Quantitation of p-aminohippuric acid in biological fluids by high-performance liquid chromatography and dual-wavelength ultraviolet detection

A modification of the method of Verbeke J. Chromatogr., 177 (1979) 69] is presented. The fatty tissue is dissolved in hexane, partitioned against methanol-sodium acetate buffer (pH 5.2) and extracted with dichloromethane. The crude extract is then purified on a disposable C18 column. The final extract together with a set of reference compounds is spotted on two opposite sides of a high-performance thin-layer chromatographic plate (10 X 10 cm), which is developed with two different eluents in two opposite directions. This mode of operation has the advantage that more reference compounds can be spotted and that the identification is based on two independent chromatographic runs. Also the total analysis time is decreased.