JFS R: Concise Reviews/Hypotheses in Food Science

R: Concise Reviews in Food Science

An Integrated Approach to Evaluate
Food Antioxidant Capacity
T. SUN AND S.A. TANUMIHARDJO

ABSTRACT: Many methods are available for determining food antioxidant capacity, which is an important topic
in food and nutrition research and marketing. However, the results and inferences from different methods may vary
substantially because each complex chemical reaction generates unique values. To get a complete and dynamic pic-
ture of the ranking of food antioxidant capacity, relative antioxidant capacity index (RACI), a hypothetical concept,
is created from the perspective of statistics by integrating the antioxidant capacity values generated from different
in vitro methods. RACI is the mean value of standard scores transformed from the initial data generated with differ-
ent methods for each food item. By comparing the antioxidant capacity of 20 commonly consumed vegetables in the
U.S. market that were measured with 7 chemical methods, we demonstrated that the RACI correlated strongly with
each method. The significant correlation of RACI with an independent data set further confirmed that RACI is a valid
tool to assess food antioxidant capacity. The key advantage of this integrated approach is that RACI is in a numerical
scale with no units and has consistent agreement with chemical methods. Although it is a relative index and may
not represent a specific antioxidant property of different food items, RACI provides a reasonably accurate rank of
antioxidant capacity among foods. Therefore, it can be used as an integrated approach to evaluate food antioxidant
capacity.
Keywords: antioxidant capacity, electron transfer, hydrogen atom transfer, vegetables

Introduction HAT methods measure the ability of an antioxidant to quench free

A ntioxidants and antioxidant capacity of food, nutraceuticals,

botanicals, and other dietary supplements have recently at-
tracted the attention of the industry, scientists, and consumers be-
radicals by hydrogen donation. In most HAT methods, antioxidants
and a probe compete for thermally generated peroxyl radicals and
the quantitation is derived from the kinetic curves after monitor-
cause of potential health benefits (Verlangieri and others 1985). An- ing the competitive reaction kinetics. HAT methods include oxy-
tioxidants in food include several classes: (1) simple phenols and gen radical absorbance capacity (ORAC), total radical-trapping an-
phenolic acids, (2) flavonoids, and (3) others such as -carotene, tioxidant parameter (TRAP), and inhibition of auto-oxidation of
lutein, -tocopherol, and ascorbic acid. Each class of antioxidants induced low-density lipoprotein (LDL). ET methods measure the
has its own unique chemical properties. Phenolic compounds have ability of a potential antioxidant to transfer 1 electron to reduce
an aromatic ring with one or more hydroxyl substituents, or func- radicals, metals or carbonyls, which use the color change of the ox-
tional derivatives (for example, esters, methyl ethers, and gly- idant as the endpoint indicator. Examples of ET methods include 2,
cosides). Flavonoids include anthocyanidins, flavonols, flavones, 2
-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferric
isoflavones, catechins, and flavanones (Herrmann 1988). The char- reducing antioxidant power (FRAP), 2, 2
-diphenyl-l-picrylhydrazyl
(DPPH), and cupric reducing antioxidant capacity (CUPRAC) meth-
acteristic structure of a flavonoid is a diphenylpropane (C 6 -C 3 -C 6 )
(Hertog and others 1992). In nature, flavonoids usually occur as gly- ods (Huang and others 2005).
cosides with the 3rd position of the C ring commonly connected to Because multiple reaction mechanisms and different phase lo-
the glycosyl group (Finotti and Di Majo 2003). Carotenoids, for ex- cations are usually involved in measuring the antioxidant capacity
ample, -carotene and lutein, are pigments found in many fruits of a complex food system, there is no simple universal method by
and vegetables. -Tocopherol, or vitamin E, is a fat-soluble vitamin which total antioxidant capacity can be measured accurately and
found in plant oil. Ascorbic acid, or vitamin C, is a water soluble quantitatively (Frankel and Meyer 2000; Niki and Noguchi 2000;
antioxidant found in citrus fruits. Niki 2002; Sanchez-Moreno 2002; Huang and others 2005; Prior and
Many evaluation methods for antioxidant capacity have been others 2005; MacDonald-Wicks and others 2006). Because many
developed. The merits and disadvantages of these methods have methods are available and are being used by different research
been fully discussed in several reviews (Halliwell and others 1995; groups, it is difficult to compare the results of food antioxidant
Frankel and Meyer 2000; Prior and others 2005; Roginsky and Lissi capacity between studies. The food and nutraceutical industries
2005; MacDonald-Wicks and others 2006). According to the chem- cannot perform strict quality control on antioxidant products. Re-
ical reaction used, methods to measure antioxidant capacity can views or critiques to standardize methods or point out problems
be mainly grouped into 2 classes: hydrogen atom transfer (HAT) have been published (Frankel and Meyer 2000; Niki 2002; Sanchez-
and electron transfer (ET) based methods (Huang and others 2005). Moreno 2002; Prior and others 2005; MacDonald-Wicks and oth-
ers 2006). The consensus is that multiple methods, based upon dif-
MS 20070537 Submitted 7/12/2007, Accepted 9/5/2007. Authors are with ferent reaction mechanisms, should be used. Given the diversity
Dept. of Nutritional Sciences, Univ. of Wisconsin-Madison, 1415 Linden of food antioxidants and complex reactions involved in the mea-
Dr., Madison, WI 53706, U.S.A. Direct inquiries to author Sun (E-mail:
tingsun@nutrisci.wisc.edu.).
surement of antioxidant capacity, multiple methods are suggested
as standard to evaluate food antioxidant capacity. However, the


