Isolation and HPLC separation of polyunsaturated species of rat brain acyl-CoA produced during decapitation - Ischemia

Acyl-CoA is a crucial metabolic intermediate for the incorporation of fatty acid into membrane phospholipid. The rat brain long chain acyl-CoAs were isolated and quantitated with a procedure involving solid phase extraction with an oligonucleotide purification cartridge, followed by separation and quantitation by HPLC with special emphasis on resolution of the polyunsaturate acyl-CoAs. This procedure is uniquely suited to the rapid analysis of all molecular species of brain acyl-CoA including the polyunsaturate molecular species which are affected by ischemia. A selective 3-4 fold increase in arachidonoyl-CoA was found in rat brain after 3 min of ischemia when compared with microwaved brain. In contrast, the concentration of all other molecular species of acyl-CoA did not change over the time course of the ischemia.