Analysis of cholesterol in mouse brain by HPLC with UV detection
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 me...
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| description | 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. |
| doi_str_mv | 10.1371/journal.pone.0228170 |
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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><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0228170</identifier><identifier>PMID: 31978159</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Acetonitrile ; Analysis ; Animals ; Biology and Life Sciences ; Biomedical research ; Blood-brain barrier ; Brain ; Brain - metabolism ; Brain research ; Calibration ; Chloroform ; Cholesterol ; Cholesterol - analysis ; Cholesterol - isolation & purification ; Chromatography ; Chromatography, High Pressure Liquid - methods ; Chromatography, Reverse-Phase ; High performance liquid chromatography ; House mouse ; Laboratory animals ; Limit of Detection ; Lipids ; Liquid chromatography ; Liquid-Liquid Extraction ; Medicine and Health Sciences ; Mice ; Mice, Inbred C57BL ; Neurophysiology ; Physical Sciences ; Research and Analysis Methods ; Spectrophotometry, Ultraviolet ; Sterols ; Ultraviolet radiation</subject><ispartof>PloS one, 2020-01, Vol.15 (1), p.e0228170-e0228170</ispartof><rights>COPYRIGHT 2020 Public Library of Science</rights><rights>2020 Paulazo, Sodero. 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metabolism</topic><topic>Brain research</topic><topic>Calibration</topic><topic>Chloroform</topic><topic>Cholesterol</topic><topic>Cholesterol - analysis</topic><topic>Cholesterol - isolation & purification</topic><topic>Chromatography</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Chromatography, Reverse-Phase</topic><topic>High performance liquid chromatography</topic><topic>House mouse</topic><topic>Laboratory animals</topic><topic>Limit of Detection</topic><topic>Lipids</topic><topic>Liquid chromatography</topic><topic>Liquid-Liquid Extraction</topic><topic>Medicine and Health Sciences</topic><topic>Mice</topic><topic>Mice, Inbred C57BL</topic><topic>Neurophysiology</topic><topic>Physical Sciences</topic><topic>Research and Analysis Methods</topic><topic>Spectrophotometry, Ultraviolet</topic><topic>Sterols</topic><topic>Ultraviolet radiation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Paulazo, María A</creatorcontrib><creatorcontrib>Sodero, Alejandro O</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Opposing Viewpoints</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>31978159</pmid><doi>10.1371/journal.pone.0228170</doi><tpages>e0228170</tpages><orcidid>https://orcid.org/0000-0001-5159-218X</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Acetonitrile Analysis Animals Biology and Life Sciences Biomedical research Blood-brain barrier Brain Brain - metabolism Brain research Calibration Chloroform Cholesterol Cholesterol - analysis Cholesterol - isolation & purification Chromatography Chromatography, High Pressure Liquid - methods Chromatography, Reverse-Phase High performance liquid chromatography House mouse Laboratory animals Limit of Detection Lipids Liquid chromatography Liquid-Liquid Extraction Medicine and Health Sciences Mice Mice, Inbred C57BL Neurophysiology Physical Sciences Research and Analysis Methods Spectrophotometry, Ultraviolet Sterols Ultraviolet radiation |
| title | Analysis of cholesterol in mouse brain by HPLC with UV detection |
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