Cadmium's Silent Sabotage: Unveiling Its Impact on Antibiotic Efficacy
Description
Raw metabolomics data, the data set is related to Figure 3
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For metabolites extraction, 100 μL serum sample from control and Cd-exposure group mice after 12 weeks of feeding were mixed with 400 μL extraction solution (MeOH:ACN, 1:1) containing deuterated internal standards. Samples were vortexed for 30 s, sonicated for 10 min at 4 °C and incubated at -40 °C for 1 h to precipitate proteins. After centrifugation (13800g for 15 min at 4 °C), the supernatants were obtained for LC-MS/MS analysis. The metabolite extracts were analyzed by a Vanquish UHPLC system (Thermo Fisher Scientific) with a Waters ACQUITY UPLC BEH Amide (2.1 mm × 50 mm, 1.7 μm) coupled to Orbitrap Exploris 120 mass spectrometer (Thermo Fisher Scientific). Water with 25 mmol/L ammonium acetate and 25 mmol/L ammonia hydroxide (pH=9.75) (A) and acetonitrile (B) were used as the mobile phase. The auto-sampler temperature was 4 ℃, and the injection volume was 2 μL. The ESI source conditions were set as follows: sheath gas flow rate as 50 Arb, Aux gas flow rate as 15 Arb, capillary temperature 320 ℃, full MS resolution as 60000, MS/MS resolution as 15000, collision energy: SNCE 20/30/40, spray voltage as 3.8 kV (positive) or -3.4 kV (negative), respectively. The raw data were converted to the mzXML format using ProteoWizard and processed with an in-house program, which was developed using R and based on XCMS, for peak detection, extraction, alignment, and integration. Then an in-house MS2 database (BiotreeDB) was applied in metabolite annotation. The cutoff for annotation was set at 0.3.