Quantitative TLC Analysis of Steroid Drug Intermediates Formed During Bioconversion of Soysterols

Abstract

A simple, rapid, and accurate method based on thin-layer chromatography (TLC) combined with image-analysis software has been developed for analysis of steroid drug intermediates formed during bioconversion of soysterols. The results obtained have been compared with those from LC. The method has been used to monitor the accumulation of widely used steroid drug intermediates androst-4-ene-3,17-dione (AD) and androsta-1,4-diene-3,17-dione (ADD), formed during the bioconversion of soysterols by Mycobacterium sp. NRRL B-3805 and Mycobacterium sp. NRRL B-3683. The percentage error between TLC and LC ranged between −0.79 to +4.50 for AD and −0.61 to +2.48 for ADD. Maximum conversion of soysterols to AD and ADD by Mycobacterium sp. NRRL B-3805 was 49.83 and 9.36 mol%, respectively, after incubation for 144 h, whereas conversion of soysterols by Mycobacterium sp. NRRL B-3683 after incubation 288 h was 41.90 mol% for AD and 37.79 mol% for ADD.