research, 30, 471–477. assessment. Circulation, 84, 1420–1425.
Maxwell, S. R. J., & Lip, G. Y. H. (1997). Free radicals and anti- Wang, H., Cao, G., & Prior, R. L. (1996). Total antioxidant capacity
oxidants in cardiovascular disease. British Journal of Clinical Phar- of fruits. J. Agric. Food Chem., 44, 701–705.
macology, 44, 307–317. Wang, M. F., Shao, Y., Li, J. G., Zhu, N. Q., Rangarajan, M.,
Michels, K. B., Giovannucci, E., Joshipura, K. J., Rosner, B. A., LaVoie, E. J., & Ho, C. T. (1998). Antioxidative phenolic com-
Stampfer, M. J., Fuchs, C. S., Colditz, G. A., Speizer, F. E., & pounds from Sage (Salvia officinalis). J. Agric. Food Chem., 46,
Willett, W. C. (2000). Prospective study of fruit and vegetable con- 4869–4873.
sumption and incidence of colon and rectal cancers. Journal of the Wayner, D. D. M., Burton, G. W., Ingold, K. U., & Locke, S. (1985).
National Cancer Institute, 92, 1740–1752. Quantitative meaurement of the total, peroxyl radical-trapping
Miller, N. J., Rice-Evans, C., Davies, M. J., Gopinathan, V., & Mil- antioxidant capacity of human plasma by controlled peroxidation.
ner, A. (1993). A novel method for measuring antioxidant capacity FEBS Letters, 187, 33–37.
and its application to monitoring the antioxidant status in pre- Whitehead, T. P., Thorpe, G. H. G., & Maxwell, S. R. J. (1992).
mature neonates. Clin. Sci., 84, 407–412. Enhanced chemoluminescent assay for antioxidant capacity in bio-
Morton, J. F. (1987). Fruits of warm climates. Miami, FL: J.F. Mor- logical fluids. Analytica Chimica Acta, 266, 265–277.
ton. Wu, J., Sugiyama, H., Zeng, L. H., Mickle, D., & Wu, T. W. (1998).
Poeggeler, B., Reiter, R. J., Hardeland, R., Sewerenek, E., Melchiorri, Evidence of Trolox and some gallates as synergistic protectors of
D., & Barlow-Waden, L. R. (1995). Melatonin, a mediator of elec- erythrocytes against peroxyl radicals. Biochemistry and Cell Biology-
tron transfer and repair reaction, acts synergistically with the chian- Biochemie ET Biologie Cellulaire, 76, 661–664.