高效液相色谱法测定人血浆样品中的椒苯酮胺  

High performance liquid chromatography method for the determination of JBAT in human plasma sample

作　　者：牟英[1] 孟志云[1] 高文静[1] 窦桂芳[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850

出　　处：《军事医学科学院院刊》2004年第1期64-67,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金资助项目 ( 3 0 3 71669)

摘　　要：目的 :建立椒苯酮胺 (JBAT)在血浆样品中的高效液相色谱测定方法以满足临床前动物药代动力学研究的需要。方法 :血浆样品用乙酸乙酯提取 ,氮气流吹干 ,用pH 3.76的 0 .2 5 3mmol L磷酸缓冲液 80 μl溶解后 ,取 5 0 μl进样分析。SymmetryShieldTM RP1 8分析柱 ,流动相为乙腈、pH 2 .8的 5mmol L磷酸缓冲液和甲醇以一定体积配比 ,流速 1ml min ;紫外检测波长 330nm。结果 :在所用HPLC条件下 ,生物样品中内源性物质不干扰JBAT的测定。用内标法测定JBAT的血浆浓度在 8～ 2 0 0 0 0ng ml范围内线性关系良好 ,最低定量限为 8ng ml,方法的准确度在- 1.0 %～ 5 .9%之间 ,日间精密度为 10 .8%～ 13.2 % ,日内精密度为 4 .5 %～ 9.6 %。方法平均回收率为 70 .2 %。结论 :本方法线性范围广、专属性强、准确、稳定。Objective:To establish a HPLC method for determination of the trace of JBAT in plasma.Methods:Following extracted by ethyl acetate, the samples were separated by a SymmetryShield TM RP 18 column (3.9 mm×150 mm, Waters Corporation) and detected at UV wavelength of 330 nm. A gradient mobile phase composed of methanol, acetonitrile and 5 mmol/L phosphoric acid buffer (pH 2.8) was run at a flow rate of 1 ml/min.Results: A good linearity was obtained over the plasma concentration ranging 8-20 000 ng/ml of JBAT with correlation coefficient from 0.9964 to 0.9986 when the weighted leastsquares method was applied and the limit of quantitation was 8 ng/ml. The precision of inter-day assay and intra-day assay was evaluated by analysis of variance with the result of 10.8%-13.2% and 4.5%-9.6%, respectively. The average extraction recovery was 70.2%.Conclusion:The merit of this method is the wide linear range, exclusive, accurate and stable.

关 键 词：JBAT 高效液相色谱法 血浆 药代动力学 

分 类 号：O657.7[理学—分析化学]