Citation: MOU Han, HONG Ming, LIU Xiao-Yun, LI Min-Chao, HUANG Ming-Zhi, CHU Ju, ZHUANG Ying-Ping, ZHANG Si-Liang. Accurate Detemination of Isotopic Abundance of Intracellular Metabolites of Saccharopolysporaerythraea Based on Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(9): 1264-1270. doi: 10.11895/j.issn.0253-3820.170205 shu

Accurate Detemination of Isotopic Abundance of Intracellular Metabolites of Saccharopolysporaerythraea Based on Ultra Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry

State Key Laboratory of Bioreactor Engnieering, East China University of Science and Technology, Shanghai 200237, China

Received Date: 30 March 2017
Revised Date: 12 June 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 21276081).

A method for measuring ¹³C isotopic abundance of intracellular metabolites of Saccharopolysporaerythraea by ultra-high performance liquid chromatography (UPLC)-triple quadrupole mass spectrometry was established. First, the chromatographic conditions of UPLC were optimized, and then the MS conditions such as unique tube lens voltage, collision energy, and ion pair were optimized. On the bases of length of the parent and daughter ions carbon chains and whether the daughter ions contain ¹³C atoms, the one-to-one method, one-to-many method and SIM method were established for measuring ¹³C isotopic abundance. Then these methods were used to measure naturally labeled intracellular metabolite standards and ¹³C labeled samples, and according to the gap between the experimental value and the theoretical value, the best method was established for each metabolite of different characteristics. The results showed that one-to-one method was most effective for measuring the metabolites of daughter ions not containing ¹³C atoms represented by sugar phosphates, one-to-many method was the best for measuring the metabolites of both parent and daughter ions containing ¹³C short carbon chains represented by carboxylic acids, SIM method could play a role in measuring the metabolites of both parent and daughter ions containing ¹³C long carbon chains represented by coenzyme A. This method had a good measurement precision and could be applied to the measurement of Saccharopolysporaerythraea intracellular metabolites, which contributed to the consequent study of metabolic mechanism and the efficient expression of erythromycin.
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