Quantitative analysis of HBY 097 and its metabolites in human serum and urine by HPLC.
Two HPLC methods were developed: one for the quantitation of HBY 097 reverse transcriptase inhibitor and its metabolites M2 and M3 in human serum, and one for the quantitation of metabolite M5 in urine. The HPLC procedure for the quantitation of HBY 097 and its metabolites M2 and M3 in human serum involved protein precipitation with acetonitrile followed by automated on-line trace enrichment. The HPLC procedure for the analysis of metabolite M5 in urine involved enzymatic hydrolysis of urine with beta-glucuronidase to convert metabolite M5 (glucuronide of M3) to M3. Reverse phase chromatographic separation with gradient elution. UV detection at 335 nm, and internal standard were used to quantitate analytes in both procedures. The lower quantitation limits were 25 ng ml-1 for HBY 097 and metabolites M2 and M3 in serum, and 0.5 microgram ml-1 for the metabolite M5 in urine measured as metabolite M3 after hydrolysis. The HBY 097 and metabolite M3 concentrations were specific but metabolite M2 was semi-specific because the two diastereomers of M2 were not resolved by the present chromatographic procedure. Both procedures were applied to the quantitation of HBY 097 and its metabolites in serum and urine of HIV positive patients who were enrolled in a clinical study of drug safety and pharmacokinetics.
Agarwal
V K
VK
Bayer Corporation, West Haven, CT 06516-4174, USA.
Krol
G J
GJ
Krone
V
V
Roberts
D
D
D000998:Antiviral Agents / Q000097:blood* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008956:Models, Chemical; D011810:Quinoxalines; D015203:Reproducibility of Results; D018894:Reverse Transcriptase Inhibitors / Q000097:blood* / Q000652:urine*; D012680:Sensitivity and Specificity
D000998:Antiviral Agents; D011810:Quinoxalines; D018894:Reverse Transcriptase Inhibitors; C093110:HBY 097