Scribd
Upload
941 views

Lab 3 Pre Lab

This document describes a procedure for using thin-layer chromatography (TLC) to separate and identify unknown compounds in mixtures. TLC uses a stationary phase and mobile phase to separate compounds based on how strongly they interact with each phase. Compounds that interact more strongly with the mobile phase will migrate farther up the TLC plate. The document provides details on performing TLC experiments to identify unknown compounds in mixtures of analgesics and in tomato paste extracts.

Uploaded by

Mina Vo
Copyright
© Attribution Non-Commercial (BY-NC)
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
941 views

Lab 3 Pre Lab

This document describes a procedure for using thin-layer chromatography (TLC) to separate and identify unknown compounds in mixtures. TLC uses a stationary phase and mobile phase to separate compounds based on how strongly they interact with each phase. Compounds that interact more strongly with the mobile phase will migrate farther up the TLC plate. The document provides details on performing TLC experiments to identify unknown compounds in mixtures of analgesics and in tomato paste extracts.

Uploaded by

Mina Vo
Copyright
© Attribution Non-Commercial (BY-NC)
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
You are on page 1/ 3

Jenna Huynh May 7, 2013 TLC-Thin Layer Chromatography Pre-lab Purpose To separate two or more compounds or ions that

can help identify how many components are in a mixture.

Introduction Many forms of chromatography include stationary phase and mobile phase. Capillary action occurs when paper is placed in a solvent that is drawn by the paper. Thinlayer chromatography is a sensitive, fast, simple and inexpensive technique that is used to carry out experiments. There are six uses of thin-layer chromatography. (1) To determine the number of components in a mixture. (2) To determine the identity of two substances. (3) To monitor the progress of a reaction. (4) To determine the effectiveness of purification. (5) To determine appropriate conditions for a column chromatographic separation. (6) To monitor column chromatography. There are two phases free phase and absorbed phase. The strength of the attraction of A and B in stationary phase structure and liquid phase depends on size, polarity, and hydrogen bonding between A and B. It also depends on polarity and hydrogen bonding ability of the stationary phase, and of the mobile phase solvent. The A molecules spend more time in the mobile phase; they will be carried through the stationary phase and is removed faster and farther. The B molecules are absorbed on

the stationary phase more than A molecules. The B molecule doesnt migrate as far in the same time. A moves ahead of B shown below. The stationary phase is polar absorbent such as alumina and silica particles shown below. The silicon or aluminum atoms are the smaller, darker spheres. Aluminum and silicon is very polar so it is hard to break its attraction at the stationary phase. Nonpolar molecules are in the mobile phase. The stronger the absorption on the stationary phase, the slower the molecules move. Nonpolar compound moves faster and a greater distance than polar compounds. Table 8.2 shows common compound move or elute on silica or alumina. Polarity increases as the number of functional groups increases. The key to chromatographic separation is the mobile phase. Table 8.3 lists solvents that are commonly used in TLC and column chromatography. Solvents for TLC and column chromatography have low boiling points, so the could evaporate easily. Structures of compounds used in the experiment are:

Procedure Experiment 1 1. 2. Use 2 chromatographic plates. Draw light pencil line 1 cm from bottom of plate. Spot aspirin, acetaminophen and unknown on line on the first plate, spot ibuprofen, caffeine, and unknown on second plate. 3. 4. Examine the plates under UV light Place 4 mL of mobile phase and a piece of filter paper in a 150 mL beaker. Cover with watch glass. 5. 6. Develop two TLC plates Remove the plates from developing beaker, mark solvent with pencil

7. 8. 9.

Allow plate to dry Examine plates under UV light, outline spots with pencil. Calculate value for all spots and identify components in unknown

Experiment 2 1. Place 1 g of tomato paste in small test tube, add 5 mL of acetone. 2. Cover test tube, mix 3. Filter mixture on Hirsh funnel, collect solution 4. Place filtrate into Erlenmeyer flask 5. Put solid residue back to test tube, filter again with funnel, add filtrate to E-flask 6. Repeat step 5 twice with dichloromethane 7. Combine extracts and pour into funnel 8. Add NaCl solution 9. Separate organic layer 10. Dry colored organic layer over Ca pellets, gravity filtration

11. Spot mix on 3 TLC plates, one spot concentrated, other lower concentration 12. Develop TLC plates with 80:20 hexane-acetone mixture, cyclohexane, toluene 13. Look at TLC plates under UV light, mark spots with pencil, calculate Reference 1. Williamson, Kenneth. Crystallization/Recrystillization. 5 . 2010. 61-85. Print. values

You might also like