ISSN : 0974 - 7435 Volume 11 Issue 9

BioTechnology
An Indian Journal
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BTAIJ, 11(9), 2015 [321-327]

Screening of different banana peels extracts for antioxidant ca-

pacity and total phenols
P.Sathish Kumar
S.T.Joseph’s College of Engineering, Sholinganallur, 600119, Chennai, (INDIA)

ABSTRACT KEYWORDS
The total phenolic content and related total antioxidant capacity of 4 types FRAP;
of banana peels were analyzed. Banana (Musa acuminata Colla AAA) peel DPPH;
extracts obtained in this work had a high capacity to scavenge 2,2-diphenyl- ABTS;
1-picrylhydrazyl (DPPH), 2,22 -azino-bis (3-ethylbenzothiazoline)-6-sulfonic NBR.
acid (ABTS+) free radicals NBR (Nitro blue tetrazolium), and they were
also good lipid peroxidation inhibitorsAcetone:water extracts were
considerably more effective (compared with methanol, ethanol, acetone,
water, methanol:water or ethanol:water) at inhibiting the peroxidation of
lipids in the â-carotene/linoleic acid system or scavenging free radicals.
However, aqueous extracts had a high capacity to protect lipids from
oxidation in the thiobarbituric acid reactive substances (TBARS) test, as
well as in the â-carotene bleaching assay. To make practical comparison of
relative antioxidant potential of phenolics extracted from selected banana
peels, the phenol antioxidant coefficient (PAC) was calculated for each
infusion. In addition, acetone:water most efficiently extracted all extractable
components (54 ± 4%), phenolic compounds (3.3 ± 0.8%), and anthocyanin
compounds (434 ± 97 ìg cyanidin 3-glucoside equivalents/100 g freeze-
dried banana peel). Banana peel contained large amounts of dopamine and
L-dopa, catecholamines with a significant antioxidant activity. However,
ascorbic acid, tocopherols or phytosterols were not detected in the different
extracts. The antioxidant activity of banana peel extracts from different
cultivars was similar. However, the impact of extraction time or temperature
should be studied in greater depth.  2015 Trade Science Inc. - INDIA

INTRODUCTION ments to be beneficial in preventing diseases. Examples

of antioxidants include vitamins C and E, selenium, and
Antioxidants are man-made or natural substances carotenoids, such as beta-carotene, lycopene, lutein,
that may prevent or delay some types of cell damage. and zeaxanthin. This fact sheet provides basic informa-
Diets high in vegetables and fruits, which are good tion about antioxidants, summarizes what the science
sources of antioxidants, have been found to be healthy; says about antioxidants and health, and suggests sources
however, research has not shown antioxidant supple- for additional information. The oxidative stress, defined
322 Screening of different banana peels extracts for antioxidant capacity BTAIJ, 11(9) 2015

