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JOURNAL OF
FOOD COMPOSITION
AND ANALYSIS
Journal of Food Composition and Analysis 19 (2006) 544– 551
Original Article
Effect of season and production location on antioxidant activity of
Moringa oleifera leaves grown in Pakistan
Shahid Iqbal
, M.I. Bhanger
Free Radical Research Laboratory, National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro-76080, Pakistan
Received 28 September 2004; received in revised form 9 May 2005; accepted 12 May 2005
Abstract
Antioxidant activity (AA) of methanolic extracts from Moringa oleifera leaves, as function of seasons and agroclimatic locations,
was investigated. Total phenolic content (TPC), total flavonoid content (TFC), ascorbic acid (AAcid) content , reducing power, AA
in linoleic acid system and scavenging power of superoxide anion radical were taken as parameters for evaluation of AA. Significant
differences were observed in the AA of the extracts from different locations and seasons. Generally, samples from Mardaan
exhibited highest AA followed by Balakot, Chakwal, Jamshoro, and Nawabshah. Overall antioxidant efficacy was greater in
December or March depending upon location, and least in June. Antioxidant potential of M. oleifera leaves from Pakistan was quite
comparable or higher than literature values for M. oleifera from other countries and some other potent antioxidants. This work
shows that season and agroclimatic locations have profound effect on the AA of M. oleifera leaves.
r2005 Elsevier Inc. All rights reserved.
Keywords: Moringa leaves; Antioxidant activity; Agroclimatic locations; Seasons; Total phenolics; Reducing power
1. Introduction
Epidemiological studies have shown that foods rich in
antioxidants provide protection against degenerative
diseases including cancer, coronary heart diseases, and
Alzheimer’s disease (Pezzuto and Park, 2002;Razali
et al., 1997;Ames et al., 1993). Therefore, it is
considered important to increase intake of antioxidants
from dietary sources (Soong and Barlow, 2004).
Previously, synthetic antioxidants like BHA and BHT
have been in use as food additives but recent reports
have expressed safety concerns about their usage (Sun
et al., 2005) and natural antioxidant have become the
focus of intense interest (Wilson, 1999). The commercial
development of plants as sources of antioxidants to
enhance health and food preservation is the focus of
current interest (Rice-Evans et al., 1997). Plants are rich
sources for natural antioxidants, the best known are
tocopherols, flavonoids, vitamin C, and different phe-
nolic compounds (Laandrault et al., 2001). Many herbs,
fruits (Ramarathnam et al., 1995), vegetables (Ismail
et al., 2004), spices, and legumes have been exploited as
sources of antioxidants.
Moringa oleifera, Lam. (Moringaceae) is grown
world-wide in the tropics and sub-tropics of Asia and
Africa (Booth and Wickens, 1988) and is one of the 14
species of genus Moringa, which are native to India,
Africa, Arabia, Southeast Asia, the Pacific and Car-
ibbean islands, and South America (Sengupta and
Gupta, 1970). The species are drought resistant and
tolerate a wide range of soil and rainfall conditions.
Various varieties of M. oleifera have been developed to
meet the tastes of local populations (Rajan, 1986).
Two species of Moringa, i.e., M. concanensis and
M. oleifera are present in Pakistan, but no compiled
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0889-1575/$ - see front matter r2005 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2005.05.001
Corresponding author. Tel.: +92 333 2656248/92 300 3079516;
fax: +92 221 771560.
E-mail addresses: ranashahid313@yahoo.com,
ranashahid313@hotmail.com (S. Iqbal).
information about these varieties is present. M. con-
canensis is found rarely, but M. oleifera is cultivated
widely throughout the temperate regions of the country
(Qaiser, 1973).
