阅读说明：本技术 一种由辣椒红分离纯化玉米黄质的方法 (Method for separating and purifying zeaxanthin from capsanthin ) 是由 刘温来 李勇 田芸 赵辰光 于正毅 于 2019-09-29 设计创作，主要内容包括：一种由辣椒红分离纯化玉米黄质的方法,属于类胡萝卜素制备技术领域。其特征在于,制备步骤为：辣椒红、醇、碱、抗氧化剂进行皂化反应；皂化物调节pH在7~8之间,用正己烷-丙酮-乙酸乙酯混合溶液进行稀释并进行解析,收集玉米黄质解析液；回收溶剂,降温析晶；过滤、洗涤、干燥得到玉米黄质晶体。本方法直接由辣椒红为原料,经过皂化处理得到玉米黄质单体,继而分离含量90%以上的玉米黄质,本产品便于规模化生产。(A process for separating and purifying zeaxanthin from capsanthin includes such steps as saponifying reaction of capsanthin, alcohol, alkali and antioxidant, regulating pH value to 7 ~ 8, diluting with n-hexane-acetone-ethyl acetate solution, analyzing, collecting the analyzed liquid, recovering solvent, cooling, crystallizing, filtering, washing and drying to obtain zeaxanthin crystal, saponifying to obtain zeaxanthin monomer, and separating zeaxanthin whose content is more than 90%.)

1. A method for separating and purifying zeaxanthin from capsanthin is characterized by comprising the following preparation steps:

1) carrying out saponification reaction on capsanthin, alcohol, alkali and an antioxidant under the condition of introducing nitrogen, and saponifying for 8h ~ 12h at 40 ℃ and ~ 60 ℃ to obtain a saponified substance, wherein the mass ratio of the capsanthin, the alcohol, the alkali and the antioxidant is 1: 0.1 ~ 0.5.5: 0.5 ~ 1: 0.01 ~ 0.1.1;

2) adding acid into the saponified substance, adjusting pH to 7 ~ 8, and diluting with n-hexane-acetone-ethyl acetate mixed solution to obtain saponified substance diluent, wherein the mass ratio of capsanthin to n-hexane-acetone-ethyl acetate mixed solution is 1: 2 ~ 5;

3) introducing the saponified substance diluent into an analytical column filled with macroporous adsorption resin, analyzing with n-hexane-acetone-ethyl acetate mixed solution, and collecting zeaxanthin analytical solution;

4) recovering solvent from zeaxanthin solution until the mass concentration of zeaxanthin is 20% ~ 60, cooling to 20 deg.C ~ 40 deg.C, maintaining the temperature for 4h ~ 6h, crystallizing, filtering to obtain crude zeaxanthin crystal, and washing with ethanol;

5) vacuum drying the zeaxanthin crystals to obtain zeaxanthin crystals;

wherein, the n-hexane-acetone-ethyl acetate mixed solution in the step 2) and the step 3) is a mixed solution prepared by n-hexane, acetone and ethyl acetate according to the volume ratio of 6 ~ 10: 0.7 ~ 2.4.4: 0.1 ~ 0.26.26.

2. The method for separating and purifying zeaxanthin from capsanthin according to claim 1, wherein the mass ratio of capsanthin, alcohol, alkali and antioxidant in step 1) is 1: 0.2 ~ 0.3.3: 0.7 ~ 0.8.8: 0.04 ~ 0.06.06.

3. The method for separating and purifying zeaxanthin from capsanthin according to claim 1 or 2, which comprises the following steps: the alcohol in the step 1) is ethanol, methanol or propylene glycol.

4. The method for separating and purifying zeaxanthin from capsanthin according to claim 1 or 2, which comprises the following steps: the alkali in the step 1) is potassium hydroxide or/and sodium hydroxide.

5. The method for separating and purifying zeaxanthin from capsanthin according to claim 4, wherein the alkali is added in the form of 30% ~ 60% by mass of aqueous solution of potassium hydroxide or/and sodium hydroxide.

6. The method for separating and purifying zeaxanthin from capsanthin according to claim 1 or 2, which comprises the following steps: the antioxidant in the step 1) is vitamin E or tert-butyl hydroquinone.

7. The method for separating and purifying zeaxanthin from capsanthin according to claim 1, which comprises the following steps: the acid in the step 2) is one or more of hydrochloric acid, phosphoric acid, oxalic acid, acetic acid and citric acid.

8. The method for separating and purifying zeaxanthin from capsanthin according to claim 1, wherein the n-hexane-acetone-ethyl acetate mixed solution in the steps 2) and 3) is a mixed solution prepared by n-hexane, acetone and ethyl acetate according to a volume ratio of 8 ~ 9: 1.3 ~ 1.7.7: 0.13 ~ 0.17.17.

