CN113582899A - Preparation method of zeaxanthin - Google Patents
Preparation method of zeaxanthin Download PDFInfo
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- CN113582899A CN113582899A CN202110963783.3A CN202110963783A CN113582899A CN 113582899 A CN113582899 A CN 113582899A CN 202110963783 A CN202110963783 A CN 202110963783A CN 113582899 A CN113582899 A CN 113582899A
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- JKQXZKUSFCKOGQ-LQFQNGICSA-N Z-zeaxanthin Natural products C([C@H](O)CC=1C)C(C)(C)C=1C=CC(C)=CC=CC(C)=CC=CC=C(C)C=CC=C(C)C=CC1=C(C)C[C@@H](O)CC1(C)C JKQXZKUSFCKOGQ-LQFQNGICSA-N 0.000 title claims abstract description 60
- QOPRSMDTRDMBNK-RNUUUQFGSA-N Zeaxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCC(O)C1(C)C)C=CC=C(/C)C=CC2=C(C)CC(O)CC2(C)C QOPRSMDTRDMBNK-RNUUUQFGSA-N 0.000 title claims abstract description 60
- JKQXZKUSFCKOGQ-LOFNIBRQSA-N all-trans-Zeaxanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2=C(C)CC(O)CC2(C)C JKQXZKUSFCKOGQ-LOFNIBRQSA-N 0.000 title claims abstract description 60
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- JKQXZKUSFCKOGQ-JLGXGRJMSA-N (3R,3'R)-beta,beta-carotene-3,3'-diol Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C[C@@H](O)CC1(C)C JKQXZKUSFCKOGQ-JLGXGRJMSA-N 0.000 title claims abstract description 57
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
- C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
- C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
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Abstract
The invention relates to the technical field of natural product extraction, in particular to a preparation method of zeaxanthin; diluting fresh marigold flower slurry with water, adjusting the pH value, performing enzymolysis by adding hydrolase, lipase and alkaline protease, pulverizing the enzymolysis product by using an ultrasonic pulverizer, introducing gas, filtering, extracting filter residues for multiple times by using a solvent in a high-pressure device, combining extract liquids, concentrating to obtain a concentrate, and drying the concentrate in vacuum to obtain a zeaxanthin extract; the invention uses fresh marigold flower as raw material, uses biological enzyme to catalyze reaction to obtain zeaxanthin extract, accelerates the reaction speed under the action of homogenizing, ultrasonic crushing instrument and knifing flash evaporation instrument, shortens the production time, reduces the treatment temperature, and improves the yield and stability of the product.
Description
Technical Field
The invention relates to the technical field of natural product extraction, and particularly relates to a preparation method of zeaxanthin.
Background
Zeaxanthin (Zeaxanthin,3, 3' -dihydroxy-beta-carotene), also known as Zeaxanthin, belongs to the isoprenoids and is a isomer of xanthophylls, which is widely used in foods and medicines. It can act on human retina, is very important for treating and preventing senile macular degeneration, and can effectively protect eyes. It also has good oxidation resistance, and has unique functions in protecting skin, enhancing immunity, reducing incidence of cardiovascular diseases, etc. Since the human body cannot synthesize zeaxanthin itself, it must be taken through the diet.
The zeaxanthin source mainly comprises natural extraction, lutein conversion and artificial synthesis. Wherein the natural extraction requires the use of large amounts of organic solvents. The artificial synthesis needs to be designed and synthesized into key intermediates through multiple steps, so that the side reactions are more, and the process control is complex. Nowadays, the planting technology of marigold is very mature, and the extraction process of lutein is also quite developed, so that the source of lutein is wide, and therefore, the preparation of zeaxanthin through the epimerization reaction of lutein is a promising method. At present, the common method is to obtain the zeaxanthin by further purifying and isomerizing marigold extract which is used as a raw material. The method needs to perform the working procedures of fermentation, drying, granulation and the like on the marigold before obtaining the marigold extract, each working procedure can cause certain loss of a target product and generate harmful substances, the cost is high, the production period is long, the obtained marigold extract also needs to be subjected to saponification reaction with strong alkali to obtain lutein, the subsequent treatment of sewage has certain difficulty, and the production cost is increased.
Chinese patent CN104447469A discloses a method for preparing zeaxanthin from lutein extract by using an ultrasonic-microwave technology, which mainly uses a large amount of propylene glycol as a solvent, saponifies strong alkali aqueous solution, and accelerates the reaction process by using ultrasonic-microwave, but the ultrasonic-microwave has longer duration and is difficult to control side reactions.
