ISOLATION AND QUANTITATION OF LONG-CHAIN ACYL-COENZYME-A ESTERS IN BRAIN-TISSUE BY SOLID-PHASE EXTRACTION

Long-chain acyl-CoA's are important intermediates in fatty acid oxidation and phospholipid metabolism. For quantitative analysis of brain acyl-CoA's, and to avoid decomposition due to high brain acyl-CoA hydrolase activity, a fast and efficient analytical method was developed for isolation and determination of acyl-CoA's. The analysis includes solid-phase extraction by an oligonucleotide purification cartridge and HPLC measurements using a synthetic internal standard. Estimates of concentration in rat brain are oleoyl-CoA (11.0 nmol/g), palmitoyl-CoA (6.0 nmol/g), stearoyl-CoA (4.0 nmol/g), and linoleoyl and arachidonoyl-CoA (2.0 nmol/g) for a total concentration of 23 nmol/g brain. (C) 1994 Academic Press, Inc.