C 2007 Institute of Food Technologists Vol. 72, Nr. 9, 2007JOURNAL OF FOOD SCIENCE R159
doi: 10.1111/j.1750-3841.2007.00552.x
Further reproduction without permission is prohibited
R: Concise Reviews in Food Science Evaluating food antioxidant capacity . . .
results from standardized methods may vary substantially as dis-
parate complex chemical reactions may induce different conclu-
sions when comparing antioxidant capacity. Indeed, a standard
relationship between methods may not exist (Prior and others
2005).
The Hypothetical Concept of Relative
Antioxidant Capacity Index
A new concept, relative antioxidant capacity index (RACI), is pro-
posed herein from the perspective of statistics by integrating
the food antioxidant capacity data determined by several methods.
This concept not only allows for the comparison of food antioxi-
dant capacity derived from different chemical methods, but also
provides a more comprehensive comparison. If measured with 1
chemical method, the relative ranking of the antioxidant capacity
between foods is only based on a single indicator. However, no sin-
gle chemical method can represent total food antioxidant capacity.
To determine the antioxidant capacity of food items and the rela-
tionship between foods for multiple assays, the most widely used
measure is central tendency (Zar 1996), where foods are ranked
based on the mean value for each food. However, it is impossible
to get a mean value for each food as the units and the scale of the
data from various chemical methods are different. If the values of
antioxidant capacity in each data set are transformed into standard
scores, the data can be compared. In statistics, a standard score
(also called z- or normal score) is a dimensionless quantity derived
by subtracting the mean from the raw data divided by the standard
deviation (Sharma 1996). The standard score is thus calculated as
follows:
x Fe2+ -TPTZ). FRAP can be affected by several factors such as inter-
where x is the raw data, is the mean, and is the standard devia-
tion (Sharma 1996).
Subtracting the mean centers the distribution and dividing the
difference by the standard deviation normalizes the distribution.
Standard scores have a mean of 0 and a standard deviation equal
to 1. The standard score represents the distance between the raw
data and the mean in units of the standard deviation, which is
negative when the raw data are smaller than the mean and pos-
itive when larger. The standard score reveals how many units a
case is smaller or larger than the mean and allows the comparison
of results with different units or with the same units but different
normal distributions (Sharma 1996). The standard score transfor-
mation is especially useful when seeking to compare the relative
standings of items from distributions with different means and/or
standard deviations. The antioxidant capacity data generated by
different methods have different units and/or scales, and there is
no standard relationship between these methods. Thus, the direct
comparison of antioxidant capacity of different foods using varying
methods is impossible (Prior and others 2005). To overcome these
difficulties, RACI of each food item was created by averaging the
standard scores transformed from the raw data generated with dif-
ferent chemical methods. After data transformation, the standard
scores (without any units) from various methods with different dis-
tributions will have similar normal distributions with a mean of 0
and variance of 1. Therefore, the standard scores with the same
scale from data generated with different methods will have simi-
lar contribution to the central tendency of the mean and can be
compared without interference from different units, scales, and dis-
