2.6.1. Sample preparation
Samples were prepared by dissolving the crude extracts in HPLC grade ethanol, followed by filtration through a 0.2 m nylon filter(Whatman, Dassel, Germany).
Prior to analysis, 500 l aliquots ofthe sample extract were centrifuged at 14,000 rpm for 5 min.
Supernatant was then diluted in the same solvent used for extraction (95%ethanol) and 300 l aliquot of the extract was transferred into auto sampler vials of (Agilent Technologies, Santa Clara, CA) for LCMS analysis.
2.6.2. Instrumentation
Agilent 1260 affinity HPLC system consisted of a quaternary pumps, auto-sampler, thermocouple column compartment and diode array detector.
The HPLC was coupled to 6540 Ultra High Def-inition (UHD) electron spray ionisation mass spectrometer (Agilent Technologies Germany).
Agilent Mass Hunter Qualitative analysis software (Version B.04.00) was used for sample analysis and Agilent Mass Hunter Workstation was used for data acquisition.