Cp sil 8 cb fused silica capillary gc column

Two-microlitre samples of the derivatised solution were injected into the GC–MS system (GC-2010 Plus gas chromatograph coupled to a 2010 Plus single-quadrupole mass spectrometer, Shimadzu Corp., Kyoto, Japan). Chromatographic separation was achieved with a 30-m 0.25-mm CP-Sil 8 CB-fused silica capillary GC column with 1.00-µm film thickness from Agilent (Agilent, J&W Scientific, Folsom, CA, USA), with helium as carrier gas. The initial oven temperature of 100 °C was maintained for 1 min, and then raised by 4 °C/min to 320 °C with a further 4 min of hold time. The gas flow was set to obtain a constant linear velocity of 39 cm/s, and the split flow was set at 1:5. The mass spectrometer was operated in electron impact (70 eV) in full-scan mode in the interval of 35–600 m/z with a scan velocity of 3333 amu/s and a solvent cut time of 4.5 min. The complete GC programme duration was 60 min. Untargeted metabolites were identified by comparing the mass spectrum of each peak with the NIST library collection (NIST, Gaithersburg, MD, USA). To identify metabolites’ identity, the linear index difference maximum tolerance was set at 10, while the minimum matching for the NIST library search was set at 85%. All measurements were performed by Theoreo srl, Montecorvino Pugliano (SA), Italy.

GC-MS analysis was performed on the derivatized extracted metabolome according to Troisi et al. [15 (link)] with a few minor changes. Briefly, 2 µL of the sample solution was injected into the GC-MS system (GC-2010 Plus gas chromatograph coupled to a 2010 Plus single quadrupole mass spectrometer; Shimadzu Corp., Kyoto, Japan) equipped with a 30-m, 0.25-mm ID CP-Sil 8 CB fused silica capillary GC column with 1.00-µm film thickness from Agilent (Agilent, J&W Scientific, Folsom, CA, USA), using He as a carrier gas. The initial oven temperature of 100 °C was maintained for 1 min and then raised by 6 °C/min to 320 °C with a further 2.33 min of hold time. The gas flow was set to obtain a constant linear velocity of 39 cm/s, and injections were performed in the splitless mode. The mass spectrometer was operated in electron impact (70 eV) in the full-scan mode in the interval of 35–600 m/z with a scan velocity of 3333 amu/s and a solvent cut-off time of 4.5 min. The complete GC analysis duration was 40 min. Untargeted metabolites were identified by comparing the mass spectrum of each peak with the NIST library collection (NIST, Gaithersburg, MD, USA).