Chinese patent CN110563625B discloses a method for separating and purifying zeaxanthin by marigold oleoresin, which takes marigold extractum as a raw material, extracts by an organic solvent, then separates and purifies the extract by macroporous resin, and then elutes by a large amount of organic solvent. The separation method is simple, the resin can be recycled, but more organic solvents are used, only the original zeaxanthin is separated, and the yield is low.
Chinese patent CN109678772A discloses a method for preparing zeaxanthin from lutein extract as reaction raw material by saponification-alkali catalytic isomerization reaction, which strengthens the saponification process of lutein extract under pressure and accelerates the isomerization reaction, but the reaction temperature is higher and has a certain damage to active substances, so that the zeaxanthin yield is lower, the reaction time is longer, and the energy consumption and cost are higher, which is not favorable for industrialization.
In summary, the development of a method for preparing zeaxanthin is still a key problem to be solved urgently in the technical field of natural product extraction.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a preparation method of zeaxanthin, which uses fresh marigold flower as a raw material and utilizes a biological enzyme to catalyze a reaction to obtain a zeaxanthin extract, accelerates the reaction speed under the actions of a homogenizing instrument, an ultrasonic crushing instrument and a scraping film flash evaporation instrument, shortens the production time, reduces the treatment temperature and improves the yield and the stability of the product.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for preparing zeaxanthin comprises diluting fresh flower pulp of marigold with water, adjusting pH, adding hydrolase, lipase and alkaline protease for enzymolysis, pulverizing the zymolyte with ultrasonic pulverizer, introducing gas, filtering, extracting the residue with solvent in high pressure device for several times, mixing the extractive solutions, concentrating to obtain concentrate, and vacuum drying to obtain zeaxanthin extract.
Preferably, the hydrolytic enzyme is a protease, a cellulase, and a pectinase.
Preferably, the solvent is one or more of ethanol, n-hexane and acetone.
Preferably, the gas is nitrogen.
Preferably, the extract is concentrated using a wiped film flash evaporator.
Preferably, the feed-liquid ratio in the extraction is 1: (2-4).
Preferably, the method comprises the following steps:
s1, taking the fresh marigold flower pulp, adding water, uniformly stirring, adding citric acid to adjust the pH value to 4.0-6.0, heating in a water bath at the temperature of 37-50 ℃, adding protease, cellulase and pectinase, uniformly stirring, and oscillating in a constant-temperature water bath in a dark place to perform enzymolysis reaction.
S2, adjusting the pH value with a sodium hydroxide solution, homogenizing for 3min, adding lipase and alkaline protease, stirring uniformly, reacting on a water bath constant temperature oscillator in a dark place, homogenizing for 2min, adding lipase twice, stirring uniformly, and reacting for 1-2h to obtain a feed liquid.
And S3, treating the feed liquid by using an ultrasonic pulverizer.
And S4, introducing nitrogen into the treated feed liquid for 20min, heating to 80-150 ℃, reacting for 4h, and filtering.
S5, extracting the filter residue in the S4 for multiple times at normal temperature in a high-pressure device by using a solvent, wherein the extraction pressure is 2-8MPa, the extraction times are 1-3 times, and the material-liquid ratio is 1: (2-4), and the extraction time is 5-10 min.
S6, combining the extracts extracted for multiple times in S5, and concentrating by a wiped film flash evaporator to obtain a concentrate.
S7, vacuum drying the concentrate for 2-4h at the vacuum drying temperature of 45-60 ℃ to obtain the zeaxanthin extract.
Preferably, in the step S3, when the feed liquid is processed by the ultrasonic crusher, the total working time is 2-10min, the working time is 15S, the interval time is 30S, and the power is 400-.
Advantageous effects
Compared with the known public technology, the technical scheme provided by the invention has the following beneficial effects:
1. the invention adopts a biological enzymolysis method to destroy cell walls, promotes the combination of target products and other substances such as protein, sugar and the like, is more beneficial to the dissolution of the target products and shortens the reaction time, is safe without generating harmful substances, and overcomes the defects of more working procedures, long time consumption and generation of partial harmful substances at high temperature of the traditional method.
2. The lutein ester hydrolysis adopts a mild biological method to replace a chemical method of strong alkali saponification, and the lipase hydrolyzes the lutein ester and the zeaxanthin ester to be lutein and zeaxanthin in free states, so that the production safety can be improved, the environmental pollution can be reduced, the use of organic solvents can be reduced, and the stability of products can be ensured.
