Accurate Determination of 13 C Isotopic Abundances of Free Intracellular Amino acids with Low Concentration by GC-MS-Selective Ion Monitoring Method
10.11895/j.issn.0253-3820.140371
- VernacularTitle: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13 C同位素丰度
- Author:
Minchao LI
;
Mingzhi HUANG
;
Yuwei LIU
;
Ju CHU
;
Yingping ZHUANG
;
Siliang ZHANG
- Publication Type:Journal Article
- Keywords:
Selected ion monitoring method;
13 C isotopic abundance;
Gas chromatography-mass spectrometry;
Free amino acids
- From:Chinese Journal of Analytical Chemistry
2014;(10):1408-1413
- CountryChina
- Language:Chinese
-
Abstract:
13 C isotopic abundance of intracellular free amino acid with a characteristic of fast- turnover can quickly reflect changes in intracellular metabolic state. But the concentration of intracellular free amino acid is low, the existed 13 C isotope detection method based on GC-MS can not satisfy the requirement with full scan mode. In this study, the selected ion monitoring method was used to detect accuracy higher likelihood of analysis of 13 C isotopic abundance of free intracellular amino acid. First, in the full scan mode we analyzed of the fracture law of different amino acids, found the feature corresponding to each amino acid fragments, and established 16 kinds of free intracellular amino acids characteristic fragment library. Then using this characteristic fragment library, only specific m/z signal was detected in sample analysis, which realized the selected ion monitoring and improved the quality of signal. The results of amino acid standards showed that the signal-to-noise ratio, measurement precision and accuracy were improved by 17, 2. 0 and 3. 8 times compared with the full scan mode. In the analysis of coenzyme Q10 producing strains of samples, this method was successfully used to detect isotopic abundance of 8 kinds of free intracellular amino acids. This method plays an important role in the detection of 13 C isotopic abundance of the intracellular free amino acid in cell metabolism research.
