Improving Effectiveness in Method Development by Using a Systematic Screening Protocol

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Margaret Maziarz, Sean M. McCarthy, and Mark Wrona
Waters Corporation
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Application Notes
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In this application, we present the development of a UPLC method, using the USP-defined assay for metoclopramide HCl and related substances as an example. Using the ACQUITY QDa Detector in conjunction with UV detection and the ACQUITY UPLC H-Class System streamlined the method development process. We demonstrate:

  • Robust UPLC method development
  • Quick and accurate identification of sample components using mass detection with the ACQUITY QDa Detector
  • Minimize the need for running individual injections of sample components to confirm the identity of peaks

A systematic protocol is employed that includes scouting, screening, and optimization steps. Results for each step are analyzed and ranked using custom calculations and reported within Empower 3 Chromatography Data Software to minimize analyst bias in decision making and ensure the overall goals are achieved.

The criteria for success with a goal of separating all nine components, achieving a resolution of ≥2.0, tailing of ≤1.5, and retention factor (k*) ≥3.0, were met.

Finally, the use of ApexTrack in Empower Software enabled consistent evaluation of chromatograms for fair comparison across the development process. Empower custom calculations and reporting allowed us to generate a scoring report to easily identify the best conditions at each step in our protocol.

Overall, using a defined systematic protocol with the UPLC system, detectors, and its column chemistries enables analytical laboratories to quickly and efficiently develop chromatographic methods. Methods developed in this manner are typically more reproducible, which allows laboratories to have a higher validation success rate.

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