Lipophilic fraction (LF) of known medicinal plants are still less studied, despite the fact that they have a unique group of biological active compounds (BAC). The main active substances of LF are chlorophylls, carotenoids, tocopherols, sterols, unsaturated fatty acids, phospholipids, and other bioactive substances that exhibit a wide spectrum of pharmacological action. In this regard, a comprehensive study of advanced plant of flora of Ukraine to increase the assortments of herbal remedies is an urgent problem. The aim of this work was to obtain and chemical research of lipophilic fractions of sickle alfalfa herbs. The alfalfa herb harvested throughout the growing season in Kharkov and Poltava regions in 2011-2012. Lipophilic fraction was obtained by extraction with chloroform in a Soxhlet apparatus. Detection of carotenoids and chlorophylls by thin layer chromatography (TLC) on plates of "Silufol" in one-dimensional and two-dimensional variants, the solvent system were: hexane: acetone (6:2) - I way, hexane-acetone (6:4) - II way. Assay of the lipophilic fraction by three-dimensional fluorescence spectroscopy (3DF-spectroscopy) was used for the analysis of mixtures containing fluorescent components. 3DF-spectres, that have the appearance of the surface, are characterized by a functions I = f (λexc, λem), recorded in the ultraviolet and visible ranges. Assay of the carotenoids and chlorophylls carried out by spectrophotometry. The reference solution was chloroform. Assay of fatty acids was performed by gas-liquid chromatography (GLC) of methyl esters of fatty acids via chromatograph with flame ionization detector "Shimadzu GC-14B". 20.0 g crushed sickle alfalfa herbs were exhaustively extracted with chloroform in a Soxhlet apparatus for produce a lipophilic fractions. The chloroform extract was evaporated in order to remove the extractant. The percentage of lipophilic fraction in the herbs was 7.5%. Quantity of the carotenoids and chlorophylls determined by spectrophotometry was 120.57 mg/% and 197.55 mg/%, respectively. By analyzing composition of the fatty acid in the lipophilic fraction of sickle alfalfa herbs were found 13 fatty acids, 6 of which were saturated: lauric, myristic, palmitic, palmitoleic, stearic, margarine and 7 - unsaturated: oleic, linoleic, α-linolenic, arachidonic, gondolic, erucic, lignoceric. Quantitatively were dominated palmitic - 34.25%, linoleic - 15.11% and α-linolenic - 17.93% acids. The total content of unsaturated fatty acids was 49.66%, what assumes F-vitamin activity of lipophilic fractions. The lipophilic fraction was first obtained of sickle alfalfa herbs. Composition and quantitative content of carotenoids (120.57 mg/%) and chlorophylls (197.55 mg/%) were detected. The percentage of lipophilic fraction was 7.5%. Assay of the fatty acids in lipophilic fractions was done by GLC. Quantitatively were dominated palmitic - 34.25%, linoleic - 15.11% and α-linolenic - 17.93% acids.