Objective: A number of Indian medicinal plants have been used for thousands of years in a traditional system of medicine. Hemidesmus indicus is an important member of the Asclepiadaceae family. It is an endemic to the southern Western Ghats, India. The aim of the study was to investigate the free radical scavenging activity of H. indicus. Methods: The aqueous and methanol leaf extracts of H. indicus were assayed for radical scavenging activity, using the stable free radical 2,2-diphenyl- 1-picryl-hydrazyl-hydrate and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid). Results: The results revealed that the IC50 values of aqueous extract of H. indicus were found to be higher than that of the other solvent extracts. The free radical scavenging activity of the plant extracts may be due to the presence of phytoconstituents. Conclusion: In all the methods, the aqueous extract has exhibited the good scavenging activity and this arises that the plant has a potential antioxidant agent

H, Abdul Kaffoor

K, Arumugasamy

K, Muthuraj

English

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Vol 10, Issue 6, 2017Online - 2455-3891 Print - 0974-2441ANTIOXIDANT ACTIVITY OF LEAF EXTRACTS OF HEMIDESMUS INDICUS (L.) R. BR. (ASCLEPIADACEAE)ABDUL KAFFOOR H*, MUTHURAJ K, ARUMUGASAMY KDepartment of Botany, Kongunadu Arts and Science College (Autonomous), Coimbatore - 641 029, Tamil Nadu, India.  Email: kaffoorparakeet7@gmail.comReceived: 10 February 2017, Revised and Accepted: 15 March 2017ABSTRACTObjective: A number of Indian medicinal plants have been used for thousands of years in a traditional system of medicine. Hemidesmus indicus is an important member of the Asclepiadaceae family. It is an endemic to the southern Western Ghats, India. The aim of the study was to investigate the free radical scavenging activity of H. indicus.Methods: The aqueous and methanol leaf extracts of H. indicus were assayed for radical scavenging activity, using the stable free radical 2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid).Results: The results revealed that the IC50 values of aqueous extract of H. indicus were found to be higher than that of the other solvent extracts. The free radical scavenging activity of the plant extracts may be due to the presence of phytoconstituents.Conclusion: In all the methods, the aqueous extract has exhibited the good scavenging activity and this arises that the plant has a potential antioxidant agent.Keywords: Hemidesmus indicus, Asclepiadaceae, Antioxidant.INTRODUCTIONIn India, traditional communities like tribal and rural population are frequently using the crude extract of local plants for medicinal and other purposes. Crude extracts and medicines manufactured on the principles of natural compounds even by pharmaceutical companies may lead to large-scale exposure of humans to natural products. The first step toward this goal in the biological and phytochemical screening of plant extracts from traditional preparations used in popular medicine [1,2]. With ever-increasing population pressure and foot depletion of natural resources, it has become extremely important to diversify the present day agriculture to meet various human needs [3]. The observed interest in the search for alternative/additional food and feed ingredients is of paramount importance mainly for two reasons, one is the low production of oilseeds and grains and another is the stiff competition between man and livestock industry for existing food and feed materials [4]. The ethnic people use a wide variety of wild plants and plant products as their food. India has one of the largest concentrations of tribal population in the world. The forest plays a key role in the economy as well as daily needs of the tribes. In times of scarcity when the staple food is in short, of supply tribal’s collected many types of wild roots and tubers to supplement their meager food available at home [5].The commercial development of plants as sources of antioxidants to enhance health and food preservation of current interest [6]. Epidemiological studies have suggested positive associations between the consumption of phenolic-rich foods or beverages and the prevention of diseases [7]. Antioxidants, which scavenge active oxygen species (free radicals) are found in a variety of foodstuffs and are commonly referred to as scavengers. Many antioxidants are placed based and play an important role in protecting plants that are exposed to strong sunlight and live under severe oxygen stress. Antioxidants also play an important role in human health because the biological defense mechanisms cannot operate under severe oxygen stress. Pigmentation from excess melanin can cause the appearance of spots and freckles on the skin. Plants are a rich source of antioxidant, which evinced through many reports on medicinal plants with antioxidant potential [8]. Almost all the sources for such phytochemical studies are from published or unpublished ethnobotanical knowledge [9].2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical method is an antioxidant assay based on electron-transfer that produces a violet solution