Different parts of this plant, i.e., leaves, flowers, roots
and fruits, have traditionally been used for dietary
purposes as vegetables (Qaiser, 1973;Siddhuraju and
Becker, 2003). Fresh leaves have been used by Indian
inhabitants for the preparation of cow and buffalo ghee
from butterfat. Its leaves are a valuable source of
vitamins A and C, they promote digestion, and they are
used in catarrhal afflictions and for treating wounds (Pal
et al., 1995). The leaves mixed with chicken soup are
used by Philippine women to enhance breast milk
production (Chopra et al., 1956). Leaves of M. oleifera
have been reported to contain flavonoid pigments such
as kaempferol, rhamnetin, isoquercitrin, and kaempfer-
itrin (Nair and Subramanian, 1962). Recently, from our
laboratory, Anwar and Bhanger (2003) exploited M.
oleifera as a non-conventional source of oil with
significantly high oxidative stability, revealing the
presence of natural antioxidants. Lalas and Tsaknis
(2002), from Greece reported the oxidative stability of
seed oil and isolated components liable for antioxidant
activity (AA) from Malawi variety. A report on
antioxidant properties of M. oleifera leaves from
different countries, suggesting wide variation, has been
exploited by Siddhuraju and Becker (2003). Recently,
Anwar et al. (2005) have reported significant variations
in oil content of Moringa seeds as function of agrocli-
matic locations.
However, no study has been conducted so far dealing
with the AA and composition of M. oleifera leaves
indigenous to Pakistan. No data describe the effect
of agroclimatic location and season on the antioxidant
potential of M. oleifera leaves. The ultimate objective
of the present study was to investigate AA and
composition of M. oleifera leaves, and the effects
of agroclimatic conditions and seasons within Pakistan
on AA.
2. Materials and methods
2.1. Materials
Fresh leaves of M. oleifera were collected in the
months of December, March, June, and September from
five different areas of Pakistan: Balakot, Chakwal,
Jamshoro, Mardaan, and Nawabshah for 2 years
w.e.f. December 2001 to September 2003. Leaves were
collected at the beginning, mid-point and at the end
of each month mentioned to ensure the reliability of
data. All reagents (analytical and HPLC) used were
from E. Merck, Fluka or Sigma Aldrich unless stated
otherwise.
2.2. Extraction of total phenolics
Extraction of Moringa leaves was carried out follow-
ing the method reported by Zuo et al. (2002). Air-dried
leaf samples (5.0 g) were ground so as to pass 1mm sieve
size and extracted with 25 mL of 80% methanol for 3 h
in an electrical shaker at room temperature. The
contents of the flasks were further extracted twice with
20 mL of 80% methanol containing 0.15% HCl under
the same set of conditions. The extracts were combined
and filtered through 0.45 mm of Nylon membrane filter.
The extracts were evaporated to dryness under reduced
pressure at 45 1C by a rotary evaporator and stored in a
freezer at 18 1C until used for further analyses.
2.3. Measurement of total phenolic content (TPC)
TPC in the methanolic extracts of M. oleifera leaves
was determined by Folin–Ciocalteu reagent assay (Folin
and Ciocalteu, 1927;Singleton and Rossi, 1965). The
reaction mixture contained 200 mL of extracts, 750 mLof
freshly prepared diluted (1:10) Folin–Ciocalteu reagent
and 2 mL of 7.5% sodium carbonate. The final mixture
was diluted to 7 mL with deionized water. Mixtures were
kept in the dark at ambient conditions for 2 h to
complete the reaction. Then the absorbance at 765 nm
was measured on a Perkin-Elmer Lambda-2 Spectro-
photometer, with a 1 cm cell. All the experiments were
conducted thrice using gallic acid as a calibration
standard, and results were recorded as gallic acid
equivalents (g/100 g of extract).
2.4. Determination of total flavonoid content (TFC)
The TFC was measured following a reported colori-
metric assay (Zhishen et al., 1999;Kim et al., 2002)with
slight modifications. Briefly, the methanolic extract of
each sample (1mL) was appropriately diluted and added
to a 10 mL volumetric flask containing 4mL of H
2
O.