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as ‘‘the imbalance between oxidants and antioxidants gin and the ability to act as efficient free radical scaven-
in favor of the oxidants potentially leading to damage’’ gers[16,22]. In last two decades the number of publica-
has been suggested to be the cause of aging and vari- tions on the potential health benefits of polyphenols,
ous disease in humans. In modern western medicine, has increased enormously[1,9,12,23,28,30,34,36]. Tea (black
the balance between antioxidation and oxidation is be- and greentea) is one of the most commonly consumed
lieved to be a critical concept maintaining a healthy bio- beveragesin the world and is rich in polyphenolic com-
logical system[1,8,9,10,34,36]. The similar concept of bal- pounds collectivelyknown as the tea flavonoids[16,22,25].
ance called yinyang has existed in traditional Chinese The currentfocus is toward natural antioxidants,
medicine for more than 2000 years. Ou, Huang, especiallyplant polyphenolics. It is of interest to investi-
Hampsch-Woodili, and Flanagan (2003) and Prior and gate theantioxidant properties of herbal infusions espe-
Cao (2000) have shown that the effective composi- cially those traditionally used in folk medicine. The aim
tions of the yin-tonic herbs are mainly flavonoids which of the present study was to examine the total phenolic
are phenolic compounds with strong antioxidant activ- content and related total antioxidant potentialIn four
ity. According to them the clear trend of antioxidant banana peel extract prepared in common way in which
activity supported the hypothesis that yin in traditional teas are prepared for human consumption. Total anti-
chines medicine refers to antioxidant process, whereas oxidant potential has been determined using ferric re-
yang relates to oxidation process. A general recom- ducing ability of plasma assay (FRAP) of Benzieand
mendation to the consumer is to increase the intake of Strain (1996). The efficiency of extracted phenolics was
foods rich in antioxidant compounds (e.g. polyphenols, evaluated using the phenol antioxidant coefficient (PAC).
carotenoids) due to their well-known healthy effects. The effect of infusion temperatures and infusion time
As a consequence these evidences accelerated the has been considered over a range similar to that en-
search for antioxidants principles, which led to the iden- countered in a domestic environment.
tification of natural resources and isolation of active
antioxidant molecules. Many plants have been identi- MATERIALS AND METHODS
fied as having potential antioxidant activities and their
consumption recommended [19,23,24,26,36,38,40] . Materials, chemicals and reagents
Bioactivephenols, especially bioflavonoids, are very The banana varieties belonging to Karpooravalli
interesting as antioxidants because of their natural ori- (Musa spp. - Karpooravalli - ABB) and those belong-

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BTAIJ, 11(9) 2015 P.Sathish kumar 323

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ing to Musa acuminata viz., Pach- ainadan (Musa spp. on their chemical reducing capacity relative to an equiva-
- Pachanadan - AABS), Poovan (Musa spp. - Mysore lent reducing capacity of (+) - catechin. Data presented
- AAB), Rasthali (Musa spp. - Rast-hali - AAB) were are average of four measurements.
collected locally from different farms in the same local- DPPH (1,1-diphenyl-2-picrilhydrazyl) radical-scav-
ity in Tiruchirapalli (Tamil Nadu, India).1,1-Diphenyl- enging assay
2-picrylhydrazyl (DPPH), 2,2-azobis-3-
DPPH scavenging activity was measured by the
ethylbenzthiazoline-6-sulphonic acid (ABTS), nitro blue
slightly modified spectrophotometric method of Brand-
tetrazolium (NBR), free radical, sodium dodecyl sul-
Williams et al.[10]. The absorbance of DPPH diluted in
phate (SDS) and ammonium molybdate were all of
methanol was considered as control. The decrease in
analytical grade.
absorbance was measured at 560 nm. The antioxidant
Preparation of peel extracts (10g) capacity to scavenge the DPPH radical was calculated
The peel of fresh naturally ripened yellow un-pig- by the fol-lowing equation: Scavenging effect (%): [(1-
mented bananas were shade-dried for about a week absorbance of sample/absorbance of control) × 100].
and then crushed to make a coarse powder. The dried Results were expressed as Mean ± SD of three experi-
powder (10 g) was weighed and solvent extraction us- ments made by triplicate. Free radical scavenging abil-
ing methanol was performed at a 10% concentration. ity by the use of a stable DPPH radical (1,1-diphenyl-
Exhaustive extraction was carried out in triplicates for 2-picrilhydrazyl). The DPPH radical scavenging activ-
about 36 h in a shaker at 37°C with a gentle shaking. ity of samples were determined using the method pro-
The extracts were then evaporated at room tempera- posed by Von Gadow, Joubert, and Hansmann (1997).
ture. The residues obtained were re-evaporated to re- Aliquot (50 lL) of the tested sample was placed in a
move impurities and stored at 4°C to carry out radical cuvette, and 2 mL of 6 · 10_5 M methanolic solution of
scavenging assays. The remaining residue was stored DPPH radical was added. Absorbance measurements
in desiccators for further use. commenced immediately. The decrease in absorbance
at 560 nm was determined after 16 min for all samples.
Free radical scavenging assays Methanol was used to zero spectrophotometer. The
(a) Total antioxidant capacity assay absorbance of the DPPH radical without antioxidant,
Aliquots of suitable working solutions (1 - 10 mg/ i.e. the control was measured daily. Special care was
ml) of the samples were mixed with 1 ml of the reagent taken to minimize the loss of free radical activity of the
solution (0.6 M sulphuric acid, 28 mM sodium phos- DPPH radical stock solution[4]. Methanolic solutions of
phate and 4 mM ammonium molybdate) and incubated pure compounds ((+)-catechin, vitamin C and querce-
at 95°C for 90 min[8]. The tubes were cooled to room tin) were tested too at different concentrations (· mol of
temperature and the absorbance was measured at 560 antioxidant/1 mol DPPH radical). All determinations
nm against a blank. Ascorbic acid was used as a stan- were performed in triplicate. The percentage inhibition
dard. Total antioxidant capacity was expressed as of the DPPH radical by the samples was calculated
equivalents of ascorbic acid. according to the formula of Yenand Duh (1994) inhibi-
tion.
(b) Total phenol concentration
% INHIBITION = {(AC(0) -AA(t)/AC(0)}*100
Total phenol concentration in selected 4 banana peel where AC(0) is the absorbance of the control at t = 0
extract were determined spectrophotometrically ac- min;and AA(t) is the absorbance of the antioxidant a.
cording to the Folin–Ciocalteu colorimetric method[35],
NBT (nitro blue tetrazolium)(2oµg)
using (+)-catechin as the standard and expressing the
results as catechin equivalents (CE). The levels of total Superoxide radical scavenging assay was carried
phenols in infusions determined according to the Folin out according to the method of zhishen et al.(1999). All
– Ciocalteu method are not absolute measurements of solutions were prepared in 0.05M phosphate buffer
the amounts of phenolic materials but are in fact based (ph7.8). The photo induced reactions were performed

