The validation of analytical methods is of crucial importance in several fields of application. A new protocol for the validation of chromatographic methods has been proposed. The overall protocol is described in a parallel paper, where the case of a multi-targeted gas chromatography – mass spectrometry (GC–MS) method for the determination of androgens in human urine is in-depth discussed. The purpose of this paper is to report the details about the GC–MS separation and detection of the target analytes, and to provide the mathematical formulas needed to perform the validation of the principal parameters. Briefly, the validation protocol foresees the repetition of three calibration curves in three different days, providing a total amount of nine replicates. Such a structured design allows to use the same experiments to • perform a rigorous calibration study, by the evaluation of heteroscedasticity, comparison of several weights and linear/quadratic calibration curves. • determine several parameters which are traditionally computed from dedicated experiments, namely intra- and inter-day accuracy and precision, limit of detection, specificity, selectivity, ion abundance repeatability, and carry over. • Finally, few further experiments are necessary to evaluate the retention time repeatability, matrix effect and extraction recovery.

Experimental and statistical protocol for the effective validation of chromatographic analytical methods

Alladio E.
First
;
Amante E.
;
Bozzolino C.;Salomone A.;Vincenti M.;
2020

Abstract

The validation of analytical methods is of crucial importance in several fields of application. A new protocol for the validation of chromatographic methods has been proposed. The overall protocol is described in a parallel paper, where the case of a multi-targeted gas chromatography – mass spectrometry (GC–MS) method for the determination of androgens in human urine is in-depth discussed. The purpose of this paper is to report the details about the GC–MS separation and detection of the target analytes, and to provide the mathematical formulas needed to perform the validation of the principal parameters. Briefly, the validation protocol foresees the repetition of three calibration curves in three different days, providing a total amount of nine replicates. Such a structured design allows to use the same experiments to • perform a rigorous calibration study, by the evaluation of heteroscedasticity, comparison of several weights and linear/quadratic calibration curves. • determine several parameters which are traditionally computed from dedicated experiments, namely intra- and inter-day accuracy and precision, limit of detection, specificity, selectivity, ion abundance repeatability, and carry over. • Finally, few further experiments are necessary to evaluate the retention time repeatability, matrix effect and extraction recovery.
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100919
100919
Chromatographic method; Effective validation protocol for chromatography mass spectrometry analytical methods; GC-MS; Multiresidual analysis; Validation protocol
Alladio E.; Amante E.; Bozzolino C.; Seganti F.; Salomone A.; Vincenti M.; Desharnais B.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2318/1763239
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