J.Jenshi roobha et al.

/ Journal of Pharmacy Research 2011,4(5),1488-1492

Research Article Available online through
ISSN: 0974-6943 http://jprsolutions.info
Antioxidant analysis of anthocyanin extracted from Musa acuminata bract
J.Jenshi roobha, M.Saravanakumar, K.M.Aravinthan and P.Suganya devi *
P.G. Department of Biotechnology,Dr. Mahalingam Centre for Research and Development,N.G.M.College, Pollachi.
Received on: 08-02-2011; Revised on: 16-03-2011; Accepted on:18-04-2011

ABSTRACT
Banana (Musa acuminata) bracts, abundant edible residues of banana production, were investigated as a potential source of natural colorant. Anthocyanins were extracted from Musa
acuminata bracts and their contents were found to be 14mg/100gm and their antioxidant property was also investigated. Anthocyanins from Musa acuminata bract exhibited free-
radicalscavenging activity against DPPH radical, superoxide anions, hydroxyl radical, metal chelating and hydrogen peroxide radical in a dose dependent manner The degradation of
deoxyribose by hydroxyl radicals were shown to be inhibited by anthocyanins acting mainly as chelators of iron ions rather than directly scavenging hydroxyl radicals. Anthocyanins
were also found to have excellent reducing power. The reducing power of anthocyanins, ascorbic acid and butylated hydroxyl toluene all at 100mg/ml were 0.138, 0.577 and 0.455,
respectively, indicating that anthocyanin from Musa acuminata bract had a strong electron donating capacity. It was therefore suggested that anthocyanins could be beneficial in
scavenging free radicals and our findings revealed that Musa acuminata bract have potential as good sources of natural antioxidant/nutraceutical compounds.

Key words: Musa acuminata bract, anthocyanin, total phenol, flavanoid content, antioxidants activity, free radicals scavenging effect.

