Identification and quantification of Shatavarin IV in five different species of the genus Asparagus through a validated HPTLC method

The steroidal saponin, Shatavarin IV is the major bioactive compound present in the root of Asparagus racemosus. Shatavarin IV contributes for a wide range of biological activities like antioxidant, antitussive, antidyspepsia, antiulcer and anticancer activity. The present investigation aimed to identify Shatavarin IV in five different species of the genus Asparagus. For the very first time, Shatavarin IV was identified in four species of Asparagus like Asparagus densiflorus, Asparagus setaceus, Asparagus plumosus and Asparagus sprengeri. The identification and quantification of Shatavarin IV was done by HPTLC analysis. The pre-coated silica gel 60 F254 plates were used as stationary phase for the development of chromatograms with ethyl acetate‒methanol‒water (7.5:1.5:1, v/v/v)) as mobile phase. The Shatavarin IV was detected at retardation factor (Rf) 0.55±0.05 and showed maximum absorption at 425nm. The method was validated for quantitative analysis and was found to be satisfactory. The six-point calibration curve of Shatavarin IV shows a standard deviation of 3.89 % with R2 value of 0.9968. The amount of Shatavarin IV were found to be 0.22%, 0.01%, 0.08%, 0.04% and 0.06% in A. racemosus, A. densiflorus, A. setaceus, A. plumosus and A. sprengeri respectively. The HPTLC Fingerprint developed in this research work could be useful for quality control and checking adulteration of all the five species of Asparagus. The presence of Shatavarin IV in other species of Asparagus would reduce the pressure on the Asparagus racemosus as theprimary source of drug thereby preserving its wild population.