Cargando…

Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach

Background: The stability of a drug or metabolites in biological matrices is an essential part of bioanalytical method validation, but the justification of its sample size (replicates number) is insufficient. The international guidelines differ in recommended sample size to study stability from no r...

Descripción completa

Detalles Bibliográficos
Autores principales: Gniazdowska, Elżbieta, Goch, Wojciech, Giebułtowicz, Joanna, Rudzki, Piotr J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8779260/
https://www.ncbi.nlm.nih.gov/pubmed/35056772
http://dx.doi.org/10.3390/molecules27020457
_version_ 1784637531629813760
author Gniazdowska, Elżbieta
Goch, Wojciech
Giebułtowicz, Joanna
Rudzki, Piotr J.
author_facet Gniazdowska, Elżbieta
Goch, Wojciech
Giebułtowicz, Joanna
Rudzki, Piotr J.
author_sort Gniazdowska, Elżbieta
collection PubMed
description Background: The stability of a drug or metabolites in biological matrices is an essential part of bioanalytical method validation, but the justification of its sample size (replicates number) is insufficient. The international guidelines differ in recommended sample size to study stability from no recommendation to at least three quality control samples. Testing of three samples may lead to results biased by a single outlier. We aimed to evaluate the optimal sample size for stability testing based on 90% confidence intervals. Methods: We conducted the experimental, retrospective (264 confidence intervals for the stability of nine drugs during regulatory bioanalytical method validation), and theoretical (mathematical) studies. We generated experimental stability data (40 confidence intervals) for two analytes—tramadol and its major metabolite (O-desmethyl-tramadol)—in two concentrations, two storage conditions, and in five sample sizes (n = 3, 4, 5, 6, or 8). Results: The 90% confidence intervals were wider for low than for high concentrations in 18 out of 20 cases. For n = 5 each stability test passed, and the width of the confidence intervals was below 20%. The results of the retrospective study and the theoretical analysis supported the experimental observations that five or six repetitions ensure that confidence intervals fall within 85–115% acceptance criteria. Conclusions: Five repetitions are optimal for the assessment of analyte stability. We hope to initiate discussion and stimulate further research on the sample size for stability testing.
format Online
Article
Text
id pubmed-8779260
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-87792602022-01-22 Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach Gniazdowska, Elżbieta Goch, Wojciech Giebułtowicz, Joanna Rudzki, Piotr J. Molecules Article Background: The stability of a drug or metabolites in biological matrices is an essential part of bioanalytical method validation, but the justification of its sample size (replicates number) is insufficient. The international guidelines differ in recommended sample size to study stability from no recommendation to at least three quality control samples. Testing of three samples may lead to results biased by a single outlier. We aimed to evaluate the optimal sample size for stability testing based on 90% confidence intervals. Methods: We conducted the experimental, retrospective (264 confidence intervals for the stability of nine drugs during regulatory bioanalytical method validation), and theoretical (mathematical) studies. We generated experimental stability data (40 confidence intervals) for two analytes—tramadol and its major metabolite (O-desmethyl-tramadol)—in two concentrations, two storage conditions, and in five sample sizes (n = 3, 4, 5, 6, or 8). Results: The 90% confidence intervals were wider for low than for high concentrations in 18 out of 20 cases. For n = 5 each stability test passed, and the width of the confidence intervals was below 20%. The results of the retrospective study and the theoretical analysis supported the experimental observations that five or six repetitions ensure that confidence intervals fall within 85–115% acceptance criteria. Conclusions: Five repetitions are optimal for the assessment of analyte stability. We hope to initiate discussion and stimulate further research on the sample size for stability testing. MDPI 2022-01-11 /pmc/articles/PMC8779260/ /pubmed/35056772 http://dx.doi.org/10.3390/molecules27020457 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gniazdowska, Elżbieta
Goch, Wojciech
Giebułtowicz, Joanna
Rudzki, Piotr J.
Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach
title Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach
title_full Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach
title_fullStr Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach
title_full_unstemmed Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach
title_short Replicates Number for Drug Stability Testing during Bioanalytical Method Validation—An Experimental and Retrospective Approach
title_sort replicates number for drug stability testing during bioanalytical method validation—an experimental and retrospective approach
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8779260/
https://www.ncbi.nlm.nih.gov/pubmed/35056772
http://dx.doi.org/10.3390/molecules27020457
work_keys_str_mv AT gniazdowskaelzbieta replicatesnumberfordrugstabilitytestingduringbioanalyticalmethodvalidationanexperimentalandretrospectiveapproach
AT gochwojciech replicatesnumberfordrugstabilitytestingduringbioanalyticalmethodvalidationanexperimentalandretrospectiveapproach
AT giebułtowiczjoanna replicatesnumberfordrugstabilitytestingduringbioanalyticalmethodvalidationanexperimentalandretrospectiveapproach
AT rudzkipiotrj replicatesnumberfordrugstabilitytestingduringbioanalyticalmethodvalidationanexperimentalandretrospectiveapproach