Robustness method validation protocol

Bosque-Sendra, J.M. Nechar, M. Cuadros Redriguez, L. Decision protocol for checking robustness with previous outlier detection in the validation of analytical methods. Fresenius J. Anal Chemie 1999, 365, 480-488. 

Once our methods have been fully developed, and procedures drafted, a protocol driven, ICH compliant method validation should be conducted. This validation should be appropriate for this early stage of development. Typically, since only one lab is running this method, and likely one chemist, intermediate precision and robustness do not need to be evaluated. Other critical attributes, such as specificity, linearity, and accuracy will need to be evaluated. 

Upon successful completion of the prevalidation audit, the analytical chemist simply performs the experiments listed in the validation protocol using the analytical method as written. Based on the results of the validation, it may be necessary to revise the method to include details such as solution stability, relative retention times, relative response factors, or cautionary statements resulting from the robustness experiments. 

The introduction of high-resolution, robust, rapid, selective, sensitive, precise, and accurate chromatographic methods is essential in food science, as well as in many other research fields, as illustrated in previous chapters of this book. The development and validation of methods to be employed in food analysis must be carried out with the greatest attention, following official protocols and guidelines (e.g., the U.S. Food and Drug Administration). The obvious reason is that foods are introduced into the human body and are, for the main part, absorbed. 

Prior to its use a method has to be validated. Validation is the formal proof that the method is suitable for the intended purpose. This requires that all steps and parameters of the method have been clearly specified in a written method description, any necessary equipment was qualified, and acceptance criteria for each validation point have been agreed upon. For quantitative methods the International Conference on Harmonization (ICH) has issued specific guidelines for setting up a validation protocol and for parameters that have to be validated for different applications. These include specificity, accuracy, precision, LOD, LOQ, linearity, and range as well as robustness. The only required validation parameter for qualitative methods is specificity. 

The following procedure is a robust calibration and validation protocol suitable for Raman spectrometers. It has been demonstrated to allow calibration transfer between different Raman spectrometer instruments. It is consistent with the recommended methods of Chapter 6, with some extensions. 

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