Measurement of plasma acetate kinetics using high-performance liquid chromatography

Previous studies suggest that plasma acetate may be an important fuel in man, accounting for approximately 10% of energy expenditure. Available methods for the determination of plasma acetate kinetics are difficult and time consuming. We describe here a procedure for the determination of plasma acetate concentration and specific activity using automated high-performance liquid chromatography that is precise and sensitive and accommodates large numbers of samples. The procedure involves extraction from plasma with diethyl ether, derivatization with bromoacetophenone, and separation on a C-18 reversed-phase column. The specific activities of D-beta-hydroxybutyrate and lactate can also be determined. Acetate turnover was measured in four dogs and was similar to that previously reported in sheep and humans. Transport of [14C]acetate into red blood cells was negligible.