Analysis was performed on an Agilent 7890B GC using a 5977A MSD mass spectrometer for compound detection. Separation was achieved on a Cp-sil 19, 25m∗0.25mm∗0.20 μm column. Temperature gradient was as follows: 60°C was held for 1.5 min after which the temperature increased to 240°C at a rate of 10°C/min. The temperature was then increased to 270, at 4°C/min. Finally the temperature increased to 300°C at 20°C/min. An injection volume of 1 μL was used in splitless mode using a splitless time of 1.5 min. The carrier gas was Helium (50 kPa). Single ion monitoring mode was used for detection of VLCFAs.
5977a msd mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The 5977A MSD mass spectrometer is a high-performance analytical instrument designed for sensitive and accurate detection and identification of chemical compounds. It utilizes electron ionization (EI) technology to generate and separate ions based on their mass-to-charge ratio, enabling the analysis of a wide range of sample types.
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5 protocols using 5977a msd mass spectrometer
1
GC-MS Analysis of Very Long Chain Fatty Acids
2
GC-MS Analysis of Compounds
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The analysis was performed on an Agilent 7890B GC system coupled to a 5977A MSD mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) and equipped with a Gerstel Multipurpose sampler (Gerstel, Mülheim an der Ruhr, Germany). For the experiments using helium and hydrogen as a buffer gas, a Peak Precision Hydrogen Trace 500cc generator (Peak Scientific Instruments, Inchinnan, Scotland, UK) was used to generate the hydrogen gas. A silica capillary column, Agilent HP-5MS (30 m x 0.25 mm i.d. 0.25 μm film thickness), was used. The operation conditions were described in Table 1 . The compounds’ spectra that were obtained were compared to the spectra of known compounds using the NIST Mass Spectral Search Program for the NIST/EPA/NIH Mass Spectral Library.
3
GC-MS Analysis of Short-Chain Fatty Acids
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The instrument used in this experiment was a GC-MS Agilent 7890B gas chromatograph and an Agilent 5977A MSD mass spectrometer in series for detection in selected ion monitoring (SIM) mode. MSD ChemStation software was used for data processing to calculate the absolute content of the target compound in the sample. (1) The required gas chromatographic parameters were as follows: chromatographic column: Agilent HP-FFAP (25 m + 0.32 μm + 0.50 mm); injection volume: 2 μL; split ratio: 20 : 1; injection port: 250°C; flow rate: 1.5 mL/min; heating program: the oven program had an initial temperature of 100°C, which was increased to 150°C at a rate of 5°C/min, and maintained at 230°C for 2 min. (2) The mass spectrometry parameters were as follows: ion source: EI+; ion source temperature: 280°C; transmission line temperature: 250°C; solvent delay: 3.5 min; scanning range: scan 35–200 m/z. The SIM mode was chosen to determine the ion mass of each SCFA, including m/z values of 41, 43, 45, 57, 60, 73, 74, 87, and 88.
4
GC-MS Analysis of Metabolites
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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 Supplementary Table S7
5
Gas Chromatography-Mass Spectrometry Analysis
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Chromatographic analyzes were performed with an Agilent 7890B gas chromatograph coupled to a 5977A MSD mass spectrometer. Samples of different extraction times were injected in triplicates, with an injection volume of 1 µL, into a non-polar capillary column composed of 5.0 % phenyl and 95.0 % methylpolysiloxane (30 m × 0.25 mm id × 1 µm df) (DB-5ms, Agilent Technologies, USA). In turn, the commercial banana aroma used as reference in this work was analyzed at a concentration of 100 ppm. Samples were injected in triplicate in splitless mode at 250 °C using a carrier gas stream of 1 mL min -1 of helium. The chromatographic conditions employed were: initial temperature of 50 °C (0.5 min) raised to 100 °C (17.17 min) at a heating rate of 3 °C min -1 ; the temperature was then raised to 250 °C (42.17 min) at a heating rate of 10°C min -1 ; finally, temperature was raised to 280 °C at a heating rate of 5 °C min -1 , remaining at this temperature for 5 min. The transfer line was set at 290 °C. The mass spectrum range was 33 -500 m z -1 and the ionization energy was 70 eV. The ion source and quadrupole were maintained at 300 °C and 150 °C, respectively.
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