Initially, 0.3 mL of 5% NaNO
2
was added to each
volumetric flask; at 5 min, 0.3 mL of 10% AlCl
3
was
added; at 6 min, 2 mL of 1 MNaOH was added. Each
reaction flask was then immediately diluted with 2.4 mL
of H
2
O and mixed. Absorbance of the mixtures was
determined at 510 nm. TFC of the samples was deter-
mined as epicatechin equivalents (mg/g). Three readings
were taken for each sample and results were averaged.
2.5. Quantitative determination of ascorbic acid
Ascorbic acid determination was carried out following
a reported indophenol titration method (Anwar et al.,
1990). Samples were homogenized in metaphosphoric
acid solution and extracted. The vitamin C was titrated
against 2,6-dichlorophenol–indophenol solution at pH
0.6 in the presence of formaldehyde, to a pink end point.
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2.6. Determination of reducing power
Reducing power was determined following the meth-
od reported by Yen and Duh (1993). Extracts (0, 1.0,
2.0, 5.0, 7.5, 10.0, and 15.0 mg) were mixed with
phosphate buffer (5.0 mL, 2.0 M, pH 6.6) and 1%
potassium ferricyanide (5 mL), and the mixtures were
incubated at 50 1C for 20 min. The quantity 5 mL of
10% trichloroacetic acid were added and the mixture
was centrifuged at 650gfor 10 min. The upper layer of
the solution (5 mL) was mixed with distilled water
(5 mL) and 0.1% ferric chloride (1 mL), and absorbance
was measured at 700 nm. The experiment was conducted
in triplicate and results were averaged.
2.7. Antioxidant activity determination in linoleic acid
system
The AA of methanolic extracts was determined
following a reported method (Osawa and Namiki,
1981). Sample extracts were added to a solution mixture
of linoleic acid (0.13 mL), 99.8% ethanol (10 mL), and
0.2 Msodium phosphate buffer (pH 7.0, 10 mL). The
total volume was adjusted to 25 mL with distilled water.
The solution was incubated at 40 1C, and the degree of
oxidation was measured according to the thiocyanate
method (Mitsude et al., 1966) with 10 mL of ethanol
(75%), 0.2 mL of an aqueous solution of ammonium
thiocyanate (30%), 0.2 mL sample solution, and 0.2 mL
of ferrous chloride (FeCl
2
) solution (20 mMin 3.5%
HCl) being added sequentially. After 3 min of stirring,
the absorption values of mixtures measured at 500 nm
were taken as peroxide contents. A control was
performed with linoleic acid but without the extracts.
Each sample was run in triplicate and results were
averaged.
2.8. Scavenging of superoxide anion radical
Superoxide anion radical scavenging activity of
methanolic extracts of M. oleifera leaves was determined
by the PMS–NADH superoxide generating system
(Nikishimi et al., 1972;Lai et al., 2001). Extracts were
added to a solution mixture that contained 200 mM
NBT, 624 mMNADH, and 80 mMPMS in 0.1 M
phosphate buffer, pH 7.4. After 2 min of incubation at
room temperature, the absorbance was measured at
560 nm. Percentage capability to scavenge the super-
oxide radical was calculated using the following
equation:
Scavenging effect ð%Þ
¼1Absorbance of sample at 560 nm
Absorbance of control at 560 nm
100
2.9. Statistical analysis
All the data is reported as mean7S.D. Data were
analyzed using two-way ANOVA with replications
following method reported by (Zar, 1996). Data was
considered significant at Po0:05. Statgraphics
1990–1991 Version 5.0 was used for statistical analysis.
3. Results and discussion
3.1. Total phenolic content
The phenolic compounds may contribute directly to
antioxidative action (Awika et al., 2003); therefore it is
necessary to investigate TPC. The TPC was determined
following a modified Follin–Ciocalteu method and
results were expressed as gallic acid equivalents
(Table 1).
Significant differences (Po0:05) in TPC were ob-
served as function of season and production location.
