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Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow
Cuprizone is an amide compound that has been wildly used in various animal studies, such as in the investigation of remyelination in mouse model. It is important to control the amount of cuprizone dosed in animals to be consistent as different amounts may lead to different clinical observations. Cup...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8429673/ https://www.ncbi.nlm.nih.gov/pubmed/34504258 http://dx.doi.org/10.1038/s41598-021-97590-z |
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author | Zheng, Fengmei Lin, Yiqing Boulas, Pierre |
author_facet | Zheng, Fengmei Lin, Yiqing Boulas, Pierre |
author_sort | Zheng, Fengmei |
collection | PubMed |
description | Cuprizone is an amide compound that has been wildly used in various animal studies, such as in the investigation of remyelination in mouse model. It is important to control the amount of cuprizone dosed in animals to be consistent as different amounts may lead to different clinical observations. Cuprizone is usually administrated as a minor component (i.e., 0.3%) of a mixture with powdered or pelleted rodent chow. Its low content, combined with the complex nature of chow, represents a significant challenge for the quantification of cuprizone in the mixture. To the best of our knowledge, no method has been reported in the literature so far. In this study, a simple, selective, and sensitive hydrophilic interaction liquid chromatographic method was developed for the quantification of cuprizone in cuprizone pre-clinical formulations. The analytical method comprises a fast ultrasound assisted extraction with acetonitrile/water as a solvent followed by gradient separation using a Waters Xbridge HILIC column with 0.1% TFA in water and acetonitrile as mobile phases and UV detection at 220 nm. The specificity, linearity, accuracy, repeatability, and limit of quantitation (LOQ) of the method were established. The method was determined to be linear in the range of 10–200 μg/mL. Accuracy was assessed by spiking a chow placebo with various amounts of a cuprizone reference standard to achieve target concentration levels and the recoveries were within the acceptance criterion of 90–110% of the target concentrations. Repeatability was demonstrated at the nominal concentration of 100 µg/mL and LOQ level of 2.5 μg/mL. This method has been demonstrated to be suitable for its intended use and has been successfully applied to the quantification of low levels of cuprizone in chow formulations. It was found that the cuprizone content in chow could varied significantly between batches and the potential causes of the variability were investigated. |
format | Online Article Text |
id | pubmed-8429673 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-84296732021-09-13 Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow Zheng, Fengmei Lin, Yiqing Boulas, Pierre Sci Rep Article Cuprizone is an amide compound that has been wildly used in various animal studies, such as in the investigation of remyelination in mouse model. It is important to control the amount of cuprizone dosed in animals to be consistent as different amounts may lead to different clinical observations. Cuprizone is usually administrated as a minor component (i.e., 0.3%) of a mixture with powdered or pelleted rodent chow. Its low content, combined with the complex nature of chow, represents a significant challenge for the quantification of cuprizone in the mixture. To the best of our knowledge, no method has been reported in the literature so far. In this study, a simple, selective, and sensitive hydrophilic interaction liquid chromatographic method was developed for the quantification of cuprizone in cuprizone pre-clinical formulations. The analytical method comprises a fast ultrasound assisted extraction with acetonitrile/water as a solvent followed by gradient separation using a Waters Xbridge HILIC column with 0.1% TFA in water and acetonitrile as mobile phases and UV detection at 220 nm. The specificity, linearity, accuracy, repeatability, and limit of quantitation (LOQ) of the method were established. The method was determined to be linear in the range of 10–200 μg/mL. Accuracy was assessed by spiking a chow placebo with various amounts of a cuprizone reference standard to achieve target concentration levels and the recoveries were within the acceptance criterion of 90–110% of the target concentrations. Repeatability was demonstrated at the nominal concentration of 100 µg/mL and LOQ level of 2.5 μg/mL. This method has been demonstrated to be suitable for its intended use and has been successfully applied to the quantification of low levels of cuprizone in chow formulations. It was found that the cuprizone content in chow could varied significantly between batches and the potential causes of the variability were investigated. Nature Publishing Group UK 2021-09-09 /pmc/articles/PMC8429673/ /pubmed/34504258 http://dx.doi.org/10.1038/s41598-021-97590-z Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Zheng, Fengmei Lin, Yiqing Boulas, Pierre Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
title | Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
title_full | Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
title_fullStr | Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
title_full_unstemmed | Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
title_short | Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
title_sort | development and validation of a novel hilic method for the quantification of low-levels of cuprizone in cuprizone-containing chow |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8429673/ https://www.ncbi.nlm.nih.gov/pubmed/34504258 http://dx.doi.org/10.1038/s41598-021-97590-z |
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