Simple and selective assay of 4-hydroxymephenytoin in human urine using solid-phase extraction and high-performance liquid chromatography with electrochemical detection and its preliminary application to phenotyping test
| Autor: | Kinuyo Kato, Sunao Kaneko, Koichi Otani, Makoto Tanaka, Kan Chiba, Kazunobu Sugawara, Hideo Hakusui, Takashi Ishizaki, Kenichi Sudo, Tadashi Ohkubo |
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| Rok vydání: | 1996 |
| Předmět: |
Detection limit
Adult Male Working electrode Chromatography Chemistry General Chemistry Urine High-performance liquid chromatography Rats Coulometry Solutions Phenotype Calibration Electrochemistry Animals Humans Anticonvulsants Indicators and Reagents Mephenytoin Solid phase extraction Gas chromatography Quantitative analysis (chemistry) Chromatography High Pressure Liquid |
| Zdroj: | Journal of chromatography. B, Biomedical applications. 676(1) |
| ISSN: | 1572-6495 |
| Popis: | A simple and selective HPLC method for the determination of 4-hydroxymephenytoin (4-OH-M) in human urine, using a controlled potential coulometric detector equipped with a dual working electrode cell of fully porous graphite, has been developed. After acid hydrolysis of urine, 4-OH-M and the internal standard (I.S.), 5-hydroxy-1-tetralone, were extracted from urine by means of a Bond Elut Certify LRC column. The extracts were chromatographed on a reversed-phase μBondapak C18 column using methanol-50 mM KH2PO4 (pH 4.0) (30:70, v/v) as the mobile phase at a flow-rate of 1.0 ml/min. Electrochemical detection at applied potential of 800 mV resulted in a limit of quantitation of 0.76 μg/ml. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity. The present method was applied to the phenotyping test in thirteen Japanese healthy volunteers who recieved an oral 10-mg racemic mephenytoin. The phenotypes determined by the present method were found to be in agreement with those obtained with the reported customary assay based on gas chromatography. |
| Databáze: | OpenAIRE |
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