Multitarget quantitative metabolic profiling of hydrophilic metabolites in fermentation broths of beta-lactam antibiotics production by HILIC-ESI-MS/MS

作者：Schiesel Simone; Laemmerhofer Michael; Lindner Wolfgang

来源：Analytical and Bioanalytical Chemistry, 2010, 396(5): 1655-1679.

DOI：10.1007/s00216-009-3432-2

• Microbial metabolomics
• Process analysis technology (PAT)
• Process control
• HILIC
• HPLC-MS/MS
• Matrix effects

The presented work deals with the development and comprehensive validation of a quantitative LC-electrospray ionization (ESI)-tandem mass spectrometry (MS/MS) method using a triple quadrupole instrument in the MRM mode for the metabolic profiling of amino acids, organic acids, vitamins, some biogenic amines, secondary metabolites of beta-lactam antibiotics biosynthesis as well as their intermediates, and degradation products in fermentation broths of beta-lactam antibiotics production (in total 57 hydrophilic compounds). A great number of chromatographic systems (22 different stationary phase/mobile phase conditions) were screened for their adequate chromatographic selectivity to cope with isobaric compounds and other critical analyte pairs. Finally, a hydrophilic interaction liquid chromatography (HILIC) method employing a zwitterionic ZIC-HILIC column was selected as best compromise. Particular focus was given on the elucidation of absolute and relative matrix effects via comparison of slopes of calibration functions of spiked matrix and standard solutions. These data as well as precision and accuracy data confirm suitability of the HILIC-ESI-MS/MS assay for metabolic profiling studies in fermentation samples. Detailed comprehensive data sets are presented which should illustrate critical issues, problems, and challenges of multitarget quantitative metabolic profiling and should outline possible strategies to circumvent pitfalls and overcome common problems.

• 出版日期2010-3