hplc

All posts tagged hplc

This image below outlines the step by step process for HPLC Method Development.

Method Development

Courtesy: Researchgate

Agilent has a few nice guides on HPLC which I’ve attached below.

Agilent HPLC Basics

Agilent HPLC Separation Fundamentals

Agilent Rapid HPLC Method Development

Below are two links to sets of videos giving very basic explanations of how various chromatographic methods work.

Chromatographic Methods:

Mr Simple Science:

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.

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.

The website below is a great resource for staying current on topics in relation to liquid chromatography and gas chromatography. Sections of the site include:

  • Publications on LC and GC systems
  • Information on techniques including:
  • LC
  • HPLC
  • UPLC
  • GC
  • Mass Spec
  • Sample Prep
  • LC/GC Television and Multimedia
  • eBooks (Free)

http://bit.ly/lcgcresource

The link below is to a really useful website explaining concisely all the terms used in speaking about HPLC/LC systems. Well worth book marking:

http://bit.ly/hplclcterms