HPLC Small Peptides -- DRAFT --

* Introduction

HPLC is High Performance (or Pressure!) Liquid Chromatography. The high pressures involved (over 1000 PSI) make this a potentially dangerous piece of equipment. Add to this the presence of volatile organic solvents and acids and you soon realize some care is needed.

* Precautions

Never leave the machine unattended while running a program
Be sure to check the system pressure regularly to avoid damage to the system.
Use only degassed liquids - gently bubble Helium gas through for 5 min before use.
Filter aqueous solutions with 0.2um filter before use.
Avoid introducing air bubbles into the system at all costs.
Ensure all liquid waste is collected in appropriate labelled containers.
Never run the system at greater than 2500 PSI.
Try and keep the pressure between 600 and 1500 PSI during normal usage.

* Basic Operation

Switch on Lamp
Switch on Mixer
Start Gilson 715 Software

* Manual Mode

Display shows:

Time: Run time
Flow: Current flow rate in ml/min
Contacts: Current system pressure in PSI (Don't go above 2000 PSI)
Pump A: Current % from Pump A
Pump B: Current % from Pump B
Chart: Chart shows currently open method (program). Blue line indicates %B

In the Manual Menu open these windows

Mobile Phase Control

Mobile phase allows direct control of liquid flow.
Time: 0 min -- time to go from current flow rate and %B to new settings
Flow: 1 ml/min -- Request a new flow rate.
%B: 2 -- Change the percentage of liquid coming from pump B

Manual Strip Chart

Check the A box to display the current absorbsion
Change the mVFS to alter the vertical scale
Change the speed to 10mm/min to alter chart speed
Autozero zeros the display - not the volatge from the recorder