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Study Notes: HPLC Handy Hints

Initially the use of the HPLC is a confusing, complicated and complex procedure. There is no substitute for hands-on experience but there are many common sense approaches that will make the use of the HPLC easier with fewer mistakes. We call these ‘Handy Hints’.

Handy Hints for RP-HPLC

  1. Print out a copy of the SOP.
  2. Read the SOP very carefully before commencing.
  3. When working with the ‘virtual’ SimuLab HPLC read the screen instructions and other information very carefully and make notes of important points.
  4. Follow the on-screen instructions exactly as stated.
  5. Check that the correct column is installed (C18).
  6. Be sure to use all of the parameters listed in the SOP and adjust the HPLC to reflect these parameters. The parameters that will need to be adjusted are:
    • Turn machine ON
    • Allow 5 minutes warm up
    • Adjust wavelength
    • Set flow rate
    • Flush column as per SOP
    • Set run time
    • Set gain and baseline on chart recorder
    • Reset flow rates
    • Inject correct volume of sample
    • Press RUN button
    • Flush column
    • Switch HPLC off.
  7. Print out a copy of the chromatogram for future reference.

 

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