DETERMINATION OF A SUBSTANCE-P ANTAGONIST IN HUMAN PLASMA AND URINE USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH ULTRAVIOLET ABSORBENCY AND TANDEM MASS-SPECTROMETRIC DETECTION
J. Zagrobelny et al., DETERMINATION OF A SUBSTANCE-P ANTAGONIST IN HUMAN PLASMA AND URINE USING HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH ULTRAVIOLET ABSORBENCY AND TANDEM MASS-SPECTROMETRIC DETECTION, Journal of pharmaceutical and biomedical analysis, 15(9-10), 1997, pp. 1427-1433
A high-performance liquid chromatographic (HPLC) assay using ultraviol
et (UV) detection was developed and compared with a HPLC method with t
andem mass spectrometric (HPLC/MS-MS) detection for the determination
of a substance P receptor antagonist nyl-4-((3-oxo-1,2,4-triazol-5-yl)
methyl)morpholine (Fig. 1, Ia, L-742 694) in human plasma and urine. T
he drug was isolated from the biological matrix through liquid-liquid
extraction. In the HPLC/UV method, the samples were initially injected
onto a cyano Hypersil column, and the chromatographic region containi
ng the peaks of interest was heart-cut onto an analytical C-18 Hypersi
l column via a column switching device. The analyte was quantified by
monitoring absorbance at 205 nm. The limit of quantification for I ext
racted from 1 ml of plasma or urine was 2.5 ng ml(-1), and the assays
were validated in the concentration range 2.5-500 ng ml(-1). The HPLC/
MS-MS method was validated in the concentration range 0.2-500 ng ml(-1
). Both assays provided data with precision, measured as coefficient o
f variation, better than 10% at all points within the standard curve r
ange and with adequate accuracy. (C) 1997 Published by Elsevier Scienc
e B.V.