Linomat autosampler

TLC system; a Camag Linomat autosampler (Muttenzl, Switzerland), a Camag micro syringe (100 µL), a Camag 35/N/30319 TLC scanner with win CATS software, UV lamp with a short wavelength at 260 nm (Desaga, Wiesloch, Germany) and TLC plates (10 × 20 cm) pre-coated with silica gel GF₂₅₄ of 0.25 mm thickness (Merck, Darmstadt, Germany).

Aliquots of stock solution (0.2–10.0 µL) from the working solution were applied in triplicate on a TLC plate using CAMAG Linomat autosampler with CAMAG micro syringe. The plates were developed using the optimized mobile phase and scanned as under Section chromatographic conditions. The average peak areas were plotted against the corresponding concentrations ranging from (0.15–7.5 µg/band) and (0.05–2.5 µg/band) for MIR and TAM, respectively to get the calibration graph. The regression equations were constructed.

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).