Characterization of in-vitro antioxidant, cytotoxic, thrombolytic and membrane stabilizing potential of different extracts of Cheilanthes tenuifolia and Stigmasterol isolation from n-hexane extract

Abstract

Background: Cheilanthes tenuifolia, a member of the Pteridaceae family, is an evergreen and small fern could be abundant of bioactive compounds. The present study was designed to investigate its many therapeutic properties and isolation of bioactive compounds from extracts of Cheilanthes tenuifolia. Methods: The dried coarse plant powder was extracted with methanol and dried with rotary evaporator. The extract was further partitioned according to the increasing polarity: N-hexane < chloroform < ethyl-acetate < methanol by modified Kupchan method. Then each extract fractions were investigated for their pharmacologic properties. Compounds were isolated from n-hexane fraction through column chromatography, followed by TLC and structure was determined by analysis of sample using 1 H-NMR and matched with published phytochemistry report. Results: Methanol fraction of Cheilanthes tenuifolia showed highest amount of total phenol content (11.32 ± 0.28 mg/gm GAE) followed by chloroform fraction (9.71 ± 0.31 mg/gm GAE) > N-hexane fraction (6.69 ± 0.67 mg/gm GAE) > ethyl acetate fraction (5.36 ± 0.54 mg/gm GAE). The methanol fraction of Cheilanthes tenuifolia possessed highest amount (7.11 ± 0.52 mg/gm QE) of total flavonoid content. Our present study indicates that methanol extract was most potent (IC 50 = 9.926 μg/ml) inhibitor of DPPH free radicals. In brine shrimp lethality bio assay, all the extracts showed dose dependent increment of mortality and chloroform extract was found most cytotoxic (LC 50 = 34.493 μg/ml) compared to other plant extracts. The chloroform fraction of Cheilanthes tenuifolia was most potent in terms of thrombolytic activity. A compound was isolated (CT-2) using column chromatography followed by TCL and PTLC (35% pet ether in CHCl 3 ) and analyzed by 1 H-NMR. The structure of stigmasterol was confirmed by comparing the 1 H-NMR data with previously published phytochemistry report.Conclusion: Cheilanthes tenuifolia could be a potential candidate for bioactive compounds and further studies on isolation and characterization of its bioactive compounds are highly required.

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