HPLC Lab

Experimental:

Use the HPLC to determine the retention time(s) for the molecule(s) in the sample assigned to you. The instructor will help you get started. Put the sample name and your first names in the “Sample Name” box. Each run is set to go for 12 minutes. After the 12 minutes is done, contact the instructor to find out how to analyze the data and print a report.

Computer (See HPLC Simulation on our class webpage.):

For all of the determinations use a column length of 10.

In #1 you will compare retention times for the know substances with the retention times found for each athlete. Use a concentration of 10 for each of the substances.

In #2 through #5 you will need to generate a standard curve for the substance that you are looking at. Plot 5-6 points up to a concentration of 20 for each substance. Show the instructor the graph for number two before going to number three.

1. Determine which of the substances each athlete has.
2. Using graph paper, determine how much aspirin each athlete has.
3. Using graph paper, determine how much caffeine each athlete has.
4. Using graph paper, determine how much creatine each athlete has.
5. Determine the amount of urea in one of the athletes.

If you complete all of these tasks, find the effect of the column length on the retention time and on the areas.