血清中美西律的柱前衍生化-液相色谱荧光检测

HPLC-fluorescent spectrometric determination of serum mexiletine concentration after derivatization with fluram

目的 建立血清中盐酸美西律的柱前衍生化 液相色谱荧光检测法。方法 血清样品用丙酮液直接沉淀蛋白后 ,定量加入荧光胺丙酮溶液 ,得盐酸美西律的荧光衍生物。采用AlltimaC1 8(15 0mm× 4 6mm ,5 μm)为分析柱 ;流动相为甲醇 水 2 4mol·L- 1 二乙胺 磷酸缓冲液 (pH 4 0 ) (70∶2 8∶2 ) ;荧光检测激发波长 (Ex)为 392nm ,发射波长(Em)为 4 80nm。盐酸美西律的保留时间为 6 5min。结果 盐酸美西律在 0 10 0～ 6 4 0 0mg·L- 1 线性关系良好。本法的最低检测质量浓度为 5 μg·L- 1 ,(S N≥ 4 )。批内、批间精密度为 1 34%～ 5 31%。结论 本法操作简便、有良好的灵敏度和专一性 。

Aim To establish an HPLC fluorescent spectrometric method for the determination of mexiletine hydrochloride in plasma after derivatization with fluram. Methods Fluram acetone solution was added to the deproteinized plasma with acetone to obtain the derivative of mexiletine. The HPLC method was performed on a column of Allitima C 18 (150 mm×4 6 mm, 5 μm) with the mobile phase of methanol water diethylamine phosphoric acid buffer (2 4 mol·L -1 , pH 4 0) (70∶28∶2), and the detective wavelength were set at E x 392 nm and E m 480 nm. Results Mexiletine has a liner range over the concentration range from 0 100-6 400 mg·L -1 . The lowest detectable concentration of this method was 5 μg·L -1 ( S/N ≥4). The intra day and inter day RSDs were 1 34%-5 31%, respectively. Conclusion This method is simple, selective and can be used for therapeutic drug monitoring (TDM) and pharmacokinetic studies of mexiletine.