2.3. Identification of Bioactive Compounds by High Resolution-Liquid Chromatography Mass Spectroscopy (HR-LCMS)

Phytochemical analysis ASE was assessed using Ultra High-performance liquid chromatography Photodiode-Array Detector 323 Mass Spectrophotometer (HR-LCMS 1290 Infinity UHPLC System), Agilent 324 Technologies^®, USA. The liquid chromatographic system consisted of a HiP sampler, binary gradient solvent pump, column compartment and Quadrupole Time of Flight Mass Spectrometer (MS Q-TOF) with a dual Agilent Jet Stream Electrospray (AJS ES) ion source. 10 μL of sample was injected into the system, followed by separation in SB-C18 column (2.1 × 50 mm, 1.8 μm particle size). 1% formic acid in deionized water (solvent A) and acetonitrile (solvent B) were used as solvents. Flow rate of 0.350 mL/min was used, while MS detection was performed in MS Q-TOF. Compounds were identified via their mass spectra and their unique mass fragmentation patterns. Compound Discoverer 2.1, ChemSpider and PubChem were used as the main tools for the identification of the phytochemical constituents.