CN107552536B - Comprehensive extraction process of wine waste residues - Google Patents

Abstract

The invention relates to a comprehensive extraction process of wine waste residues, which comprises the following specific steps: firstly, drying the wine waste residue, screening wine mud and tartar sediment, crushing, and performing subcritical extraction for 60-80 min by using liquefied petroleum gas as an extracting agent at the temperature of 40-45 ℃ and the pressure of 0.2-1.0 Mpa for 0.5-0.6 kg/L to obtain degreased meal and grape grease respectively; and (3) controlling the vacuum degree of the grape oil to be 1-1.3 Pa and the temperature to be 150-160 ℃, purifying the grape oil by using a molecular distiller to obtain the grape oil, wherein short-chain free fatty acid separated by distillation can be independently recovered, and then extracting procyanidine and tannin, extracting grape pigment, extracting tartaric acid, extracting active carbon and finally extracting feed. The method has the greatest innovation points that water replaces an organic solvent for comprehensive extraction, sulfurous acid is used for depolymerizing high polymeric proanthocyanidins, ultrafiltration is used for purifying in cooperation with resin chromatography, and nanofiltration dealcoholization replaces distillation to recover ethanol.

Description

Comprehensive extraction process of wine waste residues

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a comprehensive extraction process of wine waste residues.

Background

The grape grease contains rich VA, VD, VE, VK and linoleic acid, has the effects of reducing blood fat, preventing cardiovascular and cerebrovascular diseases, removing free radicals, resisting allergy, promoting blood circulation and the like, can enhance skin elasticity, slow wrinkles, lose weight and resist radiation, is called as a noble product of food oil, is beautiful in beauty and beauty in cosmetics and blood fat scavenger in health care products, and is mainly applied to the fields of food, cosmetics and health care products;

procyanidine can effectively remove redundant oxygen free radicals in a human body, can obviously improve the anti-aging capability of the human body, improves the cardiovascular function, prevents hypertension, inhibits cancers, enhances the anti-allergic capability of the human body, has good treatment effects on arteriosclerosis, gastric ulcer, cataract, diabetes, heart disease, arthritis and the like, is widely applied to the fields of medicines, health products, cosmetics and foods, has the reputations of 'killer of aging and cancer', 'oral cosmetics' and 'skin vitamins', and becomes a wonderful market of contemporary biological raw materials;

tannin has the effects of resisting oxidation, viruses, tumors, bacteria, cardiovascular and cerebrovascular diseases and the like, is an active ingredient of the traditional medicine, and has quite wide application prospect in the fields of medicine, food, cosmetics, petroleum, leather and printing and dyeing;

grape pigment is a food natural pigment recognized in the world, has the effects of delaying aging, enhancing resistance, promoting blood circulation and the like, and is sold well because synthetic pigments are forbidden in many countries. Therefore, the optimization of the comprehensive extraction process of the wine waste residue has great significance for promoting the benign development of the derived products in the grape industry.

The research and development of procyanidine in China starts in 1986, enterprises such as Zhixing science and technology company, Tianshan mother-of-the-world biotechnology company, century Tianyun biology company and the like exist in Xinjiang at present, procyanidine and grape seed oil are extracted by utilizing grape seeds, a relatively traditional production process is adopted, and the process flow is as follows:

the traditional extraction process has the following disadvantages:

(1) the ethanol solvent is used in a large amount, so that the ethanol solvent is easy to volatilize, flammable and explosive, has potential safety hazards in production, has the loss rate of the organic solvent generally more than or equal to 20 percent, not only has high cost, but also brings pollution;

(2) the cell wall breaking speed is slow and incomplete by depending on the permeation of an ethanol solvent, and the dissociation of the combination of the procyanidine, the protein and the cellulose is incomplete, so the extraction rate is not very high;

(3) 60% ethanol contains 40% water, and inevitably has protein, pectin and other impurities dissolved in the extract, although most of the impurities can be removed by ethyl acetate purification, the ethyl acetate is easy to remain and has low toxicity;

