The Research On The Extraction Of Phytosterol From Tea Seed And Its Functional Evaluation Of Blood-lipid Lowering

The thesis is focused on the extraction of monomer phytosterol from Tea Seed and give functional evaluation on the phytosterol. The results are as follows.1. There is obvious advantage by using the sub-critical fluid extraction technology to extract and separate heat-sensitive active ingredients. This article select 3 main factors of extraction pressure, extraction temperature, extraction time that affect the subcritical butane fluid extraction, set plant sterol yield as the evaluation target, use response surface methodology to do optimization on the sub-critical fluid extraction technology process conditions. The optimum results show that the extraction temperature is 38.90℃, the extraction time is 120.0 mins and the pressure is 0.70 Mpa, the phytosterol average yield in tea seed oil is 1.86% under the conditions above.2. The extraction of total plant cholesterol from tea seed oil by using the method of the back flow method. By optimizing the concentration of ethanol solution, potassium hydroxide saponification temperature, saponification time, solid-liquid ratio of the four main impact conditions, using plant sterol yield as the evaluation target, it is concluded that the extraction of total phytosterol, solid to liquid ratio of 2.5:1,0.5mol/L KOH ethanol solution, under the temperature of 80℃ saponification 150min, saponification and refluxing extracted the total sterol extraction rate was the highest, reached 2.35%.3. The monomer of the plant was isolated by HSCCC. The optimal proportion of the solvent system water methanol hexane ratio is 1:3:10 for the separation of (3-sitosterol and stigmasterol by shake flask method. In this solvent system, using HSCCC to separate the crude plant sterol, the HPLC detection shows that the main ingredients of 145-147min collection is β-sitosterol, and the content of stigmasterol is few. The main ingredients of the substance collected in the 157-159min period is stigmasterol, but also a small amount of β-sitosterol. The high purity of β-sitosterol and stigmasterol monomers are prepared by HSCCC in this condition.4. A high triglyceride and high cholesterol cell model was established and give functional evaluation on the β-sitosterol and stigmasterol monomers prepared by HSCCC. Results show that using 10μg/mL cholesterol lipid emulsion culture L02 cells for 24 h, the model of high triglyceride and high cholesterol cells is successfully constructed. Giving functional evaluation on the β-sitosterol and stigmasterol monomers by this model. When the concentration of P-sitosterol 50ug/mL and 100ug/mL, and the model group were significantly different, there was no significant difference between the two groups. Oil red O staining results showed β-sitosterol and stigmasterol role in L02 cell model and model group compared with obvious triglyceride lowering effect, the concentration of 50ug/mL β-sitosterol, stigmasterol treatment effect best, reduce fat droplets formation effect is close to positive drug Xuezhikang group, but between the two of intracellular lipid droplets formation of no significant difference.