使用超高分辨率质谱联用直接进样的多重稳定同位素分辨代谢组学对氨基酸同位素异构体进行定量分析。
Quantification of Isotopologues of Amino Acids by Multiplexed Stable Isotope-Resolved Metabolomics Using Ultrahigh-Resolution Mass Spectrometry Coupled with Direct Infusion.
作者信息
Yang Ye, Fan Teresa W-M, Lane Andrew N, Higashi Richard M
机构信息
Urologic Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Department of Toxicology and Cancer Biology, Markey Cancer Center, University of Kentucky, Lexington, KY, USA.
出版信息
Methods Mol Biol. 2019;2030:57-68. doi: 10.1007/978-1-4939-9639-1_6.
Stable isotope-resolved metabolomics (SIRM) is increasingly used among researchers for metabolic studies including amino acid metabolism. However, the classical GC- or HPLC-based methods for amino acid quantification do not meet the needs for multiplexed stable isotope-enriched analysis by ultrahigh-resolution Fourier transform mass spectrometry (UHR-FTMS). This is due to insufficient acquisition time during chromatographic separations and large dynamic range in concentrations of analytes, which compromises detection and quantification of the numerous metabolite isotopologues present in crude extracts. This chapter discusses a modified ethyl chloroformate derivatization method to enable rapid quantitative analysis of stable isotope-enriched amino acids using direct infusion ion introduction coupled with UHR-FTMS.
稳定同位素分辨代谢组学（SIRM）在包括氨基酸代谢在内的代谢研究中越来越受到研究人员的青睐。然而，基于经典气相色谱（GC）或高效液相色谱（HPLC）的氨基酸定量方法无法满足通过超高分辨率傅里叶变换质谱（UHR-FTMS）进行多重稳定同位素富集分析的需求。这是因为色谱分离过程中的采集时间不足，以及分析物浓度的动态范围较大，这会影响粗提物中众多代谢物同位素异构体的检测和定量。本章讨论了一种改进的氯甲酸乙酯衍生化方法，该方法可使用直接进样离子导入结合UHR-FTMS对稳定同位素富集的氨基酸进行快速定量分析。
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