Seema kumari et al. IRJP 2011, 2 (12), 267-269
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY ISSN 2230 – 8407
Available online www.irjponline.com
Research Article
BIOLOGICAL ACTIVITY OF METHANOLIC AND AQUEOUS EXTRACT OF
GLYCOSMIS MAURITIANA
AND
STREBLUS ASPER
G.S Arun kumar, Seema kumari*, B.Chandrasekhar raju, M.Ramarao
Department of Biochemistry, M.V.R. College of P.G. Studies, Gajuwaka, Visakhapatnam, Andhra Pradesh, India
Article Received on: 12/10/11 Revised on: 20/11/11 Approved for publication: 18/12/11
*Email: seemakumarisingh@gmail.com
ABSTRACT
The present study focus on evaluations of the anti-oxidant property of few medicinal plants like Glycosmis mauritiana and Streblus asper and assay of bioactive compounds responsible for its activity. Among these DPPH radical scavenging activity was highest with G.mauritiana with 93%, catalase activity with 1.0mg of protein, and acid phosphatase inhibition was highest with S.asper with alcoholic extract.
Keywords: DPPH, Acid phosphatase, catalase, bio-active compounds.
INTRODUCTION
Herbal remedies are often sought by patients with diseases especially patients with cancer to provide symptom relief natural product was one of the therapies most commonly used by adults and children now a days. A growing amount of evidence indicates a role of reactive oxygen species (ROS) such as peroxyl radicals (ROO•), hydroxyl radical (НО•), superoxide anion О2•– and singlet oxygen in the pathophysiology of aging and different degenerative diseases such as cancer, cardiovascular diseases, Alzheimer’s disease and Parkinson’s disease (Davies, 2000; Fenkel & Holbrook, 2000).The antioxidants protect from the potentially damaging oxidative stress, which is a result of an imbalance between the formation of ROS and the body antioxidant defense. Antioxidants have also been used in food industry to prevent deterioration, nutritional losses and off-flavoring in various foods, especially those containing polyunsaturated fatty acids. Recently, interest has increased considerably in finding naturally occurring antioxidants for use in foods because of their potential in health promotion and disease prevention, and their high safety and consumer acceptability (Gorinstein et al., 2003).The higher intake of plant foods correlates with lower risk of mortality from these diseases (Johnson, 2001).Polyphenols are the most significant compounds for the antioxidant properties of plant raw materials. The antioxidant activity of polyphenols is mainly due to their redox properties, which allow them to act as reducing agents, hydrogen donors, singlet oxygen quenchers, metal chelators and reductants of ferryl hemoglobin (RiceEvans et al., 1995; 1997; Prior et al., 2005; Lopez et al., 2007; Ciz et al., 2008; Gebicka & Banasiak, 2009).Rapid production of free radicals can lead to oxidative damage to biomolecules and may cause disorders such as cancer, diabetes, inflammatory disease, asthma, cardiovascular diseases, neurodegenerative diseases, and premature aging (Young and Woodside, 2001). Many medicinal plants contain large amounts of antioxidants, such as polyphenols, vitamin C, vitamin E, selenium,
β-carotene, lycopene, lutein, and other carotenoids, which play important roles in adsorbing and neutralizing free radicals, quenching singlet and triplet oxygen, or decomposing peroxides (Djeridane et al., 2006).Moreover, plant secondary metabolites such as flavonoids and terpenoids play an important role in defense against free radicals Govindarajan et al., 2005).
MATERIALS AND METHOD
Plant material: The leaves of G.mauritiana, S. asper were collected from Paderu region in the month of july.
Chemicals: Chemicals were purchased from the local dealer.
Preparation of plant crude extract: The plant materials were thoroughly cleaned, shade dried and coarsely powdered in a
mechanical blender. 10gms of material was soaked in 100 ml of methanol and water for 24hrs at room temperature, filtered through Whatmann No.1 filter paper The filtrate obtained was used for further tests.
DPPH radical scavenging activity
DPPH scavenging activity was measured by the method of (Cuedet
et al., 1997.)To 5 ml of a methanolic solution of DPPH (0.004%),
50
replaced by methanol and water. The reaction mixture was incubated for 30min.
