CN113559145A - Walnut green seedcase polyphenol extraction composite agent and extraction method - Google Patents
Walnut green seedcase polyphenol extraction composite agent and extraction method Download PDFInfo
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Abstract
The invention provides a walnut green husk polyphenol extraction compound composition and an extraction method, wherein the compound composition comprises the following components in parts by weight: 20-40 parts of choline chloride serving as a hydrogen acceptor; 10 to 15 parts of a compound serving as a hydrogen donor; 15-20 parts of microencapsulated enzyme complexing agent; 6-12 parts of a coordination agent; 100 to 500 portions of ethanol. The extraction method adopts the extraction composite agent, firstly carries out enzymolysis by adopting the microcapsule embedding enzyme composite agent obtained by self preparation, then centrifugalizes to carry out super carbon dioxide extraction on the supernatant obtained by centrifugation to purify polyphenol in an enzymolysis product, and then further adopts an ethanol mixed solution with a hydrogen acceptor, a hydrogen donor and a coordination agent to further decompose plant cell walls such as walnut green husk lignin, hemilignin and the like in solid residues obtained by centrifugation, thereby improving the further polyphenol extraction in the enzymolysis product residues and effectively improving the polyphenol purification efficiency.
Description
Technical Field
The invention belongs to the technical field of chemical substance purification, and particularly relates to a compound composition for extracting walnut green husk polyphenol and an extraction method.
Background
The green peel outside the walnut nut is the green peel of the walnut. Walnut green husk (also called green dragon skin) is the outermost green skin of walnut shell, and accounts for 5/7 of the total mass of walnut. The walnut green husk is a main byproduct of walnuts, is not effectively utilized at present, and is discarded in field lands or ditches in most cases, so that the environment is polluted and resources are wasted. The exocarpium Juglandis Immaturus contains a large amount of bioactive substances such as ketones, phenols, alkaloids, etc., and has effects of relieving pain, relieving cough, relieving diarrhea, removing toxic substance and killing parasite.
The naphthoquinone compounds in the walnut epicarp mainly comprise 1, 2-naphthoquinone, hydrogenated juglone and glycoside thereof, and various quinone isomers and glycoside compounds such as 5-hydroxy-2-ethoxy-1, 4-naphthoquinone, 3' -di-juglone, 1,4, 8-trihydroxy naphthalene-1-0-beta-D-glucopyranoside and the like can be separated from the walnut epicarp. The walnut is characterized in that the walnut outer fruits contain various phenolic substances, gallic acid, rutin, quercetin, vanillic acid, caffeic acid and the like can be separated and identified from the walnut green husks, the green husks are extracted by alcohol through macroporous resin, the number of the phenols which can be separated can reach 10, and the number of flavonoid components which can reach 20 are mainly flavonol, flavanonol and glycosides thereof.
The method for extracting polyphenol from walnut green peel commonly adopted in the prior art is to extract total polyphenol by drying after obtaining extract by vacuum concentration as disclosed in CN105920089A or to extract polyphenol from walnut green peel by using resin chromatographic column as disclosed in CN105708894A, but the extraction efficiency is low, the operation flow is complicated, and the polyphenol in walnut green peel is not extracted to the maximum extent, so that the waste of raw materials of walnut green peel is caused.
Disclosure of Invention
Aiming at the defects, the invention provides a compound composition for extracting green seedcase polyphenol and an extraction method thereof, wherein the compound composition is prepared by firstly carrying out enzymolysis by using a self-prepared microcapsule embedding enzyme complexing agent, then centrifuging, carrying out super-carbon dioxide extraction on the centrifuged supernatant to purify polyphenol in an enzymolysis product, and then further adopting an ethanol mixed solution containing a hydrogen acceptor, a hydrogen donor and a coordination agent to further decompose plant cell walls such as green seedcase lignin, hemilignin and the like in the centrifuged solid residue, thereby improving the further polyphenol extraction in the enzymolysis product residue and effectively improving the polyphenol purification efficiency.
The invention provides the following technical scheme: a walnut green seedcase polyphenol extraction compound composition comprises the following components in parts by weight:
further, the complexing agent is one or more of EDTA, ethanolamine, triethylamine, oleic acid and sodium citrate.
