High-performance liquid chromatographic assay for theophylline and its major metabolites in human urine.
A highly selective automated high-performance liquid chromatographic (HPLC) method has been developed for the assay of theophylline and its major metabolites in urine. The method involves direct injection of urine on to a reversed-phase column, followed by gradient elution and ultraviolet detection. Quantitation is achieved by the peak-height ratio method with reference to an internal standard, 8-chlorotheophylline. The assay is accurate and reproducible, with a sensitivity of 1 microgram/ml in urine. In order to confirm its accuracy, theophylline and its major metabolites in urine were quantified after HPLC separation by both peak-height ratio of ultraviolet absorbance and liquid scintillation spectrometry after oral administration of [14C]theophylline to a human volunteer. The assay was also applied to the analysis of theophylline and its major metabolites in urine following the oral administration of theophylline to three healthy volunteers.
Department of Pharmacology, St. Mary's Hospital Medical School, London, U.K.
S A
SA
Hotchkiss
J
J
Caldwell
D000284:Administration, Oral; D000328:Adult; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008297:Male; D013056:Spectrophotometry, Ultraviolet; D013806:Theophylline / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000652:urine*
D013806:Theophylline