CN110590630B - Method for separating and purifying zeaxanthin from capsanthin - Google Patents
Method for separating and purifying zeaxanthin from capsanthin Download PDFInfo
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Abstract
A method for separating and purifying zeaxanthin from capsanthin belongs to the technical field of carotenoid preparation. The preparation method is characterized by comprising the following preparation steps: carrying out saponification reaction on capsanthin, alcohol, alkali and an antioxidant; adjusting the pH value of the saponified substance to be 7-8, diluting the saponified substance with a mixed solution of n-hexane, acetone and ethyl acetate, analyzing the diluted mixed solution, and collecting a zeaxanthin analysis solution; recovering the solvent, cooling and crystallizing; filtering, washing and drying to obtain the zeaxanthin crystals. The method directly takes capsanthin as a raw material, obtains a zeaxanthin monomer through saponification treatment, then separates the zeaxanthin with the content of more than 90 percent, and is convenient for large-scale production.
Description
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 the red component with larger polarity comprises capsorubin and capsorubin accounting for 50-60% of the total amount, the other is a yellow component with smaller polarity, and the main components are zeaxanthin and carotene.
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 8-12 h at the temperature of 40-60 ℃ to obtain a saponified substance; the mass ratio of capsanthin, alcohol, alkali and antioxidant is 1: 0.1-0.5: 0.5-1: 0.01 to 0.1;
2) adding acid into the saponified substance, adjusting the pH value to be 7-8, and diluting with a mixed solution of n-hexane, acetone and ethyl acetate to obtain a 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 the solvent from the zeaxanthin resolution solution until the mass concentration of the zeaxanthin is 20-60%, cooling to 20-40 ℃, and maintaining the temperature for 4-6 h for crystallization; filtering to obtain crude zeaxanthin crystals, 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 prepared by mixing n-hexane, acetone and ethyl acetate according to a volume ratio (abbreviated as V/V) of 6-10: 0.7-2.4: 0.1-0.26.
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 volume ratio is 6-10: 0.7-2.4: 0.1-0.26 of n-hexane-acetone-ethyl acetate mixed solution is used as an analytic solution and a diluent, capsorubin and carotene can be effectively separated, lutein which is difficult to remove can be effectively separated, and therefore the purity of zeaxanthin is improved. The natural materials are used as raw materials, the natural materials are directly subjected to physical analysis and separation after saponification treatment, no toxic materials are added in the process, the obtained zeaxanthin crystals have no toxic chemical substance residues, the use and the eating are safe and reliable, the components in capsanthin can be effectively separated, and the purity of the zeaxanthin crystals is over 90 percent.
Preferably, the mass ratio of capsanthin, alcohol, alkali and antioxidant in the step 1) is 1: 0.2-0.3: 0.7-0.8: 0.04 to 0.06. The saponification treatment efficiency is higher under the optimized material proportion, each monomer cannot be damaged, and the final yield of the zeaxanthin can be better ensured.
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 an aqueous solution of potassium hydroxide or/and sodium hydroxide. Under the preferred alkali addition mode, the material can be mixed more rapidly, 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 prepared by mixing n-hexane, acetone and ethyl acetate according to a volume ratio of 8-9: 1.3-1.7: 0.13-0.17. The optimal mixture ratio can accelerate the resolution rate of the zeaxanthin, enlarge the separation trend among monomers and ensure that the obtained product has higher purity.
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 zeaxanthin crystal obtained by the method is high, and the purity of the zeaxanthin is over 90 percent. The applicant has found that the lutein is not easy to be doped with zeaxanthin when the lutein is purified by the prior art, but the problem of doping other carotenoids when the zeaxanthin is purified occurs. Zeaxanthin often coexists with a plurality of carotenoids with similar molecular weights in nature, which causes difficulty in separating zeaxanthin, and other carotenoids which are difficult to remove are often mixed in the separated zeaxanthin; when the capsanthin is used for separating zeaxanthin, impurities which are difficult to separate are isomeric xanthophylls; and is different from other raw materials, when capsanthin is taken as a raw material, the extracted natural zeaxanthin can be doped with carotenoid impurities, namely capsorubin and capsorubin, which are unique to capsanthin, so that the problem is solved, and the volume ratio of the capsanthin to capsorubin is designed to be 6-10: 0.7-2.4: 0.1-0.26 of n-hexane-acetone-ethyl acetate mixed solution is used as a diluent and a resolving solution, the mixed solution can fully separate capsorubin, capsorubin and cryptoxanthin and can separate lutein which is difficult to remove, so that the purity of the zeaxanthin is obviously improved, the purity of the obtained zeaxanthin is more than 90%, and the highest purity can reach 96.9%. The selling price of the zeaxanthin with the purity of 80% w/w sold in the market at present is about 8 thousand yuan/kg, the selling price of the zeaxanthin with the purity of 90% w/w sold in the market is about 1.1 ten thousand yuan/kg, and the selling price of the zeaxanthin with the purity of more than 95% w/w sold in the market is about 1.3 ten thousand/kg.
