Capillary gas chromatography/mass spectrometry of intact fatty acyl esters of pregnane steroids.

Synthetic long-chain fatty acyl esters of pregnenolone and 3 beta-hydroxy-5 alpha-pregnan-20-one (5 alpha-pregnanolone) were submitted to gas chromatography (GC) and GC/mass spectrometry (GC/MS). Analyses were readily performed using short (8-12 m) flexible fused-silica capillary columns coated with thin films of immobilized apolar stationary phase (e.g. OV-1 type). The electron impact (EI) mass spectra were found to be of limited value for structure investigations, as molecular ions (M+.) were either weak or absent. EI spectra also lacked characteristic fatty acyl fragment ions. In contrast, negative ion ammonia chemical ionization produced characteristic [M-H]- ions and diagnostic fragment ions. The application of the above methods to biochemical investigations is exemplified by GC/MS analysis of the steroidal fatty acyl esters in extracts of bovine adrenal tissue, and the metabolites produced by incubating starfish (Asterias rubens) testicular tissue with exogenous progesterone.