摘要
本研究基于自主研制的QLIT-6610MD液相色谱-四极杆-线形离子阱-质谱联用仪,建立了准确测量血清中甲氨蝶呤的同位素稀释质谱法。采用甲醇沉淀血清样品的蛋白后测定上清液。通过基质匹配标准曲线定量,在10~5000μg/L浓度范围内,甲氨蝶呤的线性系数R^(2)>0.9990,定量限为10μg/L,质控品精密度优于6.94%,血清样本添加回收率为86.39%~97.84%,变异系数(CV)≤8.04%。应用QLIT-6610MD与AB QTRAP 6500+液相色谱-质谱联用仪同时测定70例临床血清样本,并采用Pearson系数与BLand-ALtman法进行比较。结果表明,2台仪器所测结果一致,QLIT-6610MD可达到三重四极杆质谱仪定量测定血清中甲氨蝶呤的水平,能够满足临床诊断需求,为临床质谱分析提供了一种新的国产仪器选择。此外,本研究还初步探讨了多级碎裂技术(MSn)测定血清中甲氨蝶呤的可能性,为后续研究奠定基础。
Methotrexate(MTX)is a therapeutic drug that is widely used in clinic for treatment of a variety of cancers.However,MTX has limited selectivity and serious cytotoxicity,at the same time,there are problems such as narrow therapeutic window and large individual variability in metabolism and excretion of MTX.Therefore,blood concentration monitoring is required for MTX administration in clinical practice.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)has the advantages of strong specificity,good separation characteristics,high sensitivity and fast detection speed,which has become the internationally recognized“gold standard”for blood concentration monitoring of drugs.In this study,an isotope dilution mass spectrometry method was developed for accurate measurement of MTX in serum by the house-made QLIT-6610MD quadrupole-linear ion trap liquid chromatography-mass spectrometer.Unlike the traditional triple quadrupole QqQ tandem mass spectrometer,the quadrupole-linear ion trap(Q-LIT)mass spectrometer adopts the quadrupole-linear ion trap direct axial coupling design,i.e.,a single linear ion trap is used to replace the QqQ tandem mass spectrometer’s back-end fragmentation cell and the quadrupole,and no other components are installed between the quadrupole and linear ion trap,which greatly streamlines the quality and efficiency of the mass spectrometer while ensuring the original performance of the instrument.With this unique configuration,the QLIT-6610MD instrument uses a simultaneous fragmentation and accumulation technique,which remarkedly reduces matrix-induced interferences and effectively reduces space charge effects.Serum samples were pretreated with methanol to precipitate proteins and the supernatants were measured by QLIT-6610MD spectrometer.The linear correlation coefficient(R^(2)>0.9990)is obtained for MTX in the range of 10-5000μg/L.The limit of quantification is 10μg/L.The precision of the quality control is better than 6.94%,and the recoveries of spiked serum samples are 86.39%-97.84%with a c
作者
刘粮泽
谢洁
屈子裕
易可可
张谛
黄泽建
鄢丹
戴新华
方向
时正媛
江游
俞晓平
LIU Liang-ze;XIE Jie;QU Zi-yu;YI Ke-ke;ZHANG Di;HUANG Ze-jian;YAN Dan;DAI Xin-hua;FANG Xiang;SHI Zheng-yuan;JIANG You;YU Xiao-ping(College of Life Sciences,China Jiliang University,Hangzhou 310018,China;Technology Innovation Center of Mass Spectrometry for State Market Regulation,Center for Advanced Measurement Science,National Institute of Metrology,Beijing 100029,China;Beijing Shijitan Hospital,Capital Medical University,Beijing 100038)
出处
《质谱学报》
EI
CAS
CSCD
北大核心
2024年第3期364-374,I0002,共12页
Journal of Chinese Mass Spectrometry Society
基金
国家重点研发计划--重大科学仪器设备开发专项(2020YFF01014600)
国家自然科学基金_国家重大科研仪器研制项目(21927812)。
