Isolation, purification and characterization of antioxidative steroid derivative from methanolic extract of Carissa carandas (L.) leaves

Present study evaluates antioxidative and erythrocyte membrane stabilizing potential of methanol extract of Carissa carandas leaves Cc(L)M and its purified fraction CM 1. Phytochemically Cc(L)M revealed presence of alkaloid, steroids, saponins and tanins. The results of antioxidant assays of Cc(L)M and CM 1 found to be dose dependent. However, in TLC bioautography, CM 1 showed yellow coloured scavenged band of DPPH reagent. The results revealed that CM 1 has significant (p≤0.05) free radical scavenging potential compared to Cc(L)M. The purified fraction CM 1 was found to be most potent with lowest EC50 values for DPPH (546.4 µg/ml), OH- (498.5 µg/ml) radical scavenging and 57.48 µg/ml for reducing power assay. It also erythrocytes membrane stabilization effect with lowest EC50 1.31 mg/ml as compared to Cc(L)M (2.56 mg/ml) and standard diclofenac sodium (1.00 mg/ml). The FT-IR and GC-HRMS analysis of CM 1 reveal the presence of steroid derivative 20-hydroxypregnan 18-oic acid.