| Literature DB >> 18373963 |
Jaroslav Zelenka1, Martin Lenícek, Lucie Muchová, Milan Jirsa, Michal Kudla, Peter Balaz, Marie Zadinová, J Donald Ostrow, Ronald J Wong, Libor Vítek.
Abstract
Unconjugated bilirubin (UCB) exhibits potent antioxidant and cytoprotective properties, but causes apoptosis and cytotoxicity at pathologically elevated concentrations. Accurate measurement of UCB concentrations in cells, fluids and tissues is needed to evaluate its role in redox regulation, prevention of atherosclerotic and malignant diseases, and bilirubin encephalopathy. In the present study, we developed and validated a highly sensitive method for tissue UCB determinations. UCB was extracted from rat organs with chloroform/methanol/hexane at pH 6.2 and then partitioned into a minute volume of alkaline buffer that was subjected to HPLC using an octyl reverse phase (RP) column. Addition of mesobilirubin as an internal standard corrected for losses of UCB during extraction. Recoveries averaged 75+/-5%. The detection limit was 10pmol UCB/g wet tissue. Variance was +/-2.5%. When used to measure UCB concentrations in tissues of jaundiced Gunn rats, this procedure yielded UCB levels directly comparable to published methods, and accurately determined very low tissue bilirubin concentrations (</=40pmol UCB/g tissue) in non-jaundiced rats.Entities:
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Year: 2008 PMID： 18373963 DOI： 10.1016/j.jchromb.2008.03.005
Source DB: PubMed Journal: J Chromatogr B Analyt Technol Biomed Life Sci ISSN： 1570-0232 Impact factor: 3.205