Journal of Research in Biology
An International Scientific Research Journal
Study of FRAP and TBA in the ethanol and methanol extracts of Prunus persica
var. compressa (Loud), Persica vulgaris Miller and Prunus persica var.
nucipersica in west Azarbijan
Authors:
Journal of Research in Biology
Nasrin Veghar Moosavi,
Siavash Hosseini Sarghein
and Rashid Jamei
Institution:
Department of Biology,
Faculty of Science,
University of Urmia, West
Azarbaijan, Iran.
Corresponding author:
Nasrin Veghar Moosavi
Web Address:
http://jresearchbiology.com/
documents/RA0629.pdf
Journal of Research in Biology
An International
Scientific Research Journal
ISSN No: Print: 2231 –6280; Online: 2231- 6299
Short Communication
ABSTRACT:
The objectives of this test is to detect the antioxidant activity (FARP and TBA)
of ethanolic and methanolic extracts of Persica vulgaris Miller, Prunus persica var.
compressa (Loud), Prunus persica var. nucipersica leaves and fruits. A direct
correlation between phenol content with FRAP and TBA in the ethanolic and
methanolic extracts of fruits and leaves of these plants were noticed. The results
showed that, the largest amount of ferric reducing antioxidant power was in the
ethanolic extract of Prunus persica var. compressa (Loud) fruit and large amounts of
TBA were found in the ethanolic extracts of Persica vulgaris Miller leaves. We can
conclude that these plants have the highest ability to collect free radicals for having its
high antioxidant power.
Keywords:
Prunus persica var. compressa (Loud), Persica vulgaris Miller, Prunus persica
var. nucipersica, antioxidants, FRAP and TBA.
Article Citation:
Nasrin Veghar Moosavi, Siavash Hosseini Sarghein and Rashid Jamei
Study of FRAP and TBA in the ethanol and methanol extracts of Prunus persica var.
compressa (Loud), Persica vulgaris Miller and Prunus persica var. nucipersica in west
Azarbijan
Journal of Research in Biology (2018) 8(5): 2511-2517
Dates:
Received: 26 April 2018 Accepted: 22 May 2018 Published: 10 July 2018
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/
licenses/by/4.0), which gives permission for unrestricted use, non-commercial, distribution and
reproduction in all medium, provided the original work is properly cited.
2511-2517| JRB | 2018| Vol 8 | No 5
www.jresearchbiology.com
 Moosavi et al., 2018

INTRODUCTION methanol and mixed for three hours on shaker in a dark

Reactive oxygen species are effective in driving room. The extracts were filtered. One milligram of
out malignant diseases such as cancer, diabetes, gout, sodium azide was used as a preservative and extracts
cardiovascular disease (Kris-Ethertonmet et al., 2002). were stored in the refrigerator at four degrees until the
Phenolic compounds are secondary metabolites that tests get completed.
often synthesized through Shikimic acid pathway. Ferric Reducing Antioxidant Power (FRAP)
Depending on the measuring method used, the vegetable In order to perform FRAP evaluation on the
extract in different courses showed different antioxidant samples, method of Benzie and Strain, (1996) was used.
abilities (Gardner et al., 2000). Gardner et al. 2000 used FRAP reagent 20 mL of solution, TPTZ 10 mM in HCL
FRAP to measure the amount of ascorbic acid in orange 40 mm, 2.5 mL FeCl3 (20 mm) and 25 mL acetate
juice and showed that the letter possess antioxidant buffer (3.0 mm) (PH = 3/6) was prepared. 10 µl of
properties. extract was mixed with 3 ml of the FRAP reagent and
the absorption of reaction mixture was measured at 595
MATERIALS AND METHODS nm. Certain concentration (10-6 / 0 micro molar) of iron
Collection, preparation and extraction of samples sulfate (FeSO4) was used for the calibration curve.
Fruits and leaves of Persica vulgaris Miller, Inhibition of lipid peroxidation measured by
Prunus persica var. compressa (Loud.) and Prunus ThioBarbituric Acid (TBA)
persica var. nucipersica in July 2014 were collected The ThioBarbituric Acid method (TBA) was
randomly Urmia gardens. Fruits and leaves were freezed used to measure the capacity of lipid peroxidation. At
and stored at -80°C until extraction. During the temperatures above 100°C and at low PH,
extraction, fruit and leaf samples were crushed MalonDiAldehyde (MDA) joins with TBA and forms a
separately, at room temperature to extract; 30 g of red complex that can be determined at 532 nm (Chawla
extracts was dissolved in, 100 mL of ethanol and et al., 1976). 2 mL of 3, chloroacetic acid of 20%