(4) the organic solvent is recovered by heating and distilling twice, so that the energy consumption is high, and the oligomeric proanthocyanidins are heated for a long time and are difficult to be degraded more, so that the content of the oligomeric proanthocyanidins is influenced;

(5) only grape seed oil and procyanidine are extracted, the maximum utilization of raw material resources is not achieved, and other components are discharged as wastes, so that the environment is polluted, and the additional value is reduced;

(6) and impurities are not completely removed, so that the product purity is not high, and the product price cannot be sold in the market, so that developed countries often import crude products from China and sell the crude products after the crude products are processed into fine products.

Disclosure of Invention

The invention aims to provide a safe, efficient and low-cost comprehensive extraction process for wine waste residues.

A comprehensive extraction process of wine waste residues comprises the following specific steps:

the first step is as follows: extraction of grape oil

a. After drying the wine waste residue, screening wine mud and tartar sediment, crushing and weighing the wine waste residue by a crusher, and performing subcritical extraction for 60-80 min at the temperature of 40-45 ℃ and the pressure of 0.2-1.0 Mpa by using liquefied petroleum gas as an extracting agent for controlling the stacking degree to be 0.5-0.6 kg/L to obtain degreased meal and grape grease respectively;

b. controlling the vacuum degree of the grape oil to be 1-1.3 Pa and the temperature to be 150-160 ℃, and purifying the grape oil by using a molecular distiller to obtain the grape oil, wherein short-chain free fatty acid separated by distillation can be independently recovered;

the second step is as follows: extracting procyanidin and tannin

a. Adding 3-5 times of water into the degreased meal in a microwave extraction tank, adjusting the pH = 2-5, performing microwave treatment for 90s at the power of 650W and the frequency of 2450MHz, adding 0.4-1% of complex enzyme, controlling the temperature at 40-60 ℃, performing enzymolysis for 1.5h, heating to 80 ℃, preserving the temperature for 15min, and performing filter pressing to collect filtrate;

b. adding 0.5 time of water into filter residues, controlling the temperature at 40-60 ℃, depolymerizing for 10-30 min, adding 3-5 times of water, adjusting the pH to be 2-5, performing microwave treatment for 60s at the power of 650W and the frequency of 2450MHz, performing filter pressing to collect filtrate, and washing the filter residues until the water is nearly colorless;

c. adding 3-5 times of water into filter residues, adjusting the pH to be = 2-5, performing microwave treatment for 60s at power of 650W and frequency of 2450MHz, performing filter pressing to collect filtrate, and combining the filtrate and washing water for 3 times;

d. adding 0.03% of ZTC-A, dissolving in water at a ratio of 1%, and stirring for 5 min; adding 0.06% of ZTC-B, dissolving in water for 1%, and stirring for 30 min; centrifuging to remove precipitates, cooling the clear liquid to normal temperature, performing ultrafiltration by 10-30 ten thousand D, collecting a permeate, performing nanofiltration by 1-2Kd, concentrating to 1/5, and collecting a concentrated solution and a permeate respectively;

e. adjusting the pH of the concentrated solution to be 2-5, controlling the flow rate of 3BV/h, flowing through an ADS-8 resin chromatographic column, concentrating the column passing solution to 20%, washing with deionized water until the effluent is colorless, and combining the effluent with the column passing solution for later use; controlling the flow rate of 2BV/h, carrying out gradient elution by using 20-80% ethanol, and collecting a procyanidine elution peak and a tannin elution peak in a grading manner;

f. respectively carrying out nanofiltration on a procyanidin elution peak and a tannin elution peak through dealcoholization membranes to recover ethanol, concentrating until the concentration is less than or equal to 10%, adjusting the pH value to be = 2-5, controlling the temperature to be 180/70 ℃, and directly carrying out spray drying to obtain procyanidin dry powder and tannin dry powder respectively, wherein the ethanol can be repeatedly reused for elution;