9. The method for separating and purifying zeaxanthin from capsanthin according to claim 1, which comprises the following steps: the resin in the step 3) is one or more of XR15C, XR919LZ, XR921H and XR 910S.

Technical Field

A method for separating and purifying zeaxanthin from capsanthin belongs to the technical field of carotenoid preparation.

Background

Capsanthin, a capsanthin pigment, is a tetraterpenoid orange red pigment present in mature red pepper fruits, and belongs to carotenoid pigments, wherein red components with higher polarity are capsorubin and capsorubin accounting for 50% ~ 60% of the total amount, and yellow components with lower polarity are zeaxanthin and carotene as main components.

Zeaxanthin (Zeaxanthin, 3, 3' -dihydroxy-beta-carotene), also known as Zeaxanthin, has a molecular formula of C40H56O2 and a molecular weight of 566.88, belongs to the isoprenoids, is an oxygen-containing carotenoid, and is an isomer of lutein. Has good oxidation resistance, has unique functions in the aspects of relieving asthenopia, preventing age-related macular degeneration, protecting skin, enhancing immunity, reducing the incidence of cardiovascular diseases and the like. The macular pigment in the adult retina macula is mainly zeaxanthin, and the ratio of zeaxanthin to lutein is 2.4: 1. zeaxanthin has health promoting and coloring effects. The zeaxanthin source comprises natural extraction, lutein conversion and artificial synthesis. In the prior art, lutein is obtained by naturally extracting marigold, and the lutein is mainly converted into zeaxanthin by epimerization and conversion of the lutein at a higher temperature.

The applicant found in the study that: firstly, a method for efficiently extracting natural zeaxanthin is urgently needed at present; with the continuous development of toxicology and analytical techniques, research shows that although the purity of the zeaxanthin synthesized by a chemical method 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 zeaxanthin extracted from the natural source can occupy the active position in the market with the continuous improvement of understanding. Secondly, the natural zeaxanthin obtained by the existing separation method has low purity and high selling price; in the field of extracts, the purity is in direct proportion to the price, and after the purity of a 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; 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. Thirdly, due to the problem of impurities, in the prior art, high-purity zeaxanthin can be obtained only by separating and removing impurities for many times, and each separation process must have the loss of zeaxanthin, which causes the undesirable yield, the waste of raw materials and the 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 capsanthin by using natural components.

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

1) carrying out saponification reaction on capsanthin, alcohol, alkali and an antioxidant under the condition of introducing nitrogen, and saponifying for 8h ~ 12h at 40 ℃ and ~ 60 ℃ to obtain a saponified substance, wherein the mass ratio of the capsanthin, the alcohol, the alkali and the antioxidant is 1: 0.1 ~ 0.5.5: 0.5 ~ 1: 0.01 ~ 0.1.1;

2) adding acid into the saponified substance, adjusting pH to 7 ~ 8, and diluting with n-hexane-acetone-ethyl acetate mixed solution to obtain saponified substance diluent, wherein the mass ratio of capsanthin to the mixed solution is 1: 2 ~ 5;

3) introducing the saponified substance diluent into an analytical column filled with macroporous adsorption resin, analyzing with n-hexane-acetone-ethyl acetate mixed solution, and collecting zeaxanthin analytical solution;

4) recovering solvent from zeaxanthin solution until the mass concentration of zeaxanthin is 20% ~ 60, cooling to 20 deg.C ~ 40 deg.C, maintaining the temperature for 4h ~ 6h, crystallizing, filtering to obtain crude zeaxanthin crystal, and washing with ethanol;

5) vacuum drying the zeaxanthin crystals to obtain zeaxanthin crystals;

wherein, the n-hexane-acetone-ethyl acetate mixed solution in the step 2) and the step 3) is a mixed solution prepared by n-hexane, acetone and ethyl acetate according to a volume ratio (abbreviated as V/V) of 6 ~ 10: 0.7 ~ 2.4.4: 0.1 ~ 0.26.26.

The capsanthin has complex components, wherein the capsanthin comprises a plurality of monomer components such as capsorubin, lutein, zeaxanthin and the like, but a method which can effectively separate each monomer component and ensure the purity of the separated monomer is lacked;

the normal hexane-acetone-ethyl acetate mixed solution with the volume ratio of 6 ~ 10: 0.7 ~ 2.4.4: 0.1 ~ 0.26.26 is used as the analytic solution and the diluent, capsorubin and carotene can be effectively separated, and lutein which is difficult to remove can be effectively separated, so that the purity of the zeaxanthin is improved.

