Research Of Sea Buckthorn Seed Oil Fat Emulsion Injection

Intravenous fat emulsions have been developed to provide utilizable energy to many tissues and to prevent essential fatty acids (FAs) deficiency. There are two series of polyunsaturated fatty acids (PUFAs) that are regarded as essential fatty acids (EFAs):theω-3 and co-6 series, and the balance betweenω-3 andω-6 EFAs may markedly affect physiological function. Sea buckthorn seed oil (SBSO) had been reported to have the protective effects of gastric ulcers, blood fat,thrombus,atopic dermatitis and have the effects of increase the specific and nonspecific immune functions. It is known that SBSO is rich in the two essential fatty acids:LA and ALA.The ratio ofω-3/ω-6 EFAs is 1:1～1:2.Therefore, it is worthwhile developing SBFE as a new intravenous fat emulsions to compensate for the inadequacy in the ratio ofω-3/ω-6 EFAs in natural material without incorporating other oils.The refinement processes are established as follows:degumming-deacidification decoloring-deodorizing-dehydration-filtering under low temperature. The result showed that the relative density (0.9206), refractive index(1.478), iodine value(153),saponification number(197), fatty acids (36.45% linoleic acid,30.20%α-linolenic acid,20.83% oleic acid, 8.87% palmitic acid,2.29% stearic acid) in SBSO for injection had no significant difference between crude SBSO.The quality specification such as acid number (≤0.1 mg KOH/g), peroxide value (<3 meq/kg), unsaponifiable matter (≤1%)and water content and volatile material (≤0.3%)of SBSO for injection complied with the quality specification of soybean oil for injection in the Chinese Pharmacopoeia. Therefore the method of refinement can effectively reduce acid value and peroxide value and give the material an excellent appearance.High pressure homogenization was used to prepare sea buckthorn seed oil fat emulsion (SBFE).Taking physical appearance, pH, particle size distribution (PSD),ζ-potential, peroxide value as index, the final formulation of SBFE was SBSO 10%(w/v),soya lecithin 1.2%(w/v), F-68 0.2%(w/v), Tween80 0.2%(w/v), glycerin 2.5%(w/v), EDTA 0.005%(w/v) and Na2SO3 0.025%(w/v).Taking into account the safety of clinical applications and basing on the research of nutrition fat emulsion and the requirements of injection in SFDA, the final formulation was using egg lecithin as single emulsifier to prepare SBFE. Taking physical appearance and peroxide value as the main control index, preparation process was carefully investigated. The results showed that using high-shear mixer to short the lecithin dissolution time in SBSO can improve the emulsifying capacity of egg lecithin. Using the EDTA-2Na aqueous solution to clean the equipment ahead of time can increase the physical stability of SBFE. The final formulation and preparation process for SBFE were as follows:0.1% EDTA-2Na aqueous wash the high-shear mixer and the high pressure homogenization first; The oil phase (egg lecithin 1.2%(w/v) and SBSO 10%(w/w)) was preheated at 70℃,then with high-shear mixer at 8000-10000rpm for 3 min to pound the egg lecithin to pieces, then dissolved at 70℃to form the oil phase. Glycerol 2.5%(w/v) were dispersed in the sterile water for injection and stired at 70℃to obtain the water phase. The oil phase was added into the water phase with high-shear mixer at 8000-10000rpm, 10min-15min to repare a coarse emulsion. Then the total volume was adjusted to 100 ml with sterile water for injection. After adjusting the pH to 8.5 with 0.1 mol/L NaOH solution, the coarse emulsion was put through a high pressure homogenizer at 800Pa for 7 cycles. The temperature of the entire homogenization process was maintained at 40℃using an ice-water bath. Then, the emulsion was transferred to vials under nitrogen gas and then sterilized in a 121℃rotating water bath for 15 min.Quality control item was established, including physical appearance, pH, particle size distribution (PSD),ζ-potential, free fatty acids, anisidine value, peroxide value, fatty acids content, glycerol content, egg lecithin content. In addition, the results of long term stability test at 25±2℃and4±2℃for 12 months showed that the characteristics of SBSE were nearly unchanged, and therefore, SBFE using egg lecithin as single emulsifier had the potential for industrial production and clinical application.In this paper, Allergic substance detection method was established using ELISA. Study shows that three bathes of SBFE had antigen activity impurities.Based on the animal model of chemothe-rapy injury in mice, Intralipid group, Omegaven group and SBFE group all had the effects of recovering the injury body, but SBFE group recovered earlier. SBFE group can facilitate the expression of Bcl-XL in BMC and enhance the activity of SOD and MAO.