质谱学报2025，Vol.46Issue(4)：430-439,10.DOI:10.7538/zpxb.2024.0194
基于四极杆-线形离子阱串联质谱研究髓母细胞瘤氨基酸生物标志物定量分析方法
Quantitative Analysis of Amino Acid Biomarkers for Medulloblastoma Based on Quadrupole-Linear Ion Trap Isotope Dilution Mass Spectrometry
摘要
Abstract
A method of liquid chromatography-tandem mass spectrometry(LC-MS/MS)was developed based on the self-developed QLIT-6610MD quadrupole-linear ion trap liquid chromatography-mass spectrometry for the accurate quantification of amino acid biomarkers including leucine,phenylalanine,tyrosine,and cystine in plasma for medulloblastoma research.Plasma samples were processed by proteins precipitation with isopropanol,and the supernatant was quantified using a QLIT-6610MD quadrupole-linear ion trap tandem mass spectrometer after amino acid derivatization with 6-aminoquinolinyl-N-hydroxysuccinimidylformate(AQC).Leucine,phenylalanine and tyrosine were quantified by an isotope internal standard method,and the linearity is good in the range of 1-400 μmol/L with the determination coefficients(R2)of 0.9964,0.9935,0.9995,respectively,and the limit of quantification(LOQ)is 1 μmol/L.The precision of the quality control samples is less than 15%,and the accuracies are within the range of±14%.Cystine was quantified by an external standard method,the LOQ is 1 μmol/L,and R2 of the target is not less than 0.9979 in the range of 1-400 μmol/L,and the precision of the quality control samples ranges from 1.2%to 11.6%,the accuracy is within the range of-7.4%to 10.8%.Two liquid chromatography-mass spectrometry instruments,QLIT-6610MD and AB QTRAP 6500+,were used to analyze 60 clinical plasma samples,in which the concentrations of leucine,phenylalanine,tyrosine,cystineis are in the range of 17.16-127.06,13.21-57.89,11.14-60.97,4.70-22.97 μmol/L,respectively.Data correlation was analyzed using the Pearson coefficient,and the correlation coefficient(R)between QLIT-6610MD and AB QTRAP 6500+is≥0.996,indicating a significant positive correlation(P<0.01).Data consistency was analyzed using Bland-Altman,and the percentage of data out-of-bounds points is≤10%,indicating that the data consistency is good and meets the actual clinical monitoring needs.关键词
液相色谱-串联质谱(LC-MS/MS)/四极杆-线形离子阱质谱/生物标志物/髓母细胞瘤/氨基酸Key words
liquid chromatography-tandem mass spectrometry(LC-MS/MS)/quadrupole-linear ion trap mass spectrometry/biomarkers/medulloblastoma/amino acids分类
化学化工引用本文复制引用
刘粮泽,谢洁,江游,时正媛,屈子裕,易可可,刘文科,张谛,叶子弘,戴新华,方向..基于四极杆-线形离子阱串联质谱研究髓母细胞瘤氨基酸生物标志物定量分析方法[J].质谱学报,2025,46(4):430-439,10.基金项目
国家重点研发计划-重大科学仪器设备开发专项(2021YFC2401100) （2021YFC2401100）
国家自然科学基金-国家重大科研仪器研制项目(21927812) （21927812）
中国计量科学研究院基本业务费项目(AKYZZ2325) （AKYZZ2325）
