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The present study was carried out to evaluate the quality control analysis and antiproliferative effects of petroleum ether, chloroform, ethanol and aqueous extracts of Swertia alata (family Gentianaceae) on Leishmania donovani. A basic, exact, quick and reproducible high performance thin layer chromatography (HPTLC) has been created for synchronous analysis of Oleanolic acid and Swertiamarin from S. alata. The method was validated for linearity, precision, specificity, accuracy and robustness. Densitometric scanning of the plates directly at 254nm was used for analysis of Swertiamarin, while as for the analysis of Oleanolic acid; plates were scanned for 520 nm after spraying with anis aldehyde-sulphuric acid reagent. For the extract of plants and Amphotericin B, the concentration response curve was plotted and IC50 values were determined and also MTT assay was carried out for comparative study. Compact bands of Oleanolic acid and Swertiamarin were obtained at Rf 0.62±0.001 and 0.66±0.002, linearity was satisfactory for oleanolic acid and swertiamarin. The developed and validated HPTLC methods were having Linearity range 200-2000 and 100-2000 ng/spot for oleanolic acid and swertiamarin respectively with r2 value 0.989±0.003 and 0.999±0.03, limit of detection (11.25 and 6.19 ng/spot), limit of quantification (37.51 and 20.64 ng/spot), recovery (98.4-101.0 % and 98.2-100.8%) and precision with % RSD < 2%. Various dried extracts were found to contain oleanolic acid in the range of 0.009%-0.012% and swertiamarin in the range of 0.113-0.135% (w/w). Cytotoxicity assay of extracts over promastigotes showed that ethanolic extract was found to be most cytotoxic (IC5031.43±1.90 μg/mL) after 96 h in comparison to other extracts. The ethanolic extract showed significant reduction in promastigotes which may be due to presence of oleanolic acid as compared with the other groups. The present HPTLC method is being reported for the first time and can be used for routine quality control.

HPTLC Analysis and Antiproliferative Effect of Various Extracts of Swertia alata on Growth of Leishmania donovani Promastigotes in vitro