GC–MS data was recorded with Single Quadrupole Mass Spectrometer and GC (Trace 1,310; Thermo Fisher Scientific, United States). The column was 30 m long dimethyl (95%)/diphenyl polysiloxane (5%) RTX-5MS with 0.25 mm ID, and guard column (10 m; Restek). The initial temperature of GC oven was 60°C for 60 s and the temperature increased to 325°C (@10°C/min). Metabolites were identified using NIST library with Xcalibur software.
Guard column
Manufactured by Restek
Sourced in United States
A guard column is a short, inexpensive column placed before the analytical column in a chromatographic system. Its primary function is to protect the analytical column from contamination and wear, thereby extending the column's lifetime and maintaining the system's performance.
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Lab products found in correlation
4 protocols using guard column
1
GC-MS Analysis of Metabolites
2
Sensitive Biphenyl HPLC Separation
3
Phytotoxin Identification by LC-MS
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The identification of main phytotoxins was done with a LC-MS system. The chromatographic separation of the compounds was carried out on a Restek Ultra C18 column (250 × 4.5 mm, 5 μm) with a Restek guard-column (10 × 4.0 mm). Two solvent mixtures were used as mobile phases. Solvent A was composed of water/methanol (99.2:0.8, v/v and acidified with formic acid 20 μl L−1) while 100% methanol was used as solvent B. Samples were dissolved in minimum quantity of methanol. The sample injection volume was set at 20 μl. A solvent gradient was adopted for a total run time of 60 min at a flow rate of 1 ml min−1, with all the toxin standards eluting over 20–60 min. The solvent gradient was as follows: equilibration at 5% B for 2 min, from 5 to 35% B in 43 min., from 35 to 100% B in 5 min., 100% B for 10 min., return to initial condition in 2 min.
4
GC-MS Analysis of Volatile Compounds
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The chromatographic conditions from GC-FID were adopted to GC-MS analysis. The GC analysis was performed using a Thermo Fisher Scientific Trace Ultra gas chromatograph equipped with a BGB-wax capillary column (30 m × 0.25 mm i.d., film thickness 0.25 μm, Restek, Bellefonte, PA, USA) fitted with a guard column (1 m × 0.25 mm i.d, deactivated, Restek). The temperature of the PTV injection was 220 °C. The injection volume was 1 μl (TriPlus autosampler, Thermo Fisher scientific) using a split ratio of 1:50. Helium was used as carrier gas at a constant flow rate of 1.5 mL min−1. The oven temperature was kept at 65 °C for 10 min and then heated to 220 °C with 5 °C min−1 and kept constant at 220 °C for 9 min. The MS analysis was carried out on a Thermo DSQ II mass spectrometer detector operated in positive EI mode at 70 eV. Transfer line and ion source temperatures were set to 250 °C. Mass spectra were acquired in the full scan mode (mass range 40–300 m/z). Peak identification was performed using different libraries: NIST (version 2.2, 2014), Adams (fourth edition, 2007) and in-house libraries [58 ,63 ]. Retention indices (RI) were calculated according to the van den Dool and Kratz equation [64 (link)]. The used software was Thermo Xcalibur (Thermo Fisher scientific, version 2.2 SP1.48).
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