Among locations, highest TPC was observed for
samples from Mardaan followed by Balakot, Chakwal,
Jamshoro, and Nawabshah, respectively. Samples from
Nawabshah and Jamshoro, with almost identical
agroclimatic conditions and environmental temperature
(e.g., up to 51 1C during June) exhibited non-significant
differences in TPC and were the lowest among the
locations investigated. This might be attributed to the
strong effect of temperature on TPC. It is supported by
report of Wang and Zheng (2001), showing that
environmental temperature strongly alters antioxidant
properties in strawberry. Furthermore, the observation
of Yu et al. (2003) also supports the findings of the
present work, stating that solar radiation reflects the UV
exposure, which may be associated with free radical
formation and singlet oxygen production. Recently,
Anwar et al. (2005) described wide variation in oil
content of M. oleifera seeds from different agroclimatic
locations due to diversity in natural soil texture and
climatic constraints.
A survey of the literature reveals that TPC in M.
oleifera leaves from Mardaan was higher than those
reported from India, Niger, and Nicaragua (Siddhuraju
and Becker, 2003) throughout the year. The samples
collected from Chakwal and Balakot had TPC that was
higher than India but lower than Niger and Nicaragua.
Generally, the trend of TPC as a function of the
season was as: December4March4September4June
for all the locations investigated. However, for samples
from Mardaan, TPC in March was higher than in
December, perhaps due to extreme cold weather in
December in Mardaan. Results for December and
March were statistically non-significant among them-
selves, while for June and September TPC was
statistically significant (Po0:05). Temperature ranges
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S. Iqbal, M.I. Bhanger / Journal of Food Composition and Analysis 19 (2006) 544 –551546
for all the locations were determined from Pakistan
Meteorological Department, and an inverse relation of
environmental temperature with TPC was observed.
Highest TPC was observed in the month of December
(coldest temperatures) while the lowest was in June
(hottest temperatures). This may be due to the fact that
Moringa leaves grow in the month of June and mature
from December to March, and phenolic content is
lowest in newly opened leaves, increasing gradually with
the maturity of leaves. Previous reports suggest that
phenolic content increases with the increase in leaf-age
and is lower in early stages of leaf growth, gradually
increasing with the maturity of leaves (Julkunen-Tiitto,
1989;Wiermann, 1981). Furthermore, the activity of the
plants is at its maximum during hot seasons (June in
Pakistan), leading to a loss of phenolic content.
However, this activity reduces with the decrease in
seasonal temperature resulting in a retention of pheno-
lics (Lavola, 1998). The report by Sanders (1982),
describing that an increase in unsaturated fatty acids is
generally associated with cooler climates leading to
production of antioxidants for a self-defense system
against environmental stress, supports our present
findings suggesting pronounced effects of environmental
temperature on AA. Some literature reports (Wang and
Stretch, 2001) reveal that an increase in TPC occurs with
the increase in storage temperature up to an optimized
level, while environmental temperature exhibited con-
troversial behavior than storage temperature in the
present study.
Another observation was made concerning the effect
of altitude on TPC. Balakot and Mardaan, at an
altitude ranging from 400–2700 ft with respect to other
locations and with strong stormy wind throughout the
year, exhibited higher TPC than those of other
locations, which are flat and have no stormy winds or
rains. Although no previous findings support this
observation directly, eastern practitioners generally
believe that plants from hilly areas are more important
from a nutritional and dietary point of view (Anwar
et al., 2004).
3.2. Total flavonoid content (TFC)
Flavonoid content was determined by a reported
method (Zhishen et al., 1999) and results were expressed
as epicatechin equivalents (Table 2). TFC was found in
appreciable amounts.
Exactly the same effects of agroclimatic location on
TFC were observed as for TPC, perhaps due to the
phenolic nature of flavonoids. However, the seasonal
effects on TFC were different from TPC, and no
significant differences among seasons for TFC could
be observed. Only the results of September versus
March and December were statistically different
(Po0:05). TFC was comparable to those of Moringa
samples from India, Niger, and Nicaragua (Siddhuraju
and Becker, 2003) and in mulberry leaves (Zhishen et al.,
1999) for all samples. However, samples from Mardaan
exhibited higher TFC than the literature values (Sid-
dhuraju and Becker, 2003). These results suggest that
Moringa leaves may be exploited as an important source
in nutraceutical and functional food industries.
