阅读说明：本技术 一种由万寿菊油树脂分离纯化玉米黄质的方法 (Method for separating and purifying zeaxanthin from marigold oleoresin ) 是由 刘温来 李勇 杜明花 殷东东 赵辰光 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玉米黄质的方法,属于天然有效成分提取技术领域。其特征在于,制备步骤为：将万寿菊油树脂与乙酸乙酯-乙醇溶解液按质量比1：4~10混合、溶解；然后转入到装填有大孔吸附树脂的解析柱内,利用体积比5~10：1乙酸乙酯-乙醇解析液进行解析；解析得到含玉米黄质酯解析液；然后将含玉米黄质酯解析液经2)浓缩、3)皂化、4)析晶、5)研磨精制,最终获得高纯度玉米黄质。本发明的一种由万寿菊油树脂分离纯化玉米黄质的方法所得玉米黄质为天然玉米黄质,纯度能达到95%以上,且收率较高,便于规模化生产。(a method for separating and purifying zeaxanthin by marigold oleoresin belongs to the technical field of natural active ingredient extraction and is characterized by comprising the steps of mixing and dissolving the marigold oleoresin and ethyl acetate ~ ethanol dissolving liquid according to a mass ratio of 1: 4 ~ 10, transferring the mixture into an analytical column filled with macroporous adsorption resin, analyzing the mixture by using ethyl acetate ~ ethanol analytical liquid with a volume ratio of 5 ~ 10:1 to obtain zeaxanthin ester ~ containing analytical liquid, and then carrying out 2) concentration, 3) saponification, 4) crystallization and 5) grinding and refining on the zeaxanthin ester ~ containing analytical liquid to finally obtain the high ~ purity zeaxanthin.)

1. a method for separating and purifying zeaxanthin by marigold oleoresin is characterized by comprising the following preparation steps:

1) mixing and dissolving marigold oleoresin and ethyl acetate ~ ethanol dissolving solution according to the mass ratio of 1: 4 ~ 10, then transferring into an analytical column filled with macroporous adsorption resin, and analyzing by using ethyl acetate ~ ethanol analytical solution to obtain analytical solution containing zeaxanthin ester;

2) concentrating the analytic solution containing zeaxanthin ester to obtain zeaxanthin ester extract, wherein the residual volatile matters in the zeaxanthin ester extract are controlled to be 5-10% by mass;

3) carrying out hydrolysis reaction on the zeaxanthin ester extract, alcohol and alkali liquor in a reaction kettle filled with carbon dioxide, wherein the temperature of the hydrolysis reaction is 70 h ~ 90 ℃, and the time of the hydrolysis reaction is 2h ~ 8h, so as to obtain a zeaxanthin saponificate;

4) adding an acid into the zeaxanthin saponificate to adjust the pH value to 7 ~ 9, adding 30 ~ 50% of ethanol solution by mass percent, wherein the mass ratio of the ethanol solution to the zeaxanthin ester extract in the step 3) is 1 ~ 10:1, standing for 1 ~ 5h, and filtering to obtain a crude zeaxanthin crystal;

5) adding deionized water into the crude zeaxanthin crystals for wetting, grinding the crude zeaxanthin crystals at 70-80 ℃, dissolving, filtering and drying to obtain the zeaxanthin crystals;

the mass ratio of the marigold oleoresin to the ethyl acetate ~ ethanol resolution solution in the step 1) is 1: 2 ~ 10;

the ethyl acetate ~ ethanol solution in the step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 5 ~ 10: 1;

the ethyl acetate ~ ethanol analysis solution in the step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 5 ~ 10: 1.

2. the method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the mass ratio of marigold oleoresin to ethyl acetate ~ ethanol solution in step 1) is 1: 7 ~ 9.

3. The method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the method comprises the following steps: the macroporous adsorption resin in the step 1) is AB-8 type macroporous adsorption resin.

4. the method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the alcohol in step 3) is propylene glycol, glycerol or butanol, and the mass ratio of the alcohol to the zeaxanthin ester extract is 0.1 ~ 1: 1.

5. the method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, which is characterized in that the alkali solution in step 3) is 30 ~ 60% by mass of potassium hydroxide or/and sodium hydroxide aqueous solution, and the mass ratio of the alkali solution to the zeaxanthin ester extract is 0.3 ~ 1: 1.

6. The method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the method comprises the following steps: the acid in the step 4) is one or the combination of more than two of hydrochloric acid, phosphoric acid, oxalic acid, acetic acid and citric acid.

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

8. the method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the mass ratio of the marigold oleoresin in step 1) to the ethyl acetate ~ ethanol solution is 1: 6 ~ 8.

