Techniques for the isolation and identification of steroids in insects and algae

Analytical techniques, methods and instrumentation employed for the extraction, isolation separation, purification and identification of steroids from algae and insects are presented. The techniques include adsorption and argentation column chromatography and counter-current distribution for separating the individual steroids, and thin layer chromatography and gas liquid chromatography for monitoring the purification process. Double bond, steric and alkyl substituent separation factors and relative retention times are reported for a large number of sterols on 4 different column systems. Their use permits the tentative structural assignment of sterols. Ultraviolet, infrared, nuclear magnetic resonance (NMR) and mass spectral analyses are discussed in light of their significance in the isolation and identification of steroids from insects and algae. Numerous examples are presented, including the use of 220 MHz NMR spectrometry, which permits the differentiation and characterization of C-24 epimetric sterols and allows for a semiquantitative estimate of the 24α- and 24β-epimers present in a mixture.