Research On Hydroxytyrosol Extraction From Olive Leaves And Olive Wine Brewing

The research object of this thesis is hydroxytyrosol which has important psychologicalactivities such as anticancer and antibacterial and also has a broad market prospect. The rawmaterial is olive leaves which are from Kaijiang county, Dazhou city, Sichuan province.Thorough and systematic researches had done to the extraction and purification of thehydroxytyrosol in olive leaves. This research confirmed the method of measurement,extraction, separation, purification process for the content of hydroxytyrosol and the bestolive wine brewing technology. This study can not only make for olive pruning and recoveryof a large number of full use of the olive leaves, it can also make the health care function ofthe hydroxytyrosol develop well.Efficient analytical methods of functional ingredients in olive leaves were developed.Using methanol–phosphate solution as mobile phase, a method was developed for thedetermination of hydroxytyrosol in olive leaves–high performance liquid chromatographywith diode array detector (HPLC–DAD) analysis method. This method is rapid, accurate andsimple for raw material processing. Reversed phase high performance liquid chromatography(RP–HPLC) was established to determination of oleuropein in olive leaves, it can be used tomeasure oleuropein content in the different raw material source. And analysis of flavonoids inolive leaves by ultraviolet spectrophotometer（UV） method was eatablished, the method wasavoided to the use of toxic reagents such as acetonitrile. So they can lay the foundation forlater study.The extraction process of the hydroxytyrosol in olive leaves was also studied. Ashydroxytyrosol in olive leaves mainly exists in the form of oleuropein, the experiment usedthe β–glycosidase enzymes to hydrolysis oleuropein as the best condition of enzymaticextraction of hydroxytyrosol. Compared with the method of the oleuropein inorganic acidhydrolysis, enzymatic extraction of hydroxytyrosol content was3.71mg/g, it was a bit of lessto phosphoric acid hydrolysis extraction hydroxytyrosol, but it did not need ultrasound–assisted, mild reaction conditions, less impurity of dissolution and better keep the functionof hydroxytyrosol.Macroporous adsorption resin (MAR) purification of hydroxytyrosol in enzymolysisliquid was studied. The inactivated enzyme with methanol and removing the chlorophyll andvolatile impurities, by macroporous adsorption resins with different polarity to concentrate onthe static adsorption stripping, hydroxytyrosol adsorption ratio, stripping ratio as examiningindex, determine the best adsorption resin was DA–201macroporous adsorption resin.Through the dynamic adsorption–stripping experiment, the best purification processconditions were determined. Accordance with the above optimum experimental conditions, 1.00g olive leaveas can obtain3.85mg hydroxytyrosol solid total saponin, the purity ofhydroxytyrosol could reach to87.01%. The purity of hydroxytyrosol was improved, it couldlay foundation for industrialization pruducte of hydroxytyrosol.The wine brewing process of olive leaves was studied. There is no report of wine makingby olive leaves. In olive wine brewing process, the brewing method which was first gauze andthen fermentation were determined. The fermentation time was6d. Then done experiment onthe main fermentation temperature, initial pH value, initial sugar, yeast and SO2content todetermine the biggest impact of the four factors. Through orthogonal test, the best brewingprocess was that the initial sugar yeast addition amount was4%, the initial sugar fermentationwas10%, thefermentation temperature was25℃, the initial pH value was3.6. Under thiscondition of brewing,the wine had a special flavor and the wine was good, impurities toprecipitate, pure to taste.This paper put enzymatic technology and macroporous adsorption resin technologytogether to extract hydroxytyrosol in olive leaves efficiently. At the same time brewing olivewine by olive leaves. This study does not only increase thevariety of wine, but alsorealizes thecomprehensive utilization of raw materials, this study opens up new economic value-addedpoints for olive industry.