Evaluation of antioxidant, antimicrobial, antitumour and ... Kaviya, Thanga Priya and Priya Department of Biochemistry, Dr. N.G.P. Arts and Science College, Coimbatore Department of

SIRJ-APBBP Volume 1 Issue 4 (2014) ISSN 2349 – 0128 www.scrutinyjournals.com

Scrutiny International Research

Journal of Agriculture, Plant

Biotechnology and Bio Products

(SIRJ-APBBP)

Evaluation of antioxidant, antimicrobial, antitumour and antinociceptive activities of Musa acuminata and Bauhinia

variegata

C.Sowmia, Kaviya, Thanga Priya and Priya

Department of Biochemistry, Dr. N.G.P. Arts and Science College, Coimbatore

Department of Clinical Laboratory Technology, Dr. N.G.P. Arts and Science College, Coimbatore, India

Article history: Submitted 18 August 2014; Accepted 18 September 2014; Published 28 October 2014

Abstract India is the richest source of medicinal plants and is rightly called the “Botanical garden of the World".

The plants Musa acuminata and Bauhinia variegate were selected and were evaluated for Antioxidant,

Antimicrobial, Antitumour and Antinociceptive activities. The phytochemical analysis test indicates the presence

of proteins, carbohydrates, alkaloids, flavanoids and phenols. Flavanoids have been demonstrated to have

antioxidant, antimicrobial, antitumour and anti nociceptive activity. The level of enzymic and non-enzymic

antioxidants, phenols, total proteins and radical scavenging activity present in the extracts of Musa acuminata,

Bauhinia variegata and their formulation were evaluated. The formulation of Musa acuminata and Bauhinia

variegata have higher antimicrobial activity towards Staphylococcus aureus, the Bauhinia variegata have higher

activity towards Escherichia coli and Klebsiella aeruginosa is highly sensitive to the Musa acuminata extract. The

chemo preventive potential of formulation of Musa acuminata and Bauhinia variegata has shown significant

reduction of the tumors. The flavanoid which are present in the extract have been reported to possess

antimutagenic and anticarcinogenic activity. The extracts of Musa acuminata, Bauhinia variegate and their

formulation exhibits antinociceptive activity.

Key words: Antioxidant, Antimicrobial, Antitumour, Antinociceptive, Musa acuminata, Bauhinia

variegate, flavanoid

Corresponding author

Dr. C. Sowmia,

Associate Professor, Department of Biochemistry,

Dr. N.G.P. Arts and Science College,

Coimbatore, Tamilnadu,

India

Introduction

Plants have formed the basis for the treatment of diseases in traditional medicine systems for thousands of years and continue to play a major role in the primary health care

www.scrutinyjournals.com Sowmia et al. / SIRJ-APBBP 1:4 (2014)

16

of about 80% of the world’s inhabitants (Agrawal et al., 2012). The compounds important in maintaining human health are referred to as “bioactive compounds”. They are flavonoids, glucosinolates, carotenoids, organosulfides, sterols, and peptides (Sonam pandey et al., 2012). Plant-derived antioxidants, especially, the phenolics have gained considerable importance due to their potential health benefits (Farooq et al., 2009). Oxidative process is one of the most important routes for producing free radicals in foods, drugs and even in living systems. Antioxidants are those substances which possess free radical chain reaction breaking properties (Prakash Veeru et al., 2009). The increasing prevalence of multidrug resistant strains of bacteria and the recent appearance of strains with reduced susceptibility to antibiotics raises the spectra of untreatable bacterial infections and adds urgency to the search for new infection fighting strategies (Sangeetha et al., 2010). Cell growth and cell multiply process is known as cell division. (Dhanamani et al., 2011). A localized proliferation of tissue forming a swelling or outgrowth, commonly with a characteristic shape and unlike any organ of the normal structure is known as tumor (Mohammed mannan et al., 2013). Cancer (malignant tumour) is an abnormal growth and proliferation of cells. It is a frightful disease because the patient suffers pain, disfigurement and loss of many physiological processes. Cancer may be uncontrollable and incurable, and may occur at any time at any age in any part of the body. It is caused by a complex, poorly understood interplay of genetic and environmental factors. Cancer kills annually about 3500 per million populations around the world (Umadevi et al., 2013). Musa species is one of the valuable plant species carries a number of advantageous pharmacological effect and comes with a set of variety (Sourav et al., 2012). Bauhinia variegata is a medium sized deciduous tree found on the rocky hills of Circars, Deccan, and Carnatic regions of South India (Sonam pandey et al., 2012). Bauhinia variegata Linn. stem is reported to have antitumour, antimicrobial, anti-inflammatory, hepatoprotective, antihyperlipidemic and immunomodulatory activities (Sharma et al., 2011).

