A sensitive and efficient method was developed for the determination of carvedilol and its metabolites in human urine by gas chromatography-mass spectrometry (GC-MS). Urine samples were hydrolyzed with beta-glucuronidase/arylsulfatase (from Helix pomatia) and the target compounds were extracted with liquid-liquid extraction. The extracts were completely derivatized with MSTFA and MBTFA and analyzed by GC-MS using an Ultra-2 column. The linearity of the assay ranges were 0.75-75 ngmL(-1) for carvedilol and o-desmethyl carvedilol (o-DMC), and 3.0-75 ngmL(-1) for 4-hydroxyphenyl carvedilol (4-HPC) and 5-hydroxyphenyl carvedilol (5-HPC). The absolute recovery of carvedilol and its metabolites added to a blank urine sample was 80.1-97.8%. The limits of detection (LOD) and quantitation (LOQ) of carvedilol and o-DMC were 0.30 and 0.75 ngmL(-1), and its of 4-HPC and 5-HPC were 0.75 and 3.0 ngmL(-1), respectively. The reproducibilities were 1.86-11.5% for the intra-day assay, and 0.70-1.71% for the inter-day assay precision and the degree of inaccuracy was -3.0 to 3.9% at the concentration of 75 ngmL(-1). The proposed GC-MS method was effective for the determination of carvedilol and its three metabolites in human urine.
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| http://dx.doi.org/10.1016/j.jchromb.2005.05.023 | DOI Listing |
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