SUCCESSFUL CHROMATOGRAPHY STARTS AT THE BEGINNING
Understanding the basics of any chromatographic technique is the first step towards achieving your analytical goals and allowing correct decision making and effective pharmaceutical method development and troubleshooting. Isocratic HPLC methods are the ideal place to start.
Separation Science has teamed up with HPLC expert Tom Jupille to create an introductory online tutorial that explains some standard vocabulary and terminology, including:
- Separation Chemistry (An overview of the different chemical mechanisms used in HPLC (reversed phase, normal phase & HILIC, ion exchange, ion pair, and size exclusion)
- Retention and Selectivity (A number of parameters are used to describe an HPLC separation. In this section we will define and explore the significance of retention and selectivity)
- Efficiency and Resolution (A number of parameters are used to describe an HPLC separation. In this section we will define and explore the significance of efficiency and resolution)
- Controlling Resolution (Resolution is the key parameter that describes the quality of a separation. In this section we will look at how Rs can be controlled by using k’, alpha, and N)
- Peak Tailing and Peak Fronting (The Gaussian peak shape is an idealization that is seldom seen in practice. We will explore the standard measurements used to describe deviations from that ideal: the USP “Tailing Factor” and the ASTM “Asymmetry Factor”)