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An improved ultracentrifugation method for the separation of cholesterol carriers in bile
Author(s) -
Ayyad Nariman,
Cohen Bertram I.,
Ohshima Akira,
Mosbach Erwin H.
Publication year - 1996
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/bf02523838
Subject(s) - chromatography , ultracentrifuge , chemistry , size exclusion chromatography , cholesterol , centrifugation , elution , density gradient , filtration (mathematics) , vesicle , sucrose gradient , biochemistry , membrane , physics , statistics , mathematics , quantum mechanics , enzyme
Vesicles and micelles, the major carriers of cholesterol in bile, play a role in the formation of cholesterol gallstones. A simple and rapid ultracentrifugation method was developed to isolate these biliary cholesterol carriers when only microliter amounts of bile were available. The proposed method employs a 46 to 0% sucrose density gradient, a NVT90 near‐vertical rotor, and a centrifugation time of one hour. As little as 25 μL of bile can be used with no disruption of the carriers. The method was validated by comparison with gel filtration column chromatography using 6 mM taurocholate in the elution buffer. The sucrose linear density gradient ultracentrifugation procedure described here is simple, fast, and compares favorably with the gel filtration chromatography method for the separation of cholesterol carriers from bile.