Determination of orotic acid in urine

Abstract

Orotic acid was separated from other urinary constituents by ion-pair formation with tetrabutylammonium, and isocratic elution from a reversed-phase column. Absorbance at 280 nm was recorded for quantitation. Owing to the better column characteristics the separations are somewhat faster, and the sensitivity of the method is higher than those of analogous methods using anion-exchange columns. The method was used for the determination of orotic acid in human urine, in urine of rats with portacaval shunts and in small (30 μl) urine samples from sparse fur mice. Shunted rats excreted ca. 100% more orotic acid per 24 h than sham-operated controls, in spite of their considerably lower body weight. Excessive orotic acid in urine indicates a conditional deficiency of ornithine. Sparse fur mice are congenitally hyperammonemic because of a defective hepatic ornithine carbamoyltransferase. Determination of orotic acid in the urine is a suitable method to identify those animals among litter mates which have the hereditary enzyme defect.