阅读说明：本技术 一种由万寿菊油树脂分离纯化叶黄素的方法 (Method for separating and purifying lutein from marigold oleoresin ) 是由 刘温来 李勇 殷东东 赵辰光 田芸 于 2019-09-29 设计创作，主要内容包括：一种由万寿菊油树脂分离纯化叶黄素的方法,属于天然有效成分提取技术领域。其特征在于,制备步骤为：将万寿菊油树脂溶解于乙酸乙酯,然后利用正己烷-乙酸乙酯混合液进行解析,解析得到叶黄素酯解析液进行浓缩得到叶黄素酯浸膏；将叶黄素酯浸膏与醇、碱液和抗氧化剂在通入二氧化碳的反应釜内进行水解反应；将叶黄素皂化物用乙醇稀释、静置、过滤得到粗品叶黄素晶体；润湿、研磨破碎、溶解、过滤、干燥后得到叶黄素晶体。本产品内部几乎不含有玉米黄质等其它类胡萝卜素,是纯天然产物,同时便于规模化生产。(A method for separating and purifying lutein from marigold oleoresin belongs to the technical field of natural active ingredient extraction. The preparation method is characterized by comprising the following preparation steps: dissolving marigold oleoresin in ethyl acetate, resolving by using a normal hexane-ethyl acetate mixed solution to obtain a lutein ester resolved solution, and concentrating to obtain a lutein ester extract; carrying out hydrolysis reaction on the lutein ester extract, alcohol, alkali liquor and an antioxidant in a reaction kettle filled with carbon dioxide; diluting the lutein saponified product with ethanol, standing, and filtering to obtain crude lutein crystal; wetting, grinding, crushing, dissolving, filtering and drying to obtain lutein crystals. The product contains almost no other carotenoids such as zeaxanthin, is a pure natural product, and is convenient for large-scale production.)

1. A method for separating and purifying lutein from marigold oleoresin is characterized by comprising the following preparation steps:

1) dissolving marigold oleoresin in ethyl acetate; then transferring the mixture into a silica gel analytical column, and analyzing by using a normal hexane-ethyl acetate mixed solution as an analytical solution to obtain a lutein ester analytical solution;

2) concentrating the lutein ester analysis solution to obtain lutein ester extract, wherein the residual volatile matters in the lutein ester extract are controlled to be 5% ~ 10% by mass;

3) carrying out hydrolysis reaction on the lutein ester extract, alcohol, alkali liquor and antioxidant in a reaction kettle filled with carbon dioxide, wherein the temperature of the hydrolysis reaction is 50 ℃ and ~ 65 ℃, and the time of the hydrolysis reaction is 2h and ~ 8h, so as to obtain a lutein saponified product;

4) adding an acid into the lutein saponified product to adjust the pH value to 5 ~ 7, adding an ethanol water solution with the mass percentage concentration of 5% and ~ 25%, wherein the mass of the added ethanol water solution is 1 ~ 10 times of that of the lutein ester extract, standing for 6h and ~ 15h, and filtering to obtain a crude lutein crystal;

5) adding deionized water into the crude xanthophyll crystal for wetting, grinding and crushing the crude xanthophyll crystal by a grinder at the temperature of 70 ℃ and ~ 80 ℃, dissolving, filtering and drying the crude xanthophyll crystal to obtain the xanthophyll crystal;

the volume ratio of n-hexane to ethyl acetate in the n-hexane-ethyl acetate mixed solution in the step 1) is 5 ~ 10: 1;

the mass ratio of the marigold oleoresin to the n-hexane-ethyl acetate mixed solution in the step 1) is 1: 2 ~ 8.

2. The method for separating and purifying lutein from marigold oleoresin according to claim 1, wherein the mass ratio of marigold oleoresin to ethyl acetate in step 1) is 1: 4 ~ 10.

3. The method for separating and purifying lutein from marigold oleoresin according to claim 1, wherein the volume ratio of n-hexane to ethyl acetate in the n-hexane-ethyl acetate mixed solution in step 1) is 6 ~ 7: 1.

4. The method for separating and purifying lutein from marigold oleoresin according to claim 1, wherein the silica gel type of step 1) is 60 Å column chromatography silica gel.

5. A process for the isolation of purified lutein from marigold oleoresin according to claim 1, characterized by: the alcohol in the step 3) is propylene glycol, glycerol or butanol.

6. The method for separating and purifying lutein from marigold oleoresin according to claim 1, wherein the alkaline solution in step 3) is 30% ~ 60% by mass of potassium hydroxide or/and sodium hydroxide aqueous solution.

