LC-MS/MS法测定人血浆中福大赛因的浓度  

Determination of photocyanine in human plasma by LC-MS/MS

作　　者：郑昕 张瑞琪 崔馨戈 杨屹[2] 江骥 ZHENG Xin;ZHANG Rui-qi;CUI Xin-ge;YANG Yi;JIANG Ji(Clinical Pharmacology Research Center, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing Key Laboratory of Clinical PK and PD Investigation for Innovation Drugs, Beijing 100032, China;College of Science, Beijing University of Chemical Technology, Beijing 100029, China)

机构地区：[1]中国医学科学院北京协和医院临床药理研究中心创新药物临床PK/PD研究北京市重点实验室,北京100032 [2]北京化工大学理学院,北京100029

出　　处：《中国新药杂志》2018年第10期1130-1136,共7页Chinese Journal of New Drugs

基　　金：国家"十三五"重大新药创制专项资助项目(2018ZX09734006-001);国家自然科学基金青年基金(81503164)

摘　　要：目的:建立LC-MS/MS法测定人血浆中福大赛因(photocyanine)的浓度,用于晚期食管癌患者的药动学研究。方法:以Zn Pc-COOH作为内标,用甲醇进行蛋白沉淀处理后,使用稀释液[甲醇-二甲基甲酰胺(3∶2)]进行稀释,吸取10μL质谱进样分析。色谱柱为XBridge BEH 130 C_(18)柱(50 mm×2.1 mm,3.5μm),流动相为甲醇-0.1%氨水溶液,梯度洗脱,流速0.8 m L·min^(-1),柱温为40℃。使用电喷雾离子源进行负离子多反应监测扫描分析。福大赛因和内标化合物的检测离子对的质荷比分别为:526.0→146.0和621.2→579.0。对该方法的特异性、标准曲线与定量下限、准确度和精密度、萃取回收率和基质效应、残留效应和稳定性等进行了全面的考察。结果:血浆中福大赛因在20~4 000 ng·mL^(-1)呈良好的线性关系(r>0.990),定量下限为20 ng·mL^(-1)。在目标化合物出峰时间处,未观察到基质的内源性成分干扰。福大赛因的日内、日间的相对标准差(RSD)均小于15%;萃取回收率为87.2%~90.2%;基质效应为130.5%~138.7%。结论:LC-MS/MS法快速、灵敏、准确、选择性强、重复性好,方法考核结果均符合FDA和CFDA指导原则的要求,适用于人血浆中福大赛因的浓度测定,可以满足临床药物浓度监测以及药动学研究的需要。Objective： To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS） method for determining the concentrations of photocyanine in human plasma for a phase I clinical pharmacokinetic study in Chinese advanced esophageal cancer patients. Methods： The samples were purified by protein precipitation and Zn Pc-COOH was added as the internal standard(IS）. The optimal chromatographic behavior was performed on an XBridge BEH 130 C18(50 mm × 2. 1 mm,3. 5 μm） column under a gradient elution. The column temperature was set to be 40 ℃. The mobile phase consisted of 0. 1% ammonia aqueous solution(A） and methanol(B） and the flow rate was set to be 0. 80 mL·min^-1. The analytes were detected on a tandem mass spectrometer equipped with an electrospray ionization source and the data were acquired in thenegative multiple reaction monitoring mode(MRM）,using the transitions of m/z 526. 0→m/z 146. 0 for photocyanine and m/z 621. 2→m/z 579. 0 for IS. The selectivity,standard curve,precision and accuracy,extraction recoveries,matrix effect,carry-over effect and stability were fully validated. Results： The calibration curve exhibited excellent linearity over the concentration range of 20 4 000 ng·mL^-1(r 〉 0. 995）,and the lowest limit of quantitation(LLOQ） was 20 ng·mL^-1. No endogenous interference was observed at the retention time of the analytes and the carry-over effect from previous concentrated samples was found to be negligible. The intra-day and inter-day relative standard deviations(RSD） of the LLOQ,low,medium and high concentrations of photocyanine were all less than15%. The average extraction recovery rates of the low,medium and high concentration of quality control samples were 90. 2%,89. 4% and 87. 2%,respectively. The average matrix effects were 138. 7%,129. 8% and 130. 5%,respectively. Conclusion： The validation results indicated that the method,which meets all the requirements of FDA and CFDA guidelines,is convenient,sensitive,accurate and su

关 键 词：福大赛因 LC-MS/MS 食管癌 

分 类 号：R969[医药卫生—药理学]