PHYTOCHEMICAL EVALUATION AND IN VITRO ANTIOXIDANT AND ANTI-INFLAMMATORY EFFECTS OF CLERODENDRUM SERRATUM ROOTS
Keywords:
Clerodendrum serratum, Quality issues, Phytochemical analysis, Phenolics, antioxidant, Anti-inflammatoryAbstract
Objective: To evaluate in vitro free radical scavenging and anti-inflammatory effects of extract and fractions of Clerodendrum serratum roots along with its phytochemical analysis.
Methods: The crude 70% methanolic extract (MECSR) and ethyl acetate (EFCSR), n-butanol (BFCSR) and residual aqueous fractions (AQFCSR) of C. serratum roots were prepared and analyzed for qualitative and quantitative phytochemical study using reported methods. The in vitro anti-inflammatory effects were studied using protein denaturation and proteinase inhibitory assays whereas in vitro free radical scavenging effects were established using DPPH (1,1-diphenyl-2-picrylhydrazyl), modified 2,2'-azinobis (3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) and trolox equivalent antioxidant capacity (TEAC) assays.
Results: The qualitative and quantitative phytochemical studies of crude extract and fractions showed the presence of phenolics, flavonoids, saponins, and carbohydrates. All analyzed samples showed dose-dependent anti-inflammatory and free radical scavenging effects in the studied in vitro assays. The IC50 values of MECSR, EFCSR, BFCSR and AQFCSR for scavenging the DPPH and ABTS radical ranged from 12.52±2.21 to 200.47±2.84 µg/ml and 18.12±1.76 to 216.08±1.90 µg/ml, respectively, while that of protein denaturation and proteinase inhibition assays were ranged from 70.76±1.40 to 110.13±1.63 µg/ml and 76.66±2.02 to 116.55±2.19 µg/ml.
Conclusion: Among the analyzed samples, the EFCSR showed significantly higher free radical scavenging and anti-inflammatory effects in studied assays. The observed activities might be attributed to the higher content of polyphenols present in EFCSR fraction of roots. The present study will provide scientific data to reinforce the traditional claims of roots for treating inflammation, pain and other oxidative stress related diseases.
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Patel JJ, Acharya SR, Acharya NS. Clerodendrum serratum (L.) Moon-a reviews on traditional uses, phytochemistry, and pharmacological activities. J Ethnopharmacol 2014;154:268-85.
Zalke AS, Kulkarni AV, Shirode DS, Duraiswamy B. In vivo anticancer activity of Clerodendrum serratum (L) Moon. Res J Pharm Biol Chem Sci 2010;1:89-98.
Chinchali JF, Sanakal RD, Kaliwal BB. Evaluation of the anti-carcinogenic activity of Clerodendrum serratum leaf extract on liver and kidney of 7, 12-dimethylbenz[a]anthracene (DMBA) induced skin carcinogenesis in mice. Eur J Exp Biol 2011;1:130-41.
Vidya SM, Krishna V, Manjunatha BK, Mankani KL, Ahmed M, Singh SDJ. Evaluation of hepatoprotective activity of Clerodendrum serratum L. Indian J Exp Biol 2007;45:538-42.
Narayanan N, Thirugnanasambantham P, Viswanathan S, Rajarajan S, Sukumar E. Comparative antibacterial activities of Clerodendrum serratum and Premna herbacea roots. ‎Indian J Pharm Sci 2004;66:453-4.
Vidya SM, Krishna V, Manjunatha BK, Rao RB. Antibacterial activity of Clerodendrum serratum L. Electron. J Environ Agric Food Chem 2010;9:1059-63.
Narayanan N, Thirugnanasambantham P, Viswanathan S, Vijayasekaran V, Sukumar E. Antinociceptive, anti-inflammatory and antipyretic effects of ethanol extract of Clerodendrum serratum roots in experimental animals. J Ethnopharmacol 1999;65:237-41.
Bhujbal SS, Kewatkar SMK, More LS, Patil MJ. Antioxidant effects of roots of Clerodendrum serratum Linn. Pharmacogn Res 2009;1:294-8.
