20-HYDROXYECDYSONE DETERMINATION FROM ACHYRANTHES ASPERA LINN BY HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHIC METHOD

20-Hydroxyecdysone a phytoecdysteoroid having diverse biological activities, commonly occurs in roots of Achyranthes aspera L.,family Amaranthaceae, which have high commercial, economical and medicinal value. In the present study, a simple method was developed for the isolation of 20-hydroxyecdysterone from roots of Achyranthes aspera Linn. A validated HPTLC method has been established for the determination of 20-Hydroxyecdysone in roots and seeds of Achranthes aspera. The methanol extracts of root, seeds its formulation and standard solution were applied on silica gel F254 HPTLC plates. The plates were developed in twin chamber using mobile phase chloroform: methanol: ammonia (8:2:0.2 V/V) and scanned at 254 using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area) and amount of 20-hydroxyecdysone in the range of 600-2100ng/spot; the correlation coefficient was 0.9984 respectively. Sharp, well resolved peak 20-hydroxyecdyosne spot resolved at Rf 0.41±0.03 from other components of the sample extracts. The LOD and LOQ were found to be 50 and 152.3ng/spot respectively. The HPTLC method showed good linearity, recovery and high precision of 20-hydroxyecdysone. Amount of 20-hydroxyecdysone in roots and seeds was found to be 0.176% and 0.069% w/w.