Abstract
The chromatographic behaviour of eight ecdysone 22-long-chain fatty acyl esters differing in fatty acyl chain length and degree of unsaturation and seven 22-long-chain fatty acyl ecdysone 2,3-acetonides has been investigated on normal-phase (silica) and reversed-phase (C 18, C 8, C 6, C 1 and CN) high-performance liquid chromatography (HPLC). On silica HPLC, the ecdysone 22-long-chain acyl esters co-elute, but are resolved from ecdysone, ecdysone acetates and ecdysone 2-stearate. The ecdysone 22-long-chain acyl esters separate on the reversed-phase columns according to fatty acyl chain length and degree of unsaturation. The different phases show differing relative selectivities for the unsaturated acyl esters. Thus, sequential analysis of samples on three of the phases (C 18, C 8 or C 6, CN) permits the unequivocal confirmation of the identity of putative ecdysone 22-long-chain fatty acyl esters. Ecdysone mono-, di- and tri-long-chain fatty acyl esters can be separated and identified using non-aqueous reversed-phase HPLC on a C 18-column with a gradient of dichloromethane in methanol. The application of these methods to the identification of apolar ecdysone metabolites in biological extracts is discussed.
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