HPLC法检测水牛血浆中吡喹酮的研究

A high-performance liquid chromatographic method for determination of praziquantel in water buffalo plasma

关键词:  吡喹酮  高效液相色谱  水牛

Key words:  Praziquantel  HPLC  Water buffalo

摘要:

【摘要】:建立了一种简便、灵敏、专属、可重复的高效液相色谱法检测水牛血浆中的吡喹酮含量。吡喹酮在C18 (250mm × 4.0 mm, 5μm particle size ) 柱中可以很好的与杂质峰分离开。其保留时间、紫外检测波长、流动相、流速、色谱柱温分别为10.6min、220nm、甲醇：水(70:30, v/v)、0.8 ml/min和40℃。水牛血浆样品（0.5ml）经乙酸乙酯（3 ml）两步提取后，采用旋转蒸发仪蒸干，1 ml流动相溶解定容、取上清滤液20 μl进样作HPLC分析。吡喹酮的血浆药物浓度在0.0625-8 μg/ ml范围内，呈现良好的线性 关系，相关系数r>0.99。在0.0625、1、32μg/ ml的3个药物浓度水平下，PZQ的平均回收率均在90%以上。日内和日间变异系数小于5%。PZQ的LOD可达0.01μg/ ml。该方法的专属性强、灵敏度高，适用于水牛血浆中PZQ的药动学检测。

Abstract:

【Abstract】: A simple, sensitive, selective and reproducible method based on a reversed-phase chromatography was developed for the determination of praziquantel in water buffalo plasma. Praziquantel was separated from the impurities on a C18 column (250mm × 4.0 mm, 5μm particle size),with retention times of 10.6min. Ultraviolet detection was set at 220nm. The mobile phase consisted of methanol and distilled water (70:30, v/v), running through the column at a flow rate of 0.8 ml/min. The chromatographic analysis was operated at 40 ℃. Sample preparation (0.5ml plasma) was done by a single step liquid–liquid extraction with the ethyl acetate. Calibration curves in plasma at the concentrations ( 0.0625~8μg/ ml) were all linear with correlation coefficients better than 0.999. The precision of the method based on within-day repeatability and reproducibility (day-to-day variation) was below 5%.Limit of detection (LOD) was accepted as 0.01μg using 1ml samples. The method is applicable to water buffalo plasma pharmacokinetics of PZQ . 【Keywords】:Praziquantel; HPLC; Water buffalo