High-performance liquid chromatography of coenzyme A esters formed by transesterification of short-chain acylcarnitines: Diagnosis of acidemias by urinary analysis

✍ Scribed by Richard E. Dugan; Mary J. Schmidt; George E. Hoganson; John Steele; Bruce A. Gilles; Austin L. Shug

• Publisher: Elsevier Science
• Year: 1987
• Tongue: English
• Weight: 475 KB
• Volume: 160
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(87)90047-9

✦ Synopsis

A protocol for the identification and estimation of short-chain esters of carnitine is described; it is useful for the diagnosis of acidemias. By this method, camitine esters in urine are converted to coenzyme A esters enzymatically with camitine acetyltransferase (CAT): short-chain acylcamitine + CoA CAT * short-chain acyl-CoA + camitine.

The coenzyme A esters are separated by high-performance liquid chromatography using a radial compression system with a C8 Radial-Pak cartridge and a mobile phase containing 0.025 M tettaethylammonium phosphate in a linear gradient of I to 50% methanol. Coenzyme A esters are quantitated by integrator determination of the area under the 254-nm absorption peaks. Enzymatic conversion approaches 100% for acetyl and propionyl esters except in the presence of high levels of free camitine, which lowers the proportion of ester as acyl-CoA at equilibrium. However, since acidemia patients produce urine low in free camitine, this problem is minimized. The method is rapid and simple and identifies propionic, methylmalonic, and isovateric acidemias. 0 1987 Academic PW. 1~.