title: Simultaneous assessment of lipid classes and bile acids in human intestinal fluid by solid-phase extraction and HPLC methods.
abstract: The purpose of the study reported here was to develop a method for the determination of lipid classes in intestinal fluids, including bile acids (BAs). A solid-phase extraction (SPE) method using C18 and silica columns for the separation of BAs, phospholipids (PLs), and neutral lipids (NLs), including free fatty acids, 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 before SPE to maximize lipid recoveries. BAs, PLs, and NLs were isolated using lipophilic and polar solvents to promote elution from the SPE columns. The different lipid classes were subsequently analyzed 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 baseline separation of nearly all lipid classes investigated. In conclusion, these methods provide a means of lipid class analysis of NLs, PLs, and BAs in human fed-state small intestinal fluid, with potential use in other fluids from the intestinal tract and animals.
authors.affiliation: Department of Pharmacy, Uppsala University, S-751 23 Uppsala, Sweden.
authors.forename: Eva
authors.initials: E
authors.lastname: Persson
authors.forename: Lars
authors.initials: L
authors.lastname: Löfgren
authors.forename: Göran
authors.initials: G
authors.lastname: Hansson
authors.forename: Bertil
authors.initials: B
authors.lastname: Abrahamsson
authors.forename: Hans
authors.initials: H
authors.lastname: Lennernäs
authors.forename: Ralf
authors.initials: R
authors.lastname: Nilsson
mesh_terms: D001647:Bile Acids and Salts / Q000302:isolation & purification*; D001826:Body Fluids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D007422:Intestines / Q000737:chemistry*; D008055:Lipids / Q000302:isolation & purification*; D012542:Scattering, Radiation
chemical_list: D001647:Bile Acids and Salts; D008055:Lipids