Procedure

• A stock solution (0.1 mg/ml) of standard cane sugar and lactose was prepared in water: methanol (1:9) and sonicated for 10 minutes over an ultrasonic bath.
• The solution was filtered through Whatman No. 42 filter paper and filtrate was used as sample solution.
• A 20cm × 10cm HPTLC plate coated with silica gel 60 F 254 was used for analysis.
• The samples were applied at 10 mm from the base of HPTLC plate by means of a Camag (Switzerland) Linomat V sample applicator using a syringe (100µL, Hamilton, Bonaduz, Switzerland).
• A linear calibration curve was obtained on applying the increasing concentrations of standard cane sugar and lactose in the range (200-1000 ng).
• HPTLC analysis was performed on a computerized densitometer scanner 3, controlled by WinCATS planar chromatography manager version 1.4.4. (CAMAG, Switzerland).
• Plate was developed to a distance of 80 mm, in a Camag twin-trough chamber with mobile phase [n-Butanol: Acetic acid: Water:: 2:1:1 (v/v)].
• Plates were derivatized using aniline diphenylamine phosphoric acid.
• Plates were evaluated by densitometry at 520 nm with a Camag Scanner 3 for quantification.