The link below will take you to a very useful bit of software created using excel that allows you to explore the effects of various parameters on the elution profile of a product. You can play around with column length, elution method (gradient or isocratic), injection volume, temperature and much more and see their effect on resolution and retention time. The first link gives a good overview of the software’s capabilities and the second link is to the program itself which you can download for free and run on PC or Mac.

• Software download:

• http://bit.ly/hplcapp

Among the offered possibilities, it allows you to:

1. Illustrate the concept of chromatographic resolution including the impact of retention, selectivity and efficiency on resolution.
2. Understand the van Deemter equation and kinetic performance in HPLC.
3. Recognize the importance of analytes lipophilicity (log P) on retention and selectivity in reversed phase HPLC mode (RPLC).
4. Handle the RPLC retention, taking into account the acido-basic properties (pKa) of compounds and mobile phase pH.
5. Simulate the impact of mobile phase temperature on HPLC separations
6. Understand the behavior of a mixture of diverse compounds in both isocratic and elution gradient modes.
7. Show the influence of instrumentation (injected volume and tubing geometry) on kinetic performance and sensitivity in HPLC.
8. Demonstrate the impact of analytes molecular weight on thermodynamic (retention and selectivity) and kinetic (efficiency) performance.