A validated method for the determination of paracetamol and its glucuronide and sulphate metabolites in the urine of HIV+/AIDS patients using wavelength-switching UV detection

Paracetamol is a safe drug which has been used as an in-vivo probe to determine phase II metabolism in a HIV+/AIDS population. Due to the biohazard nature of HIV-infected samples, a high performance liquid chromatography (HPLC) assay which offers minimal sample manipulation and maximal specificity was developed. This reverse-phase HPLC method uses wavelength-switching UV detection for the simultaneous determination of paracetamol and its glucuronide and sulfate metabolites in HIV-infected urine samples. The solvent systems involves a simple isocratic elution with a composition of 50 mM sodium acetate buffer, pH adjusted to 3.5; acetonitrile (96:4 v/v) modified with 0.35% trifluroacetic acid. The validated method is highly reproducible with an inter-assay variation of <7%. This method also shows good precision and sensitivity, making it an ideal assay for phenotyping studies to determine the extent of glucurondiation and sulfation activities. (C) 1998 Elsevier Science B.V. All rights reserved.