Synapt g2 s qtofms system

In order to identify the difference of metabolites between the two varieties of A. bisporus, we selected 12 samples (six in each group, two groups) for metabolomics analysis. The untargeted metabolomics was carried out by ultra-performance liquid chromatography-quadrapole time of flight mass spectrometry (UPLC-QTOFMS) according to the previously established methods [11 (link)–13 (link)]. The freeze-dried samples were crushed using a mixer mill (MM 400, Retsch) with a zirconia bead for 1.5 min at 30 Hz. Then 100 mg powder was weighed and extracted overnight at 4°C with 1.0 ml 70% aqueous methanol containing 0.1 mg/L lidocaine for internal standard. Following centrifugation at 10000 g for 10min, the supernatant was analyzed using an LC-ESI-MS/MS system. Briefly, 2 μl of samples were injected onto a Waters ACQUITY UPLC HSS T3 C18 column (2.1 mm*100 mm, 1.8 μm) operating at 40°C and a flow rate of 0.4 mL/min. The mobile phases used were acidified water (0.04% acetic acid) (Phase A) and acidified acetonitrile (0.04% acetic acid) (Phase B). Compounds were separated using the following gradient: 95:5 Phase A/Phase B at 0 min; 5:95 Phase A/Phase B at 11.0 min; 5:95 Phase A/Phase B at 12.0 min; 95:5 Phase A/Phase B at 12.1 min; 95:5 Phase A/Phase B at 15.0 min. The effluent was connected to an the Synapt G2-S QTOFMS system (Waters, Milford, MA).