EXPERIMENT NO: 10. TO STUDY THIN LAYER CHROMATOGRAPHY (TLC) PATTERN OF ISOLATED SENNOSIDES. T.Y B. PHARM SEM - V PHARMACOGNOSY & PHYTOCHEMISTRY – II

 

T.Y B. PHARM SEM - V

PHARMACOGNOSY & PHYTOCHEMISTRY – II

 

EXPERIMENT NO: 10

AIM: To Study Thin Layer Chromatography (Tlc) Pattern Of Isolated Sennosides.

 

REFERENCE:

1.     Dr. Anup K. Chakraborty, Sushma M. Rathod. “Practical in pharmacognosy & phytochemistry – II. Edition 2019, published by S. Vikas & company (Medical publisher), Page no. 43-44

 

REQUIREMENTS:

Apparatus: TLC plates, Glass chamber, Iodine chamber, Hot air oven, Beaker, Capillary etc.

Chemical: Isolated sennosides, Ethyl acetate, n-propanol, water, Ammonia.

PROCEDURE:

Preparation of extract: Dissolved 1mg of isolated sennosides in 1ml solvent containing equal volume of Ethyl acetate, n-propanol, and water.

Adsorbent (Stationary Phase): Make 15 g silica gel slurry with 20 ml of distilled water. The plates are coated with slurry of silica gel to a width of 0.4mm, allowed to dry at room temperature, activated oven at 120°C for 20 min.

Solvent system (Mobile Phase): The solvent system Ethyl acetate, n-propanol, water (4:4:3). is poured in the TLC chamber to a depth of 3cm and allow to saturate. The chamber is lined with filter paper on three sides to maintain equilibrium of mobile phase

Application of spots: The above sample applied as two spot on each plate with the help of capillary.

Development of chromatogram: The chromatogram is established by ascending technique, allowing solvent system to reach a height of 10 cm. The plates are withdrawn from the chamber, air-dried and visualize the dried TLC plate by exposure to ammonia vapor for 5 minutes till the color develop. Cover the plates with glass and heated at 110^oC for 5-10 minutes. Sennoside A & B develop two prominent spots..

Rf Value:

The Rf value is calculated by using below formula

 

RESULT: The TLC of sennoside was performed and the Rf value of sennoside was found to be…………………….