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Method of using crude extract of olive leaf oleuropein to prepare hydroxytyrosol

A technology of crude oleuropein and hydroxytyrosol, which is applied in the fields of biology and chemistry, can solve the problems of hyperthermia in reaction conditions, expensive enzymes, etc., and achieve the effect of mild method conditions

Active Publication Date: 2015-09-23
INST OF CHEM IND OF FOREST PROD CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzymes used in this method are expensive and the reaction conditions must be thermophilic

Method used

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  • Method of using crude extract of olive leaf oleuropein to prepare hydroxytyrosol
  • Method of using crude extract of olive leaf oleuropein to prepare hydroxytyrosol
  • Method of using crude extract of olive leaf oleuropein to prepare hydroxytyrosol

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] HPLC Analysis and Calculation Method of Oleuropein and Hydroxytyrosol

[0038] Accurately weigh 4.5mg and 5.7mg of oleuropein and hydroxytyrosol reference substances and place them together in a 10mL volumetric flask, dissolve them with methanol and shake well, dilute to volume and seal tightly to obtain concentrations of 0.45mg / mL and 0.57mg / mL The reference substance is mixed with the standard mother solution. Precisely draw 1, 2, 4, 6, 8, 10, 12, 14, and 16 μL of this standard solution respectively, inject samples, repeat the injection 3 times for each sample, and take the HPLC peak areas of oleuropein and hydroxytyrosol respectively average value. Then, with the HPLC peak area as the ordinate, and the mass (μg) of oleuropein and hydroxytyrosol as the abscissa, respectively, the HPLC standard curve was drawn to obtain the equation.

[0039] The liquid conditions are as follows: the chromatographic column is a C18ODS chromatographic column (250×5mm, 5 μm), the colum...

Embodiment 2

[0042] Acid Hydrolysis of Crude Extract of Olive Leaf Oleuropein

[0043] Weigh 1 g of olive leaf extract into a 250 mL round bottom flask, add 50 mL of 0.2 mol / L HCl bath solution, and place in a constant temperature water bath shaker at 80 °C. After hydrolysis for 6 hours, it was neutralized to neutral, filtered through a 0.45 μm organic membrane, and analyzed by HPLC. Each test was carried out 3 times and the average value was calculated. It is calculated that the HT content can be 7.41% after acid hydrolysis. (OE degradation rate=1-enzymatic hydrolyzate OE mass / extract OE mass (38.6%); HT content=enzymatic hydrolyzate HT mass / extract mass.)

Embodiment 3

[0045] Alkaline Hydrolysis of Crude Extract of Oleuropein from Olive Leaves

[0046] Alkaline hydrolysis conditions: Weigh 1g of olive leaf extract into a 250mL round bottom flask, add 50mL of 0.2mol / L NaOH solution, and place in a constant temperature water bath shaker at 80°C. After hydrolysis for 6 hours, it was neutralized to neutral, filtered through a 0.45 μm organic membrane, and analyzed by HPLC. Each test was carried out 3 times and the average value was calculated. It is calculated that the HT content can be 4.09% after alkali hydrolysis. (OE degradation rate=1-enzymatic hydrolyzate OE mass / extract OE mass (38.6%); HT content=enzymatic hydrolyzate HT mass / extract mass.)

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Abstract

The invention discloses a method of using crude extract of olive leaf oleuropein to prepare hydroxytyrosol. Compared with acid hydrolysis and alkali hydrolysis, the enzymolysis process for crude extract of olive leaf oleuropein is mild in method condition, and consumption of acid and alkali catalysts is reduced in the whole process. Hemicellulase is selected from ten enzymes as an enzyme best in oleuropein degrading effect by taking oleuropein degrading rate and hydroxytyrosol content as indexes, optimal process conditions include 60 DEG C of temperature, 5.5 of pH value, 40mg of enzyme amount and 6h of time, obtained HT content is 6.07%, and OE degrading rate is 85.28%.

Description

1. Technical field [0001] The invention relates to the fields of biology and chemistry, in particular to a method for preparing hydroxytyrosol from olive leaf oleuropein crude extract. 2. Background technology [0002] Olive (Olea europaea L.) belongs to Oleaceae (Oleaceae), Oleaceae (Olea) evergreen trees, is a world-renowned woody oil tree species. Olive oil can greatly reduce the incidence of cardiovascular and lung cancer, and its nutritional value and economic value have been recognized by everyone. Biologists further found that the antioxidant active ingredients contained in olive leaves are more prominent than those in olive oil, mainly including polyphenolic compounds such as oleuropein, hydroxytyrosol, flavonoids, lignans, and caffeoylphenylethanol glycosides. . Especially hydroxytyrosol (Hydroxytyrosol, HT; the chemical name is 3,4-dihydroxyphenylethanol), which has a strong ability to scavenge free radicals and exhibits unique biological activities, such as anti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C39/11C07C37/68C07C37/00
CPCC07C37/002C07C37/004C07C37/68C07C39/11
Inventor 王成章原姣姣叶建中陈虹霞周昊陶冉张宇思
Owner INST OF CHEM IND OF FOREST PROD CHINESE ACAD OF FORESTRY
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