the third step: extraction of grape pigment

a. Combining the 1-2 KDr permeate liquid from which the procyanidins are extracted with the column passing liquid, adjusting the pH to be = 2-5, performing reverse osmosis at normal temperature, concentrating to 1/5, and respectively collecting the concentrated liquid and the permeate liquid, wherein the permeate liquid is pure water and can be reused for extraction of the procyanidins and the tannin;

b. controlling the flow rate of the concentrated solution at 1.5BV/h, flowing through an XAD-7 resin chromatographic column, concentrating to 20%, and washing with deionized water until the effluent is transparent; controlling the flow rate to be 1.5BV/h, carrying out gradient elution by using 30-90% ethanol, and collecting grape pigment elution peaks;

c. carrying out nanofiltration on grape pigment elution peaks through a dealcoholization membrane to recover ethanol, concentrating until the ethanol content is less than or equal to 10%, adjusting the pH of a concentrated solution to be = 2-5, controlling the temperature to be 180/70 ℃, and directly spraying the concentrated solution to obtain grape pigment dry powder, wherein the ethanol can be reused for re-elution;

the fourth step: extraction of tartaric acid

a. Mixing the lees and the tartaric acid precipitate, adding 5 times of water, adjusting the pH to be =3, controlling the temperature to be 40-60 ℃, stirring and extracting for 0.5h, centrifuging, collecting filtrate, and combining the filtrate with the chromatography waste liquid for extracting the grape pigment;

b. adjusting the pH of the combined solution to be 2-5, controlling the flow rate to be 4-5 BV/h, allowing the combined solution to flow through a D301R resin chromatographic column, adsorbing the combined solution to the end point, and washing the combined solution with water until the pH is 2-5; controlling the flow rate to be 2-3 BV/h, carrying out gradient elution by using 0.1-1% sodium hydroxide solution, and collecting a tartaric acid elution peak;

c. adjusting the pH of a tartaric acid elution peak to be 2-5, carrying out 200-500D nanofiltration, concentrating to be less than or equal to 10%, controlling the temperature to be 180/70 ℃, and directly spraying to dry to obtain tartaric acid dry powder;

the fifth step: extraction of activated carbon

a. Adding 0.5-1 mol/L sodium hydroxide solution which is 2 times of the grape skin seed stalk residue, controlling the temperature to be 90-95 ℃, stirring for alkaline hydrolysis for 1h, filtering, and washing with water to be neutral; adding 1-5 times of 1-5% acid solution, controlling the temperature to be 40-60 ℃, stirring and modifying for 0.5h, and filtering to collect filter residues;

b. controlling the power of 720W and the frequency of 2450MHz, carbonizing and activating filter residue by microwave equipment for 12min, washing the filter residue with deionized water to be neutral, filtering, drying at 105 ℃ and drying by a dryer to obtain activated carbon dry powder with the particle size of less than or equal to 80 meshes;

a sixth step: extracted feed

Mixing various waste liquids, and performing 200-500D nanofiltration desalination; concentrating to no more than 1/10, adding protein precipitate obtained by ZTC clarifier, adjusting pH to =7, controlling temperature to 200/70 deg.C, and directly spray drying to obtain feed dry powder.

The invention carries out optimization design aiming at the defects of the prior art, and the key points are that subcritical extraction degreasing is firstly carried out, deacidification is carried out by a molecular distillation technology, and then microwave strengthening is carried out to break cell walls and enzymolysis strengthening is carried out to dissociate protein; the innovation points are that water is used for replacing an organic solvent for comprehensive extraction, sulfurous acid is used for depolymerizing high polymeric proanthocyanidins, ultrafiltration is used for purifying in cooperation with resin chromatography, and nanofiltration is used for dealcoholizing to replace distillation to recover ethanol; the invention adopts comprehensive extraction, changes waste into valuable, greatly increases the utilization rate and the added value of raw materials, and reduces the pollution emission.