tributions. The following describes how to use RACI to compare
antioxidant capacity of vegetables.
R160 JOURNAL OF FOOD SCIENCEVol. 72, Nr. 9, 2007
Demonstration of the Concept with Published Data
on Vegetable Antioxidant Capacity
Antioxidants and antioxidant capacity of vegetables
Vegetables contain several types of antioxidants. Examples in-
clude anthocyanins, apigenin, apigenin glycoside, ascorbic acid, -
carotene, p-coumaric acid, caffeic acid, chlorogenic acid, cinnamic
acid, ferulic acid, p-hydroxybenzoic acid, kaempferol, kaempferol
glycoside, lutein, luteolin, luteolin glycoside, myricetin, quercetin,
quercetin glycoside, and -tocopherol (Sun and Powers 2007). Both
the types and amounts of antioxidants differ among vegetables.
Several methods have been used to measure the antioxidant capac-
ity of vegetables. The data on antioxidant capacity of 20 commonly
consumed vegetables in the U.S. market were collected from pub-
lished reports (Table 1) (Cao and others 1996; Vinson and others
1998; Ou and others 2002; Proteggente and others 2002; Pellegrini
and others 2003; Wu and others 2004; Eberhardt and others 2005;
Sun and others 2007). The antioxidant capacity of these vegeta-
bles was determined using 7 HAT and/or ET methods: ABTS, FRAP,
ORAC (Cu2+ ), ORAC (OH r), ORAC (ROO r), phenol antioxidant in-
dex (PAOXI), and TRAP (Table 1).
Brief review of methods
The ABTS method measures the decrease of the absorbance at
734 nm of the ABTS radical, which is produced by the reaction
of ABTS, metmyoglobin, H 2 O 2 , and peroxidase, or between ABTS
and persulfate. The ABTS method can determine both hydrophilic
and lipophilic antioxidants in the same sample (Arnao and oth-
ers 2001). The FRAP method measures the reduction of ferric 2, 4,
6-tripyridyl-s-triazine (TPTZ) to a colored product (Fe3+ -TPTZ to
ference from compounds that absorb at the wavelength of analy-
sis and reaction kinetics. In the ORAC (ROO r) method, the peroxyl
radical 2,2
-azobis-(2-amidinopropane) dihydrochloride (AAPH)
reacts with a fluorescent compound (fluorescein) to form a non-
fluorescent product, but antioxidants can scavenge the peroxyl
radicals and retard the loss of the fluorescence; thus the area
under the timefluorescence curve can be used to quantify food an-
tioxidant capacity (Cao and others 1993). ORAC (Cu2+ ) and ORAC
(OH r) methods use Cu2+ and OH r as the radical, respectively.
PAOXI is calculated by dividing the total phenol concentration of
a vegetable (mol/kg) from the FolinCiocalteau assay by the half
maximal inhibitory concentration (IC 50 ) in M from the inhibi-
tion assay of LDL oxidation. The FolinCiocalteau method deter-
mines the total phenolic content of the sample by monitoring the
absorbance change during a color reaction. The LDL method mea-
sures the antioxidants ability to inhibit the auto-oxidation of LDL
caused by Cu2+ or the AAPH radical (Handelman and others 1999).
The TRAP method evaluates the power of antioxidants to interfere
with the reaction between peroxyl radicals generated by AAPH or 2,
2-azobis (2-amidinopropane) dihydrochloride (ABAP) and a target
probe (Ghiselli and others 1995).
Development and validation of the
relative antioxidant capacity index
The antioxidant capacity of common vegetables determined by
7 chemical methods in published references was used to develop
RACI. For a vegetable with a range of data values, a mean value
was calculated to represent the antioxidant capacity determined
by each method. The units and scales of the raw data were differ-
ent (Table 1). The ranking of antioxidant capacity of vegetables dif-
fered between methods. Data from FRAP and ORAC (ROO r) meth-
ods had significant correlation with the data of most methods,
Evaluating food antioxidant capacity . . .

