摘要
结合外标法或同位素稀释质谱法,建立UPLC-Q-TOF快速定量氨基酸分析方法。在无离子对试剂及衍生化情况下,实现对样品中氨基酸的快速测定。采用外标法测定时,14种常见氨基酸在其相应线性范围内,r2为0.953~0.999,LOD为0.001~0.080 mg/g,CV为1.4%~9.8%;采用Q-TOF外标法定量时,CV为1.5%~8.0%,误差为-9.1%~40%;采用同位素稀释质谱法测定各氨基酸浓度时,CV为1.4%~3.8%,误差为-3.4%~3.7%。本方法可在2min之内完成14种氨基酸的分离和定量,大大缩短分析时间。同时由于采用高分辨的飞行时间质谱,避免检测时离子间的相互干扰,提高测定结果的准确性,可用于蛋白质标准物质均匀性、稳定性检验及定量。
In this work,a new method of UPLC-Q-TOF had been proposed for the rapid and accurate detection of common amino acids. Without derivatization and using ion-pair regent,amino acids were analyzed fast and accurately. This study could be applied with both external standard method and ID-MS. For external standard method,the r2 of each amino acid was from 0.953 to0.999,the LOD was from 0.001 mg/g to 0.080 mg/g and the CV was from 1.4% to 9.8%. For the quantitation of 14 amino acids with the external standard method,the CV was from 1.5% to 8.0%and the bias was from-9.1% to 40% compared with standard values. For ID-MS,the CV was from1.4% to 3.8% and the bias was from-3.4% to 3.7% compared with standard values. 14 common amino acids could be analyzed in 2 minutes,which reduced the analysis time greatly. With highresolution TOF mass spectrometry,this method avoids the interference of ion overlapping,which can improve the accuracy of measurement results,and will be well applied in homogeneity and stability testing of protein certified reference materials.
出处
《中国测试》
CAS
北大核心
2014年第5期70-73,共4页
China Measurement & Test
基金
国家科技支撑计划项目(2012BAK26B04)
国家质检总局科技项目(2013QK048
201310008)
