Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow

Abstract 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 observat...

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Autores principales:	Fengmei Zheng, Yiqing Lin, Pierre Boulas
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4aa87ecfba6a4e32919bcd945112a89a
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spelling	oai:doaj.org-article:4aa87ecfba6a4e32919bcd945112a89a2021-12-02T17:19:15ZDevelopment and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow10.1038/s41598-021-97590-z2045-2322https://doaj.org/article/4aa87ecfba6a4e32919bcd945112a89a2021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97590-zhttps://doaj.org/toc/2045-2322Abstract 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.Fengmei ZhengYiqing LinPierre BoulasNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Fengmei Zheng Yiqing Lin Pierre Boulas Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow
description	Abstract 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	article
author	Fengmei Zheng Yiqing Lin Pierre Boulas
author_facet	Fengmei Zheng Yiqing Lin Pierre Boulas
author_sort	Fengmei Zheng
title	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_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_sort	development and validation of a novel hilic method for the quantification of low-levels of cuprizone in cuprizone-containing chow
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/4aa87ecfba6a4e32919bcd945112a89a
work_keys_str_mv	AT fengmeizheng developmentandvalidationofanovelhilicmethodforthequantificationoflowlevelsofcuprizoneincuprizonecontainingchow AT yiqinglin developmentandvalidationofanovelhilicmethodforthequantificationoflowlevelsofcuprizoneincuprizonecontainingchow AT pierreboulas developmentandvalidationofanovelhilicmethodforthequantificationoflowlevelsofcuprizoneincuprizonecontainingchow
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Development and validation of a novel HILIC method for the quantification of low-levels of cuprizone in cuprizone-containing chow

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