SIMPLE AND HIGHLY SENSITIVE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR SEPARATING ENANTIOMERIC DIACYLGLYCEROLS BY DIRECT DERIVATIZATION WITH A FLUORESCENT CHIRAL AGENT, -TERT-BUTYL-2-METHYL-1,3-BENZODIOXOLE-4-CARBOXYLIC ACID())

A simple and highly sensitive HPLC method was developed for the determ ination of the absolute configuration and the optical purity of diacyl glycerols. The method involves direct fluorescent labelling of diacylg lycerols with (S)-TBMB-carbonyl chloride in pyridine and HPLC separati on of the derived diastereomeric (S)-TBMB-carboxylated diacylglycerol derivatives [(S)-TBMB = (S)-(+)-2-tert.-butyl-2-methyl-1,3-benzodioxol e]. Complete separation of the diastereomeric (S)-TBMB-carbonyl-diacyl glycerol derivatives and thus indirectly of the enantiomers of the par ent diacylglycerols and of the sn-1,3-regioisomer was achieved using n ormal-phase silica gel HPLC within 30 min for every saturated single a cid (R(1) = R(2)) 'diacylglycerol (C-12:0-C-22:0) examined.