INTRODUCTION
Banana is an important fruit crop of the world. They form an integral component of the farming
system in humid agro-ecolological zones of the tropics. Bananas are produced for local
consumption and trading mostly in the tropical and subtropical regions of the world. Banana
belongs to the plant family Musaceae. Musa is the largest genus in the family and was classified
into five sections based on the chromosome number and morphological characters. In many
southerneast Asian locations male buds of wild and cultivated bananas are consumed as
vegetables and bract color is a potential source of food colorants (Kasipong 2008)[1].
Most bananas have the bract colour of red, purple, or violet bract while cyanic green or yellow
are rare. In Musa acuminata the variation in bract colour is correlated with composition of
pigment mixtures. Preliminary results by paper chromatography and by acid extraction indi-
cated the glycoconjugated anthocyanin are the major pigments in bract colour. Interest in
anthocyanins has increased significantly due to their bright attractive colours, water solubility
that facilitates incorporation into aqueous systems and beneficial health effects (Timberlake &
Henry, 1988[2]; Lauro, 1991[3]; Henry, 1996[4]; Cao, Sofic and Prior, 1997[5]; Wang, Cao
and Prior, 1997[6]). New sources of anthocyanins with high tinctorial power, stability and low
cost are desired as natural food colourants (Francis, 1982[7]; Henry, 1996)[8].
Anthocyanin from edible fruits were effective antioxidants in vitro (Einbond, Reynertson, Luo,
Basile, Kennelly, 2004)[9]. Oxidation is essential to many living organisms for the production
of energy necessary for biological processes. Oxygen – centered free radicals, also known as
reactive oxygen species (ROS), including superoxide, hydrogen peroxide, hydroxyl (HO-),
peroxyl (ROO-) and alkoxyl (RO-), are produced in vivo during oxidation(Bloknina, Virolainen
and Fagerstedt, 2003)[10]. ROS are not only strongly associated with lipid peroxidation,
leading to food deterioration, but are also involved in development of a variety of diseases,
including cellular aging, mutagenesis, carcinogenesis, coronary heart disease, diabetes, and
neurodegenaration (Halliwell and Gutteridge, 1999[11]; Moskovitz, Yim and Choke 2002[12]).
Although almost all organisms possess antioxidant defence and repair systems to protect
against oxidative damage, these systems are insufficient to prevent the damage entirely (Simic,
1988)[13].
Antioxidative properties of anthocyanins arise from their high reactivity as hydrogen or
electron donors, and from the ability of the polyphenol-derived radicals to stabilize and
delocalize the unpaired electron, and from their ability to chelate transition metal ions termina-
tion of the (Rice-Evans, miller, and Panganga, 1997)[14]. Thus, anthocyanins may play a role
in to antioxidant ability. So an attempt was made to extract anthocyanin and to analyses its
antioxidant potential from Musa acuminata bract.
2. OBJECTIVE
The main aim of the paper is to extract and evaluate anthocyanin content from Musa acuminata
bracts and to evaluate in vitro antioxidant property of Musa acuminate bracts extracts using
DPPH,hydroxyl and superoxide scavenging, reducing power, hydrogen peroxide scavenging,
metal chelating, anti-ferric chloride hydrogen peroxide system, and deoxyribose degradation.
3. MATERIALS AND METHODS
3.1 Sample collection: Musa acuminata bract was collected from the field of vellore in
Udmalpet.
*Corresponding author.
P.Suganya devi,
Assistant Professor,
P.G. Department of Biotechnology,
Dr.Mahalingam Centre for Research and Development,
N.G.M.College, Pollachi.
3.2 Extraction: 0.5 gm of Musa acuminata bract were treated with 10 ml acidified methanol.
And the mixture was centrifuged at 10,000 rpm for 10 min and supernatant was taken for
analysis (Lachman et.,al 2003)[15]
3.3Analytical procedures
3.3.1 Flavanoids conformation test (Harbone-1998)[16]
A. Fecl3:1 ml of sample extraction was added with a small amount of FeCl3, and results were
observed.
B. Alcl3: 1 ml of sample extraction was added with 5% of AlCl3 solution, and results were
observed.
3.3.2 Total phenolic assay
Total phenolic compounds in anthocyanin samples were quantified by using Foliciocalteu’s
method described by Ronald et.al., (1998)[17]. 50 µl of Folin-ciocalteu’s reagent (50% v/v)
were added to 10µl of sample extract. It was incubated for 5 min. After incubation 50µl of 20
% (w/v) sodium carbonate and water was added to final volume of 400 µl. Blank was prepared
by replacing the reagent by water to correct for interfering compounds. After 30 min of
incubation, the absorbance was measured using spectrophotometer at 760 nm.
3.3.3 Stability at variable pH
The anthocyanin stability was tested by treating 1 ml of sample with 1 ml of pH 1.0 and 4.5
solutions. The color change was observed. (Strack, 1989)[18].
3.3.4 Determination of total anthocyanin
The total amount of anthocyanin content was determined by using pH differential method. A
spectrophotometer was used for the spectral measurements at 210 nm and 750 nm.(Fuleki &
Francis,1968)[19]. The absorbance of the samples (A) was calculated as follows:
A= (Absorbance λ vis-maxA 250-A750) pH 1.0-(Absorbance λ vis-max A250-A750) pH
4.5
Anthocyanin pigment content (mg/liter) = (A X MW X DF X 1000)/ (e X 1).
Where,
Molecular weight of anthocyanin (cyd-3-glu) = 449, Extraction coefficient (e) = 29,600, DF
=Diluted factor.
3.3.5 Total flavonoid content
The flavonoid content was determined according as the aluminum chloride colorimetric
method described by Chang, Yang and Chern (2002)[20]. Briefly, aliquots of 0.1g of Musa
acuminata bract sample was dissolved in 1 ml of deionized water. This solution (0.5 ml) was
mixed with 1.5 ml of 95% alcohol, 0.1 ml of 10 % aluminium chloride hexahydrate (AlCl3),
0.1 ml of 1 M potassium acetate (CH3COOK), and 2.8 ml of deionized water. After incubation
at room temperature for 40 min, the reaction mixture absorbance was measured at 415 nm
against a deionized water blank on a spectrophotometer. Quercetin was used as a standard.
Using a seven point standard curve (0-50mg/l), the levels of total flavonoid contents in Musa
acuminata bract was determined in triplicate, respectively. The data was expressed as milli-
gram quercetin equivalents (QE)/100 g fresh matter from fresh Musa acuminata bract.
3.4 ANTIOXIDANT ASSAYS
3.4.1. Scavenging activity of DPPH radical
Scavenging activity of Anthocycanins against DPPH radicals was assessed according to the
method of Larrauri, Sanchez-Moreno, and Saura-Calixto (1998) [21] with some modifications.
Briefly, 0.1 mM DPPH-methanol solution was mixed with 1 ml of 0.1mM DPPH methanol
solution. After the solution was incubated for 30 min at 25º C in dark, the decrease in the
absorbance at 517nm was measured. Control contained methanol instead of antioxidant
solution while blanks contained methanol instead of DPPH solution in the experiment.

