Valine oxidation: the synthesis and evaluation ofl-[3-3H]valine as a tracer in vivo

Abstract

The suitability of l-[3-3H]valine for measuring valine oxidation was studied by comparing its oxidation rate with that of l-[1-14C]valine in rats and pigs. l-[3-3H]valine was synthesized by removal of the tritium on carbon-2 of l-[2,3-3H]valine by acetylation. The acetyl group was removed enzymatically using pig renal acylase 1 (EC3.5.1.14) and the product was purified by ion-exchange and paper chromatography. For the first rat experiment l-[3-3H]valine was synthesized in our laboratory; for the subsequent experiments it was produced by Amersham International plc. In the first experiment in rats the two tracers were given by injection and14CO2was collected for 2 h. The oxidation of tritiated valine was significantly higher than that of l-[1-14C]valine. In a second experiment there was no difference. This was probably due to the higher purity of the labelled valine which, for the second experiment, was shown by nuclear magnetic resonance to contain only one tritium atom. In a study with pigs in which the two tracers were given by continuous infusion there was no significant difference between them in flux or oxidation. The results of this experiment were used to evaluate a model to estimate amino acid requirements. With pigs given a methionine-limiting diet a reduction in methionine intake, by reducing protein accretion, increased valine oxidation by the same proportion.