3. The invention applies the ultrasonic crusher, the high pressure device and the film scraping flash evaporator to be beneficial to improving the yield of the target product, reducing the product loss and improving the product stability.
Drawings
FIG. 1 is a flow chart of a process for the preparation of zeaxanthin.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The present invention will be further described with reference to the following examples.
Example 1
Referring to fig. 1, the preparation method of zeaxanthin in this embodiment includes the following steps:
step one, taking 1kg of fresh marigold flower pulp (subjected to crushing and pulping), adding 500g of water in a certain proportion, uniformly stirring, adding citric acid to adjust the pH value to 4.0, putting the mixture into a 37 ℃ water bath constant-temperature oscillator for preheating, adding a certain amount of hydrolase (the dosage of cellulase is 600U/g, the dosage of pectinase is 500U/g and the dosage of acid protease is 300U/g) after the temperature is reached, uniformly stirring, and vibrating in a dark place to perform enzymolysis reaction for 4 hours.
And step two, adjusting the pH to 7.0 by using a sodium hydroxide solution (2moL/L), homogenizing for 3min, adding lipase (the addition of 800U/g) and alkaline protease (the addition of 800U/g), stirring uniformly, reacting for 2h on a water bath constant temperature oscillator, homogenizing for 2min, adding the lipase (the addition of 1200U/g) twice, stirring uniformly, and reacting for 1 h.
And step three, treating the enzymatic hydrolysate by using an ultrasonic pulverizer, wherein the total working time is 5min, the working time/interval time is 15s/30s, and the power is 500W.
And step four, introducing nitrogen into the feed liquid for 20min, heating to 80 ℃, reacting for 4h, and filtering.
And step five, extracting filter residues with ethanol at normal temperature in a high-pressure device, wherein the extraction pressure is 2MPa, the extraction is carried out for 1 time, and the material-liquid ratio is 1: 3(V/V) and the extraction time is 5 min.
And step six, concentrating the combined extract liquor by a wiped film flash evaporator, and concentrating the material at the working temperature of 60 ℃.
Seventhly, vacuum drying the concentrate at 50 ℃ for 4 hours to obtain 1.8g of the zeaxanthin extract.
And (3) carrying out HPLC detection on the obtained zeaxanthin extract, wherein the HPLC detection method comprises the following steps: GB26405-2011 shows that the zeaxanthin content in the extract is 70.35% by detection.
Example 2
Referring to fig. 1, the preparation method of zeaxanthin in this embodiment includes the following steps:
step one, taking 1kg of fresh marigold flower pulp (subjected to crushing and pulping), adding 1kg of water in a certain proportion, uniformly stirring, adding citric acid to adjust the pH value to 4.5, putting the mixture into a water bath constant-temperature oscillator at 40 ℃ for preheating, adding a certain amount of hydrolase (the dosage of cellulase is 400U/g, the dosage of pectinase is 600U/g and the dosage of acid protease is 400U/g) after the temperature is reached, uniformly stirring, and vibrating in a dark place to perform enzymolysis reaction for 3 hours.
And step two, adjusting the pH to 7.5 by using a sodium hydroxide solution (1moL/L), homogenizing for 5min, adding the lipase (the addition amount is 700U/g) and the alkaline protease (the addition amount is 800U/g), stirring uniformly, reacting for 2h on a water bath constant temperature oscillator, homogenizing for 2min, adding the lipase (the addition amount is 1000U/g) twice, stirring uniformly, and reacting for 1 h.
And step three, treating the enzymatic hydrolysate by using an ultrasonic pulverizer, wherein the total working time is 3min, the working time/interval time is 15s/30s, and the power is 500W.
And step four, introducing nitrogen into the feed liquid for 20min, heating to 80 ℃, reacting for 4h, and filtering.
And step five, extracting filter residues in a high-pressure device by using acetone, wherein the extraction pressure is 4MPa, the extraction is carried out for 2 times, and the material-liquid ratio is 1: 2(V/V), the extraction time is 6min, and the extracts are combined.
And step six, concentrating the extract liquor by a wiped film flash evaporator, and concentrating the material at the working temperature of 55 ℃.
Seventhly, vacuum drying the concentrate at 45 ℃ for 3 hours to obtain 2.5g of the zeaxanthin extract.
And (3) carrying out HPLC detection on the obtained zeaxanthin extract, wherein the HPLC detection method comprises the following steps: GB26405-2011 shows that the content of zeaxanthin in the extract is 75% by detection.