IC 50 = Absorbancy of control- Absorbancy of sample X 100 Absorbancy of control
Total flavonoids determination
Aluminum chloride colorimetric method was used for flavonoids determination (Chang et al., 2002). O.5 ml of each plant extracts were separately mixed with 1.5 ml of methanol, 0.1 ml of 10% aluminum chloride, 0.1 ml of 1 M potassium acetate and 2.8 ml of distilled water. It incubated at room temperature for 30 min; the absorbance of the reaction mixture was measured at 415 nm with a. The calibration curve was prepared by preparing quercetin solutions at concentrations 20, 40, 60, 80,100μg in methanol and results represented as mg/gm of dry weight.
Total phenols determination
Total phenols were determined by Folin Ciocalteu reagent (McDonald et al., 2001). A dilute plant extract (0.5ml) or gallic acid (standard phenolic compound) was mixed with Folin Ciocalteu reagent (5 ml, 1:10 diluted with distilled water) and aqueous Na2CO3 (4ml, 1 M). The mixtures were allowed to stand for 15 min and the total phenols were determined by colorimetry at 765 nm. The standard curve was prepared using 10-100mg L-1 solutions of gallic acid in methanol: water (50:50, v/v).
Assay of catalase
2.5ml of 0.1 phosphatase buffer ,ph7.5 and 2.5ml of 0.9%hydrogen peroxide (v/v) in the same buffer were taken and 0.5ml of the plant extract was added & incubated at room temperature for 3min the reaction was then arrested by adding 0.5ml of 2Nsulphuric acid and the residual hydrogen peroxide was titrated with 0.1N permanganate solution. A balnk was carried out similarly with boiled enzyme extract Units of enzyme activity were expressed as ml of 0.1N potassium permanganate equivalent of hydrogen peroxide decomposed/ min/mg of protein.
Assay of acid phosphatase
Seema kumari et al. IRJP 2011, 2 (12), 267-269
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011
10% TCA and mix well keep the test tube for another 10min and filter the centrifuge or contents. Take 0.5ml of filterate for the determination of inorganic phosphorus by Fisky subbarow method Concentrations are in (6.2,12.4,18.6,24.8,31.8 μg) and results are expressed in mg/ml.
Statistical analysis
The values are expressed as the means +S.D of three determinants
Fig:-1 Data regarding the anti-radical by DPPH method of methanol and acqueous extracts
Fig:-2 Data regarding total flavonoid in methanol and acqueous extracts
Fig 3: Data regarding total phenolics present in the methanol and acqueous extracts
Fig 4: Data regarding catalase activity form the methanol and acqueous extracts.
Fig 5: Percentage of inhibition of acid phosphatase in presence of methanol and acqueous extracts
RESULTS AND DISCUSSION
The antioxidant ability of medicinal plants G.mauritiana and S.asper
were determined by preparing two extracts. Methanolic and acqueous extracts were prepared by soaking the material in different solvent. The biological activity was determined by DPPH method. It gives the radical scavenging ability of plant extracts. Fig 1: Suggests that the methanolic extracts of G.mauritiana (93.04%) and S.asper
(89.3%). which was comparatively higher than acqueous extract. Fig 2: Gives the data regarding total flavanoids in the methanolic & acqueous extracts of G.mauritiana and S.asper. The aqueous extract gives higher amount of total flavanoids. G.mauritiana (0.028mg/gm)
S.asper (0.02mg/gm).This is important as these bioactive
compounds may be responsible for radical scavenging activity.Fig 3: Explains the data regarding total phenolics in the methanolic and acqueous extracts of G.mauritiana and S.asper. The aqueous extract gives higher amount of total flavanoids. G.mauritiana (0.077mg/gm)
S.asper (0.056mg/gm).Fig4: Gives the data regarding total catalase
activity in the methanolic and acqueous extracts of G.mauritiana
and S.asper. the methanolic extract gives higher amount of total
catalase activity G.mauritiana (1mg of protein) S.asper (0.3mg of protein). Fig 5: Shows inhibition of acid phosphatase in presence of methanolic and acqueous extracts .Such that in the methanoli extracts shows low acid phosphatase value of G.mauritiana with 32mg/ml and S.asper aqueous extract with 14mg/ml. Elevation in level of acid phosphatase related with various diseases hence extract has shown potential decrease in value of enzyme.
CONCLUSION
Different pattern of results were obtained with different plant species(G.Mauritiana, S.Asper) among these species G.Mauritiana
has shown potential radical scavenging activity further studies on this plant species could provide valuable information.