Detailed Description

The technical solution of the present invention is described in detail below.

First, design basis

1. Ingredients of raw materials

The waste residues of the prepared grape wine are different according to the grape types and different processes, and are about 20-23% of the grape wine, wherein 7.5-9% of stems removed in grape crushing, 2.5-3.5% of squeezed peel and seed residues, 10-12% of settled wine lees obtained by pouring wine (containing seeds, peel and lees) and 0.05-0.06% of tartaric precipitation in a stable stage are obtained.

After the waste residues of the grape wine are screened, 28-30% of grape seeds, 42-45% of grape peel residues, 23-25% of grape stem stems, and 5-8% of tartar sediment and wine lees can be obtained respectively;

the dry grape seeds contain 12,25% of water, 11.44% of protein, 13-17% of fat, 35.6% of fiber, 3.86% of free procyanidine and 6.73% of protein and cellulose combined procyanidine;

the dry grape skin contains 6.53% of water, 12.55% of protein, 3.93% of fat, 27.9% of fiber, 1.83% of free procyanidine, 7.24% of protein and cellulose combined procyanidine and 5-8% of grape pigment (mainly in red grape skin);

the stem of the dried grape contains 12.44% of water, 10.58% of protein, 5.94% of fat, 29.8% of fiber, 1.52% of free procyanidin and 6.27% of protein and cellulose combined procyanidin;

most of the free procyanidin is oligomeric procyanidin, most of the combined procyanidin is polymeric procyanidin, and about 40% of the free procyanidin in the fermented grape seeds and grape skins is dissolved in the wine in the process of making the wine.

2. Main properties

Grape oil and fat: the main components are unsaturated acids such as linoleic acid and linolenic acid, sterol, resveratrol and the like, wherein the content of linoleic acid is 65-75%, the content of free fatty acid is 2-3%, the density is less than that of water, and the boiling point is low;

dissolving in petroleum ether, diethyl ether, anhydrous ethanol, acetone, ethyl acetate, dichloroethane, propane + butane fluid, supercritical CO2 fluid, etc.;

hydrolytic rancidity can occur under the action of acid, alkali and enzyme, wherein acid hydrolysis is reversible, and alkali hydrolysis is irreversible, namely saponification;

is polluted by microorganisms, is unstable when heated or is contacted with air for a long time, is easy to oxidize and rancid, and generates low-grade fatty acid; the flavor is deteriorated and the nutritive value is lowered.

Procyanidin: procyanidin (PC) structural unit is a polyphenol mixture polymerized by 3 aromatic rings and different amounts of catechol, epicatechol and gallic acid, wherein Oligomeric Procyanidin (OPC) with polymerization degree of less than or equal to 4, oligomeric procyanidin (PPC) with molecular weight of less than or equal to 5000 and Polymeric Procyanidin (PPC) with polymerization degree of more than or equal to 5 have molecular weight of more than or equal to 5000;

the oligomeric proanthocyanidins in the grapes are less, but the activity is higher and the water solubility is good; high polymeric proanthocyanidins are more, but the activity is lower, and the water solubility is poor; the free procyanidine is less, and the procyanidine combined with protein, fiber and monosaccharide by hydrogen bonds is more;

has strong polarity, is easily soluble in polar solvents such as water, methanol, ethanol, acetone, ethyl acetate, dilute acid, dilute alkali and the like, has solubility increased along with temperature rise, has a maximum absorption peak at visible light of 520nm, and is difficult to dissolve in non-polar solvents such as diethyl ether, petroleum ether, chloroform, benzene and the like;

because molecules contain more phenolic hydroxyl groups and alcoholic hydroxyl groups, the compound has the reaction characteristics of phenol and alcohol, such as electrophilic substitution, oxidation and complex reaction, and salting-out;

heating under acidic condition, the high polymeric procyanidin can be depolymerized into oligomeric procyanidin, and if the acidity is too strong or oxidized, the oligomeric procyanidin can be further degraded into monomers such as anthocyanidin, catechol and epicatechol, and is unstable to light, heat, alkalinity and oxidant;

dissolving in water, generally presenting the characteristics of sol, wherein the colloid is negatively charged, is stored at normal temperature for at most 3 days, at most 24h at 50 ℃ and at most 12h at 90 ℃, otherwise, the color deepens due to oxidation and the content of procyanidine is reduced.