9. the method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the ethyl acetate ~ ethanol solution in step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 6 ~ 7: 1.

10. the method for separating and purifying zeaxanthin from marigold oleoresin according to claim 1, wherein the ethyl acetate ~ ethanol analysis solution in step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 6 ~ 7: 1.

Technical Field

A method for separating and purifying zeaxanthin by marigold oleoresin belongs to the technical field of natural effective component 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.

zeaxanthin, molecular formula C40H56O2, molecular weight 566.88, belongs to isoprenoids, is an oxygen-containing carotenoid, and belongs to the genus of xanthophylls isomer. The corn yellow can be used as health promotion additive and colorant. The corn yellow can resist oxidation, relieve asthenopia, and prevent senile macular degeneration. The macular pigment in the adult retina macula is mainly zeaxanthin, and the ratio of zeaxanthin to lutein is 2.4: 1. the zeaxanthin source comprises natural extraction, lutein conversion and artificial synthesis. In the prior art, lutein is obtained by natural extraction of marigold, and the lutein conversion is mainly to obtain zeaxanthin by epimerization and conversion of lutein at a higher temperature. Zeaxanthin obtained by conversion in the prior art often contains part of unconverted lutein, is synthesized from isoprenoid or carotenoid, has complex process and is difficult to control large-scale production.

the applicant found in the study that: firstly, compared with the zeaxanthin obtained by lutein conversion and artificial synthesis, the natural zeaxanthin is easier to absorb, has better effect and smaller side effect, but the natural zeaxanthin obtained by the existing method has low purity and high selling price; with the continuous development of toxicology and analytical techniques, research shows that although the purity of the converted and artificially synthesized zeaxanthin is relatively high and the production cost is low, the product is easy to incorporate a small amount of toxic chemical substances, which can cause chromosome mutation and carcinogenesis, so that the naturally extracted zeaxanthin occupies an active position in the market with the continuous improvement of understanding. In the field of extracts, the purity of a single natural component is in direct proportion to the price, and after the purity of the single component reaches more than 80%, the selling price of the extract is doubled along with the improvement of the purity of the single component of the extract; the selling price of the zeaxanthin with the purity of 80% w/w in the current market is about 8 thousand yuan/kg, while the selling price of the zeaxanthin with the purity of 90% w/w is about 1.1 ten thousand/kg, and the selling price of the zeaxanthin with the purity of more than 95% w/w is about 1.3 ten thousand/kg; in order to obtain high-purity natural zeaxanthin, the existing methods are mostly supercritical extraction and subcritical fluid extraction, the method has extremely high cost and low yield, cannot be applied to large-scale industrial production, and has relatively low cost, unsatisfactory purity and low yield. Secondly, because of the problem of impurities, in the prior art, high-purity zeaxanthin can be obtained only by separating and removing impurities for many times, and the purity of the natural zeaxanthin is improved by adopting a supercritical or subcritical extraction and gradient elution mode and multiple separation processes. In addition, each separation process must have zeaxanthin loss, which leads to undesirable yield, waste of raw materials and increase of cost.

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 zeaxanthin 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 the zeaxanthin by the marigold oleoresin is characterized by comprising the following preparation steps:

1) mixing and dissolving marigold oleoresin and ethyl acetate ~ ethanol dissolving solution according to the mass ratio of 1: 4 ~ 10, then transferring into an analytical column filled with macroporous adsorption resin, and analyzing by using ethyl acetate ~ ethanol analytical solution to obtain analytical solution containing zeaxanthin ester;

2) concentrating the analytic solution containing zeaxanthin ester to obtain zeaxanthin ester extract, wherein the residual volatile matters in the zeaxanthin ester extract are controlled to be 5-10% by mass;

3) carrying out hydrolysis reaction on the zeaxanthin ester extract, alcohol and alkali liquor in a reaction kettle filled with carbon dioxide, wherein the temperature of the hydrolysis reaction is 70 h ~ 90 ℃, and the time of the hydrolysis reaction is 2h ~ 8h, so as to obtain a zeaxanthin saponificate;

4) adding an acid into the zeaxanthin saponificate to adjust the pH value to 7 ~ 9, adding 30 ~ 50% of ethanol solution by mass percent, wherein the mass ratio of the ethanol solution to the zeaxanthin ester extract in the step 3) is 1 ~ 10:1, standing for 1 ~ 5h, and filtering to obtain a crude zeaxanthin crystal;

5) adding deionized water into the crude zeaxanthin crystals for wetting, grinding the crude zeaxanthin crystals at 70-80 ℃, dissolving, filtering and drying to obtain the zeaxanthin crystals;

the mass ratio of the marigold oleoresin to the ethyl acetate ~ ethanol resolution solution in the step 1) is 1: 2 ~ 10;

the ethyl acetate ~ ethanol solution in the step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 5 ~ 10: 1;

the ethyl acetate ~ ethanol analysis solution in the step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 5 ~ 10: 1.

