Methods:

Set Up

1. Cutt of the exit tube of the C18 Sep-Pack chromatography cartridge.

2. Mount the Sep Pak on a syringe.

Collect Data

1. Use a 10-mL syringe to flush the C18 Sep-Pack cartridge with 10 mL of undiluted 18% isopropanol.  Our rate was 3 mL in 7 minutes for our first trial, and our rate was 3 mL in 15 minutes for our second trial.

2. Collect the eluate in a 10-mL graduated cylinder to monitor the flow rate of the isopropanol.

3. Using the 10-mL syringe, flush the Sep-Pack cartridge with 10 mL of distilled water.

4. Draw up 1 mL of the grape Kool-Aid sample with the 1-mL syringe.

5. Slowly inject the Kool-Aid sample into the Sep-Pack cartridge.

6. Collect and discard the effluent that washed out of the column as you inject the sample.

7. Fill the solvent pump (10-mL syringe) with 18% isopropanol solution.

8. Pump the solvent through the Sep-Pack cartridge at a steady rate of 5 to 10 mL per minute.

9. Collect the eluent in the 10-mL graduated cylinder.

10. Record in table 2 the volume of liquid in the graduated cylinger when the red or blue dye is first observed in the eluent from the Sep-Pake column and when that color is no longer observed.

11. Clean up according to your teacher's instructions.

Equipment:

-C 18 Sep-Pak Cartridge

-Syringe, 1-mL

-Syringe, 10-mL

-Graduated cylinder, 10-mL

-18% Isopropanol, 100 mL

-Unsweetened Kool-Aid Drink, 10 mL

-Distilled water