3.3. Ascorbic acid concentration
Ascorbic acid (AAcid), was determined following a
reported method (Anwar et al., 1990) and results are
presented in Table 3. Slight differences among locations
could be investigated; however, the differences were
non-significant among seasons. AAcid content was
highest in the month of March for Mardaan, Chakwal,
and Balakot locations (i.e., 0.044–0.046 (g/100 g)), and
in December for those from Jamshoro and Nawabshah,
i.e., 0.037–0.039 (g/100 g). AAcid remained unchanged
during March, June, and September for Jamshoro
samples, whereas for the Nawabshah samples it was
exactly the same during December and September.
AAcid content was lowest for all the locations in June.
AAcid content in the Mardaan and Balakot samples was
comparable to Moringa samples from India and
Nicaragua (Siddhuraju and Becker, 2003) and orange-
colored carrot, while it is almost three times higher than
ARTICLE IN PRESS
Table 1
Total phenolic content of methanolic extracts of Moringa oleifera leaves (g/100 g)
Nawabshah Jamshoro Mardaan Chakwal Balakot Average
Dec. 9.1770.45 9.3470.58 13.0970.38 11.8770.19 12.3970.40
a
11.1770.4
Mar. 8.9870.30 9.0270.21 13.5670.11 10.7970.70 12.0870.23
a
10.8970.31
Jun. 8.3770.33 8.4370.41 11.8270.31 9.6770.24 11.4770.10
b
9.9570.28
Sep. 8.7470.27 9.1670.10 12.6870.36 9.8370.37 11.8370.52
c
10.4570.32
Average
a
8.8270.34
a
8.9970.33
b
12.7970.29
c
10.5470.38
d
11.9470.31
All data is reported as mean7S.D.(n¼6).
F-statistics for seasons ¼26.405 at df ¼3, P¼0:000.
F-statistics for locations ¼230.496 at df ¼4, P¼0:000.
Values with same letter are statistically non-significant at 0.05 LSD.
Dec. ¼December, Mar. ¼March, Jun. ¼June, Sep. ¼September.
S. Iqbal, M.I. Bhanger / Journal of Food Composition and Analysis 19 (2006) 544 –551 547
those in yellow- and white-colored carrots (Alasalvar
et al., 2001), revealing a high nutritional value for
Moringa leaves. The overall order of AA was the same
for a specific geographic location and season, suggesting
marked influence of location and season on the AA of
Moringa leaves.
3.4. Reducing power
Literature reports (Siddhuraju et al., 2002;Yildirim et
al., 2001) show that the reducing power of bioactive
compounds is directly related to AA. A regular pattern
of increase in reducing power as a function of extract
concentration was observed (Table 4). The highest
reducing power was observed for samples from Mar-
daan followed by Balakot, Chakwal, Nawabshah, and
Jamshoro, respectively. Marked variation in the redu-
cing power was observed for all the locations and
seasons. Reducing power was lowest in June for all the
locations except from Balakot, where the lowest
reducing power was observed in September, while the
highest in December was for samples from Chakwal,
Jamshoro, and Nawabshah, contrary to those from
Balakot and Mardaan which showed the highest
reducing power in March. Results for reducing power
are comparable to those of the reducing power in Hsian-
tsao leaf gum and in case of samples from Mardaan,
reducing power is double to Hsian-tsao leaf gum (Lai
et al., 2001), which has been reported as a potent source
of antioxidant. In the light of these findings, M. oleifera
leaves may be exploited as an important source of
antioxidant.
3.5. Antioxidant activity in linoleic acid system
AA of all the samples was observed in linoleic acid
system (Fig. 1). Significant variation in AA with respect
to season and geographic location was observed. The
same order of AA was followed, i.e., Mardaan4Bala-
kot4Chakwal4Jamshoro4Nawabshah. Highest AA
for Jamshoro, Chakwal, and Nawabshah was observed
in the month of December whereas for Mardaan and
Balakot, this occurred in March. Many species of
compounds contribute to AA, therefore biological
uptake of various antioxidant compounds like Se by
Moringa may have vital influence on the AA of samples
from different locations, which in turn depends on soil
texture and other climatic conditions (Lorenz, 1978).