Materials and Methods Plant material

Collection of plant materials - The flowers of Musa acuminata and Bauhinia variegata were collected from Botanical survey of India, Southern circle, Coimbatore.

Preparation of Plants Extracts - The flowers of Musa acuminata and Bauhinia variegata were properly washed in tap water and then rinsed in distilled water. The rinsed flowers were dried in shade at room temperature. The dried flowers were subjected to size reduction to a coarse powder by using mixie. Then the powder was added with ethanol and kept in a mechanical shaker for 72 hours at 3000 rpm. After that, filter the contents using Whatmann No 1 filter paper. Then the supernatant is kept in an electric water bath at 100˚C until the solution gets evaporated. The remaining powders were taken for assay by dissolving each with 2 ml of DMSO.

Preliminary phytochemical screening - The photochemical present in the sample was analyzed by Paech and Tracy method (1955).

Antioxidant activity - Rapid evaluation of antioxidant activity of the extracts was done following a reported procedure as adopted by Metha (Metha et al., 1994). The linoleic acid system antioxidant activity of the extracts was determined according to the method of Osawa and Namiki (1981). The enzymic antioxidants assayed were Superoxide dismutase, catalase and Glutathione peroxidase. (Mishra and Fridovich, 1972; Luck, 1974; Rotruk et al.,


17

1973). The non enzymic antioxidants assayed were Vitamin C, vitamin E, Reduced Glutathione and Total phenols (Roe and Keuther, 1953; Emmerie, 1938; Moron et al., 1979; Malick and Singh, 1980). FRAP ASSAY (Benzie and Strain, 1996), DPPH ASSAY (Furusawa, 2005), superoxide scavenging activity, (Liu and Chang, 1997), hydroxyl radical scavenging activity (Halliwell et al., 1981), nitric oxide radical scavenging activity (Kumar et al., 2008) The extent of inhibition of lipid peroxidation in the presence and absence of the extracts was determined using goat liver homogenate by the method proposed by Ohkawa (Ohkawa et al., 1979).

Antimicrobial activity - The antimicrobial activities of the samples were analyzed by Kirby Bauer method (1966).

Antitumor activity - The antitumour activity of the samples was analyzed by potato disc assay and carrot disc assay (Galsky et al., 1980).

Antinociceptive activity - Antinociceptive activity was tested by Acetic-Acid-Induced Abdominal Constriction Test (Zakaria et al. 2007).

Results and Discussion

Phytochemicals are non-nutritive plant chemicals that have protective or disease preventive properties. Research suggests that phytochemicals may slow the ageing process and reduce the risk of many diseases, including cancer, heart disease, stroke, high blood pressure, cataracts, osteoporosis and urinary tract infections. (Pandey et al., 2012).

From the table.1, the alcoholic extract of samples indicates the presence of alkaloids, flavanoids and phenols. Flavanoids have been demonstrated to have anti inflammatory, antiallergic and antiviral activity. These broad therapeutic properties were widely used in pharmaceuticals. Due to the presence of alkaloids, flavanoids, proteins and phenols they have been used for therapeutic purposes (Mandal and Datta, 2008).

In TLC assay, ethanolic extracts of the samples exhibited the highest intensity of resulting orange color which might be attributed to the presence of a high amount of phenolic compounds. In TLC assay, ethanolic extracts of oat, groats and hull exhibited the highest intensity of resulting orange color which might be attributed to the presence of a high amount of phenolic compounds (Farooq et al., 2006).

Extracts of various plant organs also exhibited good antioxidant activity in the linoleic acid peroxidation system. The extracts (Musa acuminata, Bauhinia variegate and their formulation) at a concentration of 0.2 mg/mL inhibited 85-93.5% peroxidation of linoleic acid after incubation of 13 days (312 h). This percentage was quite comparable to that of the values obtained for BHT (94.5%) and BHA (86%).

