Analysis of cholesterol in mouse brain by HPLC with UV detection


Autoři: María A. Paulazo aff001;  Alejandro O. Sodero aff001
Působiště autorů: Institute of Biomedical Research (BIOMED), Pontifical Catholic University of Argentina (UCA) and National Scientific and Technical Research Council (CONICET), C1107AFF Buenos Aires, Argentina aff001
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0228170

Souhrn

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.

Klíčová slova:

Blood-brain barrier – Cell membranes – Gas chromatography-mass spectrometry – High performance liquid chromatography – Chloroform – Cholesterol – Lipids – Ultraviolet spectroscopy


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Článek vyšel v časopise

PLOS One


2020 Číslo 1