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J. Lipid Res.
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A more recent version of this article appeared on January 1, 2007

Papers In Press, published online ahead of print October 24, 2006
J. Lipid Res., doi:10.1194/jlr.D600035-JLR200
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Submitted on September 5, 2006
Revised on October 23, 2006
Accepted on October 23, 2006

Simultaneous assessment of lipid classes and bile acids in human intestinal fluid by solid phase extraction and HPLC methods

Eva M. Persson, Lars J. Löfgren, Göran I. Hansson, Bertil Abrahamsson, Hans Lennernäs, and Ralf G. Nilsson

Molecular Pharmacology, Astrazeneca R&D Mölndal, Mölndal SE431 83

Corresponding Author: Ralf.Nilsson{at}astrazeneca.com

The purpose of the study reported here was to develop a method for the determination of lipid classes in intestinal fluids, including bile acids. A solid phase extraction (SPE) method using C18 and silica columns for the separation of bile acids (BA), phospholipids (PL) and neutral lipids including free fatty acids (NL) has been developed and validated. Fed state small intestinal fluid collected from humans was treated with orlistat to inhibit lipolysis and mixed with acetic acid and methanol prior to the SPE to maximize lipid recoveries. BA, PL and NL were isolated using lipophilic and polar solvents to promote elution from the SPE columns. The different lipid classes were subsequently analysed using three separately optimized HPLC methods with evaporative light scattering detectors. High recoveries (> 90%) of all lipids evaluated were observed, with low coefficients of variation (< 5%). The HPLC methods developed were highly reproducible and allowed base-line separation of nearly all lipid classes investigated. In conclusion, these methods provide a means of lipid class analysis of neutral lipids, phospholipids, as well as, bile acids, in human fed state small intestinal fluid, with potential use in other fluids from the intestinal tract and animals.


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J. Lipid Res., December 1, 2008; 49(12): 2690 - 2695.
[Abstract] [Full Text] [PDF]


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