In order to explore reliable biomarkers of OSCC, we used GC–MS-based untargeted analysis to find the difference of potential metabolite between tumor and matched adjacent normal tissues of 8 OSCC patients. Materials and reagents for GC–MS analysis were prepared according to previous method [5 (link)]. GC–MS analysis was performed by Trace 1310 Gas Chromatograph equipped with an AS 1310 auto sampler, which connected a TSQ 8000 triple quadrupole mass spectrometer (Thermo Scientific, Waltham, MA, USA).
As 1310
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The AS 1310 is a laboratory equipment product from Thermo Fisher Scientific. It is a high-performance autosampler designed to handle a wide range of sample types and volumes. The core function of the AS 1310 is to automatically introduce samples into an analytical instrument for analysis.
Automatically generated - may contain errors
Lab products found in correlation
Trace 1310 gas chromatograph, by Thermo Fisher Scientific (3 mentions)
Tsq 8000 triple quadrupole mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Trace 1310, by Thermo Fisher Scientific (2 mentions)
Optima 17, by Macherey-Nagel (1 mentions)
Trace 1300 gas chromatograph, by Thermo Fisher Scientific (1 mentions)
Chromeleon software, by Thermo Fisher Scientific (1 mentions)
Stabilwax da, by Restek (1 mentions)
Tsq 8000 mass spectrometer, by Thermo Fisher Scientific (1 mentions)
8 protocols using as 1310
1
Untargeted Metabolite Analysis of OSCC
2
GC-MS Quantification of Acetazolamide
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GC-MS analyses were performed on a single-quadrupole mass spectrometer model ISQ directly interfaced with a Trace 1310 series gas chromatograph equipped with an autosampler AS 1310 from ThermoFisher (Dreieich, Germany). The gas chromatograph was equipped with a 15 m long fused-silica capillary column Optima 17 (0.25 mm I.D., 0.25 μm film thickness) from Macherey-Nagel (Düren, Germany). In quantitative analyses, the following oven temperature program was used with helium (at a constant flow rate of 1 mL/min) as the carrier gas: 1.0 min at 90 °C, then increased to 250 °C at a rate of 35 °C/min and to 320 °C at a rate of 35 °C/min, respectively. The column was held at 320 °C for 6 min. Interface, injector and ion-source were kept constant at 260 °C, 200 °C and 250 °C, respectively. Electron energy was set to 70 eV and electron current to 50 μA. Methane (2.4 mL/min) was used as the reagent gas for NICI. Aliquots (1 μL from the toluene phase) were injected in the splitless mode by means of the autosampler. Quantification of acetazolamide was performed in the NICI mode by selected-ion monitoring (SIM) the ions with m/z 581 and m/z 83 for d0-AZM and m/z 584 and m/z 86 for d3-AZM (IS) with a dwell-time of 50 ms for each ion. The electron multiplier voltage was set to 2025 V. Deviations from the conditions described above are mentioned in the sections Results and Discussion.
3
GC-MS Analysis of Medicinal Compounds
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GC–MS experiment is under the guidance of professor Tong Xie (Nanjing University of Chinese Medicine). Materials and reagents for GC–MS analysis were prepared according to previous method. GC–MS analysis was performed by Trace 1310 Gas Chromatograph equipped with an AS 1310 auto sampler, which connected a TSQ 8000 triple quadrupole mass spectrometer (Thermo Scientific, Waltham, MA). See references for specific steps [14] (link).
4
GC-MS Analysis of Nitrates, Nitrites, Creatinine, and Amino Acids
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Gas chromatographic-mass spectrometric analyses were performed on a single quadrupole mass spectrometer model ISQ directly interfaced with a Trace 1310 series gas chromatograph equipped with an autosampler AS 1310 from ThermoFisher (Dreieich, Germany). Different oven temperature programs were used for the separation of the derivatives of nitrate, nitrite, creatinine and MDA, on the one hand, and of the derivatives of the amino acids and their metabolites, on the other hand. Selected-ion monitoring of specific anions for unlabelled nitrate, nitrite, creatinine, MDA and for their stable-isotope labelled analogues was performed [33 (link)]. Amino acids were analysed by selected-ion monitoring of specific anions for endogenous and their stable-isotope labelled analogues as reported previously [34 (link)].