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 obtained by the method is high. Because the high-purity zeaxanthin is obtained by adopting the macroporous adsorption resin for one-time separation, the zeaxanthin obtained by the method has less loss in the process without repeated separation and impurity removal; and, the volume ratio is 6-10: 0.7-2.4: the n-hexane-acetone-ethyl acetate mixed solution of 0.1-0.26 has a high separation effect on the zeaxanthin, the loss of the zeaxanthin in the separation process is reduced, and the yield of the zeaxanthin can reach 71.9-76.8%.
Detailed Description
The invention is further illustrated by the following specific examples, of which example 1 is the best mode of practice.
Capsanthin used in examples and comparative examples: the content of total carotenoid in capsanthin is 10 percent, and the content of beta-carotene accounts for 13 percent; zeaxanthin accounts for 12%, and other carotenoids account for 75%. Capsanthin comes from Shandong Tianyin Biotech limited.
Macroporous adsorption resins used in examples and comparative examples: shanghai Sener chemical science and technology, selected from resin models XR15C, XR919LZ, XR921H, XR 910S.
Example 1
1) Carrying out saponification reaction on capsanthin, ethanol, potassium hydroxide and vitamin E under the condition of introducing nitrogen, and saponifying for 8h at 40 ℃ to obtain a saponified substance; the mass ratio of capsanthin, ethanol, potassium hydroxide and vitamin E is 1: 0.25: 0.75: 0.05; adding potassium hydroxide in the form of potassium hydroxide aqueous solution with the mass percent of 45%;
2) hydrochloric acid was added to the saponified product to adjust the pH to 7, and a mixture of n-hexane-acetone-ethyl acetate (V/V, 8.5: 1.5: 0.15) diluting to obtain a saponified substance diluent;
3) introducing the saponified substance diluent into an analytical column filled with 6 kilograms of XR15C macroporous adsorption resin, wherein the adsorption flow rate is 2 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixture (V/V, 8.5: 1.5: 0.15) at resolving flow rate of 1BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 65 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 25%, cooling to 30 deg.C, and maintaining for 4 hr for crystallization; washing the filtered crude zeaxanthin crystals with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 2
1) Carrying out saponification reaction on capsanthin, ethanol, sodium hydroxide and tert-butyl hydroquinone under the condition of introducing nitrogen, and saponifying for 8.5h at 45 ℃ to obtain a saponified substance; the mass ratio of capsanthin to ethanol to sodium hydroxide to tert-butylhydroquinone is 1: 0.2: 0.8: 0.04; adding sodium hydroxide in the form of sodium hydroxide aqueous solution with the mass percentage of 40%;
2) acetic acid was added to the saponified product to adjust the pH to 7, and a mixture of n-hexane-acetone-ethyl acetate (V/V, 8: 1.7: 0.13) diluting to obtain a saponified substance diluent;
3) introducing the saponified substance diluent into an analytical column filled with 6 kg of XR919LZ macroporous adsorption resin, wherein the adsorption flow rate is 1.5 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 8: 1.7: 0.13) at resolving flow rate of 1.5BV/h, and collecting zeaxanthin resolution solution;
4) recovering solvent from zeaxanthin solution (concentration temperature 68 deg.C, vacuum degree-0.07 Mpa) until zeaxanthin mass concentration is 60%, cooling to 25 deg.C, and maintaining for 4 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 3
1) Carrying out saponification reaction on capsanthin, ethanol, potassium hydroxide and vitamin E under the condition of introducing nitrogen, and saponifying for 8h at 55 ℃ to obtain a saponified substance; the mass ratio of capsanthin to ethanol to potassium hydroxide to vitamin E is 1: 0.3: 0.7: 0.06; adding potassium hydroxide in the form of 50% by mass of potassium hydroxide aqueous solution;
2) oxalic acid was added to the saponified material to adjust the pH to 7.5, and a mixture of n-hexane-acetone-ethyl acetate (V/V, 9: 1.3: 0.17) diluting to obtain saponified substance diluent;
3) introducing the saponified substance diluent into an analytical column filled with 6 kg of XR921H macroporous adsorption resin, wherein the adsorption flow rate is 2 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 9: 1.3: 0.17) at resolving flow rate of 1.5BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 65 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 30%, cooling to 30 deg.C, and maintaining for 3.5 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 4