ARTICLE IN PRESS
Table 3
Ascorbic acid content (g/100 g) of methanolic extracts of Moringa oleifera leaves (g/100 g)
Nawabshah Jamshoro Mardaan Chakwal Balakot Average
Dec. 0.03770.01 0.03970.01 0.04270.01 0.03970.01 0.04470.01
a
0.04070.01
Mar. 0.03670.01 0.03570.01 0.04570.02 0.04470.01 0.04670.01
a
0.04170.01
Jun. 0.03270.01 0.03570.01 0.03670.01 0.03670.01 0.04170.01
a
0.03670.01
Sep. 0.03770.01 0.03570.01 0.0470.02 0.03570.01 0.04270.00
a
0.03870.01
Average
a
0.03670.01
a
0.03670.01
b
0.04170.015
b
0.03870.01
c
0.04370.01
All data is reported as mean7S.D.(n¼6).
F-statistics for seasons ¼1.174 at df ¼3, P¼0:3292,
F-statistics for locations ¼1.804 at df ¼4, P¼0:1428.
Values with same letter are statistically non-significant at 0.05 LSD.
Dec. ¼December, Mar. ¼March, Jun. ¼June, Sep. ¼September.
Table 2
Total flavonoid content of methanolic extracts of Moringa oleifera leaves (g/100 g)
Nawabshah Jamshoro Mardaan Chakwal Balakot Average
Dec. 7.3770.26 7.1270.51 12.7970.30 9.9170.25 11.9570.28
a
9.8370.32
Mar. 7.0270.24 8.1470.21 12.6970.21 9.3970.27 12.1570.21
a
9.8870.23
Jun. 6.5970.21 6.9370.21 12.1570.43 8.8570.14 11.6770.36
b
9.2470.27
Sep. 6.7470.27 7.3470.17 12.4970.17 9.1370.18 11.8270.40
c
9.5170.24
Average
a
6.9370.25
a
7.3870.28
b
12.5370.28
c
9.3270.21
d
11.9070.31
All data is reported as mean7S.D.(n¼6).
F-Statistics for seasons ¼2.526 at df ¼3, P¼0:0680,
F-Statistics for locations ¼46.167 at df ¼4, P¼0:000.
Values with same letter are statistically non-significant at 0.05 LSD.
Dec. ¼December, Mar. ¼March, Jun. ¼June, Sep. ¼September
S. Iqbal, M.I. Bhanger / Journal of Food Composition and Analysis 19 (2006) 544 –551548
3.6. Superoxide anion radical scavenging activity
Superoxide radical scavenging activity for all the
samples was measured at a concentration of 1 mg/mL;
results are shown in Fig. 2. All the extracts exhibited an
appreciable scavenging activity. Significant differences
in scavenging activity were observed among samples
from different locations and seasons. Scavenging acti-
vity was significantly affected by agroclimatic locations
in the same pattern. For samples from Mardaan, the
ARTICLE IN PRESS
Table 4
Reducing power of methanolic extracts of Moringa oleifera leaves from different agroclimatic locations in different seasons
Samp month Conc. (mg) Nawabshah Jamshoro Mardaan Chakwal Balakot
Dec. 0 0.01670.001 0.02270.001 0.01370.001 0.01770.002 0.00970.001
1 0.2770.01 0.3370.01 0.4370.01 0.3970.01 0.4370.02
2 0.5070.02 0.5970.01 0.7870.01 0.7270.01 0.7670.01
5 1.3170.02 1.3270.03 1.7770.02 1.4970.03 1.6770.04
7.5 1.7870.03 1.8170.05 2.4770.07 2.0970.01 2.2270.02
10 2.2170.02 2.4270.02 3.0270.02 2.4770.02 2.6270.01
15 2.7470.03 2.7870.01 3.8470.09 3.1770.03 3.3270.03
Mar. 0 0.00970.001 0.02370.001 0.01370.001 0.02870.001 0.01970.001
1 0.2370.01 0.2770.01 0.4670.01 0.3470.01 0.4570.02
2 0.4270.01 0.4870.01 0.8370.02 0.6670.03 0.8270.04
