Biological & Pharmaceutical Bulletin 2006-12-01

Sensitive determination of 4-(4-bromophenyl)-4-hydroxypiperidine, a metabolite of bromperidol, in rat plasma by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole.

Yasuhiko Higashi, Shota Nakamura, Youichi Fujii

Index: Biol. Pharm. Bull. 29(12) , 2479-82, (2006)

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Abstract

The purpose of this study was to determine the level of 4-(4-bromophenyl)-4-hydroxypiperidine (BPHP), a bromperidol (BRO) metabolite, in rat plasma by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). After basic extraction of the samples with benzene, derivatization with NBD-F was conducted in borate buffer (pH 8.0) at 60 degrees C for 3 min. Mexiletine was utilized through the procedure as an internal standard (IS). Retention times of the BPHP and IS derivatives were 7.7 and 11.5 min, respectively. The regression equation for BPHP showed good linearity in the range of 0.01-1 mg/ml with the detection limit of 0.003 microg/ml. The coefficient of variation was less than 12.0%. The recovery was satisfactory. This method was applied for a pharmacokinetic study of BPHP in comparison with 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP), the corresponding haloperidol (HAL) metabolite, in rats. The ratio of the area under the plasma concentration curve (AUC) after p.o. administration of BPHP to the AUC after i.p. administration of BPHP (46%) was lower than that of CPHP (56%), indicating that intestinal absorption of BPHP is lower than that of CPHP. The ratio of BRO metabolism to BPHP (48%) was 1.8-fold higher than that of HAL metabolism to CPHP (27%); the ratio was estimated as (AUCp.o.,A-->B/AUCp.o.,B)x100, where AUCp.o.,A-->B is the AUC value of BPHP or CPHP after p.o. administration of BRO or HAL, and AUCp.o.,B is the AUC of BPHP or CPHP after administration of BPHP or CPHP, respectively. Our method provides a sensitive procedure for determination of BPHP in rat plasma and is suitable for pharmacokinetic studies of BPHP after BRO administration.