3 a a
bc ab ab
Ferric Reducing Antioxidant Power (FRAP)

ab
c
2.5 d
e
f
2
(FeSo4 mg/gFWmg)

g g

1.5

0.5

0
Prunus persica var. Persica vulgaris Prunus persica
compressa Miller var. nucipersica

Figure 1. Ferric reducing antioxidant power of ethanolic and methanolic extracts (mgFeSo 4/gFW) in Prunus
persica var. compressa (Loud), Persica vulgaris Miller, Prunus persica var. nucipersica fruit and leaf.
(3 repeat±SD) P≤0.05. (common letters are not statistically significantly different).
2512 Journal of Research in Biology (2018) 8(5): 2511-2517
Moosavi et al., 2018

Figure 2. Relationship between total phenolic content and ferric reducing antioxidant power of the ethanol ex-
tracts in Prunus persica var. compressa (Loud), Persica vulgaris Miller, Prunus persica var. nucipersica fruit
and leaf (Mean±SD) P≤0.05.

solution, 2 mL of TBA 67% was added to 1 mL of
extract and 1.9 mL of distilled water. Mixtures were
kept in a boiling water bath for 10 min and cooled to
room temperature for 20 min, and was centrifuged in the
3000 g. The supernatant was collected and absorbance
was read at 532 nm. Anti-oxidative activity was
calculated according to the following formula:
μgMDA / gFW = (absorption / 155) × 1000 × dilution
Statistical analysis
One way ANOVA was used for the statistical
analysis to find out the significance of results. Ms-Excel
was draw graphs and the SPSS version 16 was used in
perform statistical analysis. All experiments were
repeated for three consecutive times.

Figure 3. Relationship between total phenolic content and ferric reducing antioxidant power of methanol ex-
tracts in Prunus persica var. compressa (Loud), Persica vulgaris Miller, Prunus persica var. nucipersica fruit
and leaf (Mean±SD) P≤0.05.

Journal of Research in Biology (2018) 8(5): 2511-2517 2513

 Moosavi et al., 2018

RESULTS AND DISCUSSION reducing antioxidant power in both ethanol and

Figure 1 points out the ferric reducing
antioxidant power of the ethanol and methanol extracts
of Prunus persica var. compressa (Loud), Persica
vulgaris Miller and Prunus persica var. nucipersica. In
statistical analysis of the ferric reducing antioxidant
power, there is no significant difference between the
treatments in terms of ferric reducing antioxidant power
and One Way ANOVA was used for the analysis of
variance. In total, analysis in terms of ferric reducing
antioxidant power, the largest amounts of ferric
reducing antioxidant power was found in the ethanolic
extract of Persica vulgaris Miller leaves
(2.8030±0.0147 mg FeSO4/gFW) and the lowest
amounts at the ethanolic extract of Persica vulgaris
Miller fruit (1.654±0.0557 mg FeSO4/gFW) were
observed. Between ethanol and methanol there existed
statistically significant difference and in this test, the
best solution in terms of ferric reducing antioxidant
power was ethanol than methanol because of its higher
means. As can be seen from the correlation graphs, the
relationship between total phenol content and ferric
methanol extracts is direct and positive (Figure 1-2 & 1-
3). This means that existing of phenols such as
flavonoid can be responsible for reducing properties
observed in the three species.
Sumazian et al. (2010) in a study of the
vegetables of Malaysia have come to the conclusion that
the FRAP assay, along with other assays, has shown
high anti-oxidative activity. Okoro et al., 2010 have
determined anti-oxidative activities of 28 species
especially on the seed and skin of commonly used fruits
in China using FRAP assay. Results have shown that the
peel had the highest anti-oxidative activity. A study by
Pantelidis et al. (2007) showed that the anti-oxidative
capacity of phenol content, anthocyanins and ascorbic
acid in raspberry, blackberry, Redcurrant, red seedless
raisin, gooseberry and huckleberry, predicted a clear
difference the values of FRAP between 41 to 149 micro
moles ascorbic acid per gram dry weight. Espín et al.
(2000) have examined the activities of anti-oxidative
grain, buck wheat and showed the highest amount of
FRAP that was related to total phenol content. Thaipong

b
bcd bc bcd bcd bcd
bcd bcd
cd
cd d

Prunus persica var. Persica vulgaris Prunus persica var.

