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KMID : 0043319960190050396
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Archives of Pharmacal Research
1996 Volume.19 No. 5 p.396 ~ p.399
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Determination of Theophylline and its Metabolites in Human Urine by High-Performance Liquid Chromatography
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Kim Kyeong-Ho
Park Young-Hwan
Park Hyo-Kyung
Kim Ho-Soon
Lee Min-Hwa
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Abstract
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High-performance liquid chromatographic method with UV detecction was developed for the determination of theophylline and its metabolites in human urine using -hydroxyethyl theophylline as an internal standard. For extraction of urine sample, quality control sample and xanthine-free blank urine were mixed with decylamine (ion-paring reagent) and -HET. After saturation with ammonium sulfate, the mixture was then extracted with organic solvent at pH values of 4.0-4.5. All separations were performed with ion-pair chromatography using decylamine as an ion-pairing reagent and 3mM sodium acetate buffered mobile phase (pH 4.0) containing 1% (v/v) acetonitrile and 0.75 mM decylamine. The detection limits of theophylline, 1, 3-DMU, 1-MU, 3-MX and 1-MX in human urine were 0.17, 0.17, 0.39, 0.19 and 0.19 /ml, based on a signal-to-noise ratios of 3.0. The mean intraday coefficients of variation (C.V.s) of each compound on nine replicates were lower than 2.0%, while mean interday C.V.s on three days were lower than 1.6%. All separations were finished within 40miutes.
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KEYWORD
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Ion-pair chromatography, Theophylline, 1, 3-DMU, 1-MU, 3-MX, 1-MX, HPLC
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