Peuchant E, Wolff R, Salles C, Jensen R
Department of Biochemistry, Saint-André Hospital, Bordeaux, France.
Anal Biochem. 1989 Sep;181(2):341-4. doi: 10.1016/0003-2697(89)90254-6.
A one-step extraction procedure to directly quantify cholesterol, phospholipids, and fatty acids in red blood cell membranes has been developed. The method uses a single solvent, isopropanol, which extracts lipids and allows the rapid formation of isopropylic esters of fatty acids by acid catalysis. The efficiency of this new technique has been confirmed by comparing yields of cholesterol and total and individual phospholipids with yields obtained following conventional extraction procedures. Moreover, in comparison to the formation of methyl esters, we demonstrate that directly obtained isopropylic esters immediately allow the quantitative determination of fatty acids, without involving the hydrolytic degradation of fatty acids and the oxidation of unsaturated fatty acids.
已开发出一种一步提取程序,可直接定量红细胞膜中的胆固醇、磷脂和脂肪酸。该方法使用单一溶剂异丙醇,它能提取脂质,并通过酸催化使脂肪酸快速形成异丙酯。通过将胆固醇、总磷脂和各磷脂的产率与传统提取程序获得的产率进行比较,证实了这项新技术的效率。此外,与甲酯的形成相比,我们证明直接获得的异丙酯可立即对脂肪酸进行定量测定,而无需涉及脂肪酸的水解降解和不饱和脂肪酸的氧化。