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Ascorbic acid and BHT were used as positive controls. The inhibition of DPPH radicals by the
samples was calculated according to the following equation:
DPPH-scavenging activity (%) = [1-(absorbance of the sample-absorbance of blank)/
absorbance of the control] ×100
3.4.2. Hydroxyl radical scavenging activity
The hydroxyl radical scavenging activity was determined according to the methods described
by Singh, Murthy and Jayaprakash (2002)[22]. 0.1 ml of Musa acuminata bract anthocyanin
evaporated samples in the concentration of 1mg/ml, 10mg/ml, 50mg/ml, and 100mg/ml was
taken in different test tubes. 1.0 ml of iron-EDTA solution (0.1% ferrous ammonium sulfate
and0.26% EDTA), 0.5 ml of DMSO (0.85% v/v in 0.1 M Phosphate buffer, pH 7.4) were
added to these tubes, and the reaction was initiated by adding 0.5 ml 0f 0.22% ascorbic acid.
Test tubes were capped tightly and heated on a water bath at 80-90 0c for 15 min. The reaction
was terminated by the addition of 1 ml of ice cold TCA (17.5 %w/v).3 ml of Nash reagent (75
g of ammonium acetate, 3 ml of glacial acetic acid, and 2 ml of acetyl acetone were mixed and
raised to 1 L with distilled water) was added to all of the tubes and left at room temperature for
15 min for the color development. The intensity of the yellow color formed was measured
spectrophotometrically at 412 nm against the reagent blank. The percentage of hydroxyl
radical scavenging activity is calculated by using the formula:
% of hydroxyl radical scavenging activity=1-absorbance of sample/absorbance of
blank×100
3.4.3 Determination of superoxide radical-Scavenging activity
Superoxide radicals were generated by the method of Ginnopolites and Ries (1977)[23],
described by Siddhurajuna, Mohab and Beckera (2000)[24], with some modifications all
solutions were prepared in 0.05 M phosphate buffer (pH 7.8). The photo induced reactions
were performed in aluminium foil-lined box with two 30W fluorescent lamps. The distance
between the reaction solution and the lamp was adjusted until the intensity of illumination
reached about 4000 lux. A 30µL aliquot of Musa acuminata bract anthocyanin evaporated
samples in the concentration of 1mg/ml, 10mg/ml, 50mg/ml, and 100mg/ml was mixed with
3ml of reaction buffer solution (1.3 mm riboflavin, 13 mM methionine, 63 µM nitro blue
tetrazolium and 100µM EDTA, pH 7.8). The reaction solution was illuminated for 15 min at
25 º C. The reaction mixture, without sample, was used as a control. The scavenging activity
was calculated as follows:
Scavenging activity (%) = (1-absorbance of the sample/absorbance) ×100
3.4.4 Metal chelating activity
The chelation of ferrous ions by the extract was estimated by the method of Dinis et.,al.(1994)[25]
with slight modification and compared with that EDTA, BHT and that of ascorbic acid. The
chelation test initially includes the addition of ferrous chloride. The antioxidants present in the
samples chelates the ferrous ions from the ferrous chloride. The remaining ferrous combine
with ferrozine to form ferrous-ferrozine complex. The intensity of the ferrous-ferrozin complex
formation depends on the chelating capacity of the sample and the colour formation was
measured at 562 nm (Shimadzu UV-Vis 2450).
Different concentrations of standard and (100-500 µg/ml) of Musa acuminata bract anthocya-
nin evaporated samples in the concentration of 1mg/ml, 10mg/ml, 50mg/ml, and 100mg/ml
were added to a solution of 100 µl FeCl2 (1mM). The reaction was initiated by the addition
of 250 µl ferrozine (1 mM ). The mixture was finally quantified to 1.3 ml with methanol,
shaken vigorously and left standing at room temperature for 10 min. after the mixture had
reached equilibrium, the absorbance of the solution was measured spectrophotometrically. All
the test and analysis were done in duplicate and average values were taken. The percentage
inhibition of ferrous-ferrozine complex formation was calculated using the formula;
%= 1-As/Ac X 100. Where, ‘Ac’ is the absorbance of the control, ‘As’ is the absorbance of the
sample.
3.4.5. Determination of reducing power
The reducing power was determined according to the method of Oyaizu (1986)[26]. A 0.25ml
aliquot of Musa acuminata bract anthocyanin evaporated samples in the concentration of 1mg/
ml, 10mgml, 50mg/ml, and 100mg/ml was mixed with 2.5 ml of 200mM sodium phosphate
