Hp 5ms ui

The aqueous ammonia Q. suber bark extract was studied by gas chromatography–mass spectrometry (GC-MS) at the Research Support Services (STI) at Universidad de Alicante (Alicante, Spain), using a gas chromatograph model 7890A coupled to a quadrupole mass spectrometer model 5975C (both from Agilent Technologies, Santa Clara, CA, USA). Chromatographic conditions were as follows: 3 injections/vial, injection volume = 1 µL; injector temperature = 280 °C, in splitless mode; initial oven temperature = 60 °C, after 2 min, followed by an increase of 10 °C/min up to a final temperature of 300 °C, after 15 min. The chromatographic column used for the separation of the compounds was an Agilent Technologies HP-5MS UI of 30 m in length, 0.250 mm in diameter, and with 0.25 µm film. The conditions of the mass spectrometer were as follows: temperature of the electron impact source of the mass spectrometer = 230 °C and of the quadrupole = 150 °C; ionization energy = 70 eV. Test mixture 2 for apolar capillary columns according to Grob (Supelco 86501) and PFTBA tuning standards were used for equipment calibration. The identification of the components was based on a comparison of their mass spectra and retention time with those of the authentic compounds and by computer matching with the database of the National Institute of Standards and Technology (NIST11) and Adams [83 ].

A representative sample of cannabis flowers was homogenized by mortar and pestle. Then, 100 mg of the homogenized sample was weighed in triplicates. The samples were extracted with 1.8 mL of 0.001% tridecane (internal standard) in hexane, and then sonicated for 30 min at room temperature and centrifuged 10 min (21.200× g, room temperature). The supernatant was injected into a gas chromatography Agilent 7890A GC coupled with a HP 5975C MSD spectrometer (Agilent Technologies, Inc., Santa Clara, CA, USA). Chromatographic separation was done on HP-5MS UI (30.0 × 250.0 μm; 0.25 μm film) (Agilent Technologies, Inc., Santa Clara, CA, USA). The temperature program was as follows: from 60 to 180 °C, increasing 3 °C/min (run time 40 min). Finally, the column was kept at 310 °C for 10 min at post run. The flow rate of helium was 1.1 mL/min, and the injection volume 1 μL. The temperatures of injection and detector were 250 and 230 °C, respectively. Quantification was done using an external calibration curve of standard mixtures of typical cannabis terpenes (Restek, Bellefonte, PA, USA).

The analysis was done on an easy-to-use FlavourSpec (G.A.S., Dortmund, Germany) HS-GCxIMS. Samples were automatically shaken for 30 min at 80 °C. 1 mL of headspace was injected into the injector at 80 °C without split flow. Substances were separated on a 15 m × 0.53 mm × 1 μm MXT-5 column (Restek, Bellefonte, PA, USA) with a nitrogen carrier gas ramp (2 mL/min for 1 min., in 14 min. up to 150 mL/min, this held for 35 min, for a total of 50 min. for the analysis). Detection was carried out with a drift tube of 15.2 mm (diameter) and 98 mm (length) ionization by ³H-source, at 45 °C, a nitrogen drift gas flow rate of 150 mL/min, and a field strength of 500 V/cm.
Reference measurements were run on a thermal-gradient HS-GC-qMS (Shimadzu, Kyoto, Japan). Using the same vials, samples were incubated for 30 min at 120 °C, afterwards injecting 2 mL of the headspace at a split of 1:10. The separation was performed on a HP-5 MS UI (Agilent, Santa Clara, CA, USA) 30 m × 0.25 mm × 0.5 μm column at a constant flow rate of 35 cm/s helium and a temperature ramp (50 °C held for 4 min, at 5 °C/min to 150 °C, at 10 °C/min to 200 °C). Analytes were ionized using electron-impact ionization at 70 eV and 200 °C with a scan range of 32–300 m/z at an event time of 300 ms.

Water samples were taken to characterize the pulsed exposures during dosing and periodically when no test substance was being delivered to the test vessels. Samples (100 mL) were collected from mid‐depth in the exposure vessels into glass jars, acidified with 0.10 mL acetic acid, and subsequently processed and analyzed immediately. Replicate samples were frozen in plastic jars and stored at approximately –20 °C as backups.
Samples were extracted twice with toluene, and the extracts were combined and analyzed using a Hewlett‐Packard Model 5975 gas chromatograph equipped with an Agilent model 7890 mass selective detector operated in the selected ion monitoring mode, using an Agilent HP‐5MS UI (30 m × 250 μm, 0.25 µm film thickness) column. The level of quantitation was set at 0.249 µg/L.

GC-MS analyses were performed as reported previously (Suzuki et al., 2019 (link)) on a 5977A MSD mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) connected to a 7890B gas chromatograph (Agilent Technologies) with an HP-5MS UI (30 m × 0.25 mm, 0.25-μm film thickness; Agilent Technologies) capillary column for qualitative analysis. The injection temperature was set at 250°C. The column temperature program was as follows: 80°C for 1 min, increase to 300°C at a rate of 20°C/min, and hold for 28 min. The carrier gas was helium at a flow rate of 1.0 mL/min. The ion source temperature was 230°C, and the quadrupole temperature was 150°C. The derivatized sample (1 µL) was injected in splitless injection mode. Peaks were identified by comparing their Rt and mass spectra with those of authentic standards (Supplementary Figure S4). Samples were analyzed in selected ion monitoring (SIM) mode for relative quantification by extracting the mass chromatogram in respective extracted ion chromatogram (EIC) for each metabolite as listed in Supplementary Table S7.

GC-MS analysis was performed in an Agilent 7890A GC-MS system (Agilent Technologies, Santa Clara, CA, USA), equipped with a CTC autosampler and a PTV injector. Gas chromatography was performed on a 30 m HP-5 ms UI (Agilent J&W) capillary column (film thickness of 0.25 mm; I.D. of 0.25 µm), while back-flush elution was carried out in a 1.5 m deactivated column with a film thickness of 0.18 mm. The initial column temperature was set at 60 °C for 1 min and then increased to 300 °C at a 10 °C/min rate. The temperature was maintained at 300 °C for 6 min. Total run time was 30 min, followed by a 12 min back-flush run and a solvent delay at 6 min. Helium (99.999%) was used as the carrier gas at a flow rate of 3 mL/min, and injection volume was set at 1 μL. Splitless mode injection was performed at the PTV injector, where the temperature was increased from 270 °C to 350 °C. MS was operated in electron impact ionization mode (EI; 70 eV). Ion source and transfer line temperatures were set at 230 °C and 250 °C, respectively. All mass spectra were acquired in full scan mode between 50 and 600 amu.