The present study has been carried out to evaluate the antioxidant activity of different samples. The enzymic antioxidants analysed in the present study are catalase, superoxide dismutase and glutathione peroxidase.

Units: Amount of SOD that causes 50% reduction in pyrogallol oxidation/g Amount of catalasa required to decrease the absorbance at 420 nm/g Changes of glutathione peroxidase in absorbance/minute at 430nm/g.


18

Among the enzymatic antioxidants SOD, Catalase, Glutathione peroxidase was found to be higher in activity when compared with the others. In plant antioxidant enzymes namely catalase and peroxidase have been shown to increase when subjected to stress condition (Krishnaswami et al., 2000).

The non enzymic antioxidants analysed in the present study are vitamin C, Vitamin E, reduced glutathione and total phenols.

Among the non-enzymatic antioxidants the total phenolic content was significantly higher in the plant. Phenolic compounds are polyphenols constitute one of the most numerous and widely distributed groups of substances in the plant kingdom with more phenolic extracts. The polyphenols are products of secondary metabolites of the plants. The structure of natural polyphenols varies from simple phenolic acid molecules to highly polymerized compounds, such as condensed tannins. The ability of some phenolic compounds to act as antioxidants has been demonstrated in the literature (Beyer, 1994; Aqil et al., 2006; Meydani, 2009; Soobrattee et al., 2005).

The alcoholic extract of Musa acuminata, Bauhinia variegata and combination of Musa acuminata and Bauhinia variegata were screened for their possible antioxidant activity by FRAP assay. The results reveal that the combination of Musa acuminata and Bauhinia variegata exhibit higher ferric reducing ability. The ethanolic extracts of samples were subjected to screening the possible antioxidant activity by the DPPH assay. The present study reveals that the alcoholic extract has DPPH radical scavenging activity. This reveals that the combination of Musa acuminata and Bauhinia variegata exhibit higher activity than other samples.

DPPH is widely used as a convenient method for antioxidant using for bioactive compounds isolated from plant extracts. It is also used to quantify antioxidants in complex biological system. In conclusion, the results of the present study revealed that the combination of Musa acuminata and Bauhinia variegata has higher enzymic and non enzymic activity compared to other samples.

Hydroxyl radicals have been implicated as highly damaging species in free radical pathology. This radical has the capacity to join nucleotides in DNA, cause strand breakage, which contributes the carcinogenesis, mutagenesis and cytotoxicity. Hydroxyl radical scavenging capacity of an extract is directly related to its antioxidant activity. In the present study the percentage inhibition of hydroxyl radicals by the alcoholic extracts of Musa acuminata, Bauhinia variegata and formulation of Musa acuminata and Bauhinia variegata were found to be32.5±1.25, 36.1±2.25 and 48.67±1.8 respectively. Different parts of Bauhinia variegata like leaves, bark and flowers have free radical scavenging activity by hydroxyl radical scavenging method. All extracts have different level of antioxidant activity. Methanol extracts was found to be good solvent for extraction and having good antioxidant activity. IC50 value of Bauhinia variegata leaf, stem bark and floral buds are 17.9, 19.5 and 17.2 ug/ml (pandey et al., 2012).

Superoxide is highly reactive molecule that can react with many substrates, produced in various metabolic processes, including phagocytosis. It can cause oxidation of solutes depending on their reduction potential. Both aerobic and anaerobic organism possesses superoxide dismutase enzymes which catalyze the breakdown of super oxide radical. It is well known that superoxide anion damage biomolecules directly or indirectly


19

by forming H2O2, OH, peroxyl nitrite or singlet oxygen during pathophysiologic events such as ischemic reperfusion injury. In the present study the percentage of inhibition of SOD by alcoholic extract of Musa acuminata, Bauhinia variegata and formulation of Musa acuminata and Bauhinia variegata was found to be 22.38±2.07, 39.37±3.43 and 25.38±2.97 respectively.

In the present study the percentage of inhibition of Nitric oxide radical by alcoholic extract of Musa acuminata, Bauhinia variegata and formulation of Musa acuminata and Bauhinia variegata was found to be 49.3±3.03, 46.67±3.47 and 51.7±2.96 respectively.