Example 3
Referring to fig. 1, the preparation method of zeaxanthin in this embodiment includes the following steps:
step one, taking 1kg of fresh marigold flower pulp (subjected to crushing and pulping), adding 1.5kg of water in a certain proportion, uniformly stirring, adding citric acid to adjust the pH value to 4.8, putting into a water bath constant-temperature oscillator at 42 ℃ for preheating, adding a certain amount of hydrolase (the dosage of cellulase is 500U/g, the dosage of pectinase is 600U/g and the dosage of acid protease is 500U/g) after the temperature reaches, uniformly stirring, and vibrating in a dark place to perform enzymolysis reaction for 5 hours.
And step two, adjusting the pH to 7.5 by using a sodium hydroxide solution (2moL/L), homogenizing for 3min, adding lipase (the addition amount is 900U/g) and alkaline protease (the addition amount is 900U/g), stirring uniformly, reacting for 1h on a water bath constant temperature oscillator, homogenizing for 2min, adding the lipase (the addition amount is 1400U/g) twice, stirring uniformly, and reacting for 2 h.
And step three, treating the enzymatic hydrolysate by using an ultrasonic pulverizer, wherein the total working time is 6min, the working time/interval time is 15s/40s, and the power is 400W.
And step four, introducing nitrogen into the feed liquid for 20min, heating to 85 ℃, reacting for 4h, and filtering.
And step five, extracting filter residues in a high-pressure device by using a mixed solution (V/V1: 1) of normal hexane and ethanol, wherein the extraction pressure is 6MPa, the extraction is carried out for 2 times, and the material-liquid ratio is 1: 3(V/V), the extraction time is 7min, and the extracts are combined.
And step six, concentrating the extract liquor by a wiped film flash evaporator, and concentrating the material at the working temperature of less than 50 ℃.
Seventhly, vacuum drying the concentrate at 60 ℃ for 3 hours to obtain 2.5g of the zeaxanthin extract.
And (3) carrying out HPLC detection on the obtained zeaxanthin extract, wherein the HPLC detection method comprises the following steps: GB26405-2011 shows that the zeaxanthin content in the extract is 85% by detection.
Example 4
Referring to fig. 1, the preparation method of zeaxanthin in this embodiment includes the following steps:
step one, taking 1kg of fresh marigold flower pulp (crushed and ground-milled), adding 2kg of water, uniformly stirring, adding citric acid to adjust the pH value to 5.0, putting the mixture into a water bath constant-temperature oscillator at 40 ℃ for preheating, adding a certain amount of hydrolase (the dosage of cellulase is 600U/g, the dosage of pectinase is 700U/g and the dosage of acid protease is 500U/g) after the temperature is reached, uniformly stirring, and shaking in a dark place to perform enzymolysis reaction for 3 hours.
And step two, adjusting the pH to 7.5 by using a sodium hydroxide solution (2moL/L), homogenizing for 5min, adding lipase (the addition amount is 700U/g) and alkaline protease (the addition amount is 800U/g), stirring uniformly, reacting for 1.5h on a water bath constant temperature oscillator, homogenizing for 2min, adding the lipase (the addition amount is 1000U/g) twice, stirring uniformly, and reacting for 1.5 h.
And step three, treating the enzymatic hydrolysate by using an ultrasonic pulverizer, wherein the total working time is 8min, the working time/interval time is 15s/30s, and the power is 600W.
And step four, introducing nitrogen into the feed liquid for 20min, heating to 85 ℃, reacting for 4h, and filtering.
And step five, extracting the filter residue by using normal hexane in a high-pressure device for 3 times under the extraction pressure of 4MPa, wherein the material-liquid ratio is 1: 4(V/V), the extraction time is 10min, and the extracts are combined.
And step six, concentrating the extract liquor by a wiped film flash evaporator, and concentrating the material at the working temperature of 55 ℃.
Seventhly, vacuum drying the concentrate at 45 ℃ for 3 hours to obtain 2.5g of the zeaxanthin extract.
And (3) carrying out HPLC detection on the obtained zeaxanthin extract, wherein the HPLC detection method comprises the following steps: GB26405-2011 shows that the zeaxanthin content in the extract is 70% by detection.
