HPLC-MS/MS法同时测定人血清中8种酪氨酸激酶抑制剂的浓度  

Simultaneous determination of 8 tyrosine-kinase inhibitors in human serum by HPLC-MS/MS

作　　者：黄伊婷 成晓亮 李真宝 沈爱宗 HUANG Yi-ting;CHENG Xiao-liang;LI Zhen-bao;SHEN Ai-zong(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Nanjing Pinsheng Medical Technology Co.Ltd.,Jiangsu Nanjing 211500,China;Department of Pharmacy,First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,University of Science and Technology of China,Anhui Hefei 230001,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]南京品生医疗科技有限公司,江苏南京211500 [3]中国科学技术大学附属第一医院(安徽省立医院)药剂科,安徽合肥230001

出　　处：《中国医院药学杂志》2022年第3期235-241,共7页Chinese Journal of Hospital Pharmacy

基　　金：2020年度重大新药创制科技重大专项(编号:2020ZX09201004-007)。

摘　　要：目的:建立一种高效液相色谱-质谱法(HPLC-MS/MS)同时检测人血清中8种酪氨酸激酶抑制剂(TKI)和1种活性代谢产物的浓度,并应用于临床检测。方法:以甲醇为沉淀剂将被分析物与生物基质分离,采用Kinetex XB-C_(18)色谱柱(3.0 mm×50 mm,2.6μm)进行色谱分离,0.1%甲酸水溶液与乙腈溶液进行梯度洗脱,流速为0.3 mL·min^(-1),柱温40℃,进样量5μL;采用电喷雾正电离模式多反应监测进行检测和定量。结果:本方法色谱分离和质谱检测的单针分析时长为6.5 min,特异性良好,待测化合物阿法替尼、阿帕替尼、克唑替尼、吉非替尼、埃克替尼、伊马替尼、厄洛替尼、舒尼替尼以及N-去乙基舒尼替尼在0.4~200,0.8~400,1~500,4~2000,2~1000,4~2000,8~4000,0.4~200,0.4~200 ng·mL^(-1)范围内线性关系良好,相关系数均大于0.999。精密度和准确度、基质效应、回收率和稳定性均符合2020年版《中国药典》要求。结论:该方法专属性强、灵敏度高,可用于上述8种TKI药物和1种活性代谢产物的临床血药浓度测定。OBJECTIVE To establish and validate an HPLC-MS/MS method to simultaneously detect 8 tyrosine-kinase inhibitors(TKIs)and one of their metabolites in human serum,so as to apply it to clinical detection.METHODS The analytes were separated from the biological matrix by protein precipitation.Gradient elution was performed on a Kinetex XB-C_(18) chromatographic column(3.0mm×50 mm,2.6μm)with formic acid aqueous solution(0.1%)and acetonitrile as the mobile phase at a flow rate of 0.3 ml min^(-1),column temperature of 40℃and injection volume of 5μL.Mass spectrometry analysis was carried out in an electrospray(ESI+)interface and multiple reaction monitoring mode.RESULTS The total operation time for the chromatography separation and mass spectroscopy detection was 6.5 min per injection.Calibration ranges of afatinib,apatinib,czetinib,giefitinib,ecetinib,imatinib,erlotinib,sunitinib and N-desunitinib were 0.4-200,0.8-400,1-500,4-2000,2-1000,4-2000,8-4000,0.4-200 and 0.4-200 ng mL^(-1),respectively,and each was of good linearity with a correlation coefficient greater than 0.999.The precision,accuracy,matrix,effect,recovery rate and stability of all those TKIs complied with the official guideline.CONCLUSION This analytical method for the 8 TKIs and one of their metabolites was demonstrated to be fast,simple to operate,and highly capable for blood concentration monitoring of corresponding tyrosine-kinase inhibitors,and may greatly improve the clinical treatment effects.

关 键 词：高效液相色谱-串联质谱 酪氨酸激酶抑制剂 血药浓度检测 血清 

分 类 号：R917[医药卫生—药物分析学]