Metabolite data of Trametes Robiniophila Murr in rats
Description
Rat metabolites identified by liquid chromatography-mass spectrometry (LC-MS) after gavage of trametes robiniophila murr。
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The sample was accurately weighed into a 2 mL EP tube, 600 µL of methanol (containing 2-chloro-L-phenylalanine (4 ppm)) was added, vortexed for 30 s. Then steel beads were added, and the sample was placed into a tissue grinder (50 Hz, 120 s), sonicated at room temperature for 10 min, and centrifuged at 4°C (12,000 rpm for 10 min). Finally, filtered and analysed by liquid chromatography-mass spectrometry (LC-MS). The chromatographic separation was performed on an ultra-high performance liquid cTRMomatography (UPLC) system (Thermo Vanquish (Thermo Fisher Scientific, USA)) and an ACQUITY UPLC® HSS T3 column (100 mm×2.1 mm, 1.8 μm, Waters, Milford, MA, USA). For chromatographic separation: column temperature 40 °C, flow rate 0.3 mL/min, injection volume 2 μL. The mobile phases were positive ionic 0.1% formic acid acetonitrile (B2) and 0.1% formic acid in water (A2), and negative ionic acetonitrile (B3) and 5 mM ammonium formate in water (A3). A Thermo Q Exactive mass spectrometer (Thermo Fisher Scientific, USA) was used for the acquisition of primary and secondary spectra of the samples. The scans were performed in both positive and negative modes with the following electrospray ion source conditions: positive ion spray voltage of 3.50 kV, negative ion spray voltage of -2.50 kV, sheath gas of 40 arb, and auxiliary gas of 10 arb. The capillary temperature was 325 °C, and the primary full scans were performed at a resolution of 70,000, with primary ion scans in the range of m/z100~1000, and secondary cleavage was performed using HCD. with a collision energy of 30 eV and a secondary resolution of 17500. The first 10 ions of the acquired signal were fragmented, and dynamic exclusion was used to remove unnecessary MS/MS information. The faecal metabolites were determined at Genesky Biotechnologies Inc. The metabolite distribution of all faecal samples was evaluated using principal-component analysis (PCA). Differences between the two groups were analysed using orthogonal partial least-squares discriminant analysis (OPLS-DA). Identification of differential metabolites was based on a volcano plot (P<0.05) and VIP analysis (VIP>1.0, P<0.05).