Example 5
Referring to fig. 1, the preparation method of zeaxanthin in this embodiment includes the following steps:
step one, taking 1kg of fresh marigold flower pulp (subjected to crushing and pulping), adding 800g of water according to a certain proportion, uniformly stirring, adding citric acid to adjust the pH value to 5.5, putting into a water bath constant-temperature oscillator at 45 ℃ for preheating, adding a certain amount of hydrolase (the dosage of cellulase is 600U/g, the dosage of pectinase is 700U/g and the dosage of acid protease is 500U/g) after the temperature is reached, uniformly stirring, and vibrating in a dark place to perform enzymolysis reaction for 3 hours.
And step two, adjusting the pH to 7.5 by using a sodium hydroxide solution (2moL/L), homogenizing for 5min, adding the lipase (the addition amount is 700U/g) and the alkaline protease (the addition amount is 500U/g), uniformly stirring, reacting for 2h on a water bath constant temperature oscillator, homogenizing for 2min, adding the lipase (the addition amount is 1000U/g) for the second time, uniformly stirring, and reacting for 1 h.
And step three, treating the enzymatic hydrolysate by using an ultrasonic pulverizer, wherein the total working time is 3min, the working time/interval time is 15s/30s, and the power is 500W.
And step four, introducing nitrogen into the feed liquid for 20min, heating to 90 ℃, reacting for 4h, and filtering.
And step five, extracting the filter residue by using ethanol in a high-pressure device, wherein the extraction pressure is 6MPa, the extraction is carried out for 1 time, and the material-liquid ratio is 1: 2(V/V), the extraction time is 5min, and the extracts are combined.
And step six, concentrating the extract liquor by a wiped film flash evaporator, and concentrating the material at the working temperature of 55 ℃.
Seventhly, vacuum drying the concentrate at 50 ℃ for 2 hours to obtain 2.5g of the zeaxanthin extract.
And (3) carrying out HPLC detection on the obtained zeaxanthin extract, wherein the HPLC detection method comprises the following steps: GB26405-2011 shows that the zeaxanthin content in the extract is 72 percent through detection.
Example 6
Referring to fig. 1, the preparation method of zeaxanthin in this embodiment includes the following steps:
step one, taking 1kg of fresh marigold flower pulp (subjected to crushing and pulping), adding 2kg of water in a certain proportion, uniformly stirring, adding citric acid to adjust the pH value to 4.0, putting into a water bath constant-temperature oscillator at 50 ℃ for preheating, adding a certain amount of hydrolase (the dosage of cellulase is 600U/g, the dosage of pectinase is 700U/g and the dosage of acid protease is 500U/g) after the temperature is reached, uniformly stirring, and then vibrating in a dark place to perform enzymolysis reaction for 3 hours.
And step two, adjusting the pH to 7.5 by using a sodium hydroxide solution (2moL/L), homogenizing for 5min, adding lipase (the addition amount is 700U/g) and alkaline protease (the addition amount is 500U/g), uniformly stirring, reacting for 2h on a water bath constant temperature oscillator, homogenizing for 2min, adding the lipase (the addition amount is 1000U/g) twice, uniformly stirring, and reacting for 1 h.
And step three, treating the enzymatic hydrolysate by using an ultrasonic pulverizer, wherein the total working time is 3min, the working time/interval time is 15s/30s, and the power is 500W.
And step four, introducing nitrogen into the feed liquid for 20min, heating to 80 ℃, reacting for 4h, and filtering.
And step five, extracting filter residues by using ethanol and acetone (V/V1: 1) in a high-pressure device at the extraction pressure of 6MPa for 2 times, wherein the material-liquid ratio is 1: 4(V/V), the extraction time is 8min, and the extracts are combined.
And step six, concentrating the filtrate by a wiped film flash evaporator, and concentrating the material at a working temperature of 55 ℃.
Seventhly, vacuum drying the concentrate at 60 ℃ for 4 hours to obtain 2.5g of the zeaxanthin extract.
And (3) carrying out HPLC detection on the obtained zeaxanthin extract, wherein the HPLC detection method comprises the following steps: GB26405-2011 shows that the content of zeaxanthin in the extract is 80% by detection.
In examples 1-6: (1) and protease: the method is used for hydrolyzing the protein in marigold, and can hydrolyze macromolecular protein into products such as amino acid, polypeptide chain and the like at a certain temperature and a certain pH value, so that zeaxanthin ester and lutein ester are separated from the protein, and later-period reaction is facilitated.
(2) Cellulase and pectinase: is used for hydrolyzing beta-1, 4-glucosidic bond of cellulose in cell wall to generate soluble polymer and D-glucose, thereby destroying cell wall and being beneficial to dissolving out target product.
