Micro syringe

The Camag Linomat 5 autosampler and Camag micro syringe were employed (CAMAG, Muttenz, Switzerland). Camag TLC scanner 3 model 3 densitometer outfitted with WINCATS software for densitometric assessment, a short wavelength UV lamp producing at 250.0 nm (VILBER LOURMAT, France), slit dimension: 60.3 mm; scanning speed: 20 mm/s; data resolution: 100 m/step; bandwidth: 3 mm. Thin-layer chromatographic plates with high performance, pre-coated with silica gel 60 F254, 20 × 20 cm, 0.25 mm thickness, Merck, Darmstadt.

The thin-layer chromatography (TLC) system consists of 100 μL Camag micro syringe (Muttenz, Switzerland) as well as Camag Linomat autosampler (Camag, Switzerland). TLC was performed utilizing 0.25 mm thickness TLC plates pre-coated with silica gel G.F254 20X20 cm (Merck, Darmstadt, Germany), winCATS software, and a short wavelength (254 nm) ultraviolet (UV) lamp (USA).
The HPLC system consists of Waters Alliance 2695 Separations Module using inline vacuum degassing with different flow rates, an autosampler, and programmable temperature control. A heated column compartment provides temperatures from 5 degrees above ambient to 65 °C. The wavelength range of model 2996 photodiode array detector is 190–800 nm. Stationary phase was Kromasil^® MS C₁₈ column (150 mm × 4.6 mm, 5 µm) (Nouryon, Bohus, Sweden).Whatman syringe filter (Fisher, Nederland) was utilized. For IR and mass spectrometric analysis, an infrared (IR) spectrometer (Shimadzu, Japan) and an Ultra-performance liquid chromatography-mass spectrometer (LC/MS/MS) (Waters, Milford, USA) were used. Design-Expert software was used for data analysis (version 12) (Godward St NE, Minneapolis). Hot plate and pH meter were used (Jenway, Stone, UK).