RAPID METHOD FOR THE SEPARATION AND DETECTION OF TISSUE SHORT-CHAIN COENZYME-A ESTERS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A simple and rapid method for the separation and identification of tis sue levels of short chain coenzyme A (CoA) esters by a reversed-phase high-performance liquid chromatography with ultraviolet-visible adsorb ance detection is described. Samples of liver, heart and kidney tissue s were homogenised in 5% sulfosalicylic acid containing 50 mu M of dit hioerythritol in 1:9 w/v proportion. Following centrifugation, 20 mu l of the supernatant were directly injected onto a 3-mu m ODS C-18 colu mn (100 x 4.6 mm I.D.). The separation of acetyl-CoA, malonyl-CoA, met hylmalonyl-Coa, succinyl-CoA, propionyl-CoA and free CoASH was achieve d in less than 20 min using gradient elution with sodium phosphate, so dium acetate and methanol at a constant flow-rate of 1.5 ml/min. The l owest detection limit was 3 pmol.