Determination of Phenytoin and Its Major Metabolites in Human Serum by High-Performance Liquid Chromatography with Fluorescence Detection.
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- HARA Shuuji
- Faculty of Pharmaceutical Sciences, Fukuoka University
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- HAGIWARA Jun
- Faculty of Pharmaceutical Sciences, Fukuoka University
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- ONO Nobufumi
- Faculty of Pharmaceutical Sciences, Fukuoka University
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- KURODA Takeshi
- Faculty of Pharmaceutical Sciences, Fukuoka University
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Abstract
A highly sensitive fluorometric high-performance liquid chromatographic method was developed for the simultaneous determination of phenytoin and its major metabolites [5-(3-hydroxyphenyl)-5-phenylhydantoin and 5-(4-hydroxyphenyl)-5-phenylhydantoin]. After extracting these compounds and 5-(4-methylphenyl)-5-phenylhydantoin (MPPH) as an internal standard from serum (50 μl) with ethyl acetate, they were further converted into the corresponding fluorescent derivatives by a reaction with 3-bromomethyl-6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone in the presence of potassium hydrogen carbonate and dibenzo-18-crown-6 in acetonitrile. The derivatives were separated by reversed-phase chromatography on a YMC-Pack ODS-A column with a mixture of acetonitrile?50 mM phosphate buffer (pH 7.0) (4:6, v/v) as a mobile phase, and were then detected spectrofluorometrically at 448 nm with excitation at 365 nm. The detection limits for phenytoin, 5-(3-hydroxyphenyl)-5-phenylhydantoin and 5-(4-hydroxyphenyl)-5-phenylhydantoin added to serum were 0.6, 3.0 and 0.8 ng (2.4, 11 and 3.1 pmol) ml-1 serum at a signal-to-noise ratio of three. The method was applied to determine the unbound- and total-phenytoin and the metabolites levels in the serum obtained from two healthy volunteers, after oral administration of the drug.
Journal
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- Analytical Sciences
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Analytical Sciences 15 (4), 371-375, 1999
The Japan Society for Analytical Chemistry
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Details
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- CRID
- 1390282679232051200
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- NII Article ID
- 10002419445
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- NII Book ID
- AA10500785
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- COI
- 1:CAS:528:DyaK1MXisVSntLk%3D
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- ISSN
- 13482246
- 09106340
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- NDL BIB ID
- 4712465
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- Web Site
- https://ndlsearch.ndl.go.jp/books/R000000004-I4712465
- https://link.springer.com/content/pdf/10.2116/analsci.15.371.pdf
- https://link.springer.com/article/10.2116/analsci.15.371/fulltext.html
- http://www.jstage.jst.go.jp/article/analsci/15/4/15_4_371/_pdf
- https://search.jamas.or.jp/link/ui/1999181190
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed