Analysis of cholesterol in mouse brain by HPLC with UV detection
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. Li...
Guardado en:
| Autores principales: | , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
Public Library of Science
2020
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| Acceso en línea: | https://repositorio.uca.edu.ar/handle/123456789/10232 |
| Aporte de: |
| Sumario: | 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. |
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