高效液相色谱荧光检测法测定人血浆中溴酸加兰他敏浓度

Determination of Galanthamine in Plasma by High-performance Liquid Chromotography with Fluorescent Detection

目的：建立高效液相色谱荧光法测定人血浆中氢澳酸加兰他敏浓度的方法。方法：采用ｈｙｐｅｒｓｉｌ－Ｃ８色谱柱，以乙腈：水：三乙胺（２５：７４：ｌ）作为流动相（ＰＨ＝７．０）。碱化血浆后经无水乙醚提取，挥干乙醚后，残渣经溶有参照标准品舒必利的流动相溶解后进行测定。荧光检测的激发波长为２９０ＮＭ，发射波长为３２０ｎｍ。结果：本法测定氢溴酸加兰他敏的线性范围为５～２５０μｇ．Ｌ－１（ｒ＝０．９９９），最低检测限１ｎｇ，最低检测浓度为５μｇ．Ｌ－１。日内变异系数为８．９２％，日间变异系数为８．７９％，萃取回收率和方法回收率均＞９０％。结论：本方法精密、准确、稳定，适用于加兰他敏的临床检测和药代动力学研究。

OBJECTIVE: To establishe the valid detection method with the high- performance liquid chromatography with fluorescent detection for the determination of concentration of galanthaniine in human plasma. METHODS: sulpiride was used as the reference compound. The extracts of samples were chromatographed on a hypersil C8 reversed-phase column after one-step liquid-liquid extraction by Diethyl ether absolute and monitered by fluorescence-detection and the EX290 and EM320nm. The mobile phase consists of acetonitrile: 20: triethylamine(25:74:1, pH=7.7) at a flow -rate of lml.min^(-1) RESULTS: The calibration curve was linear over the range of 5~25Oμg.L^（-1）（r=0.999). The intra-day RSD was less than 8.92%, and the inter-day was less than 8.79% . The recovery was more than 90% and the limit of quantificaiton was 5μg.L^（-1）CONCLUSION: The method has good selevtivity, sensitivity and reproducibility and has be proved to be suitable for pharmacokinetic studies of galanthamine in humans, and the clinical monitoring the plasma level of drug.