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Qualitative threshold method validation and uncertainty evaluation: A theoretical framework and application to a 40 analytes liquid chromatography–tandem mass spectrometry method


Qualitative threshold method validation and uncertainty evaluation: A theoretical framework and application to a 40 analytes liquid chromatography–tandem mass spectrometry method

Camirand-Lemyre, Félix ORCID: https://orcid.org/0000-0003-3277-2729, Desharnais, Brigitte ORCID: https://orcid.org/0000-0001-7373-656X, Laquerre, Julie, Morel, Marc-André, Côté, Cynthia, Mireault, Pascal and Skinner, Cameron D. (2020) Qualitative threshold method validation and uncertainty evaluation: A theoretical framework and application to a 40 analytes liquid chromatography–tandem mass spectrometry method. Drug Testing and Analysis, 12 (9). pp. 1287-1297. ISSN 1942-7611

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Official URL: https://doi.org/10.1002/dta.2867


Qualitative methods hold an important place in drug testing, filling central needs in screening and analyses, among others, linked to per se legislation. Nevertheless, the bioanalytical method validation guidelines do not discuss this type of method or describe method validation procedures ill‐adapted to qualitative methods. The output of qualitative methods are typically categorical, binary results, such as presence/absence or above cut‐off/below cut‐off. As the goal of any method validation is to demonstrate fitness for use under production conditions, qualitative validation guidelines should evaluate performance based on discrete, binary results instead of the continuous measurements obtained from the instrument (e.g. area).

A tentative validation guideline for threshold qualitative methods was developed by in silico modelling of measurements and derived binary results. This preliminary guideline was applied to a liquid chromatography–tandem mass spectrometry method for 40 analytes, each with a defined threshold concentration. Validation parameters calculated from the analysis of 30 samples spiked above and below the threshold concentration (false negative rate, false positive rate, selectivity rate, sensitivity rate and reliability rate) showed a surprisingly high failure rate. Overall, 13 out of the 40 analytes were not considered validated. A subsequent examination found that this was attributable to an appreciable shift in the standard deviation of the area ratio on a day‐to‐day basis, a previously undescribed and unaccounted‐for behaviour in the qualitative threshold method validation literature. Consequently, the developed guideline was modified and used to validate a qualitative threshold method, based on the binary results for performance evaluation and incorporating measurement uncertainty.

Divisions:Concordia University > Faculty of Arts and Science > Chemistry and Biochemistry
Item Type:Article
Authors:Camirand-Lemyre, Félix and Desharnais, Brigitte and Laquerre, Julie and Morel, Marc-André and Côté, Cynthia and Mireault, Pascal and Skinner, Cameron D.
Journal or Publication:Drug Testing and Analysis
Date:31 May 2020
  • National Sciences and Engineering Research Council of Canada
  • Fonds de recherche du Québec - Nature et technologies
  • Canada First Research Excellence Fund
  • Australian Research Council DP #140100125
Digital Object Identifier (DOI):10.1002/dta.2867
Keywords:method validation, qualitative, threshold, uncertainty of measurement
ID Code:987501
Deposited On:16 Oct 2020 19:10
Last Modified:31 May 2021 01:00
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