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In Vitro Antioxidant Activity of Phyllodium Pulchellum L. Desv - an Threatened Medicinal Plant

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In Vitro Antioxidant Activity of Phyllodium Pulchellum L. Desv - an Threatened Medicinal Plant Online - 2455-3891 Vol 10, Issue 10, 2017 Print - 0974-2441 Research Article IN VITRO ANTIOXIDANT ACTIVITY OF PHYLLODIUM PULCHELLUM L. DESV - AN THREATENED MEDICINAL PLANT GOPAL VELMURUGAN*, SUBRAMANIAM PARVATHI ANAND Department of Botany, National College (Autonomous), Tiruchirappalli - 620 001, Tamil Nadu, India. Email: [email protected] Received: 13 May 2016, Revised and Accepted: 30 June 2017 ABSTRACT Objectives: In this study, we determined the in vitro antioxidant capacity of Phyllodium pulchellum of aqueous, ethanol, and chloroform leaf extracts. Methods: In this context, the in vitro ethylbenzothiazolone-6-sulfonic acid) (ABTS+) radical scavenging assay, the total antioxidant activity of phosphomolybdenum assay and hydroxyl radical scavenging activity in different leaf extracts antioxidant of P. pulchellum activity was. The demonstratedantioxidant activity by 2,2-diphenyl-1-picrylhydrazylof the extracts was compared to(DPPH), standard 2,2′-azinobis(3- ascorbic acid. Results: All the four methods of antioxidant showed good reducing power and reducing capacity with increasing concentration again taking the ethanol leaf extract to the top position. Remarkable of antioxidant activity was observed in ethanol leaf extract on the hydrogen peroxide scavenging Conclusion:activity with Thesethe lowest results inhibitory suggest concentrationthat the leaf of 50P. pulchellum values of (155.40 could be μg/ml) a valuable followed source by of DPPH new antioxidant(432.90 μg/ml) properties, and ABTS+ from (524.40 the above μg/ml). results it seen that this plant exhibits pharmaceutical activity. Keywords: Phyllodium pulchellum, Inhibitory concentration 50. Fabaceae, Leaf extracts, 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azinobis(3-ethylbenzothiazolone-6-sulfonic acid), © 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.v10i10.19919 INTRODUCTION sequentially with aqueous, ethanol, and chloroform. All the solvent L. Desv. (Vellalothi in Tamil) is a flowering Phyllodium pulchellum extracts(30 g/300 were ml) evaporated were extracted to remove in a theSoxhlet final tracesextractor of thefor respective8-10 hrs, plant species and widely distributed in South India as a threatened solvents. Dried extracts were kept at 20°C until further test was carried medicinal plant belonging to the family of Fabaceae. It is a Subshrub out. P. pulchellum (Syn. Desmodium pulchellum (L.) Benth) is used to treat various diseases such In vitro antioxidant activity 1(3) m; branchlets downy pubescent to tomentose. as anti-inflammatory, analgesic, antioxidant, hemorrhage, diarrhea, Free radical scavenging activity of the various solvent of leaf extracts poisoning, and eye diseases [1]. The genus relative plants are reported in P. pulchellum was determined using various in vitro assays such as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, anti-inflammatory [5], and antidiabetic [6]. The international union to have anthelmintic [2], anti-hepatic fibrotic [3], antidiarrheal [4], for conservation of natural and national resource has a long time ago scavenging assay, total antioxidant activity of phosphomolybdenum listed P. pulchellum (L.) Desv. as a threatened species [7]. The genus assay,2,2′-azinobis(3-ethylbenzothiazolone-6-sulfonic and hydroxyl radical scavenging activity. acid) (ABTS) radical distributed in tropical and subtropical zones of the worldwide. However, DPPH radical scavenging activity noDesmodium, studies to ofdate the have Fabaceae been able family, to reveal includes the antioxidantabout 350 effectspecies of any Desmodium species other than D. gangeticum and D. triflorum. Hence, for there is no scientific report on the antioxidant activities in extractsFree radical and scavenging taken in test activity tubes was in triplicates. measured usingThen, DPPH5 ml of method a 0.1 mM[8]. the particular species. Therefore, the present study was undertaken to ethanolDifferent solution concentrations of DPPH (1000, (1,1,diphenyl-2-picrylhydrazyl) 800, 600, 400, and 200 μg/ml) was of addedcrude evaluate and compare the antioxidative activities of different solvent to each of the test tubes and was shaken vigorously. They were then leaf extracts of P. pulchellum in different methods. without any extracts. Ethanol was used for baseline corrections in MATERIALS AND METHODS absorbanceallowed to stand(optical at density37°C for [OD]) 20 minutes.of the sample The controlmeasured was at prepared 517 NM. Ascorbic acid is used as reference antioxidant compound. A radical Plant material scavenging activity was expressed as 1% scavenging