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Quantitation of promethazine and metabolites in urine samples using on-line solid-phase extraction and column-switchingA chromatographic method for the quantitation of promethazine (PMZ) and its three metabolites in urine employing on-line solid-phase extraction and column-switching has been developed. The column-switching system described here uses an extraction column for the purification of PMZ and its metabolites from a urine matrix. The extraneous matrix interference was removed by flushing the extraction column with a gradient elution. The analytes of interest were then eluted onto an analytical column for further chromatographic separation using a mobile phase of greater solvent strength. This method is specific and sensitive with a range of 3.75-1400 ng/ml for PMZ and 2.5-1400 ng/ml for the metabolites promethazine sulfoxide, monodesmethyl promethazine sulfoxide and monodesmethyl promethazine. The lower limits of quantitation (LLOQ) were 3.75 ng/ml with less than 6.2% C.V. for PMZ and 2.50 ng/ml with less than 11.5% C.V. for metabolites based on a signal-to-noise ratio of 10:1 or greater. The accuracy and precision were within +/- 11.8% in bias and not greater than 5.5% C.V. in intra- and inter-assay precision for PMZ and metabolites. Method robustness was investigated using a Plackett-Burman experimental design. The applicability of the analytical method for pharmacokinetic studies in humans is illustrated.
Document ID
20040088640
Acquisition Source
Johnson Space Center
Document Type
Reprint (Version printed in journal)
Authors
Song, Q.
(Wyle Laboratories, Systems and Services Life Sciences, Houston, TX 77058, United States)
Putcha, L.
Harm, D. L.
Date Acquired
August 21, 2013
Publication Date
November 5, 2001
Publication Information
Publication: Journal of chromatography. B, Biomedical sciences and applications
Volume: 763
Issue: 1-2
ISSN: 1387-2273
Subject Category
Life Sciences (General)
Distribution Limits
Public
Copyright
Other
Keywords
NASA Center JSC
NASA Discipline Regulatory Physiology

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