Seasonal dynamics of Shatavarin-IV, a potential biomarker of Asparagus racemosus by HPTLC: Possible validation of the ancient Ayurvedic text.

Abstract

The medicinal property of Asparagus racemosus is primarily attributed to its constituent steroidal saponins, particularly the major component, shatavarin-IV. Thus, it can serve as a biomarker and its level can decide of the utility of the plant cultivar as a drug. Hence, a sensitive, reliable and quantitative High Performance Thin Layer Chromatography (HPTLC) method has been established for quantification of shatavarin-IV in the methanolic extracts of the roots collected in both summer and rainy seasons. The extracts of the powders of dried roots were applied to silica gel 60 F₂₅₄ aluminum-supported precoated TLC plates and developed with n-hexane: ethyl acetate: methanol, 80:10:10 (v/v), as the mobile phase. Shatavarin-IV was detected and quantified by densitometry at λ = 336 nm. The accuracy of the method was checked by conducting recovery studies at three different levels of shatavarin-IV. The average recovery was found to be 101% and 107% for summer and rainy seasons respectively. The shatavarin-IV contents, as estimated by the proposed method were 12.5 μg gm^-1 and 10.9 μg gm^-1 in summer and rainy roots respectively. The entire method was performed six times (n=6) to check the repeatability. The proposed HPTLC method for quantitative monitoring of shatavarin-IV in A. racemosus roots collected in different seasons strictly adhered to the validation issues laid down by the ICH guidelines. The method is reliable reproducible and highly precise and selective.