Standard Operating Procedure

For Varian 5000 Liquid Chromatograph

Turn the pump and detector on

Set the pump flow rate at 2.3 mL/min and detector sensitivity at 0.08 AUFS
Turn on the Kipp and Zonen flatbed recorder an set the slow speed rate
Flush the column with 20%methanol: 80%H20 for 5 to 10 minutes
o Record the response to make sure the column is clean
Inject 10 l of solution (if it is a standard, then from the least concentrated first)
into the HPLC while in the LOAD position.
Push the PRIME button and the valve will turn to the inject position
o The detector will provide a record of the sample injected
To shut down HPLC
o Clean syringe with methanol (HPLC grade)
o Run 100% methanol (HPLC grade) as a solvent for 5-10 minutes
o Turn off pump, detector, and recorder