7. A process for the isolation of purified lutein from marigold oleoresin according to claim 1, characterized by: the acid in the step 4) is one or the combination of more than two of hydrochloric acid, phosphoric acid and oxalic acid.

8. A process for the isolation of purified lutein from marigold oleoresin according to claim 1, characterized by: the antioxidant in the step 3) is tert-butyl hydroquinone or vitamin E.

9. The method for separating and purifying lutein from marigold oleoresin according to claim 1, wherein the lutein ester extract in step 4) is added with alcohol, alkali solution and antioxidant in a mass ratio of 1: 0.1 ~ 1: 0.3 ~ 1: 0.001 ~ 0.7.7.

10. The method for separating and purifying lutein from marigold oleoresin according to claim 1, wherein the mass ratio of the crude lutein crystals in step 5) to the deionized water is 1: 2 ~ 5.

Technical Field

A method for separating and purifying lutein from marigold oleoresin belongs to the technical field of natural active ingredient extraction.

Background

The marigold oleoresin is a yellow or brown solid or viscous liquid, and is prepared by drying flower of marigold of Compositae (usually in dark place), pulverizing, extracting with organic solvent (usually n-hexane), and volatilizing to remove solvent to obtain extract containing carotenoid ester.

The lutein is carotenoid, has good oxidation resistance, has unique functions in the aspects of relieving visual fatigue, preventing age-related macular degeneration, protecting skin, enhancing immunity, reducing the incidence rate of cardiovascular diseases and the like. The xanthophyll has health promoting and coloring effects. One source of lutein is extracted from marigold flowers.

The applicant finds the following problems in research, namely, the lutein obtained by the existing lutein extraction method is low in purity and contains a certain proportion of zeaxanthin, the existing extraction method is to directly saponify marigold oleoresin to obtain the lutein which contains more zeaxanthin, the proportion of the zeaxanthin is 3% ~ 9%, the purity of the lutein is insufficient and the zeaxanthin is wasted, the zeaxanthin is an isomer of the lutein and needs to be removed by a separation process, and the increase of the separation process causes the lutein loss.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: overcomes the defects of the prior art, and provides a method for separating and purifying lutein from marigold oleoresin, which has high purity and is convenient for large-scale production.

The technical scheme adopted by the invention for solving the technical problems is as follows: the method for separating and purifying lutein from marigold oleoresin is characterized by comprising the following preparation steps:

1) dissolving marigold oleoresin in ethyl acetate; then transferring the mixture into a silica gel analytical column, and analyzing by using a normal hexane-ethyl acetate mixed solution as an analytical solution to obtain a lutein ester analytical solution;

2) concentrating the lutein ester analysis solution to obtain lutein ester extract, wherein the residual volatile matters in the lutein ester extract are controlled at 5% ~ 10% by mass (abbreviated as% w/w);

3) carrying out hydrolysis reaction on the lutein ester extract, alcohol, alkali liquor and antioxidant in a reaction kettle filled with carbon dioxide, wherein the temperature of the hydrolysis reaction is 50 ℃ and ~ 65 ℃, and the time of the hydrolysis reaction is 2h and ~ 8h, so as to obtain a lutein saponified product;

4) adding an acid into the lutein saponified product to adjust the pH value to 5 ~ 7, adding an ethanol water solution with the mass percentage concentration of 5% and ~ 25%, wherein the mass of the added ethanol water solution is 1 ~ 10 times of that of the lutein ester extract, standing for 6h and ~ 15h, and filtering to obtain a crude lutein crystal;

5) adding deionized water into the crude xanthophyll crystal for wetting, grinding and crushing the crude xanthophyll crystal by a grinder at the temperature of 70 ℃ and ~ 80 ℃, dissolving, filtering and drying the crude xanthophyll crystal to obtain the xanthophyll crystal;

the volume ratio of n-hexane to ethyl acetate in the n-hexane-ethyl acetate mixed solution in the step 1) is 5 ~ 10: 1;

the mass ratio of the marigold oleoresin to the n-hexane-ethyl acetate mixed solution in the step 1) is 1: 2 ~ 8.

The mass ratio (abbreviated as w/w) of the marigold oleoresin to the ethyl acetate in the step 1) is 1: 4 ~ 10.

The volume ratio (abbreviated as V/V) of the n-hexane to the ethyl acetate in the n-hexane-ethyl acetate mixed solution in the step 1) is 6 ~ 7: 1.

The silica gel type in the step 1) is 60 Å type chromatographic silica gel, and preferably, the mesh number of the silica gel is 100 ~ 600 meshes.

The alcohol in the step 3) is propylene glycol, glycerol or butanol. The preferred alcohol has a better hydrolysis effect, and the monomers are more easily separated.