1) Carrying out saponification reaction on capsanthin, methanol, potassium hydroxide and vitamin E under the condition of introducing nitrogen, and saponifying for 11h at 60 ℃ to obtain a saponified substance; the mass ratio of capsanthin to methanol to potassium hydroxide to vitamin E is 1: 0.1: 1: 0.01; adding potassium hydroxide in the form of 60% by mass of potassium hydroxide aqueous solution;
2) hydrochloric acid was added to the saponified material to adjust the pH to 7, and XR910S was replaced with a 3-fold amount by mass of capsanthin of an n-hexane-acetone-ethyl acetate mixture (V/V, 7: 2: 0.1) diluting to obtain a saponified substance diluent;
3) introducing the saponified substance diluent into an analytical column filled with 6 kg of XR15C macroporous adsorption resin, wherein the adsorption flow rate is 1.5 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 7: 2: 0.1) at resolving flow rate of 1BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 68 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 20%, cooling to 20 deg.C, and maintaining for 4.6 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 5
1) Capsanthin, propylene glycol, potassium hydroxide and sodium hydroxide (molar ratio 1: 1) carrying out saponification reaction on tert-butyl hydroquinone under the condition of introducing nitrogen, and saponifying for 12h at 55 ℃ to obtain a saponified substance; capsanthin, propylene glycol, potassium hydroxide, sodium hydroxide and tert-butyl hydroquinone are 1: 0.5: 0.5: 0.1; adding potassium hydroxide and sodium hydroxide in the form of aqueous solution with the total mass percentage of 30%;
2) hydrochloric acid was added to the saponified product to adjust the pH to 8, and a mixture of n-hexane-acetone-ethyl acetate (V/V, 10: 1: 0.2) diluting to obtain a saponified substance diluent;
3) the saponified material dilution was passed into a batch containing 6 kg of macroporous adsorption resins XR15C and XR921H (XR 15C and XR921H mass ratio 1: 2) the adsorption flow rate in the desorption column is 1 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 10: 1: 0.2) at resolving flow rate of 1BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 68 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 45%, cooling to 40 deg.C, and maintaining for 6 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 6
1) Carrying out saponification reaction on capsanthin, ethanol, potassium hydroxide and vitamin E under the condition of introducing nitrogen, and saponifying for 8h at 40 ℃ to obtain a saponified substance; the mass ratio of capsanthin, ethanol, potassium hydroxide and vitamin E is 1: 0.25: 0.75: 0.05; adding potassium hydroxide in the form of potassium hydroxide aqueous solution with the mass concentration of 45%;
2) hydrochloric acid was added to the saponified product to adjust the pH to 7.5, and an n-hexane-acetone-ethyl acetate mixture (V/V, 7.5: 1.5: 0.2) diluting to obtain a saponified substance diluent;
3) introducing the saponified substance diluent into an analytical column filled with 6 kg of XR15C macroporous adsorption resin, wherein the adsorption flow rate is 1.5 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 7.5: 1.5: 0.2) at resolving flow rate of 1BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin resolution solution under reduced pressure to recover solvent (concentration temperature 64 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 55%, recovering solvent from the zeaxanthin resolution solution, cooling to 30 deg.C, and maintaining for 4 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 7
1) Carrying out saponification reaction on capsanthin, ethanol, sodium hydroxide and tert-butyl hydroquinone under the condition of introducing nitrogen, and saponifying for 8.5h at 45 ℃ to obtain a saponified substance; the mass ratio of capsanthin to ethanol to sodium hydroxide to tert-butylhydroquinone is 1: 0.2: 0.8: 0.04; sodium hydroxide is added in the form of an aqueous solution of sodium hydroxide with the mass concentration of 40%;
2) acetic acid was added to the saponified product to adjust the pH to 7.5, and a mixture of n-hexane-acetone-ethyl acetate (V/V, 7: 1.7: 0.18) diluting to obtain saponified substance diluent;
