| 摘要 |
The invention relates to field of medicine, in particular to clinical-laboratory studies and can be used for determination of fat acid composition of lipids in biological samples of different origin. Method for chromatographic determination of total fat acid composition of biological liquids prescribes separation of lipids from matrix material, at that liquid sample of biological material chosen from group: blood-serum, suspension of cells homogenizate of tissue is dried to obtain sample of de-watered biological material in apparatus of sublimation drying or in rotary vacuum concentrator of type “Savant SpeedVac”, at that aliquote of sample of liquid biological material under investigation in amount of 20-500 ml is de-watered directly in test-tube for derivation, and lipids of dried sample of biological material in that very test-tube are transformed to respective methyl esters with heating with solution of 0.5 N of sodium methylate in methanol in calculation of 150 mkl of solution of sodium methylate oer 100 mkl of blood or 15 % of homogenizate, and free fat acids present are heated with 10 % methanol solution of tri-fluoric boron in calculation of 0.8-1.0 ml per 100 ml of blood or 15 % homogenizate, at that after the end of methylation alcohol solution of sample of biological material is dissolved with distillated water in two times, after that one extracts methyl ethers of fat acids obtained to heptane non-polar phase, at that somewhat delayed stratification of water-alcohol and non-polar heptane phase at stage of extraction of ethers from reaction nix is accelerated by adding drop of propyl alcohol or by shirt-term (1-2 minutes) centrifugation in low-speed centrifuge, after that extracted to heptane phase methyl ethers of fat acids are determined quantitatively by method of capillary gas chromatography in column with polar immobilized liquid phase. Main advantage of the method proposed by us is in principal simplification and speed-up of preparation of biological samples for chromatographic analysis. |