buffer (pH 6.6) and 2.5 ml of 1% potassium ferricyanide. The mixture was then incubated at 50
ºC for 20 min. After 2.5 ml of 10% trichloroacetic acid (w/v) were added, the mixture was
centrifuged at 650g for 10 min. A 5ml aliquot of the upper layer was mixed with 5ml of distilled
water and 1ml of 0.1% ferric chloride at 700nm was measured. A higher absorbance a higher
reducing power.
ml), 1ml of thiobarbituric acid (TBA)(1%), and 1ml of trichloro acetic acid (TCA)(10%) were
added to the mixture, which was heated for 30 min in a boiling water both. After cooling, 5ml
of chloroform was added, and the mixture was centrifuged at 1000 x g to give a supernatant.
Absorbance of the supernatant was measured using spectrophotometer at 532 nm.
4. Molecular antioxidant analysis
4.1 Determination of inhibitory effect on deoxyribose degradation
Inhibitory effect of the anthocyanins on deoxyribose degradation was determined by measuring
the reaction activity between either antioxidants or hydroxyl radicals (referred to as non-site-
specific scavenging assay) or antioxidants and iron ions (referred to as site-specific scavenging
assay) described by Lee et. al.,(2002)[29]. For non-site-specific scavenging assay, a 0.1 ml
aliquot of different concentration of anthocyanin was mixed with 1ml of reaction buffer (100 µM
FeCl3, 104 µM EDTA, 1.5mM H 2O2, 2.5mM deoxyribose, and 100µM L-Ascorbic acid, pH
7.4) and incubated for 1 h at 370 C. A 1ml aliquot of 0.5% 2-thiobarbituric acid in 0.025 M
NaOH and 1 ml of 2.8% trichloroacetic acid were added to the mixture and it was heated for 30
min at 800 C. The mixture was cooled on ice and the absorbance was measured at 532nm. Site-
specific scavenging activity, which represented the ability of anthocyanins to chelate iron ions
and interfere with hydroxyl radical generation, was measued using the same reaction buffer
without EDTA. Percent inhibition of deoxyribose degradation was calculated as (1-absobance
of sample/absorbance of control) ×100. Control-without sample.
5. RESULTS AND DISCUSSION
5.1 ANTHOCYANIN EXTRACTION
The total anthocyanin was extracted by using acidified methanol as solvent system. Acidified
methanol resulted significantly higher values of total anthocyanin (table 1). Alexandra et.al,
2001[30] extracted anthocyanin from Musa paradisiaca bracts extracted by using acidified
methanol as solvent systems.
Table 1:Phenolic composition, flavonoid and anthocyanin content of Musa
accuminata bract in acidified methanol solvent extraction:
Musa acuminata Total phenols (mg Total flavanoid (mg Anthocyanin (mg
bract solvent gallic acid Equ/g) queracitin Equ/g) glucoside Equ/g)
1.Acidified Methanol 286±0.30 342±0.2 14.4±0.01
Values are mean (n=3)± SD (n=3, p<0.05)
5.2 CONFIRMATORY TEST FOR ANTHOCYANIN:
The anthocyanins extracted from acidified methanol were confirmed by ferric chloride test,
aluminium chloride test and also by testing the extracts with 1M hydrochloric acid and 1M
sodium hydroxide.
5.3 EVALUATION OF TOTAL FLAVONOID:
The total flavonoid content was evaluated to be 342 mg/g (table 1). A positive correlation was
observed between total phenol and flavonols suggest that total phenols are contributed mostly
by flavonols in bracts of Musa acuminata (table 1). Similar results were reported by Linda
dykes et.al., 2005[31].
5.4 DETERMINATION OF TOTAL PHENOLIC CONTENT:
The total phenolic content was observed is 2.869 mg/g in acidified methanol extracts. Awika
et.al., 2005[32] reported the highest concentration of phenols in sorghum bran by using
acidified methanol as solvent. He also reported that the acidified methanol extracts can preserve
a wide range of compounds mainly phenolic compounds. Phenolic compounds are commonly
found in both edible and non edible plants. The phenolic content and composition in plants
are depended on genetic and environmental factors, as well as post – harvest processing and
storage conditions (Majer et.al., 1979[33]; Loomis et.al., 1996[34]). Additionally, plants vary
in content and structure of phenolic compounds such as number of phenolic rings, aromatic
substitution, glycosylation and conjugation with other phenolic compounds or organic acid
will vary in their antioxidant properties and also reported the presence of phenols in Musa
species (Noradlin et.al., 2008[35]).
Total flavonoid and Total phenolic content

