The monosaccharide composition of EMOS-1a was analyzed by gas chromatography–mass spectrometry (GC–MS) [33 (link)] with some modifications. Briefly, EMOS-1a (10 mg) was hydrolyzed to monosaccharides using trifluoroacetic acid (2 mL, 4 mol/L) at 110 °C for 6 h. The hydrolyzed solution was evaporated to dryness, added to hydroxylamine hydrochloride (10 mg) and pyridine (1 mL) for reaction for 30 min at 90 °C, and then acetylated with acetic anhydride (1 mL) for 30 min at 90 °C. Thus, the monosaccharides were derivatized as acetylated aldononitriles. The final product was analyzed by GC–MS using an Agilent 6890 GC instrument (Agilent Technologies Co., Ltd., Colorado Springs, CO, USA) equipped with an HP-1701 column and an Agilent 5973 MS detector. Nitrogen was used as the carrier gas (1 mL/min). The temperature of the column was initially set at 100 °C, then increased to 280 °C at 10 °C/min, followed by 280 °C for 15 min. The injection temperature was 280 °C and the temperature of the mass spectrometer ion source was 230 °C. Seven monosaccharides (rhamnose, ribose, fucose, arabinose, xylose, mannose, and galactose) were converted into their acetylated derivatives as standards to identify the composition of the oligosaccharides.
Agilent 5973 ms detector
Manufactured by Agilent Technologies
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The Agilent 5973 MS detector is a mass spectrometer designed for analytical applications. It provides accurate mass measurements and structural information for the identification and quantification of chemical compounds.
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4 protocols using agilent 5973 ms detector
1
GC-MS Analysis of EMOS-1a Monosaccharides
2
GC-MS Analysis of PAE Congeners
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The extracted PAE congeners were analyzed using gas chromatography fixed to a mass spectrometer (GC-MS), Agilent model 6890N GC–5973 MSD (Agilent Technologies, Inc. Santa Clara, CA, USA). Extracted samples were injected into the GC equipped with an HP-5 MS fused silica capillary column (30 m × 0.25 mm × 0.25 μm film thickness) and an Agilent 5973 MS detector, operating in the selective ion monitoring mode. The column temperature was initially set at 80 °C for 1 min, then ramped at 15 °C for 1 min to 300 °C and held constant for 10 min. The transfer line and the ion source temperature were maintained at 280 and 250 °C, respectively. Helium was used as the carrier gas at a flow rate of 1 mL/min. Automated samplers injected the liquid extracts of 1.0 μL in splitless mode with a venting time of 1.15 min with an inlet temperature of 300 °C. The concentrations in the water were normalized to a dry-weight basis.
3
GC-MS Analysis of Volatile Compounds in Beer
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Volatile compounds in samples were analyzed through gas chromatography with mass spectrometry detection and previous stir bar sorptive extraction (SBSE) [23 (link)]. Briefly, 50 mL of beer, with 25% NaCl (w/v) added, were extracted for 180 min at 1000 rpm employing PDMS stir bars (Gerstel, Mülheim an der Ruhr, Germany). After the extraction procedure, the stir bars were thermally desorbed in a thermal desorption unit (TDS-2, Gerstel) for their later chromatographic analysis. An Agilent 6890 GC (Agilent Technologies, Palo Alto, CA, USA) was employed, equipped with a DB-Wax (J&W Scientific, Folsom, CA, USA) capillary column (60 m × 0.25 mm I.D., 0.25 μm coating) coupled to an Agilent 5973 MS detector (Agilent Technologies). Each analysis was performed in duplicate.
4
Trichoderma Volatile Metabolite Analysis
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The volatile metabolites were analysed by gas chromatography with a single quadrupole mass spectrometer detector (GC-MS) analysis [34, 35] (link). Prior to the analysis, dried crude extracts obtained from Trichoderma species were dissolved in ethyl acetate and placed in a 1 mL glass vial. The GC system was equipped with an HP-5MS (30 m × 0.25 mm and 0.25 µm 5% diphenyl/95% dimethylpolysiloxane) capillary column (J and W Scientific, Folsom, CA, USA). The instrument was programmed to start at 40°C (held for 2 min) and to increase to 160°C by 6°C /min, after which it increased to 260°C by 10°C/min (held for 4 min). Helium (99.9%) was used as the carrier gas, and the flow rate was 1 mL/min. The flow was then transferred from the column to an Agilent 5973 MS detector (Agilent Technologies, Palo Alto, CA). The ion source temperature was set at 230°C, the ionizing electron energy was 70 eV, and the mass range was 40-450 Da in full-scan acquisition mode [33] (link). The spectra of identified metabolites were compared with the spectra of known compounds in the GC-MS NIST database. The threshold for identification was ≥90% similarity. The analysis was carried out in triplicate to monitor the repeatability of the analysis. The instrumental responses obtained were interpreted using mass hunter ChemStation software.
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