Agilent 7890b 7000d gc ms

Fecal samples collected from the mice and tissues collected from the CRC patients were mixed with pre-cooled methanol/acetonitrile/water solution in the ratio of 2:2:1 (v/v), followed by vortex mixing, low-temperature ultrasonic processing for 30 min, freezing at −24°C for 10 min, and then centrifugation at 14,000 rpm for 20 min at 4°C. Later, the supernatant was extracted and dried in vacuum. Subsequently, 100 μL of acetonitrile aqueous solution (acetonitrile: water = 1:1, v/v) was added for mass spectrometry, followed by vortex mixing and centrifugation at 14,000 rpm at 4°C for 15 min. Finally, the samples were sent with clear liquid for analysis by Shanghai Applied Protein Technology Co., Ltd. (Shanghai, China). Mouse feces and human tissues were detected by Agilent 7890B/7000D GC-MS (Agilent, United States) for targeted detection of SCFA. In addition, mouse feces were also tested for untargeted metabolites by UHPLC-Q-TOF MS (Agilent, United States; AB SCIEX, United States).

Three flower buds of every repeat at five chilling points were crushed by a crusher (MM 400, Retsch) containing zirconia beads at 30 Hz. A total of 20 mg powder was added into 500 μL of methanol: isopropanol: water (3: 3: 2, V/V/V) solution. The supernatant (50 μL) were taken and evaporated in nitrogen after being vortexed (3 min) and sonicated (30 min), with adding internal standard. After evaporated in a nitrogen stream and freeze-drying, the residue was further derivated as follows: Firstly, the small molecule carbohydrate was mixed with methoxine hydrochloride solution (100 μL) in the 1.5 mL tubes. Secondly, bistrifluoroacetamide (100 μL) was added to the solution at 37 °C for 2 h. After vortexed, the mixture was incubated at 37 °C for 30 min. N-Hexane was used as dilution fusion. Then, mixture was detected by MetWare (http://www.metware.cn/) based on the Agilent7890B-7000D GC-MS/MS platform, and the parameters were set as reported by Gómez-González et al. [65 (link)] .

The volatile compounds of Chinese chive were analyzed using an Agilent 7890B/7000D GC-MS Agilent, Santa Clara, CA, USA) under the following conditions: capillary column, DB-WAX (30 m × 0.25 mm, 0.25 μm) with He (≥99.999% purity) as the carrier gas at a flow rate of 1 mL/min and splitless mode; initial temperature 40 °C held for 1 min, raised to 80 °C at 8 °C/min, then raised to 130 °C at 2 °C/min, and finally raised to 220 °C at 6 °C/min held for 3 min; total analysis time, 49 min; MS ionization, EI, 70 eV; MS source, 230 °C, scan area, 30–660 amu.