Maxis impact hd lc q tof mass spectrometer

Ethyl acetate extracts were obtained as described above, dried in vacuo, and re-dissolved in 200 μL of methanol before analysis. 1 μL of methanol solution was separated on C18 Phenomenex columns (100×2.1 mm, 1.7 μm) using Dionex Ultimate 3000 HPLC-DAD system coupled to MaXis Impact HD LC-Q-TOF mass-spectrometer (Bruker Daltonics). The LC runtime was 22 min, at a flow rate of 0.6 mL/min. Solvent system: water + 0.1% formic acid (HFo; solvent A), acetonitrile + 0.1% HFo (solvent B); from 95% A to 0% A in 12 min, then 3 min at 0% A, then reversion to 45% A within the remaining 7 min. Ionization was performed in positive and negative modes. Analysis of LC-MS data was performed using the program Compass Data Analysis 4.2 (Bruker Daltonics). The quantitative analysis of Clo production was done by comparing areas of extracted Clo mass-peak (697.2 Da, [M + H]⁺) present in NRRL 3504 ethyl acetate extracts to the areas of 697.2-Da mass peak produced by the known amounts of authentic Clo run through the machine. A reference calibration curve was created using the values of areas for 697.2-Da mass peaks obtained when 6 different amounts of Clo were injected and run under conditions mentioned above (see Table S1, Fig. S1). See ESM for the access to raw LC-MS data.