(3) And lipase: is a kind of fat hydrolase used for hydrolyzing zeaxanthin ester and lutein ester to obtain zeaxanthin, lutein and fatty acid, and can substitute for traditional strong alkali saponification reaction.
(4) The ultrasonic crushing instrument mainly utilizes the cavitation effect generated by sound waves in liquid, is beneficial to the crushing of cell tissues, has the functions of homogenization, emulsification and defoaming, and can disperse and accelerate the dissolution of substances.
(5) The film scraping flash evaporation instrument utilizes the technical advantages of falling film and film scraping, so that material liquid flows downwards in a film shape along the wall of the heating pipe under the action of the scraper to transfer heat and evaporate, the heat transfer efficiency is high, the evaporation speed is high, the material retention time is short, and the rapid concentration equipment with vacuum direct current, continuous liquid inlet and instant evaporation can be formed.
From examples 1 to 6, it can be seen that the preparation method of zeaxanthin of the present invention adopts a biological enzymolysis method to destroy cell walls, promote the combination of target products and other substances such as proteins, saccharides, etc., is more beneficial to the dissolution of target products and shortens the reaction time, is safe and free from harmful substances, overcomes the disadvantages of more processes, long time consumption and partial harmful substances generated at high temperature in the traditional method, adopts a mild biological method to replace a chemical method of strong alkali saponification, adopts lipase to hydrolyze lutein esters and zeaxanthin esters as free lutein and zeaxanthin, the production safety can be improved, the environmental pollution is reduced, the use of organic solvents is reduced, the stability of the product is ensured, and the application of the ultrasonic pulverizer, the high-pressure device and the wiped film flash evaporator is favorable for improving the yield of the target product, reducing the product loss and improving the product stability.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (8)
1. A preparation method of zeaxanthin is characterized in that fresh marigold flower pulp is diluted by adding water and the pH value is adjusted, hydrolysis is carried out by adding hydrolase, lipase and alkaline protease, zymolyte is crushed by adopting an ultrasonic crusher, gas is introduced and then filtration is carried out, filter residue is extracted for multiple times by using a solvent in a high-pressure device, extract liquor is combined and concentrated to obtain concentrate, and the concentrate is dried in vacuum to obtain the zeaxanthin extract.
2. The method of claim 1, wherein the hydrolytic enzyme is selected from the group consisting of protease, cellulase and pectinase.
3. The method of claim 1, wherein the solvent is one or more of ethanol, n-hexane and acetone.
4. The method of claim 1, wherein the gas is nitrogen.
5. The method of claim 1, wherein the extract is concentrated by a wiped film flash evaporator.
6. The method for preparing zeaxanthin of claim 1, wherein the feed-liquid ratio during extraction is 1: (2-4).
7. The method of claim 1, comprising the steps of:
s1, adding water into the fresh marigold flower pulp, uniformly stirring, adding citric acid to adjust the pH value to 4.0-6.0, heating in a water bath at the temperature of 37-50 ℃, adding protease, cellulase and pectinase, uniformly stirring, and oscillating in a constant-temperature water bath in a dark place to perform enzymolysis reaction;
s2, adjusting the pH value with a sodium hydroxide solution, homogenizing for 3min, adding lipase and alkaline protease, stirring uniformly, reacting on a water bath constant temperature oscillator in a dark place, homogenizing for 2min, adding lipase twice, stirring uniformly, and reacting for 1-2h to obtain a feed liquid;
s3, treating the feed liquid by using an ultrasonic pulverizer;
s4, introducing nitrogen into the treated feed liquid for 20min, heating to 80-150 ℃, reacting for 4h, and filtering;
s5, extracting the filter residue in the S4 for multiple times at normal temperature in a high-pressure device by using a solvent, wherein the extraction pressure is 2-8MPa, the extraction times are 1-3 times, and the material-liquid ratio is 1: (2-4), extracting for 5-10 min;
s6, combining the extraction liquids extracted for many times in the S5, and concentrating by a wiped film flash evaporator to obtain a concentrate;
s7, vacuum drying the concentrate for 2-4h at the vacuum drying temperature of 45-60 ℃ to obtain the zeaxanthin extract.
8. The method as claimed in claim 7, wherein in step S3, when the liquid is treated by the ultrasonic pulverizer, the total working time is 2-10min, the working time is 15S, the interval time is 30S, and the power is 400-.
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| CN110590629A (en) * | 2019-09-29 | 2019-12-20 | 山东天音生物科技有限公司 | Method for separating and purifying lutein from marigold oleoresin |
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