R: Concise Reviews in Food Science

while data from the PAOXI method had no significant correlation Data from the TRAP method showed significant correlation with
with any other method. Data from ABTS and ORAC (OH r) methods the ABTS, FRAP, and ORAC (ROO r) methods (Table 2). The standard
had significant correlation with 2 other methods. The ORAC (Cu2+ ) scores (without units) used to develop RACI from different chemi-
method only significantly correlated with data from ORAC (ROO r). cal methods had the same mean of 0 and variance of 1 (Table 3).

Table 1 --- Antioxidant capacity of vegetables from published references.

ABTS FRAP ORAC (OH r) ORAC (ROO r) TRAP
(mol Trolox (mol Fe ORAC (Cu2+ ) (mol Trolox (mol Trolox (mol Trolox
(equivalent/g equivalent/g ( 103 units/g equivalent/g (equivalent/g) PAOXI equivalent/g
Vegetables wet weight) wet weight) wet weight) wet weight) wet weight) (wet weight) wet weight)
Asparagus 3.90 10.60 N.A. N.A. 30.20 10000.00 9.70
Bean 1.30 2.30 0.20 1.70 3.53 30050.00 0.70
Beet 5.20 13.10 0.20 3.10 17.90 5300.00 2.70
Broccoli 4.75 11.70 1.60 2.40 13.08 4100.00 3.10
Cabbage 5.48 7.00 0.30 1.50 9.75 1600.00 2.60
Carrot 0.40 1.10 0.50 0.80 6.83 2300.00 0.70
Cauliflower 2.05 3.45 0.20 1.10 5.78 2200.00 1.60
Celery 0.50 1.20 0.20 0.30 3.15 1700.00 0.50
Corn N.A. N.A. N.A. 2.20 5.65 2800.00 N.A.
Cucumber 0.40 0.70 1.00 0.30 0.85 1100.00 N.A.
Garlic N.A. N.A. 2.70 1.10 19.40 31500.00 N.A.
Lettuce 1.50 3.05 0.25 1.05 5.03 1300.00 2.30
Mushroom 4.90 16.40 N.A. N.A. N.A. 2000.00 6.30
Onion 3.55 4.50 0.60 0.50 9.05 11200.00 2.40
Pepper (bell) 8.40 21.00 0.40 0.60 7.63 2700.00 5.50
Potato 0.80 3.70 0.50 1.00 7.50 4600.00 0.90
Squash N.A. N.A. 0.20 1.10 1.50 1600.00 N.A.
Spinach 8.50 26.90 1.60 2.80 16.17 2000.00 5.80
Sweet potato N.A. N.A. 0.30 1.00 6.00 4100.00 N.A.
Tomato 2.05 5.60 N.A. N.A. 4.00 3300.00 1.50
Mean 3.35 8.27 0.67 1.33 9.11 6272.50 3.09
Variance 7.18 60.29 0.51 0.72 54.01 77459861.84 6.88
N.A. = not available.

Table 2 --- Correlation coefficients between different methods for analyzing antioxidant capacity.
Methods ABTS FRAP ORAC (Cu2+ ) ORAC (OH r) ORAC (ROO r) PAOXI TRAP
ABTS 1.00

FRAP 0.93 1.00
ORAC (Cu2+ ) 0.37 0.49 1.00
ORAC (OH r) 0.53
0.54 0.20 1.00
ORAC (ROO r) 0.51
0.91
0.64
0.60 1.00
PAOXI 0.15 0.21 0.43 0.03 0.29 1.00

TRAP 0.65 0.74 0.54 0.39 0.85 0.10 1.00

P < 0.01 and P < 0.05.

Table 3 --- Standard scores of antioxidant capacity of vegetables.

