The total concentrations of palmitic acid (16:0), stearic acid (18:0), myristic acid (14:0), behenic acid (22:0), arachidic acid (20:0), gondoic (20:1), oleic (18:1), and linoleic (18:2) were determined from tissues by gas chromatography–mass spectrometry73 (link). A known quantity of tissue was hydrolyzed and extracted after adding a known amount of heptadecanoic acid (17:0). Fatty acids were analyzed as their trimethylsilyl derivatives under electron impact ionization mode using an Agilent 5973N-MSD equipped with an Agilent 6890 GC system and a DB17-MS capillary column (30 m × 0.25-mm internal diameter × 0.25-μm film thickness).
Db 17ms capillary column
Manufactured by Agilent Technologies
Sourced in United States
The DB-17MS capillary column is a high-performance gas chromatography (GC) column designed for the separation and analysis of a wide range of analytes. The column features a 50% phenyl-50% methylpolysiloxane stationary phase, which provides excellent selectivity and separation for a variety of compounds. The column is inert, thermally stable, and suitable for use in a wide range of applications.
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Lab products found in correlation
6890 gc system, by Agilent Technologies (3 mentions)
5973n msd, by Agilent Technologies (3 mentions)
Agilent 5973n msd, by Agilent Technologies (2 mentions)
Agilent 6890 gc system, by Agilent Technologies (2 mentions)
7000 triple quadrupole gc ms system, by Agilent Technologies (2 mentions)
Db 5ms, by Agilent Technologies (2 mentions)
Db 1ms, by Agilent Technologies (2 mentions)
Msd chemstation, by Agilent Technologies (1 mentions)
5975c mass spectrometer, by Agilent Technologies (1 mentions)
7890a gas chromatograph, by Agilent Technologies (1 mentions)
Tsq quantum ultra, by Thermo Fisher Scientific (1 mentions)
Hp 5ms capillary column, by Agilent Technologies (1 mentions)
Agilent 5975c quadrupole mass spectrometer, by Agilent Technologies (1 mentions)
14 protocols using db 17ms capillary column
1
Fatty Acid Profiling by GC-MS
2
Screening for Unknown Substances in Nicotine Pouches
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Screening for unknown substances contained in nicotine pouches was performed in part 1 of this study where the procedure is described in more detail (Mallock-Ohnesorg et al. 2023 (link)). In brief, a method using liquid–liquid extraction (LLE) and gas chromatography with mass spectrometric detection (GC/MS) was adapted from Hutzler et al. (2014 (link)). Nicotine pouches were submersed in ultra-pure water and extracted with ethyl acetate under acidic conditions (after addition of 0.1 M hydrochloric acid) and basic conditions (after addition of 0.2 M ammonia). A 2 µl aliquot of the organic phase was injected into the GC/MS system and separated on a DB-17 ms capillary column (30 m × 0.25 mm I.D., 0.25 µm film thickness; Agilent Technologies, Waldbronn, Germany). Peaks were identified using the software Mass Hunter Qualitative Analysis version 10.0 (Agilent Technologies, Waldbronn, Germany) and MSD ChemStation version F.01.03.2365 (Agilent, Technologies, Waldbronn, Germany) and three different spectra libraries: NIST spectral library version 11, Flavor & Fragrance Natural & Synthetic Compounds 3 (FFNSC3) library, and an in-house aroma library created with solutions of standard substances. Nicotine was included as a reference to calculate relative retention times (RRTs). For substances that were included in the in-house library, identification was verified using the RRTs (± 0.05).
3
Quantitative Palmitic Acid Synthesis Assay
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DNL was measured as previously described (49 (link)). Total palmitic acid labeling assay in the liver was assayed by GC–MS. Briefly, 20 mg liver tissue was homogenized in 1 ml KOH/EtOH (EtOH 75%) and incubated at 85 °C for 3 h, and 200 μl of 1 mM [13C16]palmitate was added to samples as IS after cool down. Extracted palmitate acid was mixed with 50 μl N-t-butyldimethylsilyl-N-methyltrifluoroacetamide (TBDMS) at 70 °C for 30 min, and the TBDMS-derivatized samples were analyzed with an Agilent 5973N-MSD equipped with an Agilent 6890 GC system, and a DB-17MS capillary column (30 m × 0.25 mm × 0.25 μm). The mass spectrometer was operated in the electron impact mode (70 eV). The temperature program was as follows: 100 °C initial, increase by 15 °C/min to 295 °C, and hold for 8 min. The sample was injected at a split ratio of 10:1 with a helium flow of 1 ml/min. Palmitate–TBDMS derivative eluted at 9.7 min, and the m/z at 313, 314, and 319 were extracted for M0, M1, and M16 palmitate quantification.
Stable isotope labeling was corrected for natural isotope abundance (96 (link)). Newly synthesized palmitic acid was calculated as: percent of newly synthesized palmitic acid labeling = total palmitic acid labeling/(plasma 2H2O labeling × 22) × 100.
