A systematic approach to development of analytical scale and microflow-based liquid chromatography coupled to mass spectrometry metabolomics methods to support drug discovery and development

•The systematic optimization of global profiling metabolomics methods was conducted.•Development of microflow-based global profiling methods using 0.3 mm i.d. columns.•Microflow methods showed increases in peak volume for a biological matrix.•Reduced sample volume shows potential applicability to limited biomedical samples.•Development of a standard mixture for evaluation of global metabolomics methods.