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Journal of Lipid Research, Vol. 43, 654-660, April 2002
Copyright © 2002 by Lipid Research, Inc.

Methods

Separation of micelles and vesicles within lumenal aspirates from healthy humans: solubilization of cholesterol after a meal

Correspondence to: Laura A. Woollett, To whom correspondence should be addressed., laura.woollett{at}uc.edu (E-mail)

Understanding the physico-chemical relationship of lumenal lipids to one another is critical when elucidating the mechanism of components known to impact cholesterol absorption. Presently, there are no studies that describe the proportion of cholesterol carried as micelles or vesicles within human lumenal contents. Part of the reason for the scarceness of data is because of the lack of appropriate methodology required for reproducible sample collection and analysis. Thus, the object of the present studies was to develop a method to measure the amount of cholesterol carried as micelles or vesicles in human lumenal samples. The method includes the collection of lumenal samples from the ligament of Trietz through a Fredrick Miller tube, separation of the aqueous subphase from the nondigested lipids, separation of micelles and vesicles on Sepharose^® 4B columns within 48 h of collection using elution buffers consisting of the intermicellar bile acid composition, and finally quantitation of cholesterol eluted off of the columns.

The distribution of cholesterol between micelles and vesicles obtained under different concentrations of bile acids and various lipids was comparable to results obtained from phase diagrams using the lumenal molar percentages of lipids obtained from the same samples.—Yao, L., J. E. Heubi, D. D. Buckley, H. Fierra, K. D. R. Setchell, N. A. Granholm, P. Tso, D. Y. Hui, and L. A. Woollett. Separation of micelles and vesicles within lumenal aspirates from healthy humans: solubilization of cholesterol after a meal. J. Lipid Res. 2002. 43: 654–660.

Supplementary key words: bile acid, absorption, lipid

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