The Preparation Of Dilinoleoylphospholipids

Dilinoleoylphospholipids is a kind of phospholipids with special physiological function, which is approved by some medical experts in the realm of hepatinica.The purpose of the research is to prepare a kind of active pharmaceutical ingredient, the dilinoleoylphospholipids, to enhance the curative effect and to provide good raw material for preparing the dilinoleoylphosphatidylcholine with higher purity.The safflower oil as raw material, the 98.32% linoleic acid was prepared by saponification, acidification and urea adduction fractionation. Based on the documents, five influence factors chosen were the substrate ratioSr (safflower oil/phospholipids, mol/mol), reaction time (Ti, h), reaction temperature (Te,℃), lipase dosage (Ld, wt.% based on two substrates) and water addition (Wd, wt.% based on lipase). Under the condition of solvent-free and lipase TLIM, two kinds of method of preparing the dilinoleoylphospholipids, safflower oil and high purity linoleic acid as raw materials respectively, were studied. On the basis of one-facor analysis, the orthogonal experimental design and response surface methodology (RSM) were used to optimize the condition of reaction. The results indicated that the quality score of linoleic acid was equal to 72.28% with the safflower oil as raw materials, however, the quality score of linoleic acid was 83.56% with the high putity linoleic acid as raw materials. So the optimal process condition was that under the condition of solovent-free system, Sr 8.52:1,Ti 55.65h,Te 56.88℃and Ld 29.37%, the transesterification between linoleic acid and phospholipids was catalyzed by Lipase TLIM.Secondly, the kinetics and thermodynamics of the transesterification between linoleic acid and phospholipids were researched. Gas chromatography -internal standard curve method, the method to quantify the linoleic acid in kinetics research was developed. The index of methodology, RSD and average recovery, were 2.15% and in the range of 95.6%-104.7%, respectively. R2 was 0.9991, and the linear range was 1μg/μL -20μg/μL. The kinetic model of the acidolysis was developed. And the conformance test of kinetic model illustrated that the transesterification reaction between linoleic acid and phospholipids was in accordance with the ping-pong Bi-Bi mechanism. The thermodynamic index, activation energy and reaction order, were 14.17kJ/mol and in the range of 1.3705~1.8244, and it was approximate second-order-reaction.Finally, some physical and chemical nature of the product, such as moisture, iodine value, acetone insoluble matter, acid value and peroxide value, were studied. And the comparison of five indexs between the raw material and product indicated that IV and AV were fluctuant, and the other indexes were constant. The residue of actone was 42.24μg/g<50μg/g detected by GC, and the results was in accordance with SB/T 10206-1994. The product was composed of two kinds of phospholipids chiefly, PC and PE, and the content of PC was 54.6%. The six phosphatidylcholine species were 18:3/18:3-PC, 18:2/18:3-PC, 18:2/18:2-PC, 16:0/18:2-PC, 18:1/18:2-PC and 18:0/18:2-PC, respectively. The content of 8:2/18:2-PC was 72% under the condition of 205nm, so the 18:0/18:2-PC was the main molecular-type.