ACKNOWLEDGMENT
The authors are thankful to M.V.R college management for providing the chemicals and lab and members of the department of Biochemistry.
REFERENCES
1. Davies KJ (2000) Oxidative stress, antioxidant defenses and damage removal, repair and replacement systems. IUBMB Life 50: 279–289.
2. Gorinstein S, Yamamoto K, Katrich E, Leontowicz H, Lojek A, Leontowicz M, Ciz M, Goshev I, Shalev U, Trakhtenberg S (2003) Antioxidative properties of Jaffa sweeties and grapefruit and their influence on lipid metabolism and plasma antioxidative potential in rats. Biosci Biotechnol Biochem 67: 907–910
3. Rice-Evans C, Miller N, Bolwell P, Bramley P, Pridham J (1995) The relative antioxidant activities of plant derived polyphenolic flavonoids. Free Radic Res 22: 375–383.
4.Rice-Evans C, Miller N, Paganga G (1997) Antioxidant properties of phenolic compounds. Trends Plant Sci 2: 152–159.
5. Young IS, Woodside JV (2001). Antioxidants in health and disease. J. Clin. Pathol., 54: 176-186
6.Govindarajan R, Vijayakumar M, Pushpangadan P (2005). Antioxidant approach to disease management and the role of ‘Rasayana’ herbs of Ayurveda. J. Ethnopharmacol., 99: 165-178.
Seema kumari et al. IRJP 2011, 2 (12), 267-269
INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011
8. Chang C, Yang M, Wen H, Chern J, Estimation of total flavonoid content in propolis by two
complementary colorimetric methods. J. Food Drug Analysis, 2002, 10: 178-182. 9. McDonald S, Prenzler PD, Autolovich M, Robards K, Phenolic content and antioxidant activity
of olive extracts. Food Chemistry, 73:73-84.
10. Rauscher FM, Sanders RA, Watkins JB III. Effects of coenzyme Q10 treatment on antioxidant pathways in normaland streptozotocin-induced diabetic rats. J Biochem Mol Tox 2001;15:41–46.
11.Sanders RA, Rauscher FM, Watkins JB III. Effects ofquercetin on antioxidant defense in streptozotocininduced diabetic rats. J Biochem Mol Tox 2001;15:143–149 12.Stefek M, Sotnikova R, Okruhlicova L, Volkovova K,Kucharska J, Gajdosik A, Gajdosikova A, Mihalova D,Hozova R, Tribulova N, Gvozdjakova A. Effect of dietary supplementation with the pyridoindole antioxidant stobadine on antioxidant state and ultrastructure of diabetic rat myocardium. Acta Diabetologica2000;37(3):111–117.
13.A. Chevallier, “The Encyclopedia of Medicinal Plants,” Dorling Kindersley, London, UK, 1996.
14.L. Tripathi, and N.J. Tripathi, “Role of biotechnology in medicinal plants,” Tropical Journal of Pharmaceutical Research, vol. 2 (2), pp. 243-253, 2003.
15.D. Ivanova, D. Gerova, T. Chervenkov, and T. Yankova, “Polyphenols and antioxidant capacity of Bulgarian medicinal plants,” Journal of Ethnopharmacology, vol. 97, Issues 1-2, pp. 145-150, 2005.
16.B. Djeridane, M. Yousfi, N. Vidal, N. Nadjemi, J.F. Lesgards, and P. Stocker,
“Screening of some Algerian medicinal plants for the phenolic compounds and their antioxidant activity,” European Food Research and Technology, vol. 224, N: 6, pp. 801-809, 2007.
17.M.P. Kahkonen, A.I. Hopia, H.J. Vuorela, J.P. Rauha, K. Pihlaja, and T.S. Kujala,
“Antioxidant activity of plant extracts containing phenolic compounds,” Journal of Agricultural and Food Chemistry, vol. 47, pp. 3954-3962, 1999.
18. J.B. Harborne, and T.J. Mabry, “The Flavonoids, Advances in Research,”
Chapman and Hall Ltd., London, UK, 1982.
19.Yen GC, Lai HH, Chou HY (2001). Nitric oxide-scavenging andantioxidant effects of Uraria crinita root. Food Chem., 74(4): 471-478.
