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.
Di Girolamo
A
A
Département de Pharmacologie, Faculté de Médecine, Université de Montréal, Quebec, Canada.
O'Neill
W M
WM
Wainer
I W
IW
D000082:Acetaminophen / Q000378:metabolism / Q000652:urine*; D000097:Acetonitriles / Q000737:chemistry; D000163:Acquired Immunodeficiency Syndrome / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005965:Glucuronates / Q000652:urine*; D006679:HIV Seropositivity / Q000652:urine*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010641:Phenotype; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019346:Sodium Acetate / Q000737:chemistry; D013431:Sulfates / Q000652:urine*; D014269:Trifluoroacetic Acid / Q000737:chemistry; D014466:Ultraviolet Rays
D000097:Acetonitriles; D005965:Glucuronates; D013431:Sulfates; D000082:Acetaminophen; D019346:Sodium Acetate; D014269:Trifluoroacetic Acid; C032159:acetonitrile