Further, the raw materials for preparing the microencapsulated enzyme complexing agent comprise the following components in parts by weight:
further, the enzyme complexing agent in the microcapsule enzyme complexing agent is a complex agent formed by cellulase, pectinase and xylanase in a mass ratio of 0.3-0.6: 0.2-0.5: 0.1-0.4.
Further, the thermosensitive polymer includes one or more of poly (N-isopropylacrylamide), polyvinyl alcohol methyl ether, and poly (isopropylacrylamide-acrylic acid).
Further, the compound of the hydrogen donor is one or more of urea, citric acid and lactic acid.
Further, the silane modifier is one or more of dichlorodimethylsilane, octyltrimethoxysilane and methyltrimethoxysilane.
Further, the preparation method of the microencapsulated enzyme complex comprises the following steps:
s1: dissolving the parts by weight of thermosensitive polymer in the parts by weight of acetone to form a thermosensitive polymer oil phase;
s2: mixing the nano silicon dioxide particles with the particle size of 50-60 nm in parts by weight with the silane modifier with the particle size of 20-50 nm in parts by weight, and stirring at 30-40 ℃ for 30-45 min to obtain a silane modified nano silicon dioxide solution;
s3: mixing the heat-sensitive polymer oil phase obtained in the step S1, the silane modified nano-silica solution obtained in the step S2 and the enzyme complexing agent in parts by weight, adding the hyperbranched polyethoxysilane in parts by weight for crosslinking, then adding a mixed solution of water and ethanol in a volume ratio of 1:3, and cleaning for three to five times to obtain the nano-silica/heat-sensitive polymer microcapsule embedding the enzyme complexing agent; in the cleaning process, the crosslinking agent hyperbranched polyethoxysilane is hydrolyzed and becomes amphiphilic from hydrophobic, so that the stable enzyme complexing agent embedded nano silicon dioxide/heat-sensitive polymer microcapsule is formed while the redundant acetone and the crosslinking agent are removed;
s4: and then drying at 25-28 ℃ to obtain the microencapsulated enzyme complexing agent.
The invention also provides a walnut green husk polyphenol extraction method adopting the composite component, which comprises the following steps:
1) picking walnut with green peel, taking the green peel, washing and cleaning the green peel on the surface under clear water, drying at 50-60 ℃, and crushing into walnut green peel powder;
2) dissolving the microcapsule complexing agent in parts by weight in 300-500 ml of distilled water to form a microcapsule complexing agent aqueous solution, placing the walnut green husk powder obtained in the step 1) in the microcapsule complexing agent aqueous solution, standing and reacting at 30-40 ℃ for 1-1.5 h, and then centrifuging at normal temperature;
3) centrifuging the supernatant obtained in the step 2) at the temperature of between 45 and 55 ℃ and under the pressure of between 25 and 30MPa, and adding 25 to 35kg/h of CO2Extracting polyphenol in the supernatant by using ethanol with the volume fraction of 75% and the flow rate of 4-5 mL/g as an entrainer;
4) extracting the solid obtained by centrifuging in the step 2) with the mixed solution of the choline chloride serving as a hydrogen acceptor, the compound serving as a hydrogen donor, the complexing agent and 500-1000 ml of alkali liquor and the ethanol in parts by weight at 75-85 ℃ for 45 min-1 h at a solid-to-liquid ratio of 20: 1-30: 1, then rapidly cooling to room temperature, carrying out vacuum filtration at 0.08-1.5 MPa, and collecting filtrate to constant volume to obtain polyphenol obtained by secondary extraction of the centrifuged solid;
5) collecting the polyphenols obtained in the step 3) and the step 4), establishing a gallic acid standard solution curve, and calculating the extraction amount of the polyphenols in the walnut green seedcase.
Further, the particle size of the walnut green husk powder crushed in the step 1) is 30-70 meshes.
