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气相色谱-质谱联用选择离子监测方法定量分析低浓度胞内游离氨基酸的13C同位素丰度
引用本文:李敏超,黄明志,刘玉伟,储炬,庄英萍,张嗣良.气相色谱-质谱联用选择离子监测方法定量分析低浓度胞内游离氨基酸的13C同位素丰度[J].分析化学,2014,42(10):1408-1413.
作者姓名:李敏超  黄明志  刘玉伟  储炬  庄英萍  张嗣良
作者单位:华东理工大学国家生化工程技术研究中心 上海,上海,200237
摘    要:胞内游离氨基酸具有周转快的特点,其13C同位素丰度能快速反映胞内代谢状态的变化。但胞内游离氨基酸的浓度很低,现有的基于气相色谱-质谱联用全扫描模式的13C同位素丰度检测方法不能满足要求。本研究考察理论上检测精度更高的选择离子监测方法在胞内游离氨基酸13C同位素丰度分析中应用的可能性。首先在全扫描模式下分析了不同氨基酸的断裂规律,找出与每种氨基酸对应的特征碎片,建立起包含有16种胞内游离氨基酸的特征碎片库。利用此特征碎片库,在样品分析时只需检测特定m/z处的信号,从而实现选择离子监测,提高信号质量。对标准品的检测结果表明,与全扫描模式相比,本方法的信噪比、测量精度和准确性分别提高了17倍、2倍和3.8倍。在对辅酶Q10生产菌株样品的分析中,本方法成功检测出8种胞内游离氨基酸的同位素丰度。

关 键 词:选择离子监测  13C同位素丰度  气相色谱-质谱联用  游离氨基酸
收稿时间:25 April 2014

Accurate Determination of 13 C Isotopic Abundances of Free Intracellular Amino acids with Low Concentration by GC-MS-Selective Ion Monitoring Method
LI Min-Chao,HUANG Ming-Zhi,LIU Yu-Wei,CHU Ju,ZHUANG Ying-Ping,ZHANG Si-Liang.Accurate Determination of 13 C Isotopic Abundances of Free Intracellular Amino acids with Low Concentration by GC-MS-Selective Ion Monitoring Method[J].Chinese Journal of Analytical Chemistry,2014,42(10):1408-1413.
Authors:LI Min-Chao  HUANG Ming-Zhi  LIU Yu-Wei  CHU Ju  ZHUANG Ying-Ping  ZHANG Si-Liang
Institution:LI Min-Chao;HUANG Ming-Zhi;LIU Yu-Wei;CHU Ju;ZHUANG Ying-Ping;ZHANG Si-Liang;National Engineering Research Center for Biotechnology (Shanghai)East China University of Science and Technology;
Abstract:13C isotopic abundance of intracellular free amino acid with a characteristic of fast-turnover can quickly reflect the changes in intracellular metabolic state. However, the concentration of intracellular free amino acid was low and the existed 13C isotope detection method based on GC-MS could not satisfy the requirement with full scan mode. In this study, selected ion monitoring method with theoretically higher detection accuracy was used for the analysis of 13C isotopic abundance of free intracellular amino acid. The fracture law of different amino acids was investigated by full scan mode, and also a characteristic fragment library for the 16 kinds of free intracellular amino acids was established. According to this characteristic fragment library, only the specific m/z signal was detected in the sample analysis, which realized the selected ion monitoring and improved the quality of signal. The results of amino acid standards indicated that, in comparison with the full scan mode, the signal-to-noise ratio (S/N), measurement precision and accuracy were improved by 17, 2 and 3.8 times, respectively. In the analysis of coenzyme Q10 producing strains of samples, 13C isotopic abundance of 8 kinds of free intracellular amino acids were successfully detected. This method was quite feasible for the detection of 13C isotopic abundance of the intracellular free amino acids in cell metabolism research.
Keywords:Selected ion monitoring method  13 C isotopic abundance  Gas chromatography-mass spectrometry  Free amino acids
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