Tannin: tannin is a polyphenol polymerization mixture with a complex molecular structure, and is divided into two types, namely condensed tannin which is a derivative polymerized by flavanol and catechol or benzenetriol and belongs to procyanidine; secondly, the hydrolyzed tannin is gallic acid ester or tannic acid ester of glucose, and belongs to ester polyphenol;

the grape is mainly condensed tannin with the molecular weight of 500-3000, is difficult to hydrolyze, is unstable to illumination, heating, strong alkalinity and oxidizing agent, can be dehydrated and condensed into spirorubin (also called tanned red) which is not easy to dissolve in water, and has the molecular weight of more than or equal to 3000; contains phenolic hydroxyl, belongs to acidic substances, is easily dissolved in polar solvents such as water, methanol, ethanol, acetone and the like, has the solubility increased along with the temperature rise, has the maximum absorption peak at the visible light of 750nm, and is not easily dissolved in non-polar solvents such as ether, chloroform, benzene and the like; the polymer is easy to combine with protein, alkaloid, polysaccharide and gelatin to precipitate, the convergence is particularly obvious at the isoelectric point of the protein, the polymerization degree is less than or equal to 10, the convergence is increased along with the increase of the polymerization degree, the polymerization degree is more than or equal to 10, and the convergence is reduced along with the increase of the polymerization degree; can carry out complex reaction with various metal ions, has an amphiphilic structure, can carry out more derivatization reactions, and has reduction property. Can be salted out.

Grape pigment: the grape pigment has a structural unit of 3 aromatic rings, is combined with saccharides in grapes to form anthocyanin, has various isomers, belongs to flavonoid, is natural, safe, free of toxic and side effects, has a color value depending on the content of anthocyanin and a relative molecular weight of 287.2, and has a great misdistinguishing effect of using anthocyanin as procyanidine in the market; the anthocyanidin is easily dissolved in polar solvents such as water, acid and alkali, methanol, ethanol, acetone and the like, has a maximum absorption peak at 560nm of visible light, is slightly dissolved in weak polar solvents such as ethyl acetate, acetic acid and the like, and is insoluble in non-polar solvents such as diethyl ether, petroleum ether, trichloromethane, grease and the like; the anthocyanin is purple red with the pH value less than or equal to 3, is light purple red with the pH =3-6, is red with the pH =6-8, is blue purple or even blue-green with the pH value more than or equal to 8, is easy to be condensed with tannin to be tannin anthocyanin polymer, has stable color tone of the combined anthocyanin, and does not have color change reaction along with the change of the pH value. Namely the color assisting function; iron ions are easy to take brown reaction, and are easy to generate blue precipitates with alkaline lead acetate, can be adsorbed by activated carbon, are basically stable to ultraviolet rays at the temperature of less than or equal to 90 ℃, and are relatively slow to degrade although being relatively unstable to oxidizing agents and reducing agents; is stable to acidity and saccharides, wherein acid can prevent anthocyanin from hydrolysis, but peracid such as hydrochloric acid is more than or equal to 0.12mol/L can hydrolyze acylated anthocyanin.

