Mass hunter software version b 07.03 509

LC-MS analysis was carried out using an Agilent HPLC 1260 System, and an Agilent Q-TOF 6530 mass spectrometer was used as the detector for fingerprinting the extract. 1 mg of free dried sample extract was dissolved in 1 mL of methanol : water (50 : 50) by volume. The resulting mixture was transferred into a 5 mL syringe and filtered through a 0.22 μm acrosdisc syringe filter into an HPLC vial for LC-MS/MS analysis. Gradient elution with composition as follows: solvent A: (water with 0.1% formic acid), solvent B: acetonitrile with 0.1% formic acid, and injection volume: 5 μL. Column temperature was maintained at 40°C and with a total run time of 27 minutes. The LC-Q-TOF-MS data was analyzed using Agilent Technologies Mass Hunter Software Version B.07.03 (509).

The constituents of the most potent extract that were in the CZSC209213 pool were analysed using liquid chromatography-mass spectroscopy in the Department of Pharmacy of the University of Zimbabwe. An Agilent HPLC 1260 system (Agilent, USA) was used equipped with a binary pump, autosampler, and thermo-stated column compartment. Chromatographic separation was performed using Agilent Poroshell 120, C18 Column, with dimensions 50 × 2.7 mm, 3 microns. The mobile phase for the positive mode consisted of 0.1% formic acid in deionised water (A) and 0.1% formic acid in acetonitrile (B). The total run time for each sample was 25 min. The flow rate was 0.25 mL min⁻¹, and the injection volume was 5 μL. The column temperature was maintained at 40°C. An Agilent Technologies Q-TOF 6530 Mass Spectrometer was used as the detector. Agilent Technologies Mass Hunter Software version B.07.03 (509) was used for data acquisition, instrument control, and data analysis.