Simultaneous analysis of primary metabolites by triple quadrupole LC/MS/MS using pentafluorophenylpropyl column

Selected reaction monitoring (SRM) transition and some parameters for primary metabolites were optimized on the flow injection analysis using purified standard compounds. These target compounds are including hydrophilic metabolites such as amino acids, organic acids, nucleotides and coenzyme etc. SRM transition (precursor ion > product ion), Q1, CE and Q3 parameters were determined in positive or negative mode. As an example of typical choice of the polarity, amino acids were found as a peak with a high intensity in positive mode and organic acids were measured in negative mode. These target metabolites were eluted by the gradient condition using pentafluorophenylpropyl column and the retention time was determined by measurement of a standard compound and its mixture. The reproducibility of the retention time was also evaluated using tissue extracts from mice as a biological sample. The linearity was confirmed by the calibration standard mixture with different concentration. As a result, the reproducibility and the linearity were confirmed to be enough in the measurement of biological samples.
Simultaneous measurement for hydrophilic primary metabolites becomes to be a powerful tool to evaluate metabolic changes in a cultured cell and a tissue caused by the disease or the genetic modification.