The sample was analyzed by GC-MS using an Agilent 7890-5975C GC-MS solution system (Agilent, Sacramento, USA) with an hp-5 capillary column (30 m × 0.25 mm, 0.25 μm film thickness; Agilent J.W. Scientific, Folsom, USA). One microliter of sample was injected into the DB-5MS capillary column coated with 5% phenyl and 95% methylpoly siloxane in split-less mode. The column temperature was held at 70 °C for 2 min and then increased to 290 °C at a rate of 8 °C/min and held for 3 min. Helium was used as a carrier gas, and the flow was constant at 1 mL/min. The transfer line and ion source temperatures were 280 °C and 250 °C, respectively. The mass scan range was 50–600 m/z. A supervised partial least-squares discriminant analysis (PLS-DA) was subsequently performed to identify the metabolites contributing to differences between the control group and the fluconazole group. A metabolite with a variable influence on the projection value (VIP) higher than 1 indicates a significant contribution to the separation of groups in PL-SDA models.
Agilent 7890 5975c gc ms solution system
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 7890-5975C GC-MS solution system is a gas chromatography-mass spectrometry (GC-MS) instrument designed for analytical applications. It combines a 7890 series gas chromatograph and a 5975 series mass spectrometer, providing a complete solution for compound separation, identification, and quantification.
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3 protocols using agilent 7890 5975c gc ms solution system
1
GC-MS Analysis of Metabolic Differences
2
GC-MS Metabolite Derivatization Protocol
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Derivatization was performed according to previously described [26] . First, the dried pellet was dissolved in 80 µL of 20 mg/mL methoxyamine hydrochloride (Sigma, St. Louis, USA) containing pyridine and incubated for 1.5 h at 37 °C. Subsequently, 80 µL N-methyl-N-trimethyl-silyl-tri uoroacetamide (MSTFA, Sigma, St. Louis, USA) was added and incubated for 0.5 h at 37 °C. The samples were then centrifuged at 12,000 × g for 10 min at 4 °C. Chemical analysis of the samples was performed using the Agilent 7890-5975C GC-MS solution system (Agilent, Sacramento, California, USA). The injector temperature was held at 270 °C. Using the splitless model, 0.5 µL of the derivatized sample was injected into a Dodecyl Benzene Sulfonate (DBS) column of 30 m in length, 250 µm in inner diameter, and 0.25 µm in thickness. The temperature program of the GC oven is 85 °C for 5 min. The temperature is raised to 285 °C at 5 °C/min, and nally to 310 °C at a rate of 20 °C/min. Helium was used as carrier gas with a ow rate of 1 mL/min. The scanning mass range was set at 50-600 m/z, and the ionization energy of the electron impact ionization (EI) was 70 eV.
3
GC-MS Metabolite Derivatization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Derivatization was performed according to previously described [26] . First, the dried pellet was dissolved in 80 µL of 20 mg/mL methoxyamine hydrochloride (Sigma, St. Louis, USA) containing pyridine and incubated for 1.5 h at 37 °C. Subsequently, 80 µL N-methyl-N-trimethyl-silyl-tri uoroacetamide (MSTFA, Sigma, St. Louis, USA) was added and incubated for 0.5 h at 37 °C. The samples were then centrifuged at 12,000 × g for 10 min at 4 °C. Chemical analysis of the samples was performed using the Agilent 7890-5975C GC-MS solution system (Agilent, Sacramento, California, USA). The injector temperature was held at 270 °C. Using the splitless model, 0.5 µL of the derivatized sample was injected into a Dodecyl Benzene Sulfonate (DBS) column of 30 m in length, 250 µm in inner diameter, and 0.25 µm in thickness. The temperature program of the GC oven is 85 °C for 5 min. The temperature is raised to 285 °C at 5 °C/min, and nally to 310 °C at a rate of 20 °C/min. Helium was used as carrier gas with a ow rate of 1 mL/min. The scanning mass range was set at 50-600 m/z, and the ionization energy of the electron impact ionization (EI) was 70 eV.
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