改良高效液相色谱荧光检测法测定人血浆氢溴酸加兰他敏浓度

Improvement of detection limit in HPLC determination of galantamine in plasma

目的:改进高效液相色谱荧光法测定人血浆中氢溴酸加兰他敏浓度的方法,提高其灵敏度和精确度。方法:采用Hypersil-C_8色谱柱,以乙腈-水(30:70)作为流动相(pH=5.9,含三乙胺0.2％)。碱化血浆后经无水乙醚2次提取,合并2次提取的有机层,挥干乙醚后,残渣经溶有参照标准品阿替洛尔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1～50μg·L^(-1)(r=0.999),最低检测限0.2ng,最低检测浓度为1μg·L^(-1)。日内变异系数为11.86％,日间变异系数为8.40％,萃取回收率和方法回收率均>80％。1μg·L^(-1)的回收率大于60％。结论:本方法精密、准确,适用于加兰他敏的临床检测。

Objective:To improve the detection limit in HPLC determination of galantamine in plasma. Methods:The alkalized plasma sample was extracted with absolute ether for twice, combined the extracts, after evaporation of ether the residue was dissolved by mobile phase with dissolved atenolol reference standard. After treatment,the sample was injected into the hypersil-C8 reverse phase column. The mobile phase consisted of acetonitrile-H2O (30:70) with 0.2% triethylamine and its pH was adjust to 5.9 with HAC,and the fluorescence detector was set as follows;EX 290nm and EM 320nm. Results : A liner range of 1 - 50μg · L -1 ( r = 0. 999) ,a minimum detectable limit of 0. 2ng, a minimum detectable concentration of 1μg·L-1, a recovery of >80% were obtained in this analysis. The intra-day and inter-day RSD were less than 11.86% and 8.4% respectively. Conclusion: A sensitive and accurate method suitable for clinical monitoring of plasma galantamine hydrobromide concentration is developed.