Determination of the absolute configuration of alpha-hydroxyglycine derivatives by enzymatic conversion and chiral high-performance liquid chromatography

We describe an approach for facile determination of the absolute configurat ion of enantiomerically chromatographed racemates by combining enzymatic co nversion and chiral chromatography. The method involves initial rapid devel opment of chiral HPLC methods using polar organic eluents with polysacchari de chiral phases. We present here evidence for using the stereospecific pep tidylamidoglycolate lyase (PGL, E.C. 4.3.2.5) to determine the absolute con figuration of oc-hydroxyglycine derivatives. The racemic solute was incubat ed with PGL, lyophilized and then enantiomerically chromatographed using th e CHIRALPAK(R)AD(TM) column. Based on the specificity of the enzyme reactio n, the unreacted enantiomer was assigned the absolute configuration R. (C) 1998 Published by Elsevier Science BN. All rights reserved.