A one-vial method for routine extraction and quantification of free fatty acids in blood and tissue by HPLC

Department of Physiology, Medical Faculty, RWTH Aachen, Federal Republic of Germany.
Analytical Biochemistry (Impact Factor: 2.22). 06/1988; 171(1):150-7. DOI: 10.1016/0003-2697(88)90136-4
Source: PubMed


A one-vial procedure has been developed to quantify free fatty acids in human blood serum and rat heart tissue. To allow routine analysis the system has been constructed to allow simultaneous processing of nine samples (Praechromat). The free fatty acids are extracted with Freon 11 and then derivatized to coumarin esters prior to HPLC. The Freon 11 extraction of the free fatty acids is rapid and complete. Neither hydrolytic degradation of natural fatty acid esters nor oxidative damage of unsaturated fatty acids was observed. Fifteen free fatty acids (FFA) were routinely quantified by isocratic elution with high reproducibility (SD less than 4%) and good recovery (0.1 mM FFA: 98-100%, 0.02 mM: 91-105%). The free fatty acids could be determined in the range from 20 pmol to 20 nmol.

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    • "After transfection, cells were cultured in DMEM supplemented by 0.2% FBS for 24 hours to strengthen cell membrane before addition of FAs. Afterwards, cells were transactivated for 24 hours in absence or presence of omega-3 FAs in concentrations varying between 1–15 μM to reflect biological plasma or red blood cells concentration of FAs [14]. Cells were treated with either solvent (dimethyl sulfoxide (DMSO), 0.01% final concentration), or treatments of EPA and/or DHA (Sigma- Aldrich, Oakville, ON, Canada). "
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