Stable isotope labeling was corrected for natural isotope abundance (96 (link)). Newly synthesized palmitic acid was calculated as: percent of newly synthesized palmitic acid labeling = total palmitic acid labeling/(plasma 2H2O labeling × 22) × 100.
4
Pyrolysis-GC/MS for Compound Identification
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A pyrolysis–gas chromatography/mass spectrometry (Py–GC/MS) system was used to separate and identify the pyrolysis products. This system is composed of a pyrolyser (CDS 5200, Chemical Data Systems, USA) connected to an Agilent 7890A gas chromatograph (GC) mated to an Agilent 5975C mass spectrometer (MS). The valve oven and transfer lines were maintained at 300 and 285 °C, respectively. The test parameters for the pyrolyser operation are as follows: sample mass, 0.5 mg; carrier gas, N2 (99.999%) with a flow rate of 1 mL/min; pyrolysis temperature, 500 °C; heating rate, 20 °C/s; holding time, 30 s. Separation of volatile pyrolysis products was achieved on an Agilent DB-17 ms capillary column (30 m × 0.25 mm × 0.25 μm film thickness). The split ratio was 50:1 with a helium carrier gas flow of 1 mL/min. The GC oven temperature was held at 40 °C for 4 min, programmed to 230 °C (2 min) at 5 C/min, and then heated further to 280 °C (5 min) at 10 °C/min. The parameters for GC/MS operation are as follows: injector temperature, 200 °C; GC/MS interface temperature, 300 °C; mass spectrometer, EI mode at 70 eV; mass spectra, from m/z 20 to 400 with a scan rate of 500 amu/s. Peak identification was carried out according to the NIST MS library and relevant literature (Additional file 1 ).
5
GC-MS Analysis of Palmitate-TMS Derivative
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Palmitate–TMS derivative was analyzed using an Agilent 5973N-MSD equipped with an Agilent 6890 GC system, and a DB-17MS capillary column (30 m x 0.25 mm × 0.25 µm). The mass spectrometer was operated in the electron impact mode (EI; 70 eV). The temperature program was as follows: 100 °C initial, increase by 15 °C/min to 295 °C and hold for 8 min. The sample was injected at a split ratio of 10:1 with a helium flow of 1 mL/min. Palmitate–TMS derivative eluted at 9.7 min. Mass scan from 100 to 600 was chosen in the method. The m/z at 313, 314, and 327 were extracted for M0/M1 palmitate and heptadecanoic acid quantification.
All the stable isotope labeling was corrected from the natural stable isotope distribution37 (link). The newly synthesized total palmitic acid was calculated as following:36 (link) %newly synthesized palmitic acid labeling = total palmitic acid labeling /(plasma 2H2O labeling × 22) × 100.
All the stable isotope labeling was corrected from the natural stable isotope distribution37 (link). The newly synthesized total palmitic acid was calculated as following:36 (link) %newly synthesized palmitic acid labeling = total palmitic acid labeling /(plasma 2H2O labeling × 22) × 100.
6
GC-MS Protocol for Compound Analysis
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GC–MS analysis was performed on a 7000 triple quadrupole GC–MS system (Agilent, Santa Clara, CA, USA) equipped with a DB-1ms, DB-5ms or DB-17ms capillary column (30 m × 0.25 mm I.D., film thickness 0.25 μm; Agilent J&W, Folsom, CA, USA). The oven temperature was maintained at 80 °C for 1 min following the injection of each sample and then increased to 300 °C at a rate of 15 °C/min. The injection port and interface temperature were set at 250 °C. Helium was used as the carrier gas at a flow rate of 1.0 ml/min. One microliter of sample solution was injected in splitless mode.
For EI mode, the ionization energy was 70 eV, the ion source temperature was 230 °C, and the scan mass range was m/z 40–400.
For CI mode, the ionization energy was 70 eV, the ion source temperature was 250 °C, and the scan mass range was m/z 43–400. Methane was used as a reactant gas.
For EI mode, the ionization energy was 70 eV, the ion source temperature was 230 °C, and the scan mass range was m/z 40–400.
For CI mode, the ionization energy was 70 eV, the ion source temperature was 250 °C, and the scan mass range was m/z 43–400. Methane was used as a reactant gas.
7
GC-MS/MS Analysis of Eugenol
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The GC-MS/MS includes a Themo Scientific GC system (US) and a Thermo TSQ Quantum Ultra triple-quadrupole mass spectrometer with electron ionization (EI) mode. The chromatographic separation was performed with a DB-17MS capillary column (30 m × 0.25 mm × 0.25 μm) made by Agilent Technologies. Helium (99.999% pure; Air Liquide) acted as carrier gas with the flow rate of 1.0 mL min− 1. The injector and the GC-MS interface temperatures were set to 260 °C and 280 °C, respectively. The initial column temperature was 80 °C and maintained for 2 min. Then it was increased to 250 °C at a rate of 25 °C min− 1 and held for 5 min. Finally, the temperature ramped to 280 °C at a rate of 25 °C min− 1 and held for 17 min. The injection volume of the samples was 1 μL and the solvent delay time was 180 s.
