Acquity 1 class uplc tqs micro ms system

Targeted metabolomics analysis was carried out using an LC-MS/MS platform targeting 203 standard metabolites from more than 25 metabolic pathways (e.g., glycolysis, tricyclic acid cycle, amino acid metabolism, glutathione, etc.). LC-MS/MS experiments were performed on a Waters Acquity I-Class UPLC TQS-micro MS system. Each sample was injected twice, 2 μL and 10 μL, for analysis using positive and negative ionization modes, respectively. Both chromatographic separations were performed in hydrophilic interaction chromatography mode.
The flow rate was 0.3 mL min -1 , autosampler temperature was kept at 4 o C, and the column compartment was set at 40 o C. The mobile phase was composed of solvents A (5 mM ammonium acetate in H2O + 0.5% acetic acid + 0.5% acetonitrile) and B (acetonitrile + 0.5% acetic acid + 0.5% water). The LC gradient conditions were the same for both positive and negative ionization modes. After an initial 1.5-min isocratic elution of 10% A, the percentage of solvent A was increased linearly to 65% at time (t) = 9 min, then remained the same for 5 min (t = 14 min), and then reduced to 10% at t = 15 min to prepare for the next injection. After chromatographic separation, MS ionization and data acquisition was performed using an electrospray ionization source. A pooled study sample was used as the quality control (QC) and run once for every 10 study samples.