Determination of benzoic acid and hippuric acid in human plasma and urine by high-performance liquid chromatography

Abstract

For patients with inborn errors of urea synthesis, oral administration of sodium benzoate is the usual treatment to increase the nitrogen excretion. Thus, monitoring hippuric acid and benzoic acid simultaneously in human biological fluids is considered to be clinically important. We developed a simple and accurate high-performance liquid chromatographic method for the simultaneous determination of hippuric acid and benzoic acid in human plasma and urine. This method requires no extraction step. Aliquots of urine and plasma are added to a solution of internal standard (o-chlorobenzoic acid) in acetonitrile and directly injected onto a reversed-phase column using an acidic (pH 2.7) eluent and ultraviolet detection at 235 nm. The preliminary plasma concentration-time and urinary excretion rate-time profiles of hippuric acid and benzoic acid from a healthy subject receiving small, medium and large doses of sodium benzoate are reported.