Preparation And In Vitro And In Vivo Evaluation Of EPA-Enriched Fish Oil Emulsion

Recent studies show that eicosapentaenoic acid(EPA)have effects of regulating blood lipid,thrombosis inhibition and cardiovascular health.Given the relevance to public health,EPA,as a nutrient,has attracted the most attention because it is more acceptable than other lipid-regulating drugs.Among the various drug delivery routes,the oral pathway is the most common way of administering EPA,so that it needs to be soluble in gastric fluid and intestinal fluid for a better absorption in the small intestine.Furthermore,digestion and absorption of EPA is thought to be highly dependent on fat meal content,which would stimulate the activity of digestive enzyme and thereby enhance absorption.Consequently,hypertriglyceridemia patients treated with EPA have to take the medication with meal to avoid minimized absorption.In order to overcome the deficiencies of commercially available formulations,the EPA-enriched fish oil emulsion(FO-ME)was prepared in this study,which can effectively improve the bioavailability of EPA so that the dosage can be reduced and the absorption of the EPA would not be restricted by the fat intake.The research contents of this thesis are as follows:1.Screening the formulation,preparation technology and quality evaluation of FOME.First of all,the detection methods and standards of indicators for emulsion quality test such as the emulsion appearance,content of the active substance,particle size,p H,free acid content,peroxide value,methoxyaniline value were established.Then the FOME was prepared with 10%(w/w)fish oil,1.2%(w/w)soybean lecithin PC-X1,0.1%(w/w)sucrose ester S1,0.04%(w/w)vitamin E and water.The preparation process and sterilization process were determined.Finally,three batches of emulsions were prepared through pilot scale-up.The emulsions met the requirements in both semi-finished product testing and finished product testing,and the differences between batches were small,indicating that the process was suitable for subsequent production.2.Stability and in vitro simulated digestion study of FO-ME.The microscopic morphology of the emulsion was observed by electron microscope as a spherical structure.The zata potential of the FO-ME was-44.5 ± 0.6 m V.The results of the stability test showed that no delamination appeared after centrifuge at 4000 rpm for 15 min,which indicates that the emulsion was stable.Furthermore,the original structure was maintained in isotonic solution(PBS)and simulated gastric fluid(SGF)for 48 h.And the FO-ME remained stable for 3 months in the accelerated stability test and 6months under the room temperature,with no abnormality in all indicators.Finally,the p H-stat method was used to simulate the digestion process of the preparation in the intestinal tract in vivo.Compared with the fish oil,the emulsion significantly increased the degree and rate of digestion and narrowed the difference in fasted and fed digestion.3.The pharmacokinetic characteristics of FO-ME in vivo.First,a method for the detection of EPA in vivo was established.Taking commercially available soft capsule as a reference,the results of pharmacokinetic experiments in rats show that FO-ME can increase the bioavailability to about 2.53 times that of soft capsule.And the bioavailability increased by 3.32 times in dogs,with no significant difference(p =0.5185)in the fasted and postprandial state.4.The efficacy of FO-ME.To evaluate the effect of FO-ME treatment on the regulating blood lipid in vivo,the hyperlipidemia rats were continuously fed with the Western diet.After a treatment of 30 days,FO-ME significantly reduce triglycerides and total cholesterol concentrations in the plasma compared to the control group at half or one-third the dose of commercially available soft capsule,and with an insignificant effect on other lipoproteins,helping to reduce the adverse effects of large doses of EPA.In summary,the preparation,quality evaluation,oral absorption and efficacy of FO-ME were mainly studied in this thesis.The emulsion promotes the absorption of EPA significantly,reduced the oral dose and fully exerted its hypolipidemic effect.FOME reduced the difference in bioavailability when taken before or after meals,the significant advantages in absorption and utilization of FO-ME could play an important role in clinical application.