Marker Based In-vitro Antioxidant Potential, Microscopy and Validated High Performance Thin Layer Chromatographic Method for Saffron

Abstract

In vitro antioxidant potential of methanolic extract of saffron and crocetin was assessed along with development of a high performance thin layer chromatography method for the quantitative estimation of crocetin. The anatomical characters of saffron stigma were also assessed and compared for the identification, prior to antioxidant and analytical studies. A simple solvent system hexane: ethyl acetate: formic acid (5:4:1, v/v/v) was developed and a compact, well resolved peak of crocetin with Rf value 0.67 ± 0.02 was observed. Standard calibration curve revealed a good linear relationship with r2 value of 0.995 between the peak area and concentration in the range of 34–850 ng spot−1. The proposed method was validated as per the International Conference on Harmonization guidelines. The considerable results of total phenolic contents by Folin–Ciocalteau method, scavenging efficacy against 2, 2-diphenyl-1-picrylhydrazyl and nitric oxide free radical were observed. The statistical analysis proved that high performance thin layer chromatography method was precise, reproducible, selective and accurate for the analysis of crocetin.