The invention has the beneficial effects that:
1. after the silicon dioxide particles are modified by the silane modifier, the hydrophilic silicon dioxide particles can be changed into hydrophobic silicon dioxide particles to form modified nano silicon dioxide particles, and then the modified nano silicon dioxide particles, the heat-sensitive polymer and the enzyme complexing agent are subjected to polycondensation under the action of the hyperbranched polyethoxysilane as a cross-linking agent to form the nano silicon dioxide/heat-sensitive polymer microcapsule embedding the enzyme complexing agent. The surface-hydrophobic modified nano silica particles can be prepared only by simply regulating and controlling the surface wettability of the solid particle surface, the interaction between oil-water interface solid particles is enhanced by methods such as chemical crosslinking and the like, and the microcapsule with certain mechanical strength is prepared.
2. Cellulase, pectinase and xylanase catalyze and degrade cellulose and colloid in walnut skin, and release polymer phenolic substances tightly combined with the cellulose and the colloid; meanwhile, the permeability of the epidermis is increased by the action of the enzymes, the dissolution of polyphenol substances is facilitated, and the catalysis and dissolution actions are enhanced by the synergistic action of the two enzymes, so that the polyphenol yield is greatly improved.
3. In the process of extracting polyphenol, firstly, the crushed and dried walnut green husk is subjected to enzymolysis by using a microcapsule enzyme complexing agent to extract polyphenol, then the obtained mixture is centrifuged, most of enzymolysis liquid containing polyphenol obtained by enzymolysis is centrifuged to supernatant, then the polyphenol in the enzymolysis liquid is extracted by using a supercritical carbon dioxide extraction mode, for solid obtained by centrifugation, mixed solvent containing hydrogen acceptor, hydrogen donor and ligand is used again to extract solid residue obtained by centrifugation under the condition of alkali liquor, and lignin, hemilignin and cellulose of plant cell walls are further decomposed under the alkali condition through the formed mixed solution with good permeability, so that the defect that the plant cell walls which are targeted and cannot be digested and degraded in the enzyme complexing agent are overcome, and the extraction efficiency of the polyphenol in the residue obtained by enzymolysis of the walnut green husk is further improved.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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 chemical reagents adopted by the invention are all commercially available.
Example 1
The compound composition for extracting the polyphenol from the walnut green seedcase is characterized by comprising the following components in parts by weight:
the microcapsule enzyme complexing agent comprises the following raw materials in parts by weight:
the enzyme complexing agent in the microcapsule enzyme complexing agent is a complex agent formed by cellulase, pectinase and xylanase in a mass ratio of 0.3:0.5: 0.1.
The preparation method of the microencapsulated enzyme complexing agent comprises the following steps:
s1: dissolving 60 parts of poly (N-isopropyl acrylamide) in 25 parts of acetone to form a poly (N-isopropyl acrylamide) oil phase;
s2: mixing 30 parts of nano silica particles with the particle size of 50nm with 20 parts of octyl trimethoxy silane, stirring at 30 ℃ for 45min to obtain a silane modified nano silica solution, and modifying the silica with a hydrophilic surface into hydrophobic nano silica by using the octyl trimethoxy silane;
s3: mixing the poly (N-isopropyl acrylamide) oil phase obtained in the step S1 with the silane modified nano-silica solution obtained in the step S2 and 30 parts of enzyme complexing agent, crosslinking 15 parts of hyperbranched polyethoxysilane, then adding a mixed solution of water and ethanol in a volume ratio of 1:3, and cleaning for three times to obtain a nano-silica/poly (N-isopropyl acrylamide) microcapsule embedding the enzyme complexing agent, wherein the hyperbranched polyethoxysilane serving as the crosslinking agent is hydrolyzed in the cleaning process and becomes amphiphilic from hydrophobic, so that the stable nano-silica/poly (N-isopropyl acrylamide) microcapsule embedding the enzyme complexing agent is formed while removing redundant acetone and the crosslinking agent;
s4: and then drying at 25 ℃ to obtain the microencapsulated enzyme complexing agent.
