质谱学报2024，Vol.45Issue(3)：364-374,11.DOI:10.7538/zpxb.2023.0082
自主研制四极杆-线形离子阱质谱仪测定血清中甲氨蝶呤
Determination of Methotrexate in Serum by House-Made Quadrupole-Linear Ion Trap Mass Spectrometer
摘要
Abstract
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(R2＞0.999 0)is obtained for MTX in the range of 10-5 000 μg/L.The limit of quantification is 10 μg/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 coefficient of variation less than 8.04%.We used the QLIT-6610MD and AB QTRAP 6500+ liquid chromatography-mass spectrometers to analyze 70 clinical serum samples for comparison of the performance of the house-made instrument by means of Pearson's coefficient and BLand-ALtman method.The results showed that the QLIT-6610MD mass spectrometry achieves a similar level of performance to AB ATRAP 6500+ instrument,and provides a new domestic instrument option for clinical mass spectrometry.In addition,this study explores the possibility applying the multi-stage fragmentation technique(MSn)to determine MTX in serum,gaining fundamental data for future studies.关键词
液相色谱-串联质谱(LC-MS/MS)/同位素稀释质谱/四极杆/线形离子阱/甲氨蝶呤(MTX)Key words
liquid chromatography-tandem mass spectrometry(LC-MS/MS)/isotope dilution mass spectrometry/quadrupole/linear ion trap/methotrexate(MTX)分类
化学化工引用本文复制引用
刘粮泽,时正媛,江游,俞晓平,谢洁,屈子裕,易可可,张谛,黄泽建,鄢丹,戴新华,方向..自主研制四极杆-线形离子阱质谱仪测定血清中甲氨蝶呤[J].质谱学报,2024,45(3):364-374,11.基金项目
国家重点研发计划——重大科学仪器设备开发专项(2020YFF01014600) （2020YFF01014600）
国家自然科学基金——国家重大科研仪器研制项目(21927812) （21927812）
