E-cigarette fluids were extracted from the pods and devices, and 50 μL was dissolved in 0.95 mL of isopropyl alcohol (Fisher Scientific, Fair Lawn, NJ). Chemical analysis was performed with an Agilent 5975C GC/MS system (Santa Clara, CA) using internal standard-based calibration procedures and methods previously described in detail.6 (link),33 (link) The method analyzes 180 flavor chemicals plus nicotine.
Agilent 5975c gc ms system
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 5975C GC-MS system is a gas chromatography-mass spectrometry (GC-MS) instrument. It is designed to provide high-performance analysis of complex samples by combining gas chromatography separation with mass spectrometry detection.
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8 protocols using agilent 5975c gc ms system
1
Analysis of E-Cigarette Fluid Composition
2
GC/MS Analysis of E-Liquid Flavors
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The extracted e-liquid was analyzed using previously described GC/MS methods4 (link). Each sample (50 µL) was dissolved in 0.95 mL of isopropyl alcohol and shipped overnight on dry ice to Portland State University. A 20 µL aliquot of internal standard solution (2000 ng/µL of 1,2,3- trichlorobenzene dissolved in isopropyl alcohol) was added to each diluted sample prior to analysis. Using internal-standard-based calibration procedures described elsewhere36 (link) analyses for 178 flavor-related target analytes, two synthetic coolants, and nicotine were performed with an Agilent 5975C GC/ MS system (Santa Clara, CA). A Restek Rxi-624Sil MS column (Bellefonte, PA) was used (30 m long, 0.25 mm id, and 1.4 µm film thickness). A 1.0 µL aliquot of the diluted sample was injected into the GC with a 10:1 split. The injector temperature was 235 °C. The GC temperature program for analyses was 40 °C hold for 2 min, 10 °C/min to 100 °C, then 12 °C/min to 280 °C and hold for 8 min at 280 °C, and then 10 °C/min to 230 °C. The MS was operated in the electron impact ionization mode at 70 eV in the positive-ion mode. The ion source temperature was 220 cc, and the quadrupole temperature was 150 cc. The scan range was 34 to 400 amu. Each of the 181 (178 flavor chemicals, 2 synthetic coolants, and nicotine) target analytes were quantitated using the authentic standard material.
3
GC-MS Profiling of SSME Phytochemicals
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SSME was separately dissolved in n-hexane and dichloromethane (DCM) and the mixtures were centrifuged thrice at 12000 rpm for 15 min. The two clear supernatant was used for GC-MS analysis for identification of different classes of phytochemicals. Agilent 5975C GC-MS system (Agilent Technologies, USA) attached with HP-5 ms Capillary Column (30 m × 0.25 mm i.d. × 0.25 μm film thickness) and equipped with inert MSD triple axis mass detector conditioned at ion trap 200°C, transfer line 280°C, electron energy 70 eV (vacuum pressure- 2.21e-0.5 torr) was used for analysis. The carrier gas was helium at a flow rate of 1 ml/min. 2 μl sample was injected in a splitless mode. The column temperature was set at 60°C for 1 min. followed by 5°C/ min upto 250°C. The major and essential compounds in SSME were identified by their retention times and mass fragmentation patterns using Agilent Chem Station integrator and the database of National Institute Standard and Technology (NIST) with a MS library version 2010 and by analyzing MS fragmentation patterns.
4
GC-MS Analysis of Refill Fluids
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For each refill fluid, 50 µl were dissolved in 0.95 ml of isopropyl alcohol (IPA) (Fisher Scientific, Fair Lawn, NJ). All diluted samples were shipped overnight on ice to Portland State University and analyzed using GC-MS on the day they were received. A 20 µl aliquot of internal standard solution (2000 ng/µl of 1, 2, 3-trichlorobenzene dissolved in IPA) was added to each diluted sample before analysis. Using internal standard-based calibration procedures described elsewhere52 (link), analyses were performed with an Agilent 5975 C GC-MS system (Santa Clara, CA). A Restek Rxi-624Sil MS column (Bellefonte, PA) was used (30 m long, 0.25 mm id, and 1.4 µm film thickness). A 1.0 µl aliquot of diluted sample was injected into the GC with a 10:1 split. The injector temperature was 235 °C. The GC temperature program for analyses was: 40 °C hold for 2 min; 10 °C/min to 100 °C; then 12 °C/min to 280 °C and hold for 8 min at 280 °C, then 10 °C/min to 230 °C. The MS was operated in electron impact ionization mode at 70 eV in positive ion mode. The ion source temperature was 220 °C and the quadrapole temperature was 150 °C. The scan range was 34 to 400 amu. Each of the 178 target analytes was quantitated using authentic standard material and an internal standard compound normalized multipoint calibration.
5
GC-MS Analysis of Hua-Feng-Dan Powder
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An Agilent 5975C GC-MS system (Agilent Technologies, Lexington, MA, United States) with a HP-5MS chromatography column (30 m ×0.25 mm × 0.25 μm) was used for chromatographic analysis as described previously (Xu et al., 2020 (link)).
