High-performance liquid chromatographic analysis of phenobarbital and phenobarbital metabolites in human urine.
A HPLC assay using UV detection and post-column alkalinization was developed to quantify possible urinary excretion products of phenobarbital in human urine. After filtration the urine was injected directly onto the HPLC column for analysis of phenobarbital, p-hydroxyphenobarbital, phenobarbital N-glucosides and phenobarbital N-glucuronides. The accuracy and precision of the assay were within +/- 15% and the limit of detection (LOD) was 1 microM, suitable for pharmacokinetic studies. Phenobarbital was administered orally to five male subjects and urine was collected for a period of 96-108 h. Phenobarbital, p-hydroxyphenobarbital, and phenobarbital N-glucosides were detected and quantified in the urine of all five subjects. The phenobarbital N-glucuronides were not detected in the urine. This assay provides a rapid method with improved selectivity to analyze urine for phenobarbital and its metabolites.
Paibir
S G
SG
Department of Medicinal Chemistry, School of Pharmacy, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0540, USA.
Soine
W H
WH
D000328:Adult; D002851:Chromatography, High Pressure Liquid; D005965:Glucuronates / Q000652:urine; D006801:Humans; D008297:Male; D008875:Middle Aged; D010634:Phenobarbital / Q000031:analogs & derivatives / Q000652:urine*; D012680:Sensitivity and Specificity
D005965:Glucuronates; C017414:4-hydroxyphenobarbital; C043855:phenobarbital-N-glucoside; D010634:Phenobarbital