Waters xevo g2 qtof

Analysis was performed on an UPLC system coupled to a QTOF-MS (Waters Xevo G2 QTof, Waters, Milford, MA, USA) instrument operated in electrospray ionization (ESI) mode at a mass resolution of 20,000 and controlled by MassLynx 4.1 software. An Acquity UPLC BEH C18 column (2.1 mm I.D. × 100 mm, 1.7 μm, Waters, USA) at 35 °C was used for chromatographic separation. The sample (1 μL) was injected using an autosampler. The mass spectrometer was calibrated with 0.5 mM sodium formate. Leucine enkephalin (2 µg/mL, m/z 556.2771 in positive mode) was used as lock spray at a flow rate of 10 µL/min. The collision energy equaled 6 V. The source parameters were as follows: capillary voltage 2.5 kV, sampling cone voltage 30 V, extraction cone voltage 4 V, source temperature 150 °C, desolvation temperature 500 °C, desolvation gas flow 1000 L/h, cone gas flow 50 L/h. Each compound was fragmented using a range of 25–30 V for MS/MS scans. The mobile phase contained 1% HCOOH in H₂O (A) and MeCN (B). F50 was separated using the following gradient: initial 10% B for 1.2 min, 15% B at 3.5 min, 25% B at 5.2 min, 35% B at 6.4 min, 45% B at 7.0 min, 55% B at 8.1 min, hold until 10 min. F80 was separated isocratically using 20% B for 50 min.

Data were collected using an ultra-high-performance liquid phase (Waters Acquity UPLCTM I-class system, Waters Corp, Milford, MA, USA) coupled with a high-resolution time-of-flight mass spectrometer (Waters Xevo G2 QTof, Waters Corp., Milford, MA, USA). Chromatography was performed using Waters UPLC HSS T3 at 40°C. The data were processed using Waters MassLynx 4.1. The mobile phase was water (A) and acetonitrile (B) containing 0.1% formic acid, the flow rate was 0.3 mL/min, and the elution conditions were as follows: 0–2 min, 0–5% B; 2–10 min, 5%–15% B; 10–15 min, 15%–25% B; 15–18 min, 25%–50% B; 18–23 min, 50%–100% B; 23–25 min, 100%–2% B; and 25–30 min, 2% B. The injection volume was 2 μL. The mass spectra were obtained in Fast DDA and positive ion mode, with an ESI ion source, and the following settings were used: electrospray ionization ion scan range: 50–2000 m/z; capillary voltage: 3.0 kV; ion source temperature: 100°C; desolvation gas (N₂) temperature: 500°C; desolvation gas flow rate: 1000 L/h; cone gas flow rate (N₂): 100 L/h; and collision gas: argon.

Analysis was performed on a UPLC system coupled to a QTOF-MS (Waters Xevo G2 QTof, Waters, Milford, MA, USA) instrument operated in electrospray ionization (ESI) mode at a mass resolution of 20,000 and controlled by MassLynx 4.1 software. An Acquity UPLC BEH C18 column (2.1 mm I.D. x 100 mm, 1.7 μm, Waters, USA) at 35°C was used for chromatographic separation. The sample (1 μL) was injected using an autosampler. The mass spectrometer was calibrated with 0.5 mM sodium formate. Leucine enkephalin (2 μg/mL, m/z 554.2615 in negative mode) was used as lock spray at a flow rate of 10 μL/min. The collision energy equaled 6V. The source parameters were as follows: capillary voltage 2.5kV, sampling cone voltage 30V, extraction cone voltage 4V, source temperature 150⁰C, desolvation temperature 500⁰Cgas flow 1000L/h, cone gas flow 50L/h. The mass spectrum analysis was done using Waters Xevo G2 QTof. The column used is AQUITY UPLC BEH C18 1mm ID x 100mm.