3.4.6.Hydrogen Peroxide Scavenging Activity

0 .35
A solution of hydrogen peroxide (40mM) was prepared in phosphate buffer (pH 7.4). 0.2 ml
0 .34
Concentration in mg/g

of various concentration of Musa acuminata bract anthocycanins (1, 10, 50, 100mg/ml) in
1.6ml phosphate buffer (pH 7.4) was added to 0.6ml of 40mM hydrogen peroxide solution. 0 .33
The absorbance value of the reaction mixture was recorded at 230 nm.(Lcin 2005)[27]. The 0 .32
percentage of hydrogen peroxide scavenging of Musa acuminata bract anthocyanin was 0 .31 Total fla vo vonoid
calculated using the following formula: 0.3
0 .29 Total phenolic
Hydrogen peroxide scavenging activity % =[( absorbance of the sample- absorbance of
the control)/ absorbance of the control]*100. 0 .28
0 .27
0 .26
3.4.7. Estimation of anti-FeCl2-H2O2 stimulated linoleic acid peroxidation 0 .25
The effect of anti -FeCl2-H2O2 stimulated linoleic acid peroxidation was determined by the Acidified m e thanol
method as described by Duh (1998)[28]. In brief, 0.2 ml of various concentration of Musa
acuminata bract anthocycanins (1, 10, 50, 100mg/ml) were added to a solution of 0.1 M
linoleic acid (0.2 ml), 2.0mM FeCl2. H2O (0.2ml) and 0.2M phosphate buffer (pH 7.4, 5 ml).
The reaction mixture was incubated at 370 C for 24 h. After incubation, 0.2ml of BHA(20mg/ Figure1.Phenolic composition, flavonoid content of Musa accuminata bract
in acidified methanol solvent extraction
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5.5.STABILITY AT VARIABLE PH
The sample appears red color at pH 1 and color disappeared at pH 4.5, Giusti, 2003[36] DPPH and Hydroxyl radical assay
reported that, the anthocyanins are stable in low pH. The results were found to be same in
the extracts of methanol and acidified methanol. 120
100
5.6 ANTHOCYANIN CONTENT: Standard for Hydroxyl

