1100 isocratic hplc pump

Synchronized Synechococcus 7942 wild-type cells were harvested with fast filtration and immediately suspended in the extraction solution, as described previously [29 (link)]. Metabolite concentrations were analysed with capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS), using an Agilent CE capillary electrophoresis system equipped with an Agilent 6210 time-of-flight mass spectrometer, an Agilent 1100 isocratic HPLC pump, an Agilent G1603A CE-MS adapter kit, and an Agilent G1607A CE-ESI-MS sprayer kit with an attached platinum needle (Agilent Technologies, Waldbronn, Germany). Cationic metabolites were separated with a fused silica capillary, using 1 M formic acid as the electrolyte. Anionic metabolites and nucleotides were separated with a COSMO (+) capillary (Nacalai Tesque, Kyoto, Japan), using 50 mM ammonium acetate (pH 8.5) as the electrolyte. Details of the CE-TOFMS conditions and the methods used for data analysis were described previously [30 (link)]. Absolute quantifications were performed using metabolite standards and calculated as the amount of metabolite per 10⁷ cells (i.e. pmol per 1 × 10⁷ cells) in each sample (S2 Table). We were unable to separate the TOF-MS peaks of 3-phosphoglycerate (3-PG) and 2-phosphoglycerate (2-PG). Consequently, they are depicted as the combined total amounts of 3-PG and 2-PG.

Metabolomes were profiled by CE-TOFMS using an Agilent CE Capillary Electrophoresis System equipped with a 6210 Time-of-Flight mass spectrometer, an 1100 isocratic HPLC pump, a G1603A CE-MS adapter kit and a G1607A CE-ESI-MS sprayer kit (all from Agilent Technologies, Waldbronn, Germany), and this system was controlled by G2201AA ChemStation software version B.03.01 for CE (Agilent Technologies).