Zb 1701 gc capillary column

The samples from Intracellular, extracellular, and biomass were derivatized using the methyl chloroformate (MCF) method as previously described (Smart et al. 2010 (link)). The chemical derivatives were analyzed by a system of Agilent GC7890B coupled to an MSD5977A mass selective detector (EI) set at 70 eV. The ZB-1701 GC capillary column (30 m × 250 µm id × 0.15 µm with 5 m guard column, Phenomenex) was used for metabolite analysis. The parameter analysis was previously described (Smart et al. 2010 (link)).

Metabolites in Vibrio sp. samples were extracted in cold methanol-water solution (MeOH:H₂O, 50% and 80%, sequentially), and derivatized via methyl chloroformate alkylation (MCF), as previously described^{35 (link),36 (link)}. Derivatized extracts were transferred into 2 mL amber GC glass vials fitted with inserts (Sigma-Aldrich, St. Louis, MO, USA) for GC-MS analyses which were performed on a gas chromatograph GC7890B coupled to a quadrupole mass spectrometer MSD5977A (Agilent Technologies, USA), with a quadrupole mass selective detector (EI) operated at 70 eV. The system was equipped with a ZB-1701 GC capillary column (30 m × 250 μm id × 0.15 μm with 5 m stationary phase) (Phenomenex, Torrance, CA, USA). Helium was used as the carrier gas and was held constant at the flow of 1 mL min⁻¹. The instrumental setup parameters for MCF derivatized samples were conducted according to Smart, et al.^{35 (link)}. The injection volume was 1 μL and all samples were injected randomly.
Confirmation of specific metabolites in the TCA cycle/glyoxylate shunt (citric acid, cis-aconitic acid, isocitric acid, succinic acid, fumaric acid, malic acid, glyoxylic acid and pyruvic acid) was conducted by extraction, derivatization and GC-MS measurements of the standards of these compounds (20 μL of 20 mM solutions).

The extracted samples were chemically derivatized via the methyl chloroformate (MCF) method based on the recommendations published by Smart et al. [20 (link)]. All cord plasma samples were analyzed in a single batch and derivatized compounds were examined by an Agilent GC7890B system using a ZB-1701 GC capillary column. An MSD5977A mass selective detector with the electron impact voltage set to 70 eV was applied to analyze the compounds. The GC column used for metabolite separation was the ZB-1701 GC capillary column (30 m × 250 μm id × 0.15 μm with a 5 m guard column, Phenomenex). The GC temperature was set up according to the protocol of Han et al. [21 (link)].