6220 tof ms

Analytical HPLC was performed on a Shimadzu NexeraXR 20A System with autosampler, degasser, column oven, diode array detector, and a Phenomex Luna C18 column (2.9 μm, 50 × 2.1 mm) with a gradient (in 5.5 min from 5% B to 95% B, 0.5 min 95% B and back to 5% B in 3 min, total run time 9 min) at a flow rate of 650 μl/min and column oven temperature of 40°C. HPLC solvent A consists of 99.9% water and 0.1% TFA, solvent B of 99.9% acetonitrile and 0.1% TFA.
Analytical LC-MS was performed on an Agilent 6220 TOF-MS with a Dual ESI-source, 1200 HPLC system with autosampler, degasser, binary pump, column oven, diode array detector, and a Hypersil Gold C18 column (1.9 μm, 50 × 2.1 mm) with a gradient (in 11 min from 0% B to 98% B, back to 0% B in 0.5 min, total run time 15 min) at a flow rate of 300 μL/min and column oven temperature of 40°C. HPLC solvent A consisted of 94.9% water, 5% acetonitrile, and 0.1% formic acid, solvent B of 5% water, 94.9% acetonitrile and 0.1% formic acid. ESI mass spectra were recorded after sample injection via 1200 HPLC system in extended dynamic range mode equipped with a Dual-ESI source, operating with a spray voltage of 2.5 kV.

For mass spectrometry, the Agilent 6220 TOF-MS with an electrospray ionization source (ESI) in negative mode was used. The flow rate of dying gas (N2) was set at 9 L/min. The nebulizer was set at 45 psi. The other optimal conditions were as follows: dying gas temperature of 350°C, fragment voltage of 120 V. Data were collected in the full-scan mode from m/z 50 to 1050 amu over 0–12 min. The MS data were collected in centroid mode.

Phenolic compounds contained in aqueous extract (extracts used by the traditional healers), hydroethanolic extract (most active crude extract), and ethyl acetate fraction (most active fraction) were quantified using analytical LC-MS. The analysis was performed on an Agilent 6220 ToF-MS with a Dual ESI-source, 1200 HPLC system with autosampler, degasser, binary pump, column oven, diode array detector, and a Hypersil Gold C18 column (1.9 μm, 50 × 2.1 mm) with a gradient (in 11 min from 0% B to 98% B, back to 0% B in 0.5 min, total run time 15 min) at a flow rate of 300 μL/min and column oven temperature of 40°C. HPLC solvent A consists of 94.9% water, 5% acetonitrile and 0.1% formic acid, solvent B of 5% water, 94.9% acetonitrile, and 0.1% formic acid. The mass spectra are recorded in both profile and centroid mode with the MassHunter Workstation Acquisition B.04.00 software (Agilent Technologies, Santa Clara, CA, USA). MassHunter Qualitative Analysis B.07.00 software (Agilent Technologies, Santa Clara, CA, USA) was used for processing and averaging of several single spectra.