Analysis of cholesterol in mouse brain by HPLC with UV detection

Autores
Paulazo, Maria Alejandra; Sodero, Alejandro Omar
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Fil: Paulazo, Maria Alejandra. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
Fil: Paulazo, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sodero, Alejandro Omar. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
Fil: Sodero, Alejandro Omar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Abstract: We describe a sensitive high performance liquid chromatography (HPLC)-based method for the determination of cholesterol in brain tissue. The method does not require the derivatization of the analyte and uses separation and quantification by reversed-phase HPLC coupled to UV detection. Lipids were methanol/chloroform extracted following the method of Bligh and Dyer, and separated using isopropanol/acetonitrile/water (60/30/10, v/v/v) as mobile phase. We observed lineal detection in a wide range of concentrations, from 62.5 to 2000 ng/μL, and were able to detect a significant increase in the brain cholesterol levels between postnatal days 2 and 10 in C57BL6 mice. Based on our validation parameters, we consider this analytical method a useful tool to assess free cholesterol in rodent brain samples and cell cultures.
Fuente
PLoS ONE. 2020, 15(1): e0228170
Materia
COLESTEROL
CEREBRO
CROMATOGRAFIA LIQUIDA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
Repositorio Institucional (UCA)
Institución
Pontificia Universidad Católica Argentina
OAI Identificador
oai:ucacris:123456789/10232
Acceder
id RIUCA_353cba1e0507222410105fbdd37875cc
oai_identifier_str oai:ucacris:123456789/10232
network_acronym_str RIUCA
repository_id_str 2585
network_name_str Repositorio Institucional (UCA)
spelling Analysis of cholesterol in mouse brain by HPLC with UV detectionPaulazo, Maria AlejandraSodero, Alejandro OmarCOLESTEROLCEREBROCROMATOGRAFIA LIQUIDAFil: Paulazo, Maria Alejandra. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; ArgentinaFil: Paulazo, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sodero, Alejandro Omar. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; ArgentinaFil: Sodero, Alejandro Omar. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaAbstract: We describe a sensitive high performance liquid chromatography (HPLC)-based method for the determination of cholesterol in brain tissue. The method does not require the derivatization of the analyte and uses separation and quantification by reversed-phase HPLC coupled to UV detection. Lipids were methanol/chloroform extracted following the method of Bligh and Dyer, and separated using isopropanol/acetonitrile/water (60/30/10, v/v/v) as mobile phase. We observed lineal detection in a wide range of concentrations, from 62.5 to 2000 ng/μL, and were able to detect a significant increase in the brain cholesterol levels between postnatal days 2 and 10 in C57BL6 mice. Based on our validation parameters, we consider this analytical method a useful tool to assess free cholesterol in rodent brain samples and cell cultures.Public Library of Science2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://repositorio.uca.edu.ar/handle/123456789/102321932-620310.1371/journal.pone.0228170Paulazo, M. A., Sodero, A. O. Analysis of cholesterol in mouse brain by HPLC with UV detection [en línea]. PLoS ONE. 2020, 15(1): e0228170. doi:10.1371/journal.pone.0228170 Disponible en: https://repositorio.uca.edu.ar/handle/123456789/10232PLoS ONE. 2020, 15(1): e0228170reponame:Repositorio Institucional (UCA)instname:Pontificia Universidad Católica Argentinaenginfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/4.0/2025-07-03T10:57:25Zoai:ucacris:123456789/10232instacron:UCAInstitucionalhttps://repositorio.uca.edu.ar/Universidad privadaNo correspondehttps://repositorio.uca.edu.ar/oaiclaudia_fernandez@uca.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:25852025-07-03 10:57:26.007Repositorio Institucional (UCA) - Pontificia Universidad Católica Argentinafalse
dc.title.none.fl_str_mv Analysis of cholesterol in mouse brain by HPLC with UV detection
title Analysis of cholesterol in mouse brain by HPLC with UV detection
spellingShingle Analysis of cholesterol in mouse brain by HPLC with UV detection
Paulazo, Maria Alejandra
COLESTEROL
CEREBRO
CROMATOGRAFIA LIQUIDA
title_short Analysis of cholesterol in mouse brain by HPLC with UV detection
title_full Analysis of cholesterol in mouse brain by HPLC with UV detection
title_fullStr Analysis of cholesterol in mouse brain by HPLC with UV detection
title_full_unstemmed Analysis of cholesterol in mouse brain by HPLC with UV detection
title_sort Analysis of cholesterol in mouse brain by HPLC with UV detection
dc.creator.none.fl_str_mv Paulazo, Maria Alejandra
Sodero, Alejandro Omar
author Paulazo, Maria Alejandra
author_facet Paulazo, Maria Alejandra
Sodero, Alejandro Omar
author_role author
author2 Sodero, Alejandro Omar
author2_role author
dc.subject.none.fl_str_mv COLESTEROL
CEREBRO
CROMATOGRAFIA LIQUIDA
topic COLESTEROL
CEREBRO
CROMATOGRAFIA LIQUIDA
dc.description.none.fl_txt_mv Fil: Paulazo, Maria Alejandra. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
Fil: Paulazo, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sodero, Alejandro Omar. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
Fil: Sodero, Alejandro Omar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Abstract: We describe a sensitive high performance liquid chromatography (HPLC)-based method for the determination of cholesterol in brain tissue. The method does not require the derivatization of the analyte and uses separation and quantification by reversed-phase HPLC coupled to UV detection. Lipids were methanol/chloroform extracted following the method of Bligh and Dyer, and separated using isopropanol/acetonitrile/water (60/30/10, v/v/v) as mobile phase. We observed lineal detection in a wide range of concentrations, from 62.5 to 2000 ng/μL, and were able to detect a significant increase in the brain cholesterol levels between postnatal days 2 and 10 in C57BL6 mice. Based on our validation parameters, we consider this analytical method a useful tool to assess free cholesterol in rodent brain samples and cell cultures.
description Fil: Paulazo, Maria Alejandra. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://repositorio.uca.edu.ar/handle/123456789/10232
1932-6203
10.1371/journal.pone.0228170
Paulazo, M. A., Sodero, A. O. Analysis of cholesterol in mouse brain by HPLC with UV detection [en línea]. PLoS ONE. 2020, 15(1): e0228170. doi:10.1371/journal.pone.0228170 Disponible en: https://repositorio.uca.edu.ar/handle/123456789/10232
url https://repositorio.uca.edu.ar/handle/123456789/10232
identifier_str_mv 1932-6203
10.1371/journal.pone.0228170
Paulazo, M. A., Sodero, A. O. Analysis of cholesterol in mouse brain by HPLC with UV detection [en línea]. PLoS ONE. 2020, 15(1): e0228170. doi:10.1371/journal.pone.0228170 Disponible en: https://repositorio.uca.edu.ar/handle/123456789/10232
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/4.0/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/4.0/
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv PLoS ONE. 2020, 15(1): e0228170
reponame:Repositorio Institucional (UCA)
instname:Pontificia Universidad Católica Argentina
reponame_str Repositorio Institucional (UCA)
collection Repositorio Institucional (UCA)
instname_str Pontificia Universidad Católica Argentina
repository.name.fl_str_mv Repositorio Institucional (UCA) - Pontificia Universidad Católica Argentina
repository.mail.fl_str_mv claudia_fernandez@uca.edu.ar
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score 13.001348

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