Percentage
The total anthocyanin content extracted by acidified methanol was found to be 14.4 mg/ 80
Standard for DPPH
g of bract. Several authors reported that aqueous acetone was better than various alcoholic 60
solvents for fruit procyanidins, anthocyanins and other phenols (Garcia viguera et.al., DPPH
40
1998[37]; Kallithraka et.al., 1995[38]). However acidified methanol preserves that ex- Hydroxyl radical scavenging
tracted anthocyanins in their original form better, it should be solvent choice for quanti- 20
fication and analysis of anthocyanins. Kasipong (2008) [1] reported the presence of 0
anthocyanin in M.acuminata sub species with purple, red purple, blue bract color; they BHT Ascorbic 1mg 10mg 50mg 100mg
also reported the absence of anthocyanin in M.acuminata yellow bract color group because acid
they might have problem in anthocyanin synthesis pathway probably the step of turning
leucoanthocyanidin to anthocyanidin (Grotewald, 2006[39). Concentration in mg/ml

Total anthocyanin
Concentration in mg/100g

Figure 3. Antioxidant activity of Musa acuminata bract

Acidified 6.2SUPEROXIDE ANION SCAVENGING ACITIVITY

methanol Superoxide anion radicals are produced by a number of cellular reactions, including various
enzyme system such as lipoxygenases, peroxidase, NADPH oxidase and xanthine oxidase.
Superoxide anion plays an important role in plant tissue and also involving in formations of
other cell damaging free radicals (Blokhina et.al., 2003) [10], In the present study superoxide
radical was generated by illuminating a solution containing riboflavim. The relative scaveng-
ing effect of Musa acuminata anthocyanin bract on superoxide radical are shown in table 2.
The anthocyanin extracts exhibided 71.92% scavenging acitivity at 100 mg per ml. The result
of superoxide radical anion scavenging acitivity of anthocyanin extract, butylated hydroxy
toluene and ascorbic acid were shown in figure 4. The superoxide anion on radical scavenging
Total anthocyanin
acitivity might be due to the action of phenolic compounds. Also the flavanoid molecules with
polyhydroxylated subsititution on the rings A and B and a free three hydroxyl substitution
Figure 2. Anthocyanin content of Musa accuminata bract in acidified confired superoxide anion scavenging acitivity according to Siddhuraju et.al., 2002[45].
methanol solvent extraction
6.ANTIOXIDANT ACTIVITY
Superoxide and Metal chelating assay
6.1 DPPH RADICAL SCAVENGING ACTIVITY
Free radical scavenging is one of the known mechanisms by which antioxidants inhibits lipid 100
peroxidation (Bloknina et.al., 2003[10]; Rice-Evan et.al., 1997[14]). The DPPH radical
80
scavenging activity has been extensively used for screening antioxidants from fruits and Standard for super oxide
Percentage