The embodiment also provides a walnut green seedcase polyphenol extraction method adopting the compound composition, which comprises the following steps:
1) picking walnut with green peel, taking the green peel, washing and brushing the green peel on the surface under clear water, drying at 60 ℃, and crushing into 30-mesh walnut green peel powder;
2) dissolving 20 parts of microcapsule complexing agent in 400ml of distilled water to form microcapsule complexing agent aqueous solution, placing the walnut green husk powder obtained in the step 1) into the microcapsule complexing agent aqueous solution, standing and reacting for 1h at 40 ℃, and then centrifuging at normal temperature;
3) centrifuging the supernatant obtained in the step 2) at 55 ℃ and 25MPa with 25kg/h of CO2Extracting polyphenol in the supernatant with supercritical carbon dioxide at flow rate, entrainer dosage of 5mL/g and ethanol with volume fraction of 75%;
4) extracting the solid obtained by centrifuging in the step 2) with a mixed solution of 40 parts of choline chloride serving as a hydrogen acceptor, 10 parts of lactic acid serving as a hydrogen donor, 6 parts of EDTA complexing agent, 1000ml of alkali liquor and 100 parts of ethanol at 75 ℃ for 45min at a solid-to-liquid ratio of 20:1, then rapidly cooling to room temperature, carrying out vacuum filtration at 1.0MPa, collecting filtrate, and fixing the volume to obtain polyphenol obtained by secondary extraction of the centrifugal solid;
5) collecting the polyphenols obtained in the step 3) and the step 4), establishing a gallic acid standard solution curve, and calculating the extraction amount of the polyphenols in the walnut green seedcase, wherein the unit is mg/g.
Accurately weighing 0.1g of gallic acid, dissolving in 70% methanol solution to prepare 1000 μ g/mL standard stock solution, weighing 1.0, 2.0, 3.0, 4.0 and 5.0mL respectively, adding into 100mL volumetric flask for constant volume to form 10, 20, 30, 40 and 50 μ g/mL gallic acid standard stock solutions, sucking the gallic acid standard stock solutions with different concentrations, performing blank control by using distilled water seat, adding 5.0mL of 10% Fulin phenol actual into a 1.0mL test tube to obtain polyphenol sample to be tested obtained in step 5), shaking uniformly, reacting for 5.0min at room temperature, adding 4.0mL of 7.5% sodium carbonate solution, shaking uniformly, preventing for 60min at room temperature, measuring absorbance at 765nm wavelength, taking the absorbance Y as ordinate, and taking the mass concentration of the gallic acid of 10, 20, 30, 40 and 50 μ g/mL as abscissa, the standard curve is returned to obtain the regression equation of Y-0.0015X +0.007, R2＝0.9989。
Then, calculating the mass concentration of the polyphenol obtained in the step 5) according to the obtained standard curve of the regression equation, further calculating the extraction amount H of the polyphenol in the walnut green seedcase, and calculating according to the following formula:
H＝(10-3×C×V×d)/M
wherein C is the mass concentration of polyphenol obtained according to a standard curve, and the unit is mu g/ml; v is the volume of polyphenol solution used for determination, and the unit is ml; d is the dilution multiple; m is the dry mass of the sample of the walnut green husk adopted in the step 1).
The polyphenol content of the walnut green husk extracted in the embodiment is calculated to be 29.7mg/g, and the polyphenol purity is 98.67%.
Example 2
The embodiment provides a walnut green seedcase polyphenol extraction compound composition, which comprises the following components in parts by weight:
the microcapsule enzyme complexing agent comprises the following raw materials in parts by weight:
the enzyme complexing agent in the microcapsule enzyme complexing agent is a complex agent formed by cellulase, pectinase and xylanase in a mass ratio of 0.45:0.2: 0.4.
The preparation method of the microencapsulated enzyme complexing agent comprises the following steps:
s1: dissolving 50 parts of poly (isopropyl acrylamide-acrylic acid) in 20 parts of acetone to form a poly (isopropyl acrylamide-acrylic acid) oil phase;
s2: mixing 32.5 parts of nano silicon dioxide particles with the particle size of 60nm with 15 parts of dichlorodimethylsilane, stirring at 35 ℃ for 30min to obtain a silane modified nano silicon dioxide solution, wherein the dichlorodimethylsilane modifies silicon dioxide with a hydrophilic surface into hydrophobic nano silicon dioxide;
s3: mixing the poly (isopropyl acrylamide-acrylic acid) oil phase obtained in the step S1 with the silane modified nano-silica solution obtained in the step S2 and 40 parts of enzyme complexing agent, adding 12.5 parts of hyperbranched polyethoxysilane for crosslinking, then adding a mixed solution of water and ethanol with a volume ratio of 1:3, and cleaning for five times to obtain a nano-silica/poly (isopropyl acrylamide-acrylic acid) microcapsule embedding the enzyme complexing agent, wherein the hyperbranched polyethoxysilane serving as the crosslinking agent is hydrolyzed in the cleaning process and becomes amphiphilic from hydrophobicity, so that the stable nano-silica/poly (isopropyl acrylamide-acrylic acid) microcapsule embedding the enzyme complexing agent is formed while removing redundant acetone and the crosslinking agent;
s4: and then drying at 26 ℃ to obtain the microencapsulated enzyme complexing agent.