Ismail SM, Leelavathi S. Evaluation of antioxidant activity of Anisolmeles malabarica R Br and Clerodendrum serratum L. extracts against rheumatism. Res J Pharm Biol Chem Sci 2011;2:488-95.
Mohamed AJ, Mohamed EAH, Abdalrahim FAA, Ameer OZ, Ismail Z, Ismail N, et al. Antioxidant, antiangiogenic and vasorelaxant activities of methanolic extract of Clerodendrum serratum (Spreng.) leaves. J Med Plants Res 2012;6:348-60.
Harborne JB. Phytochemical methods: a guide to modern techniques of plant analysis. Chapman and Hall: London; 1998. p. 34-86.
Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 1965;16:144-58.
Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 1999;64:555-9.
Hiai S, Oura H, Nakajima T. Color reaction of some sapogenins and saponins with vanillin and sulphuric acid. Planta Med 1976;29:116-22.
Pawar HA, D’Mello PM. Spectrophotometric estimation of total polysaccharides in Cassia tora gum. J Appl Pharm Sci 2011;1:93-5.
Williams LAD, Connar AO, Latore l, Dennis O, Ringer S, Whittaker JA, et al. The in vitro anti-denaturation effects induced by natural products and non-steroidal compounds in heat-treated (immunogenic) bovine serum albumin is proposed as a screening assay for the detection of anti-inflammatory compounds, without the use of animals, in the early stages of the drug discovery process. West Indian Med J 2008;57:327-31.
Oyedepo OO, Femurewa AJ. Antiâ€protease and membrane stabilizing activities of extracts of Fagra zanthoxiloides, Olax subscorpioides and Tetrapleura tetraptera. Int J Pharmacogn 1995;33:65â€9.
Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. Lebenson Wiss Technol 1995;28:25-30.
Nadeem AM, Zia KIU, Mukhtar IA, Ashraf QF. Modified 2,2’-azinobis (3-ethylbenzo thiazoline)-6-sulphonic acid radical cation decolorization assay for antioxidant activity of human plasma and extracts of traditional medicinal plants. Acta Chim Slov 2008;55:408-18.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. ‎Free Radical Biol Med 1999;26:1231-7.
Sharma M, Gupta A. Preliminary phytochemical investigations of methanolic roots extract of Clerodendrum serratum: antiulcer activity and histopathological study of stomach mucosa of wistar rats in ethanol-induced ulcer. J Pharm Res Opinion 2013;1:202-8.
Adesegun SA, Fajana A, Orabueze CI, Coker HAB. Evaluation of antioxidant properties of Phaulopsis fascisepala C. B. Cl. (Acanthaceae). J Evidence-Based Complementary Altern Med 2009;6:227-31.
Wong SP, Leong LP, Koh JHW. Antioxidant activities of aqueous extracts of selected plants. Food Chem 2006;99:775-83.
Garg VKR, Jain M, Sharma PKR, Garg G. Anti-inflammatory activity of Spinacia oleracea. Int J Pharm Professional's Res 2010;1:1-4.
Sakat S, Juvekar AR, Gambhire MN. In vitro antioxidant and anti-inflammatory activity of methanol extract of Oxalis corniculata Linn. Int J Pharm Pharm Sci 2010;2:146-55.
Amir M, Javed SA, Kumar H. Design and synthesis of 3-[3-(substituted phenyl)-4-piperidin-1-ylmethyl/-4-morpholin-4-ylmethyl-4,5-dihydro-isoxazol-5-yl]-1H-indoles as potent anti-inflammatory agents. Med Chem Res 2010;19:299-310.
Chu KF, Dupuy DE. Thermal ablation of tumours: biological mechanisms and advances in therapy. ‎Nat Rev 2014;14:199-208.
Anandjiwala S, Bagul MS, Parabia M, Rajani M. Evaluation of free radical scavenging activity of an ayurvedic formulation-Panchvalkala. ‎Indian J Pharm Sci 2008;70:31-6.