Pcdl manager software

A PCDL was constructed using Agilent PCDL Manager software (version B8.00). Either readily available or isolated and fully characterized in-house chemical compounds were utilized as reference standards to establish the PCDL. Data including the retention time, exact mass, and high-resolution MS fragmentation patterns were exported to the PCDL. Additional information, such as the molecular formula, compound name, and CAS number were assigned to each entry for constructing the PCDL.

Data processing for qualitative metabolic flux analysis was performed using VistaFlux software suite from Agilent Technologies. First, Pathways to PCDL software (version B.07.00, Agilent Technologies) and PCDL Manager software (version B.07.00, Agilent Technologies) was used to build a metabolite library for metabolites in both glycolysis and citric acid cycle. Then, acquired LC-MS raw data files (.d) were loaded into Profinder (version B.08.00, Agilent Technologies) for the extraction of metabolite isotopologues using the constructed metabolite library. The ion abundance criterion for data extraction was set as ‘use peak core area 20% of peak height’. The qualification parameters were set as: mass tolerance: ±15 ppm +2.00 mDa; retention time tolerance :±0.20 min; anchor ion height threshold: 250 counts; sum of ion heights threshold: 1000 counts; correlation coefficient threshold: 0.5. The final results were exported into a csv file for the subsequent comparative and statistical analyses.