Phytochemical characterization of ADPE by LC–MS

ADPE was subjected to phytochemical analysis by high-performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) on a triple quadrupole tandem mass spectrometer (ACQ-TQD#QBB1152). LC was performed on a C18 reverse-phase column (150 × 2.1, 2.6 μm) and elution was done for 30 min, using mobile phase solvent  acetonitrile/water (5:95, v/v), acetonitrile, methanol and 5 mM ammonium acetate (95:5 H₂O:Acetonitrile, pH 6.5). The mobile phase was kept at a flow ramp rate of 0.45 mL min^-1 and the sample injection volume was 5 μL. Waters Acquity PDA detector type- UPLC eLambda 800 nm was used at wavelength range 210–800 nm and resolution 1.2 nm. The spectrometer was operated in positive and negative modes. The source temperature was 120 °C, the desolvation temperature was 350 °C and the cone voltage was set at 30 V. MS data was recorded in the mass range, m/z 200–2000 from 0–30 min under ionization mode of ES⁺ and ES^-. All eluted peaks from HPLC were recorded at different retention times. The fractions were then characterized by mass spectrometry. This analysis was carried out at the Sophisticated Analytical Instrumentation Facility (SAIF), CSIR- Central Drug Research Institute (CDRI), Lucknow, India.