Esi qtof microtof qii

Mass spectrometry experiments were carried out by using ESI-Q-TOF (micrOTOF-Q II, Bruker, Bremen, Germany) mass spectrometer at negative ion mode and the data were analysed with Bruker ESI Compass Data Analysis v. 4.0 software. Optimal soft ionization conditions were obtained based on previous methods [21 (link)]. This system can measure m/z in the range of 50–3000. In order to achieve a satisfactory ionization of the DNA samples, an equal volume of 60% methanol solution was added before MS analysis. The spectrum acquisition time of each sample is 0.6 min and the sample infusing into the ion source is at a rate of 3 µl min⁻¹.

The extract was analyzed by Ultra Fast Liquid Chromatography (UFLC) (Shimadzu) coupled to Diode Array Detector (DAD) (240–800 nm, Shimadzu) and electrospray ionization time-of-flight (ESI-QTOF-micrOTOF QII) (operating in positive and negative mode, 120–1200 Da, Bruker Daltonics). A C-18 column was used (Kinetex, 2.6 μm, 150 × 2.2 mm, Phenomenex) protected by a guard column of the same material. The mobile phase was as follows: water (solvent A) and acetonitrile (solvent B) both with 0.1% of formic acid in a gradient of 0–2 min 3% B, 2–25 min 3–25% B, and 25–40 min 25–80% B followed by washing and reconditioning of the column (8 minutes). The flow rate was 0.3 mL/min and 1 μL (1 mg/mL) of extract was injected. The other micrOTOF-QII parameters were as follows: temperature, 200°C; N₂ drying gas flow rate, 9 L/min; Nebulizer, 4.0 bar; capillary voltage, −3500 V (negative) and +4500 V (positive); and internal calibration with TFA-NA injected at the end of the chromatographic analysis. The rutin and epicatechin standards were obtained from Sigma-Aldrich with a purity of ≥95%.

ESVR was analyzed in an ultrafast liquid chromatograph (UFLC, Shimadzu) coupled to a diode array detector (DAD, Shimadzu) and electrospray ionization time-of-flight mass spectrometer (ESI-QTOF-micrOTOF QII, Bruker Daltonics; operating in the positive and negative ionization modes, 120-1200 Da). A C-18 column was used (Kinetex, 2.6 μm, 150 × 2.2 mm, Phenomenex), protected by a guard precolumn of the same material. The mobile phase was water (solvent A) and acetonitrile (solvent B), both with 0.1% formic acid, in a gradient of 0-2 min 3% B, 2-25 min 3-25% B, and 25-40 min 25-80% B, followed by the washing and reconditioning of the columns (8 min). The flow rate was 0.3 mL/min, and 1 μL of the extract (1 mg/mL) was injected. The other micrOTOF QII parameters were as follows: temperature, 200°C; N₂ gas flow rate, 9 L/min; nebulizer, 4.0 bar; capillary voltage, 3500 V (negative), +4500 V (positive); and internal calibration with sodium trifluoroacetate (TFA-Na) injected at the end of the chromatographic analysis. The catechin and piceatannol authentic standards were purchased from Sigma-Aldrich with ≥95% purity. The metabolites present in ESVR were identified based on the interpretation of mass and UV absorption spectra and based on comparison with the literature. When available, the compounds were confirmed by comparison with authentic standards.