3) the saponified material dilution was passed into a chamber containing 6 kg of macroporous adsorption resins XR15C and XR921H (XR 15C and XR921H mass ratio 2: 1) the adsorption flow rate in the desorption column of the macroporous adsorption resin is 2 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 7: 1.7: 0.18) at resolving flow rate of 1.5BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 65 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 30%, cooling to 25 deg.C, and maintaining for 4 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 8
1) Carrying out saponification reaction on capsanthin, ethanol, potassium hydroxide and vitamin E under the condition of introducing nitrogen, and saponifying for 8h at 55 ℃ to obtain a saponified substance; the mass ratio of capsanthin to ethanol to potassium hydroxide to vitamin E is 1: 0.3: 0.7: 0.06; potassium hydroxide is added in the form of an aqueous solution of potassium hydroxide with the mass concentration of 50%;
2) hydrochloric acid was added to the saponified product to adjust the pH to 8, and an n-hexane-acetone-ethyl acetate mixed solution (V/V, 8: 1.4: 0.13) diluting to obtain a saponified substance diluent;
3) the saponified material dilution was passed into a batch containing 6 kg of macroporous adsorption resins XR15C and XR921H (XR 15C and XR921H mass ratio 1: 1) the adsorption flow rate in the desorption column is 2 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 8: 1.4: 0.13) at resolving flow rate of 1BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 65 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 40%, cooling to 30 deg.C, and maintaining for 3.5 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Example 9
1) Carrying out saponification reaction on capsanthin, methanol, potassium hydroxide and vitamin E under the condition of introducing nitrogen, and saponifying for 11h at 60 ℃ to obtain a saponified substance; the mass ratio of capsanthin to methanol to potassium hydroxide to vitamin E is 1: 0.1: 1: 0.01; potassium hydroxide is added in the form of an aqueous solution of potassium hydroxide with the mass concentration of 60%;
2) hydrochloric acid was added to the saponified product to adjust the pH to 8, and an n-hexane-acetone-ethyl acetate mixed solution (V/V, 9: 0.7: 0.26) diluting to obtain saponified substance diluent;
3) introducing the saponified substance diluent into an analytical column filled with 60 kg of XR910 macroporous adsorption resin, wherein the adsorption flow rate is 1.5 BV/h; after adsorption, washing the mixture with deionized water until the mixture is colorless; resolving with n-hexane-acetone-ethyl acetate mixed solution (V/V, 9: 0.7: 0.26) at resolving flow rate of 1BV/h, and collecting zeaxanthin resolution solution;
4) concentrating the zeaxanthin solution under reduced pressure to recover solvent (concentration temperature 65 deg.C, vacuum degree-0.07 Mpa) until the zeaxanthin mass concentration is 50%, cooling to 20 deg.C, and maintaining for 4.6 hr for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with 95% ethanol water solution by mass percent to obtain wet zeaxanthin crystals;
5) vacuum drying the wet zeaxanthin crystals to obtain zeaxanthin crystals.
Comparative example 1
Steps 1) to 5) are the same as in example 1, except that the resolving solution used in steps 2) and 3) is a mixed solution of n-hexane-acetone-ethyl acetate (V/V, 13: 1.5: 0.15). The amount of n-hexane used in this comparative example was greater than that used in the inventive examples.
Comparative example 2
Steps 1) to 5) are the same as in example 1, except that the resolving solution used in steps 2) and 3) is a mixed solution of n-hexane-acetone-ethyl acetate (V/V, 4: 1.5: 0.15). The amount of n-hexane used in this comparative example was less than that used in the inventive examples.
Comparative example 3
Steps 1) to 5) are the same as those in example 1, except that the resolving solution used in steps 2) and 3) is a mixed solution of n-hexane-acetone-ethyl acetate (V/V, 8.5: 4: 0.15). The comparative acetone was used in an amount greater than the inventive examples.
Comparative example 4
Steps 1) to 5) are the same as those in example 1, except that the resolving solution used in steps 2) and 3) is a mixed solution of n-hexane-acetone-ethyl acetate (V/V, 8.5: 0.3: 0.15). The comparative acetone was used in an amount less than the inventive examples.