The table 9 shows the percent inhibition of lipid peroxidation by the Musa acuminata flower extract in liver homogenate was found to be 6.68±0.28. Bauhinia variegata flower extract showed 12.91±0.21 percentage inhibition of lipid peroxidation. The Musa acuminata and Bauhinia variegata combined sample showed a better percentage inhibition of lipid peroxidation and was found to be 22.11±0.06.The fresh extracts of samples displayed a broad spectrum of inhibition of invitro lipid peroxidation in liver homogenate. The overall results revealed that the combination of Musa acuminata and Bauhinia variegata extract showed good inhibition and exhibited a broad spectrum of inhibitory activity than the other samples.

From the Table.8 the results were noticed in the ethanolic extracts of Musa acuminata against Escherichia coli (5.0±0.5), Klebsiella aerµginosa (11.7±0.2), Salmonella typhii (4.88±0.2), Staphylococcus aureus (6.25±0.4) and Aspergillus niger (14.12±0.2). These results shows that the maximum zone of inhibition was found against Aspergillus niger (fungi) Staphylococcus aureus, Klebsiella aerµginosa (bacteria) followed by Escherichia coli and Salmonella typhii.

The results of ethanolic extracts of Bauhinia variegata against Escherichia coli (9.75±0.05), Klebsiella aerµginosa (5.98±0.4), Salmonella typhii (2.83±0.41), Staphylococcus aureus (10.25±0.4) and Aspergillus niger (11.07±0.2). These results indicate that the maximum zone of inhibition was found against Aspergillus niger, Staphylococcus aureus, Escherichia coli followed by Klebsiella aerµginosa and Salmonella typhii.

The results of ethanolic extracts of combination of Musa acuminata and Bauhinia variegata against Escherichia coli (6.78±0.38), Klebsiella aerµginosa (7.25±0.6), Salmonella typhii (2.94±0.01), Staphylococcus aureus (11.65±0.2) and Aspergillus niger (14.65±0.2). These results shows that the maximum zone of inhibition was found against Aspergillus niger, Staphylococcus aureus, Klebsiella aerµginosa followed by Escherichia coli and Salmonella typhii.

Antitumor activity

The antitumour activity was done by the method of potato disc assay which was carried out using Agrobacterium tumefaciens for tumour induction. In the present study the percentage inhibition of the development of tumour was determined and the result was shown in table 9.

Maximum inhibition was observed in alcoholic extract of combination of Musa acuminata and Bauhinia variegata (52 % and 46%) than the unique samples i.e. Musa acuminata (36% and 28%), Bauhinia variegata (42% and 38%): in Potato disc assay and Carrot


20

disc assay respectively. Usually greater than 20% inhibition is considered as a significant value for plant extracts.

Table No. 1: Phytochemical analysis of alcoholic extract of samples

Tests

Musa acuminata

Bauhinia variegata

Musa acuminata + Bauhinia variegata

Alkaloids Dragendroff’s test + + + Wagner’s Test + + + Meyer’s Test + + +

Flavanoids

Dil.HCl + + +

Carbohydrates Fehling’s Test + + + Benedict’s Test + + + Molish’s Test + + +

Protein Millon’s Test + + +

Phenol Ferric Chloride Test + + + Lead acetate Test + + + Libermann’s test + + + Cardiac glycosides - + Glycosides + - +

Steroids Libermanns-burchart test

- - -

Salkowski test - - -

Tannins Ferric chloride test + + + Lead acetate test + + +

Saponins Sodium bicarbonate test + + + Thiols + + Resins - - -

Figure No.1: Thin layer chromatography


21

Figure No. 2: Linoleic acid peroxidation

Table No. 2: Level of enzymic antioxidants

Enzymic antioxidants

Sample Musa acuminata

Bauhinia variegata

Musaacuminata+ Bauhinia variegata (M.a+B.v)

Superoxide dismutase 17.5±0.51 10.5±0.52 26.4±0.46

Catalase 12.6±0.30 9.09±0.42 32.8±0.68 Glutathione peroxidase 2.7±0.78 2.91±0.52 4.6±0.68

Table No. 3: Level of non enzymic antioxidants


Bauhinia variegata

Musa acuminata+Bauhinia variegata (M.a+B.v)

Vitamin C 0.76±0.50 0.58±0.65 1.0±0.5

Vitamin E 1.77±0.1 0.918±0.4 2.25±0.3

Reduced glutathione 0.45±0.3 0.186±0.6 0.724±0.2 Total phenols 4.101±0.004 2.05±0.002 4.31±0.001

