Abstract

Ms. Jahanvi Patel, Dr. Ishita Basera

ABSTRACT

The chromatographic separation has been achieved on HPTLC silica 60 F254 plates (10.0 cm?×?10.0 cm, 0.20 mm thickness, E. Merck, Germany). The specimens were applied as bands measuring 6 mm in width using a 100 ?l sample syringe onto precoated silica gel aluminum plates 60 F254 (10×10 cm, E Merck, Darmstadt, Germany) utilizing a Camag Linomat 5 sample applicator (Switzerland). Application was conducted through a CAMAG Linomat 5 auto-sampler employing a CAMAG microliter syringe (100.0 ?L) (CAMAG, Muttenz, Switzerland). Prior to chromatography, the plates underwent methanol prewashing and were activated at 110° for 5 minutes. A constant application rate of 150 nl/sec was maintained, with a 6 mm spacing between two bands. Slit dimensions were set at 6×0.45 mm. The mobile phase, comprised of Ethyl acetate : Methanol : Toluene: Glacial acetic acid (7.5:1:1:0.5, v/v/v/v) facilitated linear ascending development in a 10×10 cm twin-trough glass chamber. The optimized chamber saturation time for the mobile phase was set at 30 minutes, at a temperature of (25±2°) and relative humidity of (60±5%). The chromatogram run length was 8 cm, and TLC plates were air-dried. Densitometric scanning was executed using a Camag TLC Scanner 3 equipped with winCATS software version 1.3.0 at 270 nm, with a deuterium lamp serving as the radiation source. Evaluation was conducted utilizing peak area with linear regression.The chromatographic separation has been achieved on HPTLC silica 60 F254 plates (10.0 cm?×?10.0 cm, 0.20 mm thickness, E. Merck, Germany). The specimens were applied as bands measuring 6 mm in width using a 100 ?l sample syringe onto precoated silica gel aluminum plates 60 F254 (10×10 cm, E Merck, Darmstadt, Germany) utilizing a Camag Linomat 5 sample applicator (Switzerland). Application was conducted through a CAMAG Linomat 5 auto-sampler employing a CAMAG microliter syringe (100.0 ?L) (CAMAG, Muttenz, Switzerland). Prior to chromatography, the plates underwent methanol prewashing and were activated at 110° for 5 minutes. A constant application rate of 150 nl/sec was maintained, with a 6 mm spacing between two bands. Slit dimensions were set at 6×0.45 mm. The mobile phase, comprised of Ethyl acetate : Methanol : Toluene: Glacial acetic acid (7.5:1:1:0.5, v/v/v/v) facilitated linear ascending development in a 10×10 cm twin-trough glass chamber. The optimized chamber saturation time for the mobile phase was set at 30 minutes, at a temperature of (25±2°) and relative humidity of (60±5%). The chromatogram run length was 8 cm, and TLC plates were air-dried. Densitometric scanning was executed using a Camag TLC Scanner 3 equipped with winCATS software version 1.3.0 at 270 nm, with a deuterium lamp serving as the radiation source. Evaluation was conducted utilizing peak area with linear regression.