pH as an indicator of free fatty acid release from adipocytes

Abstract

A convenient method of measuring initial rates of free fatty acid efflux from isolated adipocytes during triglyceride breakdown by hormone-sensitive lipase is described. The procedure is based on the dissociation of protons from carboxyl groups of free fatty acids. A recording pH meter is used to monitor H+ concentration in the medium continuously as an index of free fatty acid release. A stoichiometric relationship was demonstrated between proton release and extracellular free fatty acid concentration as determined by the 63Ni radioassay method of Ho (1970. Anal. Biochem. 36: 105-113). An acid pH (6.8) caused a reduction in proton release, which was immediately and completely reversed by raising the pH to 7.4.