An ion-pair free LC-MS/MS method for quantitative metabolite profiling of microbial bioproduction systems

Abstract Data-driven engineering of microbes has been demonstrated for the sustainable production of high-performance chemicals. Metabolic profiling analysis is essential to increase the productivity of target compounds. However, improvement of comprehensive analysis methodologies is required for the high demands of metabolic engineering. Therefore, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) based methodology was designed and applied to cover a wide target range with high precision. Ion-pair free separation of metabolites on a pentafluorophenyl propyl column enabled high-precision quantification of 113 metabolite. Then, we evaluated the method for high reproducibility and robustness. Target metabolites consisted of primary metabolites and intermediate metabolites for microbial production of high-performance chemicals. 95 metabolites could be indicated the high reproducibility of peak area (intraday data; CV