3. Principal standard

Grape oil and fat: the method can refer to the national GB/T22487-2008 standard, the market price mainly depends on the content of linoleic acid, and the main indexes are as follows:

procyanidin: the procyanidine does not have a national quality standard at present, but quite a lot of enterprises mix the procyanidine content with the oligomeric procyanidine content, in order to better meet the international market, the proposed standard proposed by Tianjin peak natural product research and development company is adopted at present, wherein the standard 1 is the trade standard of the mainstream product of the procyanidine in the international market, countries such as Europe, America, Japan and Korean mostly adopt the standard, the standard 2 is the pronoun of the low-end product of the procyanidine in the international market, which is called grape seed extract, the underdeveloped countries mostly adopt the standard, and the standard system is as follows:

detecting items	Standard 1	Standard 2	Detection method
				Appearance of the product	Light red powder	Reddish brown powder	Sense organ
Taste (0.1%)	Astringent taste	Astringent taste	Sense organ
				Polyphenol content	≥95%	——	CAE
Content of procyanidin	——	≥95%	Batc-Smith
				Content of OPC	≥80%	≤40%	HPLC
Water insoluble substance	≤5%	≤6%	1%，25℃
				Residue of red heat	≤2%	≤2%	CHP2000
Loss on drying	≤5%	≤5%	CHP2000
				pH (4%)	2.5-4.5	2.5-4.5	CHP2000
Heavy metals	≤10mg/l	≤10mg/l	CHP2000
				Total number of bacteria	≤1000CFU/g	≤1000CFU/g	CHP2000

Tannin: the standard comprises industrial, food and pharmaceutical grades, wherein China only has the industrial grade GB5308-1985 standard at present, the food grade can be selected from American food FCC-IV standard references, the pharmaceutical grade can be selected from British 1993 edition and American 1995 pharmacopoeia standard references, wherein the industrial grade standard is as follows:

detecting items	First stage	Second stage	Three-stage
				Appearance of the product	Light yellow and light brown powder	Light yellow and light brown powder	Light yellow and light brown powder
Content%	81	78	75
				Loss on drying%	9	9	9
Water insoluble matter%	0.6	0.8	1.0
				0.5% concentration color grade	≤2	≤3	≤4

Grape pigment: currently, only GB is sent to the approval standard: a purple red powder; the 1% solution A525 is more than or equal to 4; the drying weight loss is less than or equal to 12 percent; the burning residue is less than or equal to 13 percent; the PH value is more than or equal to 3; arsenic is less than or equal to 2 ppm; lead is less than or equal to 5 ppm; heavy metal is less than or equal to 20 ppm; alkaline and other acidic pigments are negative.

A comprehensive extraction process of wine waste residues comprises the following specific steps:

1. extraction of grape oil

(1) After drying the wine waste residue, screening wine mud and tartar sediment, crushing and weighing the wine waste residue by a crusher, and performing subcritical extraction for 60-80 min by using liquefied petroleum gas (propane and butane) as an extracting agent and controlling the stacking degree to be 0.5-0.6 kg/L, the temperature to be 40-45 ℃, the pressure to be 0.2-1.0 Mpa, so as to respectively obtain degreased meal and grape grease;

(2) and controlling the vacuum degree of the grape oil to be 1-1.3 Pa and the temperature to be 150-160 ℃, and purifying the grape oil by using a molecular distiller to obtain the grape oil meeting the first-level standard, wherein the short-chain free fatty acid separated by distillation can be independently recovered.

The yield of the grape oil obtained by the process is more than or equal to 60g/kg of grape wine residues, the grape oil meets the first-level oil standard, the linoleic acid content is more than or equal to 75%, and the grape oil is high-quality oil.

2. Extracting procyanidin and tannin

(1) Adding 3-5 times of water into the degreased meal in a microwave extraction tank, adjusting the pH = 2-5, performing microwave treatment for 90s at the power of 650W and the frequency of 2450MHz, adding 0.4-1% of complex enzyme, performing enzymolysis for 1.5h at the temperature of 40-60 ℃, heating to 80 ℃, preserving the temperature for 15min, and performing filter pressing to obtain filtrate;

(2) adding 0.5 time of water into filter residues, controlling the temperature to be 40-60 ℃, depolymerizing for 10-30 min, adding 3-5 times of water, adjusting the pH to be 2-5, performing microwave treatment for 60s at the power of 650W and the frequency of 2450MHz, performing filter pressing to collect filtrate, and washing the filter residues until the water is nearly colorless;