Table No. 4: Reducing power determined by frap assay


Bauhinia variegata

Musa acuminata+Bauhinia variegate(M.a+B.v)

FRAP Value (µl) 0.832 0.435 0.995

Table No. 5: Percent inhibition –DPPH assay

Sample Musa

acuminata

Bauhinia

variegata

Musa acuminata+Bauhinia

variegate(M.a+B.v)

% inhibition 48.92 45.35 55.71


22

Table No. 6: Level of radical scavenging activity of formulated extract

Sample Radicals Hydroxyl SOD Nitric oxide

Musa acuminata 32.5±1.25 22.38±2.07 49.3±3.03

Bauhinia variegata 36.1±2.25 39.37±3.43 46.7±3.47

Musa acuminata+Bauhinia variegata (M.a+B.v)

48.67±1.8 25.38±2.97 51.7±2.96

Table No. 7: Extent of inhibition of invitro lipid peroxidation

Sample Musa acuminate

Bauhinia variegata

Musa acuminata+Bauhinia variegate (M.a+B.v)

% inhibition of in vitro LPO

6.68± 0.28 12.91±0.21 22.11±0.06

Table No. 8: Antimicrobial activity of samples on selected pathogens

Test Organisms

Samples Musa acuminata

Bauhinia variegata

Musa acuminate + Bauhinia variegate (M.a+B.v)

Escherichia coli 5.0±0.5 9.75±0.05 6.78±0.38

Klebsiella aerµginosa 11.7±0.2 5.98±0.4 7.25±0.6

Salmonella typhii 4.88±0.2 2.83±0.41 2.94±0.01

Staphylococcus aureus 6.25±0.4 10.25±0.4 11.65±0.2

Aspergillus niger 14.12±0.2 11.07±0.2 14.65±0.2

Figure No. 2: Zone of inhibition of the Figure No.3: Zone of inhibition of the

extracts against Escherichia coli extracts against Klebsiella aeruginosa

Figure No. 4: Zone of inhibition of Figure No. 5: Zone of inhibition of extracts

extracts against Salmonella typhii against Staphylococcus aureus


23

Figure No. 6: Zone of inhibition of Figure No. 7: Carrot disc assay

extracts against Aspergillus niger

Figure No. 8: Antitumour activity

Table No. 9: Percent inhibition in development of tumour- potato disc assay

(PDA) and carrot disc assay (CDA)

Sample Extracts % inhibition in PDA % inhibition in CDA

Musa acuminata 36 28

Bauhinia variegata 42 38

Musa acuminata + Bauhinia variegate (M.a+B.v) 52 46

Figure No. 9: Potato disc assay


24

Table No. 10: Antinociceptive activity

Sample Writhing Inhibition

Musa acuminata 40.0±1.83

Bauhinia variegate 38.96±1.36

M.acuminata+B.variegata(M.a+B.v) 43.5±1.05

Aspirin (control) 60.00±1.37

Maximum inhibition was observed in alcoholic extract of combination of Musa acuminata and Bauhinia variegata (52 % and 46%) than the unique samples i.e. Musa acuminata (36% and 28%), Bauhinia variegata (42% and 38%): in Potato disc assay and Carrot disc assay respectively. Usually greater than 20% inhibition is considered as a significant value for plant extracts.

Results on the effect of plant extracts on antinociceptive activity were shown in table. All plant extracts significantly reduced the number of abdominal constriction in acetic acid-induced mice at different degree of inhibition at concentration tested of 200 mg/kg. The non-steroidal anti-inflammatory drug (NSAID), aspirin, manifested significant diminished number of writhes (constriction inhibition of 60.00 ± 1.37%) at concentration tested of 100 mg/kg.

Figure No. 10: Writhing inhibition

Statistical analysis

Data analysis results were expressed as mean ± S.E. (Standard Error) and differences between means were analyzed statistically using an analysis of variance (ANOVA) according with SPSS 15.0 software package in Microsoft Windows 7.0 Operating System. Differences are considered significant when P≤0.05, P≤0.01 and P≤0.001.

Conclusion

On conclusion from the present study that the ethanolic extracts of Musa acuminata and Bauhinia variegata contains phytochemicals which possess significant antioxidant, antimicrobial, antitumor and antinociceptive activity. The extracts have similar effects compared with traditional antibiotics to combat the pathogenic microorganisms studied.


