Chromatography assignment

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Chromatography assignment

Objectives:

1. To identify a unknown analgesic

2. To identify the analgesic you will be given 4 known analgesics compounds. Each of which can be separated using by Thin Layer Chromatography.

3. Using thin layer chromatography the unknown analgesic will be compared to the 4 known analgesics.

Spotting of the sample 

1. Prepare a microcapillary tube for each analgesic and each standard

2.  Immerse the small end of the tube into the sample vial until some of the sample is

      drawn into the tube.

3.  Very gently press the small end of the tube onto the plate about one cm from the

      bottom of the plate.  Keep the spots small and concentrated by applying the

      sample 5 times and allowing the spot to dry between applications. Additional        spots may be required for the ibuprofen standard in order to see its spot clearly.

4.  Repeat this procedure for each of the standards available.

5.  Two or more samples may be applied to each plate if they are kept one cm apart.

      Mark the position of the spots lightly in PENCIL and be sure to keep a record of

      which spot represents each product. 

Development of the TLC plates 

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1. Prepare a developing chamber by adding 15-20 ml of solvent

2. Place the TLC plates in the chamber so that they do not touch.  Allow the

      solvent to rise to within one cm of the top of the plates.

3.  Remove the plates, mark the solvent front using pencil, and allow them to dry.

4.  Visualize the spots by illumination under a UV lamp.

5. Trace around each spot with a pencil and then measure the distance traveled by

      each component.

6. Calculate the Rf by dividing the distance of the solvent front by the distance of the

      spot.  ...

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