activity and was P. pulchellum was collected from the Jambhudu hamlet of calculated by the following formula: Bodamalai at Namakkal District, Tamil Nadu, India, and identified following OD control – OD sample Radical scavenging activity ()% = ×100 theherbarium Botanical of the Survey Department of ofIndia Botany, and National the OD control voucherCollege (Autonomous), specimen (BSI/SRC/5/23/2012-13/Tech-1795 Tiruchirappalli - 620 001, Tamil Nadu, & India.Serial No. 2) was deposited in the ABTS radical scavenging assay Preparation of plant extracts The efficacy of plant extracts to scavenge free radicals was determined Fresh plant material was washed under running tap water, air dried using ABTS radical scavenging assay with minor modification [9]. and powdered. About 30 g of coarsely powdered plant materials Freshly prepare the ABTS radical solution by adding 5 ml of 4.9 mM Velmurugan and Anand Asian J Pharm Clin Res, Vol 10, Issue 10, 2017, 282-285 ammonium persulfate solution to 5 ml of a 14 mM ABTS solution and keep for 16 hrs under dark condition. The solution is diluted with acetone were mixed and raised to 1 L with distilled water) were added andof ammonium left at room acetate, temperature 3.0 ml offor glacial 15 minutes, acetic acid, and andthe test2 ml was of acetyl done distilled water to yield an absorbance of 0.70±0.02 at 734 nm, and the The intensity of the color formed was measured spectroscopically reactionsame is used mixture for theis vortexed assay. To for 900 10 μl seconds of the ABTS and radicalthe assay solution, was done. add at3 times.412 nm The against reaction the mixture reagent without blank. sampleThe percent was used hydroxyl as a control.radical 100 μl of the extract (1000, 800, 600, 400, and 200 μg/ml) and the scavenging activity is calculated by the following formula: Ascorbic6 minutes acid after was record used asthe reference absorbance standard. at 734 The nm radical against scavenging distilled Test sample absorbance water using (Beckman DU-530) ultraviolet-visible spectrophotometer. Hydroxyl radicalscavenging activity = ×100 activity was calculated using the formula: Blank sample absorbance Absorbance of test sample ABT scavenging activity = ×100 Absorbancce of blank sample RESULTS AND DISCUSSION DPPH radical scavenging activity Total antioxidant activity of phosphomolybdenum assay The scavenging activity of all the extracts was found to be less when Determination of total antioxidant capacity developed by method [10]. compared to that the standard ascorbic acid. Among the different extracts, the ethanol leaf extracts of showed high DPPH scavenging 0.2 ml of plant extract (1000, 800, 600, 400, and 200 μg/ml) is mixed (Table 1). From the results, it is known that the species, P. pulchellum with 1.8 ml of distilled water, 2 ml of phosphomolybdenum reagent possessactivity athydrogen 1000 μg/ml donating (81.5±0.02) capabilities followed for byethanolic aqueous leaf and extracts chloroform and asolution. reagent Incubate blank. The it attest 95°C was for performed 90 minutes. in triplicate. The mixture The isantioxidant cooled to execute scavenging free radicals. capacityroom temperature, is expressed and theas ascorbicabsorbance acid is measuredequivalent at (AAE) 695 nm and against was calculated by the following theoretical formula: ABTS radical scavenging assay ABTS radical scavenging activity of different extracts of P. pulchellum cV× is shown in Table 2. Ethanolic leaf extracts of P. pulchellum showed A = M followedmaximum by free aqueous radical and scavenging chloroform for extracts. ABTS assayAmong at the1000 all μg/mltested plantconcentration samples, (75.13%)ethanolic extractswith a respective of P. pulchellum standard value of (82.13%) in AAE, c=The concentration of ascorbic acid established from the effective radical scavenging activity. All the extracts showed an increase Where, A=Total content of antioxidant compounds, (mg/g) leaf extract, in antioxidant capacity with an increase in the amount.exhibited the majority weight of crude leaf extract (g). calibration curve, (mg/ml), V=The volume of extract (ml), and m=The Total antioxidant activity of phosphomolybdenum assay Hydroxyl radical scavenging activity Phosphomolybdate method is also a quantitative assay, while the total The scavenging activity of the leaf extract on hydroxyl radical was antioxidant capacity is expressed as AAE. The total antioxidant activity measured according to the method [11]. Various concentrations (200, of different concentrations in (200-1000 μg/ml) this method. The 400, 600, 800, and1000 μg/ml) of extracts were added to 1.0 ml of ofethanolic the extracts leaf extract was foundis higher to thethan decrease (2.069±0.002) arrange with as infollowed respective by iron-ethylenediaminetetraacetate (EDTA) solution (0.13% ferrous
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