RPLC Method Development Strategies Using Stationary Phase and Mobile Phase Screening, and Some Helpful Tools and Tips

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Event Overview:

Reversed-phase method development can be a very time-consuming and demanding task, particularly if your samples contain many analytes, and if you don’t follow a systematic approach.  In industrial and pharmaceutical research laboratories, researchers often must develop analytical methods that require the separation of tens of components including active pharmaceutical ingredients (APIs), process impurities, API degradants, formulation excipients, as well as degradants resulting from chemical reactions between APIs and excipients.  

The development of a robust and rugged separation and method that can be validated and used in multiple laboratories over period of 10–20 years can be a significant challenge. Some pharmaceutical companies dedicate a group of individuals whose main responsibility is to develop, continuously improve, and apply a designated set of stationary phases and analysis conditions, which they use to carry out column and condition screening for new drug entities.  The results of their screening efforts can then be transferred to other investigators, who then carry out further method development, refinement and validation using the learnings gained from the screening group’s experiments. Conversely, there are other situations where a method development project only entails the separation of a limited number of analytes, and the resulting method will only be used in the method developer’s laboratory for a few weeks or months. In such situations, an acceptable separation of only 5–10 compounds is necessary.  

Those developing methods for both relatively simple and more complex samples can benefit from using a stationary phase and mobile phase screening approach for reversed-phase HPLC method development. In fact, with modern computer-controlled instrumentation including column and mobile phase selection, a variety of stationary phases, organic modifiers and blends, and mobile phase buffers can be compared in automated fashion-often overnight. Even for those who must do some of this experimentation manually, it’s quite practical to compare multiple stationary phases, different organic modifiers and one or more pHs in an 8-hour working day.

In this webcast, we will discuss the usefulness of reversed-phase HPLC stationary phase and mobile phase screening and will show how it can be effective for selecting combinations for further development and refinement. We will also review some tools and calculations that are useful in experimental planning, delay volume and extra-column volume calculations.

Key Learning Objectives

• Why is it important to compare stationary phases with different retention mechanisms when developing a new reversed-phase HPLC separation?

• How do you know which stationary phase is most suitable for your sample and analytes?

• How do you decide which column geometry is appropriate for column screening and your instrumentation?

• At which point in method development and validation is it most useful to evaluate separation robustness and ruggedness?

• What are some tools and calculations that can make method development easier?

Speaker: Thomas J. Waeghe, Ph.D., Senior Scientist, MAC-MOD Analytical, Inc.

Time and Date: Wednesday, Nov. 13, 2019 at 1pm EST | 12pm CST | 10am PST

On demand available after airing until Nov. 13, 2020

Sponsor: MAC-MOD Analytical, Inc.

Register free: http://www.chromatographyonline.com/lcgc_w/rplc