in ethanol. This free radical, stable at room temperature, is reduced in the presence of an antioxidant molecule, giving rise to a colorless ethanol solution. The use of the DPPH assay provides an easy and rapid way to evaluate antioxidants by spectrophotometry, so it can be useful to assess various products at a time. Due to the lack of evidence about which solution can be more effective as an antioxidant or even if there are other solutions with equal or more capacity to eliminate free radicals from dental surfaces after bleaching procedures, the purpose of this study was to evaluate the antioxidant activity of several agents proposed for reversion of problems caused by bleaching procedures using the DPPH free radical assay [10].METHODSCollection of plantHemidesmus indicus (L.) R. Br. belongs to the family Asclepiadaceae was collected from a natural forest of Mettupalayam, Coimbatore, Tamil Nadu, India. The specimen was identified by Dr. M. Murugesan, Scientist B, Botanical Survey of India, Shillong. A voucher specimen was deposited in the Department of Botany, Kongunadu Arts and Science College, Coimbatore (KASC/BOT/0012), Tamil Nadu, India. The plants were thoroughly washed in running tap water before being shade-dried for 20 days.Preparation of plant extractsAbout 50 g of powdered plant materials were extracted in soxtron apparatus for 1-2 hrs, sequentially with the alcoholic solvents, viz., © 2017 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ajpcr.2017.v10i6.17634Research Article201Asian J Pharm Clin Res, Vol 10, Issue 6, 2017, 200-202 Kaffoor et al. petroleum ether, chloroform, methanol, and water. Then, the extract was evaporated to dryness using a vacuum rotary evaporator and stored in vials kept in 4°C for further use.DPPH free radical scavenging activityThe DPPH scavenging activity of this plant tuber extract was measured according to the method of Blois [11]. IC50 values of the extracts and concentration of the extracts necessary to decrease the initial concentration of DPPH by 50% were calculated.2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) cation radical scavenging activityThe total antioxidant activity of the samples was measured by ABTS·+ radical cation decolorization assay according to the method of Re et al. [12]. In this improved version, ABTS the oxidant is generated by per sulfate oxidation of ABTS2−. ABTS•+ radical cation (ABTS•+) was produced by reacting ABTS•+ solution (7 mm) with 2.45 mm ammonium per sulfate, and the mixture was allowed to stand in the dark at room temperature for 12-16 hrs before use. For this study different concentration that range from 25 to 100 µg/ml of aqueous extract was added to 0.3 ml of ABTS•+ solution and the final volume was made up with aqueous to 1 ml. The absorbance was read at 734 nm, and the percentage of inhibition was calculated.Statistical analysisThe data were subjected to a one-way analysis of variance, and the significance of the difference between the means was determined by Duncan’s multiple range test (p<0.05) using statistics (Stat Soft Inc., OK, USA). Values expressed are means of three replicate determinations ± standard deviation.RESULTS AND DISCUSSIONDPPH radical scavenging activityThe result showed the dose response, radical scavenging activities observed in the MeOH and water extracts of whole plans of H. indicus (Table 1). Both extracts, there was an increase in the DPPH• radical scavenging activity with increasing extract concentration. The results of DPPH• radical scavenging activity of H. indicus extracts along with the reference standards. Concentration of the sample necessary to decrease the initial concentration of DPPH• by 50% (IC50) under the experimental condition was calculated. Therefore, a lower value of IC50 indicates the highest antioxidant activity of 51.92±11.19 and 70.10±8.28 µg/ml, respectively (Table 1). Similarly, reported Samydurai and Thangapandian [13] the Decalepis hamiltonii (Asclepiadaceae) aqueous root extract showed good antioxidant activity could be used in the prevention of free radical diseases and general health tonic. The generation of free radicals can bring about thousands of reactions and thus cause extensive tissue damage. Lipids, proteins, and DNA are all susceptible to attack by free radicals [14]. According to Houghton et al. [15], free radical reactions are involved in the pathology of many diseases such as Alzheimer’s disease, cancer, and inflammation.The whole plant extracts of H. indicus showed a good radical scavenging activity as indicated by their IC50 values, which were significantly different from that of the butylated hydroxytoluene standard (Table 1). The results suggest the likely presence of assuring compounds that are possibly aggressive in their radical scavenging activities.Total antioxidant activity by ABTS•+ABTS•+ cation radical scavenging activity decolorization assay applicable to both lipophilic and hydrophilic antioxidants, including flavonoids, hydroxycinnamates, carotenoids, and plasma antioxidants. The preformed radical monocation of 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+) is generated by