vegetable juices or extract (Robards, Penzler, Tueker, Swatistang, and Glores, 1999[40], 60 Standard for metal chelating
Scanchez- Moreno, 2002[41]). DPPH radical scavenging activity of Musa acuminata bract
40 Superoxide radical activity
anthocyanin, ascorbic acid and butylated hydroxy toluene were shown in table 2. The
anthocyanin significantly inhibited the activity of DPPH radical is an dose dependant manner. Metal chelating
20
Antioxidant activity of sample extracted in acidified methanol is shown in figure 3. At 1 mg
level the scavenging effects were 37.3%. At 10 mg/ml and 50 mg/ml the scavenging effects 0
were 60.27% and 85.18 % respectively, while almost complete inhibition of DPPH radical BHT Ascorbic 1mg 10mg 50mg 100mg
activity was observed where anthocyanin were used at 100mg/ml. It shows that Musa acid
acuminata bract anthocyanin have strong hydrogen donating capacity and can effectively Concentration in mg/ml
scavenge DPPH radicals. Matook et.al., 2005[42] reported the antioxidant effects of crude
extracts from green banana and yellow peel were investigated and the results indicated that the
extract of green peel recorded high antioxidant activity measured by DPPH free radical
scavenging system. Figure 4 . Antioxidant activity of Musa acuminata bract
6.4 METAL CHELATING ACTIVITY:
Table.2 . Antioxidant assay from Musa acuminata evaporated samples Ferrozine can form complexes with Fe2+ but in the presence of chelating agent, the complex
Antioxidant assays Concentration(mg/ml) formation is disrupted with the result that the red color at the complex is decreased.
1mg 10mg 50mg 100mg Measurment of color reduction therefore allows estimation of chelating activity of the co-
1.DPPH assay 37.3% 60.27% 85.18% 99.89% existing chealtor (Yamaguchi et.al., 2000) [46]. In this assay the anthocyanins extracts
2.Hydroxyl Scavenging activity 35% 65% 91.3% 97.3% interfere with the formation of ferrous and ferrozine complex, suggesting they have chelating
3.Superoxide radical activity 10.06% 49.6% 56.42% 71.92% activity and can capture ferrous ion before ferrozine. Anthocyanins extracts registered the
4.Reducing power 0.064 0.449 0.454 0.455
5.Deoxyribose degradation(site specific) 23.7% 42.2% 78.8% 86.8%
highest metal chelating activity at 100 mg/ml which are comparably higher to the positive
6.Deoxyribose degradation(non site spesific) 19.4% 63.94% 78.67% 85.14% standards ascorbic acid and butylated hydroxyl toluene respectively (table 2). Chanda et.al.,
7.Metal chelating 83.21% 86.2% 87.5% 87.8% (2009)[47] reported the metal chelating activity in some medicinal plants for evaluating their
8.H2O2 scavenging 31.5% 43.5% 65.58% 85.74%
9.Anti-FeCl3 50.84% 70.64% 87.9% 88.94%
antioxidant activity.

6.5 REDUCING POWER:

6.2 HYDROXYL RADICAL SCAVENGING ACTIVITY
The hydroxyl radical is an extremely reactive free radical formed in biological system and has
been implicated as a highly damaging species in free radical pathology capable of damaging
almost every molecule found in living cells. This species is considered to one of the quick
initiater of the lipid peroxidation process, abstracting hydrogen atoms from unsaturated fatty
acids (Kappus, 1991[43]). Hydroxyl radical scavenging activity of anthocyanin extracts which
increases with increasing concentration (Figure 3). The extractrs exhibited the highest activity
of 97.3% at 100mg/ml where is 35% inhibition was noted at 1mg/ml respectively.This is
similar to observations of several others who have reported as dose dependent activity in
sesame coat, pomegranate peel and seeds and grape pomoce (Chang et.al., 2002[20]; Murthy,
Singh and Jayaprakash 2002[44]; Singh et.al., 2002[22]).
It has been reported that reducing power is associated with antioxidant activity and may serve
as significant reflection on the antioxidant activity (Chang, Yen, Hung, and Duh, 2002[20];
Yen and Duh, 1993[48]). Anthocyanins from Musa acuminata bract exhibited a higher
reducing power than butylated hydroxyl toluene and ascorbic acid, suggesting that strong
electron-donating capacity (table 2). The reducing power of Musa acuminata bract anthocyannin
at 1 mg/ml, 10 mg/ml, 50 mg/ml, and 100 mg/ml were 0.064, 0.449, 0.454, 0.455
respectively. The reducing properties are generally associated with the presence of reductones
(Pin-per Duh, 1998) [49], which have been shown to exert antioxidant action by breaking the
free radical chain by denoting a hydrogen atom (Gordon,1990)[50]. The reducing power of
Musa acuminata bract is probably due to the presence of phenolic compounds which might
act as electron-donor.