The alkali liquor in the step 3) is potassium hydroxide or/and sodium hydroxide aqueous solution with the mass concentration of 30% ~ 60%.

The acid in the step 4) is one or the combination of more than two of hydrochloric acid, phosphoric acid and oxalic acid.

The antioxidant in the step 3) is tert-butyl hydroquinone or vitamin E.

Adding the lutein ester extract in the step 4) with alcohol, alkali liquor and antioxidant according to the mass ratio of 1: 0.1 ~ 1: 0.3 ~ 1: 0.001 ~ 0.7.7.

The mass ratio of the crude lutein crystals in the step 5) to the deionized water is 1: 2 ~ 5, the grinding effect is better under the preferred wetting effect, and the dissolving of the higher fatty acid salt impurities can be facilitated.

Preferably, the mass ratio of the marigold oleoresin to the ethyl acetate in the step 1) is 1: 6 ~ 8, the mass ratio of the marigold oleoresin to the n-hexane-ethyl acetate mixed solution in the step 1) is 1: 4 ~ 6, the preferred mass ratio of the marigold oleoresin to the ethyl acetate can ensure that the marigold oleoresin is rapidly dissolved, then the marigold oleoresin is added into a silica gel analytical column in the form of a solution, and then the n-hexane-ethyl acetate analytical solution is added according to the mass ratio, while the n-hexane-ethyl acetate analytical solution with a specific volume ratio can help to rapidly complete the well-defined analysis and effectively separate lutein from other carotenoids, and the obtained lutein ester analytical solution and zeaxanthin ester analytical solution are ensured to have higher respective purities, so that the purity of the product is higher.

The specific operation of the step 1) is that marigold oleoresin is dissolved in ethyl acetate, then the marigold oleoresin is transferred into a silica gel analytical column, the adsorption flow rate is 1.5 ~ 2.5.5 BV/h, the temperature is 50 ℃, deionized water is used for washing until the marigold oleoresin is colorless after adsorption is finished, ethyl acetate-ethanol analytical solution is added for analysis, the analytical flow rate is 0.5 ~ 1.5.5 BV/h, n-hexane-ethyl acetate mixed solution is used as the analytical solution for analysis, and lutein ester analytical solution is obtained through analysis.

Preferably, the solvent in the lutein ester analysis solution obtained in the step 2) is condensed and recovered when being concentrated, and the lutein ester analysis solution is returned to the step 1) to be prepared again according to the volume ratio for use, so that the production cost can be reduced.

The grinding step of step 5) helps to promote dissolution of the higher fatty acid salts in the crude crystals and then the higher fatty acid salts dissolved in water are removed by filtration to increase the purity of the lutein the preferred grinding conditions are grinding at 70 ℃ ~ 80 ℃ for 10 ~ 20 minutes the equipment used for grinding in step 5) is a grinder, preferably a colloid mill, a high shear crusher or a rod nitre grinder, the filtration in step 5) is filtered using a metal rod microfilter the grinder in step 5) is a colloid mill, a high shear crusher or a rod nitre grinder.

Compared with the prior art, the method for separating and purifying lutein from marigold oleoresin has the beneficial effects that:

1. the lutein obtained by the method is high in purity, the purity can reach more than 95% and can reach 96.69% to the maximum, the method firstly separates various carotene esters in the marigold oleoresin, the lutein ester, the zeaxanthin ester and other components in the marigold oleoresin are directly adsorbed and resolved by silica gel to obtain the lutein ester and the zeaxanthin ester respectively, the lutein ester is hydrolyzed, crystallized, nano-ground, alcohol-washed and the like to obtain the high-purity lutein, the volume ratio (V/V) of n-hexane and ethyl acetate in n-hexane-ethyl acetate mixed solution is 5 ~ 10: 1, the volume ratio can accelerate the resolution rate of the zeaxanthin and the lutein, the separation trend among monomers is enlarged, and the purity of the obtained product is higher.

2. The method can remove higher fatty acid salt impurities, thereby improving the purity of lutein. The applicant finds in research that the obtained lutein crystal is doped with partial higher fatty acid salt impurities, and the impurities are difficult to remove in the crystal; in order to solve the problem, the applicant designs that the crude crystals are wetted by deionized water in the step 5), ground, and then filtered to remove the impurities of higher fatty acid salts.

3. The lutein obtained by the method has high yield which can reach 77.9 ~ 80.5.5 percent, is convenient for large-scale production, hardly contains other carotenoids such as zeaxanthin and the like in the lutein obtained by the method, is a pure natural product, and is convenient for large-scale production.

Detailed Description

The invention is further illustrated by the following specific examples, of which example 1 is the best mode.