Determination of Valproic Acid in Human Serum by High-Performance Liquid Chromatography with Fluorescence Detection

A simple and highly sensitive high-performance liquid chromatographic method is described for the determination of valproic acid in human serum. The method is based on the direct derivatization of serum sample with 6, 7-methylenedioxy-1-methyl-2-oxo-1, 2-dihydroquinoxaline-3-ylpropionohydrazide. The...

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Veröffentlicht in:Biological & pharmaceutical bulletin 1999/09/15, Vol.22(9), pp.975-977
Hauptverfasser: HARA, Shuuji, KAMURA, Motoki, INOUE, Kazuo, FUKUZAWA, Misa, ONO, Nobufumi, KURODA, Takeshi
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Sprache:eng
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Zusammenfassung:A simple and highly sensitive high-performance liquid chromatographic method is described for the determination of valproic acid in human serum. The method is based on the direct derivatization of serum sample with 6, 7-methylenedioxy-1-methyl-2-oxo-1, 2-dihydroquinoxaline-3-ylpropionohydrazide. The derivatization reaction proceeds in aqueous solution in the presence of pyridine and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at 37°C. The resulting derivatives were separated on a reversed-phase column (YMC Pack ODS-A) with isocratic elution and were fluorimetrically detected at 440 nm with excitation at 365 nm. The detection limit (signal-to-noise ratio=3) for valproic acid added to serum sample was 0.1 μg (700 fmol)/ml serum sample (2.3 fmol on column). The method was applied to determine the unbound- and total-valproic acid levels in the serum obtained from three healthy volunteers after oral administration of the drug (600 mg).
ISSN:0918-6158
1347-5215
DOI:10.1248/bpb.22.975