Isolation and Characterization of Bioactives from Roots of Achyranthes Asperal with Special Reference to Anti-Appetite and Anti-Hyperlipidemic Activity

Abstract

Achyranthes aspera is a well-known herb from Ayurveda mainly used in various ailments like Antimicrobial,anticancer,antidiabetic,hypoglycemic since ancient time. It is also well reported for anti-appetite activity and there were some clinical reports about the same. Further, the plant is a rich source of triterpenoids and saponin. Literature shows that plant has very good antioxidant potential; also it shows many other activity apart from above mentioned like antimicrobial, anticancer, antidiabetic, antihyperlipidemic etc. There are many constituents reported from Achyranthes aspera like Ecdysterone, Betaine, Saponins, Ecdystone but still further detailed investigation is needed for identification of group of constituents responsible for its potent activities. In our previous studies, we have identified aa07 (IC50= 18μg/ml) as a potent antioxidant compound when evaluated by DPPH assay. Aa07 was isolated using conventional normal phase column chromatography, with usage of gradient mobile phase (Ethylacetate:Methanol:formic acid (7:3:0.1). Hence, the present work was focused on isolation of aa07 in its purest form for complete structure elucidation and to isolate the same using direct crystallization method assisted with flash chromatography which shall be easier for separation at industrial level. Further to evaluate its antioxidant potential and to evaluate in vitro and in vivo anti lipase activity. It was also reported that the plant has good anti-appetite potential, hence we have investigated anti-appetite activity of aa07 using wistar rats. A. aspera powder was extracted gradient from low polar to high polar solvent i.e. hexane, dichloromethane, ethylacetate and methanol using continuous extraction for 8 hrs respectively. The ethylacetate and methanol fractions were separated and concentrated and subsequently cooled to isolate aa07. The compound was further purified by flash chromatography and subsequently analysed for IR, Mass, 1H-NMR for elucidation of the compound. The result of in vitro lipase activity showed IC50 150 μg/ml and found significant when compared to standard drug Sibutramine (10 μg/ml). Similarly, in vivo intra lipid test of aa07 (150mg/kg/day, p.o) showed significant lowering of triglyceride level when compared to standard drug Atovastatitns (30mg/kg/day, p.o). (p< 0.05) The result of anti-appetite activity of aa07 (150mg/kg/day, p.o) was highly significant as it has reduced total food intake when compared with normal animals. In nutshell, present research has identified aa07 as one of the responsible chemical constituent of roots of A. aspera for its antioxidant as well as anti-lipase and anti-appetite activity. There is a need for further investigations for complete structure elucidation of aa07 and toxicity study of aa07. The present research has provided a break through invention which may attribute great help to future scientists working on A. aspera.