• Application Note

Automating the Creation of Chromatographic Methods for Method Validation Using Empower Sample Set Generator

Automating the Creation of Chromatographic Methods for Method Validation Using Empower Sample Set Generator

  • Margaret Maziarz
  • Mark Wrona
  • Paul D. Rainville
  • Jade Byrd
  • Mia Summers
  • Diane M. Diehl
  • Waters Corporation

This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief demonstrates the use of Empower 3 Software’s Sample Set Generator to automate the creation of chromatographic methods and sample set methods, used in the validation of a UPLC method for metoclopramide HCl and related substances.

Benefits

Empower 3 Sample Set Generator simplifies the creation of instrument methods, method sets, and sample set methods by defining ranges of variables needed for testing.

Introduction

The validation process of an analytical method is a complex and demanding activity, consisting of many time-consuming steps. Some of these steps include acquiring, reviewing and processing data, performing calculations, approving, and final reporting of the validation results. One critical task is robustness testing, during which the effects of minor changes in chromatographic parameters on method performance are investigated to establish tolerance limits. Multiple chromatographic methods must be carefully designed and created to acquire data for each validation test. Designing, creating, and verifying these methods manually can be tedious and prone to errors.

Empower 3 Sample Set Generator simplifies the creation of instrument methods, method sets, and sample set methods by defining ranges of variables needed for testing. By automating these tasks, chromatographic method and the sample sequence generation is streamlined and transcription errors eliminated. As the tedious tasks are minimized, laboratory efficiency and productivity increases.

Here we illustrate the use of Empower 3 Sample Set Generator to automatically create chromatographic methods for robustness testing in the validation of a UPLC method for metoclopramide HCl and its USP-specified related substances.

Results and Discussion

The UPLC method for Metoclopramide HCl and its USP-specified related substances was validated using Empower 3 Software’s Method Validation Manager (MVM).1 MVM software streamlines the entire validation process in one application, from creating validation protocol method to acquiring data, reviewing, analyzing, approving, and reporting validation data.

Empower 3 Sample Set Generator was used to streamline robustness testing for creating chromatographic methods. For robustness, we assessed these parameters:

  • Column temperature: 45 ± 2.0 °C
  • Flow rate: 0.6 ± 0.05 mL/min 
  • Wavelength: 270 ± 2 nm

In the Empower 3 MVM project, we opened Sample Set Generator and loaded the robustness validation test as shown in Figure 1. The MVM experimental design, with a combination of eight different instrument conditions, was imported to Empower 3 Sample Set Generator.

Figure 1. Design of Experiments (DoE) for robustness test loaded to the Empower 3 Sample Set Generator from the Empower 3 MVM protocol.

Next, we used the Sample Set Generator to create instrument methods, method sets, and a sample set method to run the robustness experiments by completing the following steps:

1. Map factors for column temperature, flow rate, and detection wavelength to the desired settings

2. Define settings for gradient separation

3. Configure requirements for blanks/standards solutions and equilibration time

4. Generate sample set method

Using Empower 3 Sample Set Generator, we were able to automatically create instrument methods with different chromatographic conditions, method sets, and sample set method for the robustness study. The sample set method for the robustness test (Figure 2) is designed according to the experimental plan for the robustness validation test, with injections of blanks/standard solutions, experiment name, and method sets for each run. The equilibration steps are added between sample lines when there is a change in instrument condition, such as flow rate or column temperature. The instrument methods are automatically built into the methods sets.

Figure 2. Sample set method for robustness test generated using Empower 3 Sample Set Generator.

This automated generation allowed us to quickly start the chromatographic run with confidence that all the methods are correctly created. In addition, it reduced the time needed to create chromatographic methods  by about 95% compared to a manual process. The chromatographic data acquired for the robustness testing is shown in Figure 3.

Figure 3. Separation of metoclopramide and  USP-specified related substances according to the experimental design for robustness.

Conclusion

By using the Empower 3 Sample Set Generator, users are able to simultaneously and automatically create instrument methods, method sets, and sample set methods to perform chromatographic runs. As a result of automation, transcription errors that may arise during the manual process are eliminated and the time associated with generation of chromatographic methods is reduced. This improves laboratory efficiency, hence enabling increase in productivity.

Empower 3 Sample Set Generator can be adapted by any analytical laboratory to automate creation of chromatographic methods for wide range of applications performed on Waters ACQUITY UPLC Systems, including method development and validation.

References

  1. Maziarz M, McCarthy SM, Wrona M. Increasing Efficiency of Method Validation for Metoclopramide HCl and Related Substances with Empower MVM Software. Waters Application Note, 2014: 720005111en.

720005127, August 2014

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