compressa Miller nucipersica

Figure 4. Inhibition power of lipid peroxidation of ethanolic and methanolic extracts (mgFeSO 4/gFW) in
Prunus persica var. compressa (Loud) Bean, Persica vulgaris Miller, Prunus persica var. nucipersica fruit and
leaf (Mean±SD) P≤0.05 (common letters are not statistically significantly different).
2514 Journal of Research in Biology (2018) 8(5): 2511-2517
Moosavi et al., 2018

Figure 5. Relationship between total phenolic content and Inhibition power of lipid peroxidation of ethanol
extracts in Prunus persica var. compressa (Loud), Persica vulgaris Miller, Prunus persica var. nucipersica fruit
and leaf (Mean±SD) P≤0.05.
et al. (2006) have studied the anti-oxidant activity by flavonoid content. It means, that the cultivars with high
several measures, that in the meantime, FRAP assay, content of phenols and flavonoids high-power, have
showed direct and significant correlation with the high ferric reducing antioxidant power too, which was
phenolic content of the extracts. Barreira et al., 2008 coordinated with the results of the present work.
have studied anti-oxidative activity of methanol extracts Figure 4 points out the inhibition power of lipid
of almonds with 10 local and commercial varieties. peroxidation of ethanol and methanol extracts of Prunus
Results have shown that, there was the positive persica var. compressa (Loud), Persica vulgaris Miller
correlation between reducing power with phenolic and and Prunus persica var. nucipersica. In statistical

Figure 6. Relationship between total phenolic content and inhibition power of lipid peroxidation of methanol
extracts in Prunus persica var. compressa (Loud), Persica vulgaris Miller, Prunus persica var. nucipersica fruit
and leaf (Mean±SD) P≤0.05.

Journal of Research in Biology (2018) 8(5): 2511-2517 2515


analysis of the inhibition power of lipid peroxidation,
there is no significant difference between treatments in
terms of the inhibition power of lipid peroxidation and
therefore one way ANOVA method was used for the
analysis of variance. In total, in terms of the inhibition
power of lipid peroxidation, the largest amounts of the
inhibition of lipid peroxidation in the ethanolic extract
of Prunus persica var. compressa fruit (18.22± 047.0
µgMDA/gFW) and the lowest amounts in methanolic
extract of Persica vulgaris Miller fruit (7.479± 140.0
µgMDA/gFW) were observed. Between ethanol and
methanol statistically significant difference existed and
in this test, the best solution in terms of inhibition power
of lipid peroxidation was showed by ethanol than the
methanol because of its higher means. As can be seen
from the correlation graphs, the relationship between the
inhibition power of lipid peroxidation in ethanol
extracts is negative and reverse correlation in methanol
extracts is a direct and positive (Figure 1-5 & 1-6). The
direct correlation suggested that the cultivars who have
more phenolic content than others, can effectively
intercept lipid peroxidation in vivo.
Lipid peroxidation is one of the important
effects of the accumulation of Reactive Oxygen Species
(ROS), which will eventually lead to burnout biological
systems. The process can be initiated by free radicals, a
hydrogen atom from the methylene group of the fatty
acid chains with a few places of polyunsaturation. This
reaction is combined with the re-arrangement of double
bonds. Then the lipid radical to convert oxygen to proxy
species. Drugs whose formulations based on anti-
oxidative properties were usually used for the
prevention and treatment of diseases such as
atherosclerosis, diabetes, alzheimer and cancers. One of
the anti-oxidative activity assays showed the inhibition
of lipid peroxidation in which the main material used is
TBA (Marja et al., 1999).
The production of polyunsaturated fat oxidation
by the TBA caused a color complex. The results showed
2516
Moosavi et al., 2018
that two molecules of TBA reacts with a molecule of
MDA. Of course it should be noted that
malondialdehyde is not observed in all oxidized
systems. However, the reaction product with TBA, is
formed from the fatty acids that are with three or more
double bond (Fennema, 1996).
CONCLUSION
Rosacea family is a rich source of phenolic
compounds. Prunus persica var. compressa (Loud),
Persica vulgaris Miller and Prunus persica var.
nucipersica can be considered more in human nutrition
and also in the medical use. The collection of radicals,
in addition of phenolic compounds, can be regarding to
plant chemicals such as alkaloids, ascorbic acid, etc.,
and also the inhibition power of various extracts
significantly depends on the number and position of
hydroxyl groups and the molecular weight of phenolic
compounds. Thus, with knowing amounts of phenolic
compounds in the extracts of different plants.
ACKNOWLEDGEMENT
The authors would like to thank Dr. Siavash
Hosseini Sarghein and Dr. Rashid Jamei for
proofreading the manuscript and their guidance and
advice.
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