oxidation of ABTS•+ with potassium persulfate and is reduced in the presence of such hydrogen donating antioxidants. The activity of the testing sample extract is expressed as a micro molar equivalent of butylated hydroxytoluene. The effect of methanol and aqueous extract of Hemidesmus hamiltonii on ABTS•+ cation radical scavenging activity is shown in Table 1, and the plant extract revealed that the higher total antioxidant activity of 11108.9 and 2720.1 µmol/g. Polyphenols are the major plant compounds with antioxidant activity, although they are not the only ones. The antioxidant activity of phenolic compounds is reported to be mainly due to their redox properties [16,17], which can play an important role in adsorbing and neutralizing free radicals, quenching singlet and triplet oxygen, or decomposing peroxides.CONCLUSIONAntioxidant-rich plant extracts serve as sources of nutraceuticals that alleviate the oxidative stress and therefore prevent or slow down the degenerative diseases. An effort has been made to explore the antioxidant properties of the leaf extract H. indicus. This indicates the potential of the extracts, as a source of natural antioxidants or nutraceuticals with potential application to reduce oxidative stress with consequent health benefits. Aqueous extract of H. indicus was found to be higher than that of the other solvent extracts. The free radical scavenging activity of the plant extracts may be due to the presence of phytoconstituents present in the plant.REFERENCES1. Paz A, Cerdeiras C, Fernadez MP, Ferreira J, Moyna F, Soubes P, et al. Screening of Uruguayan medicinal plants for antimicrobial activity. J Ethnopharmacol 1995;20:67-9.2. Sohni YR, Kaimal P, Bhatt RM. The antiamoebic effect of a crude drug formulation of herbal extracts against Entamoeba histolytica in vitro and in vivo. J Ethnopharmacol 1995;45(1):43-52.3. Janardhanan K, Gurumoorthi P, Pugalenthi M. Nutritional potential of five accessions of a South Indian tribal pulse, Mucuna pruriens var. Utilis: II. Investigations on total free phenolics, tannins, trypsin and chymotrypsin inhibitors, phytohaemagglutinins, and in vitro protein digestibility. J Trop Subtrop Agroecosyst 2003;1:141-52.4. Siddhuraju P, Becker K, Makkar HP. Studies on the nutritional composition and antinutritional factors of three different germplasm seed materials of an underutilized tropical legume. Mucuna pruriens var. Utilis. J Agric Food Chem 2000;48(12):6048-60.5. Vidyarthi LP. Role of forest in tribal life. In: Sinha SP, editor. Tribals and Foress. Ranchi, India: Bihar Tribal Research Institute; 1987. p. 323.6. Rice-Evans CA, Miller NJ, Paganga G. Structure antioxidant activity relationships of flavonoids and phenolic acids. Free Radic Biol Med 1996;20(7):933-56.7. Scalbert A, Williamson G. Dietary intake and bioavailability of polyphenols. J Nutr 2000;130 8S Suppl:2073-85.8. Katalinic V, Milos M, Modun D, Music I, Boban M. Antioxidant effectiveness of selected wines in comparison with (+) - Catechin. Food Chem 2004;86:593-600.9. Kaushik N, Singh BG, Tomar UK, Naik SN, Satya V, Bisla SS, et al. Regional and habitat variability in azadirachtin content of Indian neem (Azadirachta indica A Jusieu). Curr Sci 2007;92(10):1400-6.10. Zahin M, Aqil F, Ahmad I. The in vitro antioxidant activity and total phenolic content of four Indian medicinal plants. Int J Pharm Pharm Sci 2011;3(1):238-42.11. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature 1958;29:1199-200.12. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation Table 1:  DPPH• and ABTS•+ free radical scavenging activities of leaf extracts of H. indicusConcentration of sample (mg/ml)Inhibition of DPPH ABTS•+ (µmol/g)Methanol Aqueous Methanol Aqueous25 34.97 59.09 13040.9 4819.550 48.70 65.86 12494.2 2632.575 62.05 74.67 8491.5 1725.3100 61.96 80.79 6409.1 1306.1IC50 (µg/ml) 51.92 70.10 11108.9 2720.1DPPH: 2,2-diphenyl-1-picryl-hydrazyl-hydrate, ABTS: 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)202Asian J Pharm Clin Res, Vol 10, Issue 6, 2017, 200-202 Kaffoor et al. decolorization assay. Free Radic Biol Med 1999;26(9-10):1231-7.13. Samydurai P, Thangapandian V. Antioxidant property and polyphenols evaluation of aqueous root extract of Decalepis hamiltonii Wight & Arn. Int Curr Pharm J 2012a;1:71-6.14. Cotran RS, Kumar V, Collins T. Pathologic Basis of Disease. Philadelphia, PA; W.B. Saunders; 1999.15. Houghton PJ, Hylands PJ, Mensah AY, Hensel A, Deters AM. In vitro tests and ethno pharmacological investigations: Wound healing as an example. J Ethnopharmacol 2005;100(1-2):100-7.16. Srinivasahan V, Durairaj B. Antioxidant and free radical scavenging effect of Morinda citrifolia fruit extract. Int J Pharm Pharm Sci 2014;6(4):55-9.17. Zheng W, Wang SY. Antioxidant activity and phenolic compounds in selected herbs. J Agric Food Chem 2001;49(11):5165-70.

ANTIOXIDANT ACTIVITY OF LEAF EXTRACTS OF HEMIDESMUS INDICUS (L.) R.BR. (ASCLEPIADACEAE).

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ANTIOXIDANT ACTIVITY OF LEAF EXTRACTS OF HEMIDESMUS INDICUS (L.) R.BR. (ASCLEPIADACEAE).

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