超高效液相色谱-串联质谱法测定Beagle犬血浆中抗肿瘤新药SM-1的方法研究及验证  

Establishment and validation of UPLC-MS/MS method for determination of an innovative antitumor drug SM-1 in plasma of Beagle dogs

作　　者：刘淑洁 于敏[1] 王宇 黄舒佳 淡墨[1] 李佐刚[1] 耿兴超[1] 张河战[1] 杨进波 刘丽[1] LIU Shu-jie;YU Min;WANG Yu;HUANG Shu-jia;DAN Mo;LI Zuo-gang;GENG Xing-chao;ZHANG He-zhan;YANG Jin-bo;LIU Li(National Center for Safety Evaluation of Drugs,National Institute of Food and Drug Control,Beijing 100176,China;Centre for Drug Evaluation,National Medical Products Administration,Beijing 100022,China)

机构地区：[1]中国食品药品检定研究院,北京100176 [2]国家药品监督管理局药品审评中心,北京100022

出　　处：《中国新药杂志》2020年第12期1349-1354,共6页Chinese Journal of New Drugs

基　　金：国家“重大新药创制”科技重大专项资助项目(2012ZX09103101-051)。

摘　　要：目的:SM-1为抗肿瘤创新药物,已经进入临床前研究。本研究建立并优化了超高效液相色谱-串联质谱法(ultra high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)测定Beagle犬血浆中SM-1的分析方法,按照《生物样品定量分析指导原则》开展系统的方法学验证。方法:本研究采用蛋白沉淀方法对Beagle犬血浆样品中SM-1有效提取,采用Thermo Accucore C18色谱柱,以80%乙腈/甲醇溶液-0.1%甲酸/2 mmol·L^-1甲酸铵水溶液为流动相进行梯度洗脱分离,流速为0.3 mL·min^-1。进而采用电喷雾离子化电离源(ESI)、正离子检测模式和选择反应监测(SRM),以m/z 407.130→203.000(SM-1)和m/z 435.200→397.300(IS)为定量离子反应进行定量分析。结果:本方法在15~10000 ng·mL^-1定量范围内线性关系良好,准确度、精密度、稀释可靠性、选择性、残留、基质效应均符合要求,SM-1 Beagle犬血浆样品在室温放置6 h、-70℃条件下冻存37 d、经过4次冻融均稳定,样品提取液在自动进样器保存26 h稳定。结论:本研究建立并验证了测定Beagle犬血浆中SM-1浓度的方法,本方法操作简便、重现性好,为SM-1药动学、毒动学研究提供了良好的生物分析方法。Objective:To establish an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of an innovative antitumor drug SM-1 in Beagle dog plasma,which is now undergoing preclinical studies.Methods:Methanol was added in the samples for protein precipitation.A Thermo Accucore C18 chromatographic column was employed for separation.The mixture of 80%acetonitrile-methanol and water(containing 0.1%formic acid and 2 mmol·L^-1 ammonium formate)was used as mobile phases gradiently eluted at a flow of 0.3 mL·min^-1.Electrospray was operated in positive ionization mode and the analytes were identified by selected reaction monitoring(SRM)mode.The monitoring ion reactions were set as m/z 407.130→203.000(SM-1)and m/z 435.200→397.300(IS),respectively.Results:The validation results showed that the linear relationship was qualified in the range of 15~10000 ng·mL^-1.The accuracy,precision,dilution reliability,selectivity,residue,and matrix effect well met the requirements for biological samples detection.The stability of SM-1 in Beagle dog plasma was proved after being placed at room temperature for 6 h,frozen at-70℃for 37 d,or treated with 4 freeze-thaw cycles,and it was also stable after being placed in autosampler for 26 h post-extraction.Conclusion:This study established and validated a method for determination of SM-1 in Beagle dog plasma.The method meets the requirements of biological sample detection and is suitable for the pharmacokinetics and toxicity study of SM-1.

关 键 词：超高液相色谱-串联质谱法 抗肿瘤创新药 BEAGLE犬 血浆 蛋白沉淀 方法学验证 

分 类 号：R932[医药卫生—生药学]