The embodiment also provides a method for extracting the walnut green seedcase polyphenol by using the compound composition as defined in any one of claims 1 to 7, which is characterized by comprising the following steps:
1) picking walnut with green husk, taking the green husk, washing and brushing the green husk on the surface under clear water, drying at 50 ℃, and crushing into 70-mesh walnut green husk powder;
2) dissolving 20 parts of microcapsule complexing agent in 300ml of distilled water to form microcapsule complexing agent aqueous solution, placing the walnut green husk powder obtained in the step 1) into the microcapsule complexing agent aqueous solution, standing and reacting for 1.2h at 35 ℃, and then centrifuging at normal temperature;
3) centrifuging the supernatant obtained in the step 2) at 48 ℃ and 30MPa with 30kg/h of CO2Extracting polyphenol in the supernatant with supercritical carbon dioxide at flow rate, entrainer dosage of 4.5mL/g and ethanol with volume fraction of 75%;
4) extracting the solid obtained by centrifuging in the step 2) with a mixed solution of 31.5 parts of choline chloride as a hydrogen acceptor, 9.5 parts of citric acid as a hydrogen donor, 10 parts of triethylamine coordination agent, 800ml of alkali liquor and 300 parts of ethanol at 85 ℃ for 55min at a solid-to-liquid ratio of 25:1, then rapidly cooling to room temperature, carrying out vacuum filtration at 0.08MPa, collecting filtrate, fixing the volume, and obtaining polyphenol by secondary extraction of the centrifuged solid
5) Collecting the polyphenols obtained in the step 3) and the step 4), establishing a gallic acid standard solution curve, and calculating the extraction amount of the polyphenols in the walnut green seedcase, wherein the unit is mg/g.
Similarly, the mass concentration of the polyphenol collected in the step 5) is calculated by adopting the standard curve obtained in the example 1, and then the extraction amount of the polyphenol in the walnut green husk is calculated to be 43.8mg/g, and the purity of the polyphenol is calculated to be 99.12%.
Example 3
The compound composition for extracting the polyphenol from the walnut green seedcase provided by the embodiment comprises the following components in parts by weight:
the microcapsule enzyme complexing agent comprises the following raw materials in parts by weight:
the enzyme complexing agent in the microcapsule enzyme complexing agent is a complex agent formed by cellulase, pectinase and xylanase in a mass ratio of 0.6:0.35: 0.25.
The preparation method of the microencapsulated enzyme complexing agent comprises the following steps:
s1: dissolving 50 parts of polyvinyl alcohol methyl ether in 23 parts of acetone to form a polyvinyl alcohol methyl ether oil phase;
s2: mixing 35 parts of nano silicon dioxide particles with 15 parts of methyltrimethoxysilane, stirring at 40 ℃ for 35min to obtain a silane modified nano silicon dioxide solution, wherein the silane modifies silicon dioxide with a hydrophilic surface into hydrophobic nano silicon dioxide;
s3: mixing the polyvinyl alcohol methyl ether oil phase obtained in the step S1 with the silane modified nano-silica solution obtained in the step S2 and 35 parts of enzyme complexing agent, adding 10 parts of hyperbranched polyethoxysilane for crosslinking, then adding a mixed solution of water and ethanol with a volume ratio of 1:3, and cleaning for four times to obtain nano-silica/polyvinyl alcohol methyl ether microcapsules embedded with the enzyme complexing agent, wherein the hyperbranched polyethoxysilane serving as the crosslinking agent is hydrolyzed during cleaning and becomes amphiphilic from hydrophobic, so that the stable nano-silica/polyvinyl alcohol methyl ether microcapsules embedded with the enzyme complexing agent are formed while removing redundant acetone and the crosslinking agent;
s4: and then drying at 28 ℃ to obtain the microencapsulated enzyme complexing agent.