(3) adding 3-5 times of water into filter residues, adjusting the pH to be = 2-5, performing microwave treatment for 60s at power of 650W and frequency of 2450MHz, performing filter pressing to collect filtrate, and combining the filtrate and washing water for 3 times;

(4) adding 0.03% of ZTC-A (dissolved in water and added at a concentration of 1%), stirring for 5min, adding 0.06% of ZTC-B (dissolved in water and added at a concentration of 1%), stirring for 30min, centrifuging to remove precipitates, cooling the clear solution to normal temperature, performing ultrafiltration by 10-30 ten thousand D, collecting a permeate, performing nanofiltration by 1-2Kd, concentrating to 1/5, and respectively collecting a concentrated solution and a permeate;

(5) adjusting the pH of the concentrated solution to be 2-5, controlling the flow rate of 3BV/h, flowing through an ADS-8 resin chromatographic column, passing through the concentrated solution of the column to 20%, washing with deionized water until the effluent is colorless (combined with the solution passing through the column for use), controlling the flow rate of 2BV/h, performing gradient elution with 20-80% ethanol, and collecting a procyanidin elution peak and a tannin elution peak in a grading manner;

(6) and (3) respectively carrying out nanofiltration on the procyanidin elution peak and the tannin elution peak through dealcoholization membranes to recover ethanol, concentrating until the concentration is less than or equal to 10%, adjusting the pH value to be = 2-5, controlling the temperature to be 180/70 ℃, and directly carrying out spray drying to obtain procyanidin dry powder and tannin dry powder respectively, wherein the ethanol can be repeatedly reused for elution.

The yield of procyanidin in the process is 25g/kg of grape wine residues, wherein the content of OPC is more than or equal to 85 percent, the content of polyphenol is more than or equal to 95 percent, and the quality of the grape wine residues can meet the international market; the yield of the tannin is more than or equal to 40 g/kg of the wine residues, wherein the purity is more than or equal to 90 percent, and the quality can reach the pharmaceutical grade.

3. Extraction of grape pigment

(1) Mixing the 1-2KD permeate liquid from which the procyanidins are extracted with the column passing liquid, adjusting the pH to be = 2-5, performing reverse osmosis at normal temperature, concentrating to 1/5, and respectively collecting the concentrated liquid and the permeate liquid, wherein the permeate liquid is pure water and can be reused for extraction of the procyanidins and the tannin;

(2) controlling the flow rate of the concentrated solution at 1.5BV/h, flowing through an XAD-7 resin chromatographic column, concentrating to 20%, and washing with deionized water until the effluent is transparent; controlling the flow rate to be 1.5BV/h, carrying out gradient elution by using 30-90% ethanol, and collecting grape pigment elution peaks;

(3) and (3) carrying out nanofiltration on the grape pigment elution peak through a dealcoholization membrane to recover ethanol, concentrating until the ethanol is less than or equal to 10%, adjusting the pH of the concentrated solution to be = 2-5, controlling the temperature to be 180/70 ℃, and directly spraying the solution to obtain grape pigment dry powder, wherein the ethanol can be reused for re-elution.

The yield of the grape pigment in the process is more than or equal to 20g/kg of grape wine residue, the purity is more than or equal to 90 percent, the color value is more than or equal to 260, and the quality can reach food grade.

4. Extraction of tartaric acid

(1) Mixing the lees and the tartaric acid precipitate, adding 5 times of water, adjusting the pH to be =3, controlling the temperature to be 40-60 ℃, stirring and extracting for 0.5h, centrifuging, collecting filtrate, and combining the filtrate with the chromatography waste liquid for extracting the grape pigment;

(2) adjusting the pH of the combined solution to be 2-5, controlling the flow rate to be 4-5 BV/h, allowing the combined solution to flow through a D301R resin chromatographic column, adsorbing the combined solution to the end point, and washing the combined solution with water until the pH is 2-5; controlling the flow rate to be 2-3 BV/h, carrying out gradient elution by using 0.1-1% sodium hydroxide solution, and collecting a tartaric acid elution peak;

(3) adjusting the pH of a tartaric acid elution peak to be 2-5, carrying out 200-500D nanofiltration, concentrating to be less than or equal to 10%, controlling the temperature to be 180/70 ℃, and directly spraying to dry to obtain tartaric acid dry powder.

