High-performance liquid chromatographic determination of forphenicine in mouse serum and muscle by pre-column fluorescence derivatization using 1,2-diamino-4,5-ethylenedioxybenzene as fluorogenic reagent.
A simple and sensitive high-performance liquid chromatographic method has been developed for the determination of forphenicine in biological samples. Forphenicine in the deproteinized sample is converted by reaction with 1,2-diamino-4,5-ethylenedioxybenzene into a fluorescent derivative. The derivative is separated on a reversed-phase column (TSK gel ODS-120T) by isocratic elution with acetonitrile-30 mM phosphate buffer (pH 6.5) (5:1, v/v) and monitored fluorimetrically. The method allows the quantification of forphenicine in serum (100 microliters) and muscle (0.1 g) of mice dosed with forphenicine or forphenicinol. The limits of detection (signal-to-noise ratio of 3) are 7.35 pmol/ml in serum and 5.36 pmol/g in muscle. The distribution of forphenicine and forphenicinol in the mouse serum and muscle after oral administration of these compounds is also described.
Chao
W F
WF
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Kai
M
M
Ohkura
Y
Y
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes; D005998:Glycine / Q000031:analogs & derivatives / Q000032:analysis / Q000097:blood; D006207:Half-Life; D006863:Hydrogen-Ion Concentration; D007651:Keto Acids / Q000032:analysis; D051379:Mice; D008810:Mice, Inbred C57BL; D009132:Muscles / Q000032:analysis; D010655:Phenylenediamines; D013050:Spectrometry, Fluorescence
D000900:Anti-Bacterial Agents; D005456:Fluorescent Dyes; D007651:Keto Acids; D010655:Phenylenediamines; C016464:forphenicine; C035946:forphenicinol; C058887:1,2-diamino-4,5-ethylenedioxybenzene; D005998:Glycine