Tqd uplc ms ms

Qualitative analysis of thapsigargins from the in vitro plants was performed using a Waters Acquity TQD UPLC/MS/MS (triple quadrupole mass spectrometer) system associated with a PDA detector using the standards mentioned above. The mass spectrums was compared with those provided by Prof. Søren B. Christensen, University of Copenhagen, Denmark. The system was directed by the software MassLynx 4.1. Extracts were filtered at 0.2 µm and 7.5 μL of each extract was separated on a C18 Luna (2)-HST (100 × 2.0 mm, 2.5 μm) column (Phenomenex, USA). The flow rate was set to 0.6 mL/min and the mobile phases consisted of water with o-phosphoric acid (0.01%) for A and acetonitrile with o-phosphoric acid (0.01%) for B. The binary gradient elution was: 90% A (0–0.5 min.), 90–0% A (0.5–7 min., linear gradient), 0% A (7–10 min.), 0–95% A (10–10.2 min., linear gradient) and 95% A (10.2–13 min.). The PDA swept wavelengths from 200 to 400 nm with a 2.4 resolution. Mass spectra were obtained in positive and negative modes, using an electrospray ionization (ESI) source on a triple quadrupole instrument (Waters Acquity) in full scan (50–2000 m/z). The conditions were as follows, capillary voltage: 3000 V, cone voltages: 30 and 60 V, desolvation temperature: 450 °C, source temperature: 150 °C, gas flow cone: 50 l/min, desolvation gas: 800 l/min.