Method
Vegetable ABTS FRAP ORAC (Cu2+ ) ORAC (OH r) ORAC (ROO r) PAOXI TRAP
Asparagus 0.20 0.30 2.87 0.42 2.52
Bean 0.77 0.77 0.66 0.44 0.76 2.70 0.91
Beet 0.69 0.62 0.66 2.10 1.20 0.11 0.15
Broccoli 0.52 0.44 1.30 1.27 0.54 0.25 0.01
Cabbage 0.79 0.16 0.52 0.21 0.09 0.53 0.19
Carrot 1.10 0.92 0.24 0.62 0.31 0.45 0.91
Cauliflower 0.49 0.62 0.66 0.27 0.45 0.46 0.57
Celery 1.07 0.91 0.66 1.21 0.81 0.52 0.99
Corn 1.03 0.47 0.39
Cucumber 1.10 0.97 0.46 1.21 1.12 0.59
Garlic 2.84 0.27 1.40 2.87
Lettuce 0.69 0.67 0.59 0.33 0.55 0.56 0.30
Mushroom 0.58 1.05 0.49 1.23
Onion 0.07 0.49 0.10 0.98 0.01 0.56 0.26
Pepper (bell) 1.88 1.64 0.38 0.86 0.20 0.41 0.92
Potato 0.95 0.59 0.24 0.39 0.22 0.19 0.83
Squash 0.66 0.27 1.03 0.53
Spinach 1.92 2.40 1.30 1.74 0.96 0.49 1.03
Sweet potato 0.52 0.39 0.42 0.25
Tomato 0.49 0.34 0.69 0.34 0.60
Mean 0.00 0.00 0.00 0.00 0.00 0.00 0.00
Variance 1.00 1.00 1.00 1.00 1.00 1.00 1.00

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R: Concise Reviews in Food Science Evaluating food antioxidant capacity . . .
The minimum, maximum, and 50% median standard scores were
1.10, 1.90, and 0.20 for ABTS; 1.00, 2.40, and 0.40 for FRAP;
0.70, 2.80, and 0.45 for ORAC (Cu2+ ); 1.20, 2.10, and 0.30 for
ORAC (OH r); 1.10, 2.90, and 0.30 for ORAC (ROO r); 0.60, 2.90,
and 0.40 for PAOXI; and 1.00, 2.50, and 0.30 for TRAP, respec-
tively. The standardized scores by definition had 100% correlation
with the corresponding raw data from each chemical method.
The RACI of each vegetable was calculated as the mean of stan-
dard scores transformed from the raw data generated with dif-
ferent chemical methods. The difference in units and variances
in the raw data had no influence on the RACI. Stepwise regres-
sion between RACI and different chemical methods revealed that
(1) each of the 7 chemical assays was selected as a significant
variable with no single chemical method being removed, (2) each
chemical method contributed the same weight (1/7) in build-
ing RACI, and (3) the regression was highly significant (r = 1,
P < 0.001). Therefore, RACI of each vegetable is a scientific com-
bination of data from different chemical methods with no unit
limitation and no variance among methods, and makes compar-
ison of food antioxidant capacity possible and perhaps more ac-
curate. Based on the RACI value of the 20 common vegetables, the
rank of antioxidant capacity (Figure 1) from the highest to the low-
est was garlic with a value of 1.71 to celery with a value of 0.88.
Using RACI, garlic, spinach, and asparagus have high antioxi-
dant capacity while celery, cucumber, carrot, and squash have low
antioxidant capacity. The trend of the RACI value matched with the
standard score from the 7 methods (Figure 2). Furthermore, the cor-
relation analysis demonstrated that the RACI value correlated with
each method; the correlation coefficients of RACI with ABTS, FRAP,
ORAC (Cu2+ ), ORAC (OH r), ORAC (ROO r), PAOXI, and TRAP were
0.79, 0.84, 0.76, 0.56, 0.86, 0.44, and 0.87, respectively. Therefore,
this novel approach is proven to be an appropriate way to quan-
tify food antioxidant capacity as it can represent each chemical
method.
Halvoren and others (2006) studied the antioxidant capacity of
16 of the 20 vegetables used in the development of RACI (ex-
cludes cauliflower, beet, spinach, and garlic). These data were in-
R162 JOURNAL OF FOOD SCIENCEVol. 72, Nr. 9, 2007
dependently used to validate RACI. In their research, a modified-
FRAP method that determines the antioxidant capacity of both
water- and lipid-soluble antioxidants was used to determine the
antioxidant capacity of vegetables and the result was expressed as
total antioxidant content. The RACI developed herein showed sig-
nificant linear relationship with the total antioxidant content (r =
0.75, P < 0.01; Figure 3). The scientific value of RACI is therefore
enhanced by its significant correlation with this independent data
set. Thus, the RACI is a valid approach to rank food antioxidant
capacity.
Method recommendations
Ideally, the more chemical methods used in developing RACI,
the closer RACI represents the ranking (high or low) of the food an-
tioxidant capacity. In many situations, due to limitations of instru-
mentation, time, and budget, it is not practical to measure antioxi-
dant capacity of food items with more than 3 chemical methods. To
choose a method to determine food antioxidant capacity, the types,
and concentrations of antioxidants in the food, the type of sub-
strates, and analysis conditions need to be considered. RACI can be
used as a reference to choose a chemical assay that can better rep-
resent the food antioxidant status. Based on the concept of RACI,
the statistically more sensitive combined methods, which had high
correlation coefficients with RACI as well as a higher percentage of
significant correlation with methods not used to develop RACI, are
recommended for determining antioxidant capacity (Table 4). RACI
developed from the combination of ORAC (ROO r), FRAP, and ABTS,
PAOXI, or TRAP, had high correlation coefficients of 0.93, 0.95, or
0.94, respectively, and correlated with other methods with the high-
est percentage (75%), compared with the other 3-method combina-
tions.
Due to a variety of reasons, 1 method may be justified. For deter-
mining antioxidant capacity using a single method, ORAC (ROO r),
the most statistically sensitive method in developing RACI, showed
the highest percentage of significant correlation with other meth-
ods (r = 0.83) and high correlation with RACI (86%). Further-
more, ORAC (ROO r) was recommended as a standard method by
Figure 1 --- Relative antioxidant
capacity index of 20 commonly
consumed vegetables.
Evaluating food antioxidant capacity . . .

