Wincats software version 1

The MBS was standardized for the content of marker compound, amentoflavone using HPTLC method. Amentoflavone and MBS were dissolved in methanol for the analysis. The sample solutions were applied on prewashed and activated precoated silica gel aluminum HPTLC plate 60F₂₅₄ (20 cm × 10 cm with 250 μm thickness) in the form of band of 6 mm width with a 100 μl syringe (Camag, Muttenz, Switzerland) using a Camag Linomat V sample applicator (Camag, Muttenz, Switzerland). HPTLC plates were then developed with 20 ml mobile phase consisting of toluene: ethyl formate: formic acid (5:4:1, v/v/v). Linear ascending development was carried out in 20 cm × 10 cm twin trough glass chamber saturated with the solvent system. The chamber saturation time for the solvent system was 15 min at room temperature of 25°C ± 2°C and relative humidity of 60% ± 5%. After chromatography, plates were dried in an air current. Densitometric scanning was performed using Camag TLC scanner III with winCATS software version 1.4.4 (Camag, Mutttenz, Switzerland) at 366 nm.[8 ]

The sample solutions were applied on prewashed and activated precoated silica gel aluminum HPTLC plate 60F₂₅₄ (20 cm × 10 cm) in the form of bands of 6 mm width with a Hamilton syringe (100 μL) using a Camag Linomat V (Switzerland) sample applicator. The slit dimension was 5 mm × 0.45 mm and 10 mm/s scanning speed was employed. HPTLC plate was then developed, at a constant temperature, with 20 mL mobile phase consisting of toluene:ethyl acetate:methanol (15.0:1.7:3.3, v/v/v). Linear ascending development was carried out in 20 cm × 10 cm twin trough glass chamber (Camag, Muttenz, Switzerland) saturated with the mobile phase. The optimized chamber saturation time for mobile phase was 15 min at room temperature (25 ± 2°C) at a relative humidity of 60 ± 5%. The length of chromatogram run was 8 cm. Densitometric scanning was performed within 10 min after chromatographic development using Camag TLC scanner III with Camag TLC scanner III with winCATS soft ware version 1.4.4 (Camag, Muttenz, Switzerland) in the reflectance mode at 317 nm.