25

This approach has opened up the possibility of the use of these plants in drug development for human consumption for future use. All these properties probably explain the use of Musa acuminata and Bauhinia variegata flower by the indigenous people to treat and heal number of infections and diseases. The plants Musa acuminata and Bauhinia variegata and their formulation are having significant Antioxidant, Antimicrobial, Antitumour and Anti nociceptive activity.

Reference

Aruna C. and P. G. Padmaja. 2009. Evaluation of genetic potential of shoot fly resistant sources in sorghum Sorghum bicolor (L.) Moench. Journal of Agricultural Science. 147:71-80.

Agrawal.R.C, Sonam Pandey and Maheshwari.S, International Journal of Scientific and Research Publications, In-Vitro Antioxidant and Free Radical Scavenging, Activity of Bauhinia Variegata Linn. Vol- 2, Issue-2, Pg- 2250-3153., (Feb-2012).

Aqil et al., 2006). Aqil F, Ahmed I, Mehmood Z (2006). Antioxidant and free radicalscavenging properties of twelve traditionally used Indian medicinal plants. Turk. J. Biol. 30:177-183.

Bauer, A. W., W. M. M. Kirby, J. C. Sherris, and M. Turck. 1966. Antibiotic susceptibility testing by a standardized single disk method. Am. J. Clin. Pathol. 36:493-496.

Beyer RE (1994). The role of ascorbate in antioxidant protection Beyer RE (1994). The role of ascorbate in antioxidant protection of biomembranes: interaction with vit-E and coenzyme. Q. J. Bioen.Biomemb. 24: 349-358.

Benzie IFF, Strain JJ (1996). Ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: The FRAP assay. Anal Biochem 239:70-76.

Dhanamani.M, Lakshmi Devi.S and Kannan.S, Hygeia.J.D.Med .,Hygeia Journal for drugs and medicines ethnomedicinal plants for cancer therapy – a review., .vol.3 (1), 1-10., (2011).

Farooq Anwar, Amer Jamil, Shahid Iqbal and Munir A. Sheikh., Antioxidant activity of various plant extracts under ambient and accelerated Storage of sunflower oil., 189-197, ISSN: 0017-3495 191., (2006).

Furusawa, T. (2006) The Roles of Western biomedicine and folk medicine in rural Solomon Islands: A quantitative analysis of villagers' response to illness. Tropical Medicine and Health 34(2):83-91.

Galsky AG.,Wilsey JP and Powell RG(1980), Crown gall biodisc bioassay. Plant Physiol.,65: 184-185.

Halliwell,B. M. Grootveld, J.M.C. Gutteridge, Methods of Biochemical Sciences, 1981, 33, 59-90.

Krishnaswami et al., in 2000 Krishnaswami.P.R, Chandalia.B and Nomani.H.K., Free radicals in diabetes: Some observation on SOD, current perspectives of food antioxidant in health., Hindusthan Lever Research Centre, Bangalore.,pp-7-14.

Kumar S, Kumar D, Manjusha, Saroha K, Singh N, Vashishta B. Antioxidant and free radical scavenging potential of Citrullus colocynthis (L.) Schrad. methanolic fruit extract. Acta Pharm. 58: 215–220, 2008.


26

Liu and Chang (1997) Liu.F.,Ooi,VE and Chang ST., (1997)Free radical scavenging activities of Mushroom polysaccharide extracts, PubMed.gov., Life sciences, 60 (10) 763-71.

Luck H (1965). Catalase. In: Methods of Enzymatic Assays. (Ed): HU Bergmeyer. New York, Academic Press: 885-894.

Malik, C.P. and M.B. Singh: In: Plant enzymology and histo-enzymology. Kalyani Publishers, New Delhi. p. 53 (1980).

Mandal, S, Datta, K.K, Hore, DK and Mohanty Suchitra (2008) Biodiversity and Organic agriculture opportunities and challenges for the north-east region of India and a model for principles involved, outlook on Agriculutre, Vol37, no.2 pp 1-8.

Mehta, P.; Wyman, J.A.; Nakhla, M.K.; Maxwell, D.P. (1994) Transmission of tomato yellow leaf curl geminivirus by Bemisia tabaci. Journal of Economic Entolomogy 87, 1291-1297.