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324 Screening of different banana peels extracts for antioxidant capacity BTAIJ, 11(9) 2015

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SAMPLES OD AT 560nm
1. Karpooravalli 0.724
2. Rasthali 0.668
3. Pachainadam 0.708
4. Poovan 0.711

SAMPLES OD AT 560nm PERCENTAGE%

1. Karpooravalli 0.411 32.95
2. Rasthali 0.127 79.28
3. Pachainadam 0.182 0.182
4. Poovan 0.209 0.209

in aluminium foil lined with two 20w fluorescent lamps.

The distance between reactant and the lamp was ad-
justed until reach the 4000 lux intensity of illumination.
The total volume of the reactant mixture was 5ml and
the concentration of the riboflavin methionine and NBT
were 3*10-6,1*10-2. The reactant was illuminated at
25c for 25min. The photo chemically reduced riboflavins
generated oxygen which reduced NBT from blue
formazon. The unilluminated reaction mixture was used
as a blank and the absorbance (A) was measured at until more than 6 h had elapsed. The radical was stable
560nm. The rhizome extract were added to the reac- in this form for more than two days when stored in the
tion mixture in which oxygen was scaveged then by in- dark at room temperature. For the study of infusion
hibiting the NBT reduction absorbance was measured samples the ABTS*+ solution was diluted with Metha-
and decrease in oxygen was represented by A-A1. The nol to an absorbance of 0.70(•}0.02) at 734 nm and
degree of scavenging was calculated by the following equilibrated at 30 _C. Reagent blank reading was taken
equation, (A0). After addition of 2.0 mL of diluted ABTS*+ so-
SCAVENGING (%) =(A-A1/A)*100 lution (A734 nm = 0.700 •} 0.020) to 20 lL of anti-
ABTS (2,20azinobis-(3-ethylbenzthiazoline -6- oxidant compounds (final concentration 0–15 lM) the
sulphonic acid) absorbance reading was taken at 30 _C exactly 6 min
after initial mixing (At). Appropriate solvent blanks were
The free radical scavenging activity of extracted run in each assay. All determinations were carried out
samples was determined by ABTS radical cation de- at least three times. The percentage inhibition of absor-
colorization assay[32]. ABTS was dissolved in water to bance at 560 nmwas calculated using BELOW for-
a 7 mM concentration. ABTS radical cation (ABTS*+) mula and decrease of the absorbance between A0 and
was produced by reacting ABTS stock solution with At
2.45 mM potassium persulfate (final concentration) and % inhibition = [(Acontrol – Asample)/Acontrol] × 100%
allowing the mixture to stand in the dark at room tem-
perature for 12–16 h before use. Because ABTS and PHENOLIC TEST
potassium persulfate react stoichiometrically at a ratio
of 1:0.5, this will result in incomplete oxidation of the Total phenol concentration in selected
ABTS. Oxidation of the ABTS commenced immedi- medicinalplant infusions were determined spectropho-
ately, but the absorbance was not maximal and stable tometrically according to the Folin–Ciocalteu colori-