Approximately 5 g of Hua-Feng-Dan powder was mixed with 200 ml water and extracted for 2 h with a volatile oil extractor. The resultant liquid was dissolved in 2 ml of petroleum ether, and the volume was fixed in a 5-ml volumetric flask. GC-MS steam distillation: the program temperature started at 40°C and maintained for 3 min, then increased to 52°C at a rate of 1°C/min, and continued to increase to 160°C at a rate of 20°C/min, and finally increased to 280°C at a rate of 5°C/min, and maintained for 3 min. The carrier gas was nitrogen (99.999%), the volume flow rate was 1 ml/min, the shunt ratio was 10:1, and the injection volume was 1 μl. The solvent was delayed by 3.5 min. The ion source of MS was electron ionization with 70 eV at 230°C. The specific conditions of MS analysis were as follows: interface temperature, 280°C; quadruple temperature, 150°C; full-scan mode, m/z 35–550 amu.
Approximately 5 g of Hua-Feng-Dan powder was mixed with 200 ml water and extracted for 2 h with a volatile oil extractor. The resultant liquid was dissolved in 2 ml of petroleum ether, and the volume was fixed in a 5-ml volumetric flask. GC-MS steam distillation: the program temperature started at 40°C and maintained for 3 min, then increased to 52°C at a rate of 1°C/min, and continued to increase to 160°C at a rate of 20°C/min, and finally increased to 280°C at a rate of 5°C/min, and maintained for 3 min. The carrier gas was nitrogen (99.999%), the volume flow rate was 1 ml/min, the shunt ratio was 10:1, and the injection volume was 1 μl. The solvent was delayed by 3.5 min. The ion source of MS was electron ionization with 70 eV at 230°C. The specific conditions of MS analysis were as follows: interface temperature, 280°C; quadruple temperature, 150°C; full-scan mode, m/z 35–550 amu.
6
GC-MS Analysis of Biofuels NOSE and NORE
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NOSE and NORE were initially bi-fractionated by dissolving in dichloromethane and n-hexane separately. The mixtures were centrifuged thrice (12,000 rpm) for 15 min. The clear supernatant was used for GC-MS analysis using Agilent 5975C GC-MS system (Agilent Technologies, USA) attached with HP-5 ms Capillary Column (30 m × 0.25 mm i.d. × 0.25 μm film thickness). The machine was equipped with inert MSD triple axis mass detector conditioned at ion trap 200 °C, transfer line 280 °C, electron energy 70 eV (vacuum pressure- 2.21e–0.5 torr) was used for analysis. Helium was used as carrier gas (1 ml/min). Sample volume was 2 μl and injected in a splitless mode. The column temperature was kept at 60 °C for 1 min followed by 5 °C/min up to 250 °C. The major and essential compounds present in samples were identified by their retention times and mass fragmentation patterns using Agilent Chem Station Integrator and the database of National Institute Standard and Technology (NIST) with a MS library version 2010.
7
GC-MS Analysis of Lipid Fraction Extract
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LFE was dissolved in n-hexane and the mixture was centrifuged thrice at 12,000 rpm for 15 min. The clear supernatant was used for GC–MS analysis. Agilent 5975 CGCMS system (Agilent Technologies, USA) attached with HP-5 ms Capillary Column (30 m × 0.25 mm i.d. × 0.25 μm film thickness) and equipped with inert MSD triple axis mass detector condition edation trap 200 °C, transfer line 280 °C, electronenergy70eV (vacuum pressure-2.21e-0.5 Torr) was used for analysis. The carrier gas, helium, was used at a flow rate of 1 ml/min. 2 ml sample was injected in a split less mode. The column temperature was set at 60 °C for 1 min followed by 5 °C/min up to 250 °C. The major and essential compounds in LFE were identified by the retention times and mass fragmentation patterns using Agilent Chem Station integrator and the database of National Institute of Standard and Technology (NIST) with a MS library version2011.
8
Analysis of Calophyllum inophyllum Seed Oil
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Calophyllum inophyllum seed oil was purchased from a Darwin-based company that produces the oil in bulk using a cold press method from seeds collected in the Darwin region of the Northern Territory (NT), Australia. Samples of CSO were derivatised to fatty acid methyl ester [21 ]. Briefly, 100 μl of CSO was mixed with 5 mL of sodium methoxide in methanol (0.5 N) in a screw cap test tube and heated in a water bath at 50 °C for 20 min. The test tube was cooled to room temperature before adding n-hexane (5 mL). The mixture was shaken for 5 min and then centrifuged at 1750 rpm for 5 min. The upper layer of the FAME sample was injected into a gas chromatography-mass spectrometry (GC-MS; Agilent 5975C GC/MS system with a DB-5 (30 m × 0.25 mm × 0.25 μm) ms capillary column coated with polysiloxane for analysis. The carrier gas was helium at a 1.5 mL/min constant flow rate. The initial temperature of the column was 70 °C and raised to 300 °C at 9 °C/min and then maintained for 4 minutes at 300 °C. A diluted FAME standard in hexane was used as a reference.
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