The embodiment also provides a walnut green seedcase polyphenol extraction method adopting the compound composition, which comprises the following steps:
1) picking walnut with green peel, taking the green peel, washing and brushing the green peel on the surface under clear water, drying at 55 ℃, and crushing into 50-mesh walnut green peel powder;
2) dissolving 15 parts of microcapsule complexing agent in 500ml of distilled water to form microcapsule complexing agent aqueous solution, placing the walnut green husk powder obtained in the step 1) into the microcapsule complexing agent aqueous solution, standing and reacting for 1.5h at 30 ℃, and then centrifuging at normal temperature;
3) centrifuging the supernatant obtained in the step 2) at 45 ℃ and 28MPa with 27.5kg/h of CO2Extracting polyphenol in the supernatant with supercritical carbon dioxide at flow rate, entrainer dosage of 4mL/g and ethanol with volume fraction of 75%;
4) extracting the solid obtained by centrifuging in the step 2) with a mixed solution of 20 parts of choline chloride serving as a hydrogen acceptor, 15 parts of urea serving as a hydrogen donor, 500ml of alkali liquor and 500 parts of ethanol at 80 ℃ for 1 hour at a solid-to-liquid ratio of 30:1, then rapidly cooling to room temperature, carrying out vacuum filtration at 1.5MPa, collecting filtrate, fixing the volume, and carrying out secondary extraction on the filtrate to obtain polyphenol as a centrifuged solid
5) Collecting the polyphenols obtained in the step 3) and the step 4), establishing a gallic acid standard solution curve, and calculating the extraction amount of the polyphenols in the walnut green seedcase, wherein the unit is (mg/g).
Similarly, the mass concentration of the polyphenol collected in the step 5) is calculated by adopting the standard curve obtained in the example 1, and then the extraction amount of the polyphenol in the walnut green husk is calculated to be 50.07mg/g, and the purity of the polyphenol is calculated to be 99.34%.
Comparative example 1
The difference between the comparative example and the example 1 is that in the extraction process of the walnut green husk, after the step 2), the supercritical carbon dioxide is not adopted to extract the polyphenol in the supernatant, the supernatant is extracted by only adopting ethanol and then passes through a resin column to extract the polyphenol in the supernatant, the other steps are the same, and the detection shows that the polyphenol content in the comparative example is 18.5mg/g, and the polyphenol purity is 73.36%.
Comparative example 2
The difference between the comparative example and the example 1 is that in the extraction process of the walnut green husk, the enzyme complexing agent adopted in the step 1) is a self-prepared microcapsule for coating, only the composite agent formed by cellulase, pectinase and xylanase with the mass ratio of 0.3:0.5:0.1 is adopted, the other steps and parameters are the same as those of the example 1, and the detection shows that the polyphenol content in the comparative example is 12.3mg/g, and the polyphenol purity is 52.16%.
Comparative example 3
The difference between the comparative example and the example 1 is that in the extraction process of the walnut green husk, the step 3) does not adopt the steps of extracting the solid obtained by centrifuging the step 2) with 40 parts of choline chloride as a hydrogen acceptor, 10 parts of lactic acid as a hydrogen donor, 6 parts of EDTA complexing agent and a mixed solution of 1000ml of alkali liquor and 100 parts of ethanol for 45min at 75 ℃ in a solid-to-liquid ratio of 20:1, and then rapidly cooling to the room temperature;
extracting the centrifuged solid by using ethanol, passing the solid through a resin column, extracting polyphenol in the extracted solution, wherein the rest steps are the same, and the polyphenol content in the comparative example is 17.2mg/g and the polyphenol purity is 65.98% through detection.
The above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Furthermore, those skilled in the art will appreciate that while some embodiments herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the invention and form different embodiments. For example, in the claims above, any of the claimed embodiments may be used in any combination. The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
Claims (10)
1. The compound composition for extracting the polyphenol from the green seedcase of the walnut is characterized by comprising the following components in parts by weight:
2. the walnut green seedcase polyphenol extraction compound composition as claimed in claim 1, wherein the complexing agent is one or more of EDTA, ethanolamine, triethylamine, oleic acid and sodium citrate.