Preferably, the mass ratio of the capsanthin, the alcohol, the alkali and the antioxidant in the step 1) is 1: 0.2 ~ 0.3.3: 0.7 ~ 0.8.8: 0.04 ~ 0.06.06.

Preferably, the alcohol in step 1) is ethanol, methanol or propylene glycol. The preferred alcohol has a better saponification effect, and the monomers are more easily separated by resolution.

Preferably, the alkali in the step 1) is potassium hydroxide or/and sodium hydroxide.

Preferably, the alkali is added in the form of 30% ~ 60% by mass of potassium hydroxide or/and sodium hydroxide aqueous solution, and under the preferable adding mode of the alkali, the materials can be mixed uniformly more quickly, and the saponification effect is better.

Preferably, the antioxidant in step 1) is vitamin E or tert-butylhydroquinone.

Preferably, the acid in step 2) is one or a mixture of two or more of hydrochloric acid, phosphoric acid, oxalic acid, acetic acid and citric acid.

Preferably, the n-hexane-acetone-ethyl acetate mixed solution in the step 2) and the step 3) is a mixed solution prepared from n-hexane, acetone and ethyl acetate according to the volume ratio of 8 ~ 9: 1.3 ~ 1.7.7: 0.13 ~ 0.17.17.

The resin in the step 3) is one or more of XR15C, XR919LZ, XR921H and XR 910S. The macroporous adsorption resin used in the invention is produced by Shanghai Sener chemical engineering science and technology Co. Preferably, the macroporous adsorbent resin is model XR 15C.

Compared with the prior art, the invention has the beneficial effects that:

1. the purity of the obtained zeaxanthin crystals is high, the purity of the zeaxanthin is more than 90%, the applicant finds that in the prior art, the zeaxanthin is not easily doped in the lutein when the lutein is purified, but other carotenoids are doped in the zeaxanthin when the zeaxanthin is purified, the zeaxanthin often coexists with a plurality of carotenoids with similar molecular weights in nature, so that the zeaxanthin is difficult to separate, other carotenoids which are difficult to remove are often doped in the separated zeaxanthin, the main impurities which are difficult to separate are the xanthophyll of an isomer when the zeaxanthin is separated from capsanthin, and the impurities are different from other raw materials, when the capsanthin is used as the raw material, the carotenoid impurities which are specific to the capsanthin, namely the capsanthin and the capsanthin are also doped in the extracted natural zeaxanthin, namely the capsanthin and the capsanthin, in order to solve the problems, a mixed solution which has the volume ratio of 6 ~ 10: 0.7 ~ 2.4.4: 0.1.1. 0.1 ~ 0.26.26, is designed as a dilution liquid which is capable of fully diluting zeaxanthin, the purity of the zeaxanthin which is more than 1.90%, and the purity of the zeaxanthin is remarkably improved, so that the purity of the zeaxanthin sold in the market is remarkably increased, the mixed solution, and the purity of the zeaxanthin of more than 1.90%, the zeaxanthin, so that the zeaxanthin obtained by more than the purity of the zeaxanthin, the purity of the zeaxanthin is remarkably increased by more than the purity of the zeaxanthin, the purity of the zeaxanthin, the zeaxanthin by more than the normal hexane.

2. The process flow of the invention is safe and reliable, and has no toxic chemical auxiliary agent residue. The invention takes natural materials as raw materials, and the natural materials are directly obtained by physical analysis and separation after saponification treatment, no toxic materials are added in the process, and the obtained zeaxanthin crystals have no toxic chemical substance residues and are safe and reliable to use and eat. The invention directly takes capsanthin as a raw material, obtains monomers such as zeaxanthin, capsanthin and the like through saponification treatment, then obtains the zeaxanthin through separation and purification, and is convenient for large-scale production.

3. The zeaxanthin obtained by the method is a natural product. The natural zeaxanthin obtained by the method has high selling price and better market prospect compared with artificially synthesized zeaxanthin.

4. The zeaxanthin yield is high, the high-purity zeaxanthin is obtained by adopting the macroporous adsorption resin for one-time separation, repeated separation and impurity removal are not needed, the loss of the zeaxanthin in the process is less, the separation effect of the n-hexane-acetone-ethyl acetate mixed solution with the volume ratio of 6 ~ 10: 0.7 ~ 2.4.4: 0.1 ~ 0.26.26 on the zeaxanthin is high, the loss of the zeaxanthin in the separation process is reduced, and the zeaxanthin yield can reach 71.9 ~ 76.8.8%.

Detailed Description

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