R: Concise Reviews in Food Science

Prior and others (2005) to be used as routine quality control the radical sources and solvent. Therefore, ORAC (ROO r) is recom-
and measurement of food antioxidant capacity. It is a HAT-based mended for analyzing the antioxidant capacity of vegetables if only
method that uses a controllable source of peroxyl radicals and can 1 method is justifiable. FRAP also had a high correlation coefficient
detect both hydrophilic and hydrophobic antioxidants by altering (r = 0.87), but a low percentage of significant correlation with the

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R: Concise Reviews in Food Science Evaluating food antioxidant capacity . . .

other 6 methods (67%). For a 2-method combination, ORAC (ROO)
and FRAP are recommended because RACI developed by these 2
methods had the highest correlation (r = 0.95) and the highest per-
centage of correlation with other methods (80%).
Limitations of the relative antioxidant capacity index
RACI correlated with each chemical assay and may be used as
a reference for measuring food antioxidant capacity. As a simple
numerical approach with a relative value, RACI has its own weak-
nesses. First, RACI is not a fixed value and is thus subject to ma-
nipulation by the methods it includes. These methods may differ
substantially in chemical mechanism and thus contribute different
proportions to RACI. It may also change as different food items or
the same food with different genotypes are added or removed from
the testing list, which affects the standard score of each food item.
Second, although RACI was developed from chemical assays, it can-
not directly reflect a single specific chemical mechanism that is in-
volved in the antioxidant reactions of a selected food item. It is a
combination of transformed data generated from several mecha-
nisms into a standard score. Food items with the same RACI may
have different antioxidants in quality and/or quantity. Third, even
though RACI was developed from 7 chemical assays, it may not
cover all the chemical aspects of antioxidant reactions. A variety of
chemical methods measuring different antioxidant properties may
reflect a more accurate RACI. Indeed, the value of RACI might be
enhanced if the number and type of antioxidant methods was de-
fined and fixed. Furthermore, measuring antioxidant capacity of
food items with multiple chemical methods is not always practi-
cal. Although one can collect data from independent experiments,
which is the case of the present analyses, assumptions are made
that experimental conditions are similar and experimental errors
are ignored for the same chemical method.
When assessing health benefits from food antioxidants in bio-
logical systems, the combined effects of complex bioactive com-
pounds need to be considered (Eberhardt and others 2000). The
RACI in the present study was developed from in vitro chemical
assays. To achieve a more powerful RACI, more tests of food an-
tioxidant capacity in real biological systems, particularly humans,
should be included to represent the bioavailability of antioxidants
and their clinical effectiveness (Collins 2005). An ideal RACI would
be developed from data generated from both in vitro and in vivo
assays with equal contribution from assays covering all antioxidant
properties. If 1 special group of antioxidants is of interest, such as
lipophilic or hydrophilic antioxidants, RACI can be developed by
methods that cover the properties of that special group.

