630b accurate mass qtof ms

The samples were also analyzed qualitatively by HPLC/ESI-QTOF-MS system in positive ion mode using a 630B accurate mass QTOF-MS (Agilent Technologies INc., Santa Clara, CA, USA) mass spectrometer with an ESI-Jet-Stream ion source. The chromatograph was equipped with binary gradient pump, autosampler, column oven (20 °C) and DAD detector. Separation of analytes was performed on Gemini 3 μm C18 110Ǻ (100 × 2.1 mm) column. Water with 0.1% HCOOH was used as mobile phase A and acetonitrile with 0.1% HCOOH as a mobile phase B. The following gradient was applied: 0–45 min 15%–80% B; 45–46 min 80%–95% B; 46–50 min 95% B; post time 10 min. Flow rate was 0.3 mL/min, injection volume was 10 µL, total time of analysis was 60 min. ESI-QTOF-MS analysis was performed with following parameters. Dual spray Jet Stream, positive ion mode, gas (N₂) temperature: 300 °C, flow rate 12 mL/min, 35 psig, sheath gas temperature: 325 °C, flow rate 12 L/min, 35 psig, fragmentor voltage 140 V, Vcap 4000 V. Collision induced cell at two energies 10 and 40 eV. Data acquisition was performed in Auto MS/MS mode at the range of 100–1000 mass units for MS and MS/MS. Mass Hunter B.07.00 software (Agilent Technologies INc., Santa Clara, CA, USA) was used for data analysis.

Analysis was performed using HPLC/ESI-QTOF-MS system in positive ion mode with a 630B accurate mass QTOF-MS (Agilent Technologies INc., Santa Clara, CA, USA) mass spectrometer and an ESI-Jet-Stream ion source. The separation was done using Gemini 100 × 2.1, 3.5 µm column, thermostated at + 40 °C. The mobile phase consisted of a mixture of H₂O + 0.1% HCOOH (A) and ACN + 0.1% HCOOH (B). The gradient elution was carried out at a constant flow of 0.4 mL/min. The gradient applied was as follows 0 min., 5%B; 15 min., 98%B; 20 min., 98%B. The return to the initial gradient compositions (95% A/5% B) was held for 5 min. The total time of analysis was 20 min. ESI-QTOF-MS analysis was performed with the following parameters, gas (N₂) temperature: 300 °C, flow rate 10 mL/min, 35 psig, sheath gas temperature: 325 °C, flow rate 10 L/min, fragmentor voltage 140 V, Vcap 4000 V. Two collision energies, 20 and 40 eV were chosen. Data acquisition was performed in Auto MS/MS mode at the range of 100–1000 mass units. Mass Hunter B.08.00 software (Agilent Technologies INc., Santa Clara, CA, USA) was used for data analysis. Mass calibration was done before analysis according to the manufacturer’s recommendations to ensure mass accuracies below 5 ppm. Data handling was performed by MassHunter Workstation Software B.08.00 and OriginPro 9.5.