Meydani.M., Nutritive Rev., Vitamin E and prevention of heart disease in high risk patients., 58, pp-278-281.

Misra, H. P. and Fridovich, I. (1972) J. Biol. Chem., 247, 3170.

Moron, Μ. S., Depierre, J. W. and Mannervik, Β. (1979) Biochim. Biophys. Acta., 582, 67.

Mohammad Abdul Mannan, Tushar Chandra Sarker, Ahmad Humayan Kabir, Md. Mostafizur Rahman and Mohammad Firoz Alam., Avicenna Journal of Phytomedicine., Antitumor properties of two traditional aromatic rice genotypes (Kalijira and Chinigura).,pp1-12.( 2013).

Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxidation in animal tissues by thiobarbituric acid reaction. Annals of Biochemistry.

Osawa, T., & Namiki, M. (1981). A novel type of antioxidant isolated from leaf wax of Eucalyptus leaves. Agricultural and Biological Chemistry, 45(3), 735–739.

Peach D and Tracey MV. Modern methods of plant analysis. 4th edn., Springer Berlin, Verlag, 1955; 373-374.

Prakash Veeru, Mishra Pankaj Kishor and Mishra Meenakshi., Journal of Medicinal Plants Research., Screening of medicinal plant extracts for antioxidant Activity., Vol. 3(8), pp. 608-612, (August, 2009).

Rajesh Kumar Sharma, Rajani G.P., Vivek Sharma and Komala N., Ind J Pharm Edu Res., Effect of Ethanolic and Aqueous Extracts of Bauhinia Variegata Linn. On Gentamicin-Induced Nephrotoxicity in Rats., Vol- 45: Issue 2 pg-192 (Apr-Jun, 2011).

Roe J.H. and Keuther.A. the determination of ascorbic acid in whole blod and Wine through 2, 4 –dinitrophenyl hydrazine derivative of dehydroascorbic acid, Journal of Biochemistry, 1953, 147: 39–404.

Rosenberg, H.R. (1992), Chemistry and Physiology of Vitamins. Interscience Publichers Inc., New York. 452-453.


27

Rotruck J. T., Pope A. L., Ganther H. E., Swanson A. B., Hafeman D. G. and Hoekstra W. G. (1973) Selenium: biological role as a component of glutathione peroxidase. Science 179, 588-590.

Sangeeta sajjan, Chetana.S.H, Padmaa M Paarakh and Vedamurthy.A.B., Indian journal of natural products and resources., Antimicrobial activity of momordica cymbalaria Fenzl aerial parts extracts.vol.1(3), pp296-300.( September 2010).

Sonam Pandey, Agrawal.R.C and Maheshwari.S, International Journal of Scientific and Research Publications, In-Vitro Antioxidant and Free Radical Scavenging, Activity of Bauhinia Variegata Linn. Vol- 2, Issue-2, Pg- 2250-3153., (Feb-2012).

Sourav ,Banerjee, Bhabatosh Halder, Nishith R. Barman and Ajoy K. Ghosh., Food and Nutrition Sciences., An overview on different variety of Musa species: Importance and its enormous pharmacological action., 2, 1128-1133., (2011).

Soobrattee MA; Bahorun T; Neergheen VS; Googoolye K, Aruoma OI. Assessment of the content of phenolics and antioxidant actions of the Rubiaceae, Ebenaceae, Celastraceae, Erythroxylaceae and Sterculaceae families of Mauritian endemic plants. Toxicol in vitro 2008; 22: 45-56.

Umadevi.M, Sampath Kumar.K.P, Debjit Bhowmik and Duraivel.S, Journal of medicinal plant studies. Vol. 1- Issue 3, Pg- 56, 57.,(2011).

Zakaria, Z. A. Mustapha, S. Sulaiman, M. R. Mat Jais, A. M. Somchit, M. N. and Abdullah, F. C. “The antinociceptive action of aqueous extract from Muntingia calabura leaves: the role of opioid receptors,”Medical Principles and Practice, vol. 16, no. 2, pp. 130–136, 2007.

Documents

Evaluation of antioxidant, antimicrobial, antitumour and ... Kaviya, Thanga Priya and Priya Department of Biochemistry, Dr. N.G.P. Arts and Science College, Coimbatore Department of