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

• Published in: Journal of Chromatography A Vol. 828; no. 1; pp. 191 - 198
• Main Authors: McIninch, Jane K., Geiser, Fiona, Prickett, Keith B., May, Sheldon W.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 18.12.1998; Elsevier
• Subjects: Aliphatic compounds; Amidine-Lyases - metabolism; Analytical chemistry; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid - methods; Exact sciences and technology; Glycine - analogs & derivatives; Glycine - chemistry; Glycolates - chemistry; Organic chemistry; Other chromatographic methods; Peptidylamidoglycolate lyase (PGL); Preparations and properties; Stereoisomerism; α-Hydroxyglycine derivatives; Peptidylamidoglycolate lyase (PGL); α-Hydroxyglycine derivatives; Enantiomer separation; Absolute configuration; Separation; Amidine-lyases; Phase composition; Enzyme; HPLC chromatography; Peptidylamidoglycolate lyase; Chiral stationary phase; Lyases; Experimental study; Mobile phase; Scission; Carbon-nitrogen lyases; α-Aminoacid; Ultraviolet detector; Optical resolution; Enantiomer; Enzymatic reaction
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(98)00651-7

We describe an approach for facile determination of the absolute configuration of enantiomerically chromatographed racemates by combining enzymatic conversion and chiral chromatography. The method involves initial rapid development of chiral HPLC methods using polar organic eluents with polysaccharide chiral phases. We present here evidence for using the stereospecific peptidylamidoglycolate lyase (PGL, E.C. 4.3.2.5) to determine the absolute configuration of α-hydroxyglycine derivatives. The racemic solute was incubated with PGL, lyophilized and then enantiomerically chromatographed using the CHIRALPAK®AD™ column. Based on the specificity of the enzyme reaction, the unreacted enantiomer was assigned the absolute configuration R.