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R e d u c ing p o w e r Deoxyribose degradation

0.7 120
0.6 100 standard for site spesific

percentage
Absorbance

0.5 80
S tandard standard for non site spesific
0.4 60
0.3 Reducing power 40 non-site spesific
0.2 20 site spesific
0.1
0
0

mg

mg

g
BH
T

1m
g

mg
mg

0m
id
id

1m
BH

0m

10

50
ac
ac

50
10

10
10

c
ic

rbi
rb

co
co

As
As

C o n c e ntra ti o n m g / m l concentration in mg/ml

Figure 5. Antioxidant activity of Musa acuminata bract
6.6 HYDROGEN PEROXIDE SCAVENGING ACTIVITY
Hydrogen peroxide scavenging activities of the anthocyanin extracts, BHT and ascorbic acid
was measured at 230nm. Hydrogen peroxide scavenging activities of the anthocyanin extracted
by using acidified methanol were 31.5% at 1 mg/ml, 43.5% at 10 mg/ml, 65.58% at 50 mg/
ml and 85.74% at 100 mg/ml respectively. BHT(94.6%) had higher hydrogen peroxide
scavenging activity than anthocyanin extracts (Table 2).
6.7 ESTIMATION OF ANTI-FECL 2 -H 2O 2 STIMULATED LINOLEIC ACID
PEROXIDATION
Iron salts are thought to react with H2O2 called the Fenton reaction, to make hydrogen radicals
which bring about peroxide reaction of lipids (Duh, 1998)[28]. The effect of anthocyanin
extracts of Musa acuminata bract on the formation of malonaldehyde (MDA) from linoleic acid
is show in table 2. As the concentration of the antioxidant extracts increased the formation of
MDA decreased. A dose dependent MDA inhibition in linoleic acid oxidation was evident.
Acidified methanol extraction showed a higher inhibition of MDA ranging from 50% to
88.94% at 1 mg/ml to 100 mg/ml than BHT and ascorbic acid (Figure 6).
Hydrogen peroxide and anti ferric chloride hydrogen perooxide
assay
100
Percentage
Figure -7 Antioxidant activity of Musa acuminata bract
8. CONCLUSION
From the results it can be concluded that Musa acuminata has high amount of anthocyanins
content on its bract. Reactive oxygen species plays a crucial role in a wide range of common
diseases and age related degenerative conditions including cardiovascular diseases, inflamma-
tory conditions and neuro degenerative diseases such as Alzheimer’s disease, mutations and
cancer. So antioxidant capacity is widely used as a parameter to characterize food or medicine
plant and their bio-active components. In this study, the antioxidant activity of the anthocya-
nins extracted from Musa acuminata bract was evaluated and it showed very strong antioxi-
dant activity.
Thus these results suggest that anthocyanin extract from Musa acuminata bract can be used as
antioxidant material, food addictives.
9. AKNOWLEDGEMENT
The authors are grateful to Principal Dr.Raj Kumar, Head of the department R.Kavitha
Krishna, Nallamuthu Gounder Mahalingam College, and Pollachi, Tamil nadu, for permitting
this research work, they also thankful to Research scholars Kathiresh, Rakkimuthi and
Vinodha of Biotechnology for their kind co-operation for complete this research work.
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7. MOLECULARS ASSAYS 10.
7.1 INHIBITORY EFFECTS OF DEOXYRIBOSE DEGRADATION
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