Qp2010 ms

In accordance with the protocol described in Loukili et al. [109 (link)] with some modifications, methyl esters of hexane fatty acids and ethyl acetate were extracted from G. bursa-pastoris. The characterization and identification of fatty acids were analyzed using the Shimadzu GC system (Kyoto, Japan), which was equipped with a BPX25 capillary column with a 5% diphenyl and 95% dimethylpolysiloxane phase (30 m × 0.25 mm × 0.25 µm) and connected to a QP2010 MS. Helium gas (99.99%) was used as the mobile phase with a flow rate of 3 L/min. The temperature of the injection, ion source, and interface was set at 250 °C, while the temperature program for the column oven was set to 50 °C for 1 min and then heated to 250 °C at a rate of 10 °C/min and maintained for an additional minute. The ionization of the sample components was performed in Electron Ionization (EI) mode at 70 eV, with a scanned mass range of 40 to 300 m/z. A 1 μL volume of each extract was injected in split mode, and each sample was analyzed in triplicate. The compounds were characterized by comparing their retention times, mass spectra fragmentation patterns, and databases, including the National Institute of Standards and Technology’s database (NIST). Data processing was carried out using LabSolutions (version 2.5, Shimadzu, Kyoto, Japan).

The extracts of CS and CM were dissolved in hexane. The separation and identification were performed on a Shimadzu GC system (Kyoto, Japan) equipped with a BPX25 capillary column with 5% diphenyl and 95% dimethylpolysiloxane phase (30 m × 0.25 mm inner diameter × 0.25 μm film thickness), coupled to a QP2010 MS. Pure helium gas (99.99%) was used as a carrier gas with a constant flow rate of 3 mL/min. The injection, ion source, and interface temperatures were all set at 250°C. The temperature program used for the column oven was 50°C (held for 1 min), heated to 250°C at 10°C/min, and held for 1 min. The ionization of the sample components was done in the EI mode (70 eV). The mass range scanned was 40–300 m/z. 1 μL of each prepared extract diluted with an appropriate solvent was injected in a splitless mode (split ratio 90 : 1). All samples were analyzed in triplicate. Finally, compounds were identified by comparing their retention times with those of authentic standards and their mass spectral fragmentation patterns with those found in databases or those stored on the National Institute of Standards and Technology (NIST) 147, 198 compounds. LabSolutions (version 2.5) was used for data collection and processing.

A Shimadzu GC system (Kyoto, Japan) with a BPX25 capillary column with a 95 percent dimethylpolysiloxane diphenyl phase (30 m 0.25 mm ID 0.25 m film thickness) and an MS QP2010 was used for separation and identification. As a carrier gas, pure helium (99.99 percent) was employed at a constant flow rate of 3 mL/min. The mass range studied was 40 to 300 m/z, and 1 L of each produced oil was fed into the chamber and diluted with an appropriate solvent. A total of 1 μL of each prepared oil diluted with an appropriate solvent was injected in the fractionation mode (fractionation ratio 90:1). Samples were evaluated three times. Finally, the compounds were recognized by comparing their retention times to verified standards, and their mass spectrum fragmentation models to those found in databases or on NIST compounds. Data were collected and processed by using Laboratory Solutions (v2.5).

The qualitative and semi-quantitative analysis of PVEO was conducted using a gas chromatograph with a mass spectrometer detector, as previously described in [46 (link)]. A Shimadzu GC system from Kyoto, Japan was utilized in combination with an MS QP2010 (Shimadzu Scientific Instruments, Kyoto, Japan) to identify and separate compounds. Separation was achieved using a BPX25 capillary column with a 95 percent dimethylpolysiloxane diphenyl phase with a 30 m length, 0.25 mm internal diameter, and 0.25 m film thickness. Pure helium (99.99 percent) was used as the carrier gas, at a constant flow rate of 3 mL/min, and the injection, ion source, and interface temperatures were fixed at 250 °C. The column furnace was programmed to increase from 50 °C (for 1 min) to 250 °C at a rate of 10 °C/min, maintaining the temperature for 1 min. Sample components were ionized in EI mode at 70 eV, and the mass range studied was 40 to 300 m/z. Each produced oil was fed into the chamber at 1 L, diluted with an appropriate solvent. Then, 1 μL of each prepared oil was injected in fractionation mode (fractionation ratio 90:1). Three evaluations were conducted per sample, and compounds were identified by comparing the retention times to verified standards and mass spectrum fragmentation models found in databases or on NIST compounds. Laboratory Solutions (v2.5) was used to collect and process data.

Volatiles were collected at different time intervals after culture inoculation (1, 4, 8, and 12 days) using polydimethylsiloxane (PDMS) sorbent [48 (link)]. Two to three 5 mm PDMS tubes were carefully mounted on sterile metal wires imbedded in the PDA. The headspace sampling time was 2 h for all experiments unless otherwise indicated. After sampling, the tubes were placed in 1.5 mL brown glass vials and stored at −20 °C for analysis.
Volatiles collected on PDMS tubes were analyzed using a GC-2010 plus gas chromatograph coupled to a MS-QP2010 quadrupole mass spectrometer equipped with a TD-20 thermal desorption unit (Shimadzu, Japan) and a GC Cryo-Trap (Tenax®). A single tube was placed in a 89 mm glass thermal desorption tube and desorbed at a flow rate of 60 mL min⁻¹ for 8 min at 200 °C under a stream of N₂ gas. The desorbed substances were focused in a cryogenic trap at −60 °C. The Tenax® adsorbent was heated to 230 °C and the analytes were injected using split mode (1:50) onto a Rtx-5MS GC column (thickness—0.25 µm, length—30 m, film diameter—0.25 µm) using helium as carrier gas. Separation, detection, and data analysis were identical to those reported for the quantitative analysis.