the mass ratio of the marigold oleoresin to the ethyl acetate ~ ethanol solution in the step 1) is 1: 7 ~ 9, and the marigold oleoresin can be rapidly dissolved under the mass ratio of the marigold oleoresin to the ethyl acetate ~ ethanol solution.

the macroporous adsorption resin in the step 1) is AB ~ 8 type macroporous adsorption resin which can promote separation and improve yield, and preferably, the mass ratio of marigold oleoresin to macroporous adsorption resin is 5 ~ 10: 1.

the alcohol in the step 3) is propylene glycol, glycerol or butanol, the mass ratio of the alcohol to the zeaxanthin ester extract is 0.1 ~ 1: 1, the preferred alcohol has a better hydrolysis effect, and each monomer is easier to analyze and separate.

the alkali liquor in the step 3) is 30% ~ 60% of potassium hydroxide or/and sodium hydroxide aqueous solution in percentage by mass (abbreviated as% w/w), the mass ratio of the alkali liquor to the zeaxanthin ester extract is 0.3% ~ 1: 1, and under the preferable alkali adding mode, the materials can be mixed more quickly and uniformly, and the hydrolysis effect is better.

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

the mass ratio of the crude zeaxanthin crystals in the step 5) to the deionized water is 1: 2 ~ 5, and the grinding effect is better under the preferred wetting effect.

the mass ratio (abbreviated as w/w) of the marigold oleoresin to the ethyl acetate ~ ethanol analysis solution in the step 1) is 1: 6 ~ 8.

the ethyl acetate ~ ethanol solution in the step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio (abbreviated as V/V) of 6 ~ 7: 1.

the ethyl acetate ~ ethanol analysis solution in the step 1) is a mixed solution of ethyl acetate and ethanol in a volume ratio of 6 ~ 7: 1, so that the analysis rate can be increased, the separation trend between zeaxanthin ester and other carotenoids can be expanded, the analysis with clear boundaries can be completed quickly, the purity of the obtained zeaxanthin ester analysis solution is high, and the purity of the obtained natural zeaxanthin is high.

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

the specific operation of the step 5) is that deionized water is added into the crude zeaxanthin crystals for wetting, the crude zeaxanthin crystals are ground by a grinding machine at 70 ~ 80 ℃ for 5 ~ 30 minutes, then the obtained solid is filtered, and the zeaxanthin crystals are obtained after vacuum drying, the grinding step at 70 ~ 80 ℃ for 5 ~ 30 minutes in the step 5) is helpful for promoting the dissolution of higher fatty acid salts, and then the higher fatty acid salts dissolved in water are removed by filtration, so that the purity of the zeaxanthin is improved.

the specific operation of the step 1) is that marigold oleoresin and ethyl acetate ~ ethanol dissolving solution are mixed and dissolved according to the mass ratio of 1: 4 ~ 10, then the marigold oleoresin and the ethyl acetate ~ ethanol dissolving solution are transferred into an analytical column filled with macroporous adsorption resin, the adsorption flow rate is 1.5 ~ 2.5BV/h, the temperature is 50 ℃, deionized water is used for washing until the adsorption is finished, ethyl acetate ~ ethanol analytical solution is added for analysis, the analysis flow rate is 1 ~ 1.5BV/h, and zeaxanthin analytical solution is collected.

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

1. the method comprises the steps of separating various carotene esters in marigold oleoresin, directly adsorbing and analyzing and separating lutein esters, zeaxanthin esters and other components in the marigold oleoresin by macroporous adsorption resin to obtain lutein crystals with purity of more than 95%, and the like, wherein the lutein esters are subjected to hydrolysis, crystallization, nano grinding, alcohol washing and the like, and the zeaxanthin crystals with purity of more than 95% are obtained.

2. the zeaxanthin yield is high, the AB ~ 8 type macroporous adsorption resin can effectively separate zeaxanthin ester, the zeaxanthin ester yield is improved, the separation effect of ethyl acetate ~ ethanol analysis liquid with the volume ratio of 5 ~ 10:1 on zeaxanthin is high, the loss of the zeaxanthin in the separation process is reduced, high ~ purity zeaxanthin ester is obtained by adopting the macroporous adsorption resin for separation in one process, other carotenoid impurities do not need to be removed by multiple separation, the loss of the obtained zeaxanthin in the process is less, and the zeaxanthin yield can reach 46.37 ~ 80.62%.

Detailed Description

The invention is further illustrated by the following specific examples, of which example 2 is the best mode.