Conclusions

G
iven the fact that food has different classes of antioxidants
and each method to measure the antioxidant capacity has its
own limitations, RACI provides an alterative approach to integrate
several chemical assays. Although RACI may not directly represent
any individual chemical mechanism, which is the traditional way to
measure and define antioxidant capacity, it does correlate with re-
sults from different chemical assays. The key advantage of RACI is
that it is a numerical scale integrating multiple chemical methods,
allowing comparison of antioxidant capacity of a large number of
foods. RACI goes beyond chemical reactions and ranks antioxidant
capacity without the restriction of mechanism. Although RACI does
not allow avoidance of performing chemical assays, it is an alterna-
tive assessment to help standardize methods. RACI is a relative food
Figure 3 --- Regression (r = 0.75, P < 0.01) between rela- antioxidant capacity ranking tool that presents the antioxidant ca-
tive antioxidant capacity index (RACI) and total antiox-
idant content measured with a modified-ferric reducing pacity of each food in a standardized format for use by the food in-
antioxidant power assay in 16 vegetables. dustry, scientists, and consumers.

Table 4 --- Recommended chemical methods used to evaluate food antioxidant capacity.
Numbers of methods used to develop RACI
1 2 3 3 3
Recommended methods ORAC (ROO r) ORAC (ROO r) ORAC (ROO r) ORAC (ROO r) ORAC (ROO r)
FRAP FRAP FRAP FRAP
ABTS PAOXI TRAP
Correlation coefficient (r )a 0.86 0.95 0.93 0.95 0.94
Methods tested ABTS ABTS ORAC (Cu2+ ) ABTS ABTS
FRAP ORAC (Cu2+ ) ORAC (OH r) ORAC (Cu2+ ) ORAC (Cu2+ )
ORAC (Cu2+ ) ORAC (OH r) PAOXI ORAC (OH r) ORAC (OH r)
ORAC (OH r) PAOXI TRAP TRAP PAOXI
PAOXI TRAP
TRAP
Methods with significant correlation ABTS ABTS ORAC (Cu2+ ) ABTS ABTS
FRAP ORAC (Cu2+ ) ORAC (OH r) ORAC (Cu2+ ) ORAC (Cu2+ )
ORAC (Cu2+ ) ORAC (OH r) TRAP TRAP ORAC (OH r)
ORAC (OH r) TRAP
TRAP
Methods without significant correlation PAOXI PAOXI PAOXI ORAC (OH r) PAOXI
Number of methods with significant correlation 5 4 3 3 3
Number of methods without significant correlation 1 1 1 1 1
Significant correlation (%) 83.3 80.0 75.0 75.0 75.0
a
Correlation with relative antioxidant capacity index (RACI) developed from 7 chemical methods.

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Evaluating food antioxidant capacity . . .
Acknowledgments
These analyses were supported by the Natl. Research Initiative of
the USDA Cooperative State Research, Education and Extension
Service, grant no. 2003-35200-05377 and Standard Process Inc.,
Palmyra, Wis. We appreciate the 2 anonymous reviewers construc-
tive comments that have improved the quality of this article.
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