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An Indian Journal
BTAIJ, 11(9) 2015 P.Sathish kumar 325

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SAMPLES OD AT 560nm Total Antioxidant activity in Total Antioxidant
1. Karpooravalli 0.558 peels of banana varie-ties. Activity mM AAE
Banana Varieties g–1
2. Rasthali 0.245
Karpooravalli 3.49b ± 0.02
3. Pachainadam 0.753
Poovan 3.59b ± 0.03
4. Poovan 0.615
Pachainadan 5.85e ± 0.11
metric method[35], using (+)-catechin as thestandard and Rasthali 3.39b ± 0.09
expressing the results as catechin equivalents (CE). The
The direction and magnitude of correlation between
levels of total phenols in infusions determinedaccording
variables was done using analysis of variance (ANOVA)
to the Folin–Ciocalteu method are notabsolute mea-
and quantified by the correlation factor ‘‘r’’. The P-
surements of the amounts of phenolicmaterials but are
values less than 0.05 were considered statistically sig-
in fact based on their chemical reducingcapacity rela-
nificant.
tive to an equivalent reducing capacityof (+)-catechin.
Data presented are average of fourmeasurements. Comparison of DPPH and ABTS
Comparison of DPPH radical and ABTS radical
cation scavenging properties. Because of their high re-
activity, most free radicals react rapidly with
oxidizablesubstrates. Methods used for evaluation of
radical-trappingproperties often utilize stable model free
radicals asindicators for radical-scavenging abilities,
among which1,1-diphenyl-2-picrilhydrazyl radical
(DPPH*) and2,20azinobis-(3-ethylbenzthiazoline-6-
sulphonic acid) radical cation (ABTS*+), have gained
the highestpopularity. From the methodological point
of view the DPPH* method is recommended as easy
OD AT mg GALLIC ACID and accurate with regard to measuring the antioxidant
SAMPLES
560nm EQUIVALENT activity of fruit and vegetable juices or extracts. The
1. Karpooravalli 0.138 0.90 results are highly reproducible and comparable to other
2. Rasthali 0.920 0.23 free radical scavenging methods such as ABTS[14]. Re-
3. Pachainadam 0.660 0.132 action kinetics between phenols and ABTS* + have
4. Poovan 0.239 0.65 been found to differfrom that between phenols and
DPPH* over a similar range of concentrations. Cam-
TOTAL ANTIOXIDANT ACTIVITY pos and Lissi (1996) have suggested that this differ-
ences can be partly result of differentequilibrium dis-
Chemicals placements in reaction (1) as a result of the fact that the
reactions of DPPH* are carried out in the absence of
All chemicals and reagents were of analytical grade
added DPPH-H (i.e. the reduced form), but the re-
and were purchased from Sigma Chemical Co. (St Louis,
duced form ABTS is always present in the systems con-
MQ, USA), Aldrich Chemical Co. (Steineheim, Ger-
taining ABTS*[20]
many), Merck (Darmstadt, Germany) and Kemika
(Zagreb, Croatia).
CONCLUSION
Spectrophotometric measurements
Spectrophotometric measurements were performed In conclusion, we might say that our results further
by UV–VIS spectrophotometer 2.2. (double-beam) support the view that different banana peels extracts
Specord 200 Analytik Jena GmbH, Germany. are promising sources of natural antioxidants. Total phe-
nol contentand total antioxidant capacity differs signifi-
Statistical analysis
BioTechnology
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326 Screening of different banana peels extracts for antioxidant capacity BTAIJ, 11(9) 2015

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