5 1.1370.04 1.1970.01 1.8670.04 1.4070.06 1.7670.05
7.5 1.6670.03 1.6170.02 2.5470.05 1.9270.03 2.3170.02
10 1.9770.02 2.0570.04 3.1970.12 2.4170.01 2.7970.03
15 2.5970.02 2.4970.03 4.0270.02 3.0770.03 3.3970.07
Jun. 0 0.00770.001 0.01470.001 0.01870.001 0.0270.001 0.01570.001
1 0.1970.01 0.2270.01 0.3970.01 0.2970.01 0.3970.01
2 0.3770.01 0.4070.01 0.6770.01 0.5370.02 0.6870.03
5 1.0470.04 0.9170.02 1.5870.03 1.2470.01 1.4770.06
7.5 1.5270.02 1.3270.03 2.2770.08 1.7370.04 2.0770.03
10 1.8070.02 1.6670.01 2.8470.04 2.0470.01 2.5470.02
15 2.4170.06 2.3170.02 3.6970.06 2.4270.03 3.2770.06
Sep. 0 0.01970.001 0.00870.001 0.02170.001 0.02470.002 0.00370.001
1 0.1970.01 0.2670.01 0.4270.01 0.3170.01 0.3970.01
2 0.4170.02 0.4670.01 0.7270.02 0.5770.01 0.7170.02
5 1.0470.03 1.0470.01 1.8170.03 1.3070.02 1.5670.02
7.5 1.5770.01 1.4570.02 2.4570.01 1.8170.03 2.0070.04
10 1.8570.03 1.8970.02 2.9670.07 2.1970.02 2.4270.01
15 2.4970.01 2.4670.03 3.7870.04 2.7670.02 3.2170.06
All data is reported as mean7S.D.(n¼6).
Statistical analysis was subjected to values of 5 mg for all locations and seasons.
F-Statistics for seasons ¼56.523 at df ¼3, P¼0:0000.
F-Statistics for locations ¼264.785 at df ¼4, P¼0:0000.
Values with same letter are statistically non-significant at 0.05 LSD.
Dec. ¼December, Mar. ¼March, Jun. ¼June, Sep. ¼September.
80
82
84
86
88
90
92
94
Nawabshah Jamshoro Mardan Chakwal Balakot
Production Locations
Inhibition of peroxidation (%)
.Dec. Mar.
Jun. Sep.
Fig. 1. Antioxidant activity of methanolic extracts of Moringa oleifera in linoleic acid system.
S. Iqbal, M.I. Bhanger / Journal of Food Composition and Analysis 19 (2006) 544 –551 549
highest scavenging activity was observed in March and
the lowest in June. For all other locations the highest
scavenging activity was in December and lowest in
March for Nawabshah, in June for Chakwal, Mardaan,
Jamshoro, and Balakot. For Nawabshah, a controversy
relative to previous results was observed, reducing
power in June was far higher than in September
and March. Overall order of scavenging was similar to
TPC, TFC, and reducing power for all the locations.
Results of scavenging effects for Jamshoro, Nawabshah,
and Chakwal were closer to one another, while
Balakot and Mardaan exhibited closer values among
themselves.
4. Conclusions
From the present work, it could be concluded that
agroclimatic locations and seasons have profound
effects on the AA of M. oleifera leaves. AA of samples
from cold areas was relatively higher than those from
temperate regions. Similarly, from all the samples, AA
was highest in December (cold month) and lowest in
June (hot month), with few exceptions. These findings
suggest that environmental temperature has a significant
effect on AA evaluation. However, there is still a need to
investigate effects of soil properties on AA of Moringa
leaves.
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