The ion source and quadrupole temperatures were 230 °C and 150 °C, respectively. The electron ionization energy of the mass selective detector was 70 eV. The major fragment ions used to quantify and qualify eugenol are shown in TableS1 .
The ion source and quadrupole temperatures were 230 °C and 150 °C, respectively. The electron ionization energy of the mass selective detector was 70 eV. The major fragment ions used to quantify and qualify eugenol are shown in Table
8
Muscle Protein Synthesis Measurement Protocol
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Plasma samples and standards were prepared and analyzed as described previously [23 (link)]. Briefly, 20 μl of plasma/standard, 4 μl of 5% acetone in acetonitrile (vol/vol), and 2 μl of 10 N NaOH were combined and permitted to settle for 24 h. Each sample/standard was thoroughly mixed with 600 μl of chloroform and 0.5 g of Na2SO4. The sample/standard was analyzed on a GC-MS (Agilent 5973 N-MSD furnished with an Agilent 6890 GC System and DB17-MS capillary column).
Approximately 30 mg of muscle (soleus and plantaris) from each sample was homogenized with 0.3 ml of 10% TCA and centrifuged for 15 min at 3800 rpm at 4 °C. The resultant pellet was then washed with 10% TCA, centrifuged, and disposed of the supernate for an additional 3 times. The pellet was then mixed with 6 N HCl and incubated for 18 h at 100 °C. The hydrolysate was then freeze dried for 24 h before 100 μl of a 3:2:1 ratio of methyl-8, methanol and acetonitrile were added to each sample and analyzed on the GC-MS.
The following equation was used to determine the mixed muscle protein FSR:
EA signifies the quantity of 2H-labeled alanine in protein (%), EBW indicates the amount of 2H2O found in body water (%), and t represents time in h [23 (link)].
Approximately 30 mg of muscle (soleus and plantaris) from each sample was homogenized with 0.3 ml of 10% TCA and centrifuged for 15 min at 3800 rpm at 4 °C. The resultant pellet was then washed with 10% TCA, centrifuged, and disposed of the supernate for an additional 3 times. The pellet was then mixed with 6 N HCl and incubated for 18 h at 100 °C. The hydrolysate was then freeze dried for 24 h before 100 μl of a 3:2:1 ratio of methyl-8, methanol and acetonitrile were added to each sample and analyzed on the GC-MS.
The following equation was used to determine the mixed muscle protein FSR:
EA signifies the quantity of 2H-labeled alanine in protein (%), EBW indicates the amount of 2H2O found in body water (%), and t represents time in h [23 (link)].
9
GC-MS Analysis of Complex Samples
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GC–MS was performed on a 7000 Triple quadrupole GC–MS system (Agilent, Santa Clara, CA, USA) equipped with a DB-1ms, a DB-5ms, or a DB-17ms capillary column (30 m × 0.25 mm I.D., film thickness 0.25 μm, Agilent J&W, Folsom, CA, USA). The oven temperature was maintained at 80 °C for 1 min following injection and then raised to 300 °C at a rate of 15 °C/min. The injection port and interface temperature were set at 250 °C. Helium was used as the carrier gas at a flow rate of 1.0 ml/min. One microliter of sample solution was injected in splitless mode.
For EI mode, the ionization energy was 70 eV, the ion source temperature was 230 °C, and the scan mass range was m/z 40–400. For CI mode, the ionization energy was 70 eV, the ion source temperature was 250 °C, and the scan mass range was m/z 43–400.
For EI mode, the ionization energy was 70 eV, the ion source temperature was 230 °C, and the scan mass range was m/z 40–400. For CI mode, the ionization energy was 70 eV, the ion source temperature was 250 °C, and the scan mass range was m/z 43–400.
10
Pesticide Residue Analysis Using GC and HPLC
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An Agilent 6890N gas chromatography device was used for the analysis of pesticide residues. The detector temperature was set to 300°C, the H2 flow rate was 3.0 mL/min, the air flow rate was 60 mL/min, and there were columns in the HP-5MS capillary column (30 m × 0.25 mm, 0.25 μm, Agilent) and DB-17MS capillary column (30 m × 0.25 mm, 0.25 μm, Agilent). The gas flow rate was 1.0 mL/min. The HPLC device used for the analysis of pesticide residue was a Hewlett Packard 1100 (Agilent, Santa Clara, CA, USA). The detection wavelength was set to 254 nm and 275 nm and the column was an InsertSustain C18 column (4.6 × 250 mm, 5 μm; GL Science, Torrance, CA, USA). The mobile phase was a mixture of water and acetonitrile: 70:30 (0–5.0 min) and 15:85 (5.0–22.0 min) with a flow rate of 1.0 mL/min.
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