Comparative example 5
Steps 1) to 5) are the same as those in example 1, except that the resolving solution used in steps 2) and 3) is a mixed solution of n-hexane-acetone-ethyl acetate (V/V, 8.5: 1.5: 1). The comparative example used a greater amount of ethyl acetate than the inventive examples.
Comparative example 6
Steps 1) to 5) are the same as those in example 1, except that the resolving solution used in steps 2) and 3) is a mixed solution of n-hexane-acetone-ethyl acetate (V/V, 8.5: 1.5: 0.05). The comparative acetone was used in an amount less than the inventive examples.
Comparative example 7
Steps 1) to 5) are the same as in example 1, except that the resolving solution used in steps 2) and 3) is a n-hexane-acetone mixed solution (V/V, 8.5: 1.5).
Comparative example 8
Steps 1) to 5) are the same as in example 1, except that the resolving solution used in steps 2) and 3) is a n-hexane-ethyl acetate mixed solution (V/V, 8.5: 0.15).
Comparative example 9
Steps 1) to 5) are the same as in example 1, except that the resolving solution used in steps 2) and 3) is an acetone-ethyl acetate mixed solution (V/V, 1.5: 0.15).
Performance testing
A method for detecting the zeaxanthin content comprises the following steps: the purity (purity namely content percent w/w) of the zeaxanthin is detected according to the method in the national standard GB 26405-2011. High performance liquid chromatograph: model LC-20AT, manufacturer Shimadzu corporation, Japan. The test data results are reported in the table below.
Secondly, a yield calculation formula: the yield of the zeaxanthin is% = the total content of the zeaxanthin/the total content of the zeaxanthin in the capsanthin is multiplied by 100%.
TABLE 1 zeaxanthin purity and yield for examples 1-9
TABLE 2 comparative examples 1-9 zeaxanthin purity and yield
As can be seen from tables 1 to 2, the purity and yield of examples 1 to 9 are significantly higher than those of comparative examples 1 to 9. In comparative examples 1-6, the ratio of n-hexane, acetone and ethyl acetate is different from that in example 1, so that the purity and yield are reduced. In comparative examples 7-9, only any two of n-hexane, acetone and ethyl acetate are used as mixed liquid, and the purity and yield of zeaxanthin are obviously reduced. The n-hexane-acetone-ethyl acetate mixed solution claimed by the invention can be proved to be capable of improving the purity and yield of the zeaxanthin.
The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention are within the protection scope of the technical solution of the present invention.
Claims (8)
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 8-12 h at the temperature of 40-60 ℃ to obtain a saponified substance; the mass ratio of capsanthin, alcohol, alkali and antioxidant is 1: 0.1-0.5: 0.5-1: 0.01 to 0.1;
2) adding acid into the saponified substance, adjusting the pH value to be 7-8, and diluting with a mixed solution of n-hexane-acetone-ethyl acetate to obtain a saponified substance diluent, wherein the mass ratio of the capsanthin to the mixed solution of n-hexane-acetone-ethyl acetate 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 the solvent from the zeaxanthin resolution solution until the mass concentration of the zeaxanthin is 20-60%, cooling to 20-40 ℃, and maintaining the temperature for 4-6 h for crystallization; filtering to obtain crude zeaxanthin crystals, and washing with ethanol;
5) vacuum drying the zeaxanthin crystals to obtain zeaxanthin crystals;
the n-hexane-acetone-ethyl acetate mixed solution in the step 2) and the step 3) is prepared from n-hexane, acetone and ethyl acetate according to a volume ratio of 6-10: 0.7-2.4: 0.1-0.26 of a mixed solution;
the antioxidant in the step 1) is vitamin E or tert-butyl hydroquinone.
2. The method for separating and purifying zeaxanthin from capsanthin according to claim 1, which comprises the following steps: the mass ratio of capsanthin, alcohol, alkali and antioxidant in the step 1) is 1: 0.2-0.3: 0.7-0.8: 0.04 to 0.06.
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, which comprises the following steps: the alkali is added in the form of 30-60% by mass of potassium hydroxide or/and sodium hydroxide aqueous solution.
6. 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.
7. The method for separating and purifying zeaxanthin from capsanthin according to claim 1, which comprises the following steps: the n-hexane-acetone-ethyl acetate mixed solution in the step 2) and the step 3) is prepared from n-hexane, acetone and ethyl acetate according to the volume ratio of 8-9: 1.3-1.7: 0.13-0.17.
8. 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.
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