5
Quantification of Short-Chain Fatty Acids in Colon Digesta
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The colon digesta samples were thawed on ice. A mixture of 500 mg of digesta sample and 500 µl of ice cold MQ-H2O was sonicated for 5 min in cold water. After mixing and centrifugation (15 min, 4 °C, 15000 g), the supernatant was transferred to a spin column (45 kDa). After another centrifugation step (15 min, 4 °C, 15000 g), supernatants were spiked with internal standard (2-methylvaleric acid). SCFA were measured by “TRACE 1300 Gas Chromatograph” equipped with a flame ionization detector and autosampler “AS 1310” (Thermo Fischer Scientific, Milan, Italy). The capillary column (Stabilwax-DA; 30-m × 0.25-mm i.d., 0,25 µm; Restek, Bellefonte, PA, USA) were operated as follows: starting temperature 90 °C (2 min); temperature increase 10 °C/min until 150 °C, 50 °C/min until 250 °C (1 min). The rate of helium flow was 3 mL/min. Concentrations of acetic, propionic acids, as well as butyric, valeric acids and their isomers were quantified against external standards using Chromeleon software (Dionex, Thermo Scientific) and reported in µmol per gram of intestinal contents.
6
GC-MS Analysis of Cellular Metabolites
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When cells were 80% to 90% confluent, cells were collected in 10-cm dishes and sent to Dr. Tong Xie (Nanjing University of Chinese Medicine). Materials and reagents for GC-MS analysis were prepared according to the previous method 22 (link). GC-MS analysis was performed by using a Trace 1310 Gas Chromatograph equipped with an AS1310 auto sampler which connected to a TSQ 8000 triple quadrupole mass spectrometer (Thermo Scientific, Waltham, MA).
7
GC-MS Analysis of n-Alkanes and PAHs
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Analyses for n-alkane and PAHs were performed on a Thermofisher Scientific Trace 1300 Gas Chromatograph coupled with Thermo TSQ8000 Mass Spectrometer with a Thermo AS 1310 auto-sampler under select ion monitoring (SIM) mode. A DB-5 column having 60-m length × 0.25-mm internal dia. with 0.25-μm film thickness with helium (1.2 mL/min) as a carrier gas was used for the sample analysis. The injector and detector temperatures were set to 260°and 300 °C, respectively, for n-alkane and PAH analyses. The oven temperature program for n-alkane analysis was initially 50 °C hold for a minute, then ramp to 140 °C at 10 °C/min, and ramp to 320 °C at 6 °C/min and finally 28 min hold at 320 °C. The oven temperature program for PAH analysis was initially 70 °C with a hold of 2 min, ramp to 150 °C at 30 °C/min then ramp to 310 °C at 4 °C/min and finally a 10 min hold at 310 °C. Typical n-alkanes and PAH chromatograms are shown in Fig. 1.
8
GC-MS Analysis of NOHA Derivatives
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GC-MS analyses were performed on a GC-MS apparatus consisting of a single-stage quadrupole mass spectrometer model ISQ, a Trace 1210 series gas chromatograph and an AS1310 autosampler from Thermo Fisher (Dreieich, Germany). Aliquots of 1 µL toluene extracts were injected in the splitless mode. The injector temperature was kept at 280 °C. Helium was used as the carrier gas at a constant flow rate of 1.0 mL/min. The oven temperature was held at 40 °C for 0.5 min and ramped up to 210 °C at a rate of 15 °C/min and then to 320 °C at a rate of 35 °C/min; finally the column was kept at 320 °C for 1 min. Interface and ion-source temperatures were set to 300 and 250 °C, respectively. Electron energy was 70 eV and electron current 50 µA. Methane (2.4 mL/min) was used as the reagent gas for both negativeion chemical ionization (NICI) and positive-ion chemical ionization (PICI). Mass spectra of NOHA derivatives were generated by scanning in the mass-to-charge (m/z) range of 50-1000. In quantitative analyses, the dwell time was 100 ms for each ion in the selected-ion monitoring (SIM) mode. The electron multiplier voltage was set to 1900 V.
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