3. The walnut green seedcase polyphenol extraction complex composition as claimed in claim 1, wherein the raw materials for preparing the microencapsulated enzyme complex comprise the following components by weight:
4. the walnut green seedcase polyphenol extraction compound composition as claimed in claim 3, wherein the enzyme complexing agent in the microencapsulated enzyme complexing agent is a compound composition formed by cellulase, pectinase and xylanase in a mass ratio of 0.3-0.6: 0.2-0.5: 0.1-0.4.
5. The walnut green seedcase polyphenol extraction compound composition of claim 3, wherein the thermosensitive polymer comprises one or more of poly (N-isopropyl acrylamide), polyvinyl alcohol methyl ether and poly (isopropyl acrylamide-acrylic acid).
6. The walnut green seedcase polyphenol extraction complex composition of claim 1, wherein the compound of the hydrogen donor is one or more of urea, citric acid and lactic acid.
7. The walnut green seedcase polyphenol extraction composite composition of claim 1, wherein the silane modifier is one or more of dichlorodimethylsilane, octyltrimethoxysilane, and methyltrimethoxysilane.
8. The walnut green seedcase polyphenol extraction complex composition as claimed in claim 3, wherein the preparation method of the microencapsulated enzyme complex comprises the following steps:
s1: dissolving the parts by weight of thermosensitive polymer in the parts by weight of acetone to form a thermosensitive polymer oil phase;
s2: mixing the nano silicon dioxide particles with the particle size of 50-60 nm in parts by weight with the silane modifier with the particle size of 20-50 nm in parts by weight, and stirring at 30-40 ℃ for 30-45 min to obtain a silane modified nano silicon dioxide solution;
s3: mixing the heat-sensitive polymer oil phase obtained in the step S1, the silane modified nano-silica solution obtained in the step S2 and the enzyme complexing agent in parts by weight, adding the hyperbranched polyethoxysilane in parts by weight for crosslinking, then adding a mixed solution of water and ethanol in a volume ratio of 1:3, and cleaning for three to five times to obtain the nano-silica/heat-sensitive polymer microcapsule embedding the enzyme complexing agent;
s4: and then drying at 25-28 ℃ to obtain the microencapsulated enzyme complexing agent.
9. The method for extracting the walnut green seedcase polyphenol by adopting the compound composition as claimed in any one of claims 1 to 7 is characterized by comprising the following steps:
1) picking walnut with green peel, taking the green peel, washing and cleaning the green peel on the surface under clear water, drying at 50-60 ℃, and crushing into walnut green peel powder;
2) dissolving the microcapsule complexing agent in parts by weight in 300-500 ml of distilled water to form a microcapsule complexing agent aqueous solution, placing the walnut green husk powder obtained in the step 1) in the microcapsule complexing agent aqueous solution, standing and reacting at 30-40 ℃ for 1-1.5 h, and then centrifuging at normal temperature;
3) centrifuging the supernatant obtained in the step 2) at the temperature of between 45 and 55 ℃ and under the pressure of between 25 and 30MPa, and adding 25 to 35kg/h of CO2Extracting polyphenol in the supernatant by using ethanol with the volume fraction of 75% and the flow rate of 4-5 mL/g as an entrainer;
4) extracting the solid obtained by centrifuging in the step 2) with the mixed solution of the choline chloride serving as a hydrogen acceptor, the compound serving as a hydrogen donor, the complexing agent and 500-1000 ml of alkali liquor and the ethanol in parts by weight at 75-85 ℃ for 45 min-1 h at a solid-to-liquid ratio of 20: 1-30: 1, then rapidly cooling to room temperature, carrying out vacuum filtration at 0.08-1.5 MPa, and collecting filtrate to constant volume to obtain polyphenol obtained by secondary extraction of the centrifuged solid;
5) collecting the polyphenols obtained in the step 3) and the step 4), establishing a gallic acid standard solution curve, and calculating the extraction amount of the polyphenols in the walnut green seedcase.
10. The method for extracting walnut green husk polyphenol as claimed in claim 9, wherein the particle size of the crushed walnut green husk powder in step 1) is 30-70 mesh.
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