The yield of tartaric acid in the process is more than or equal to 15g/kg of wine waste residue, no calcium ions and sulfate radicals are left, and the quality can reach food grade.

5. Extraction of activated carbon

(1) Adding 0.5-1 mol/L sodium hydroxide solution which is 2 times of the grape skin seed stalk residue, controlling the temperature to be 90-95 ℃, stirring for alkaline hydrolysis for 1h, filtering, and washing with water to be neutral; adding 1-5 times of 1-5% acid solution, controlling the temperature to be 40-60 ℃, stirring and modifying for 0.5h, and filtering to collect filter residues;

(2) controlling the power of 720W and the frequency of 2450MHz, carbonizing and activating filter residue by microwave equipment for 12min, washing the filter residue with deionized water to be neutral, filtering, drying at 105 ℃ and drying by a dryer to obtain the active carbon dry powder with the particle size of less than or equal to 80 meshes.

The yield of the activated carbon in the process is more than or equal to 150g/kg of the wine waste residue, meets the first-grade requirement in the national standard GB-13804-1992, and is the high-quality activated carbon special for wine decolorization.

6. Extracted feed

Mixing various waste liquids, and performing 200-500D nanofiltration desalination. Concentrating to no more than 1/10, adding protein precipitate obtained by ZTC clarifier, adjusting pH to =7, controlling temperature to 200/70 deg.C, and directly spray drying to obtain feed dry powder.

The yield of the dry powder of the feed is more than or equal to 100g/kg of the wine waste residue, and the dry powder of the feed is high-quality feed due to good water solubility and the protein content of more than or equal to 60%.

Compared with the existing process, the extraction process has the advantages of less organic solvent, low cost and good safety; the extraction yield is high, the purity is high, the utilization rate of raw material resources is high, and the best use can be made of things; changing waste into valuable, easily reaching the standard of pollutant discharge and having obvious economic benefit. The comparison is shown in the following table:

Claims (1)

1. a comprehensive extraction process of wine waste residue is characterized in that: the extraction process comprises the following specific steps:

the first step is as follows: extraction of grape oil

a. After drying the wine waste residue, screening wine mud and tartar sediment, crushing and weighing the wine waste residue by a crusher, and performing subcritical extraction for 60-80 min at the temperature of 40-45 ℃ and the pressure of 0.2-1.0 Mpa by using liquefied petroleum gas as an extracting agent for controlling the stacking degree to be 0.5-0.6 kg/L to obtain degreased meal and grape grease respectively;

b. controlling the vacuum degree of the grape oil to be 1-1.3 Pa and the temperature to be 150-160 ℃, and purifying the grape oil by using a molecular distiller to obtain the grape oil, wherein short-chain free fatty acid separated by distillation is independently recovered;

the second step is as follows: extracting procyanidin and tannin

a. Adding 3-5 times of water into the degreased meal in a microwave extraction tank, adjusting the pH = 2-5, performing microwave treatment for 90s at the power of 650W and the frequency of 2450MHz, adding 0.4-1% of complex enzyme, controlling the temperature at 40-60 ℃, performing enzymolysis for 1.5h, heating to 80 ℃, preserving the temperature for 15min, and performing filter pressing to collect filtrate;

b. adding 0.5 time of water into filter residues, controlling the temperature at 40-60 ℃, depolymerizing for 10-30 min, adding 3-5 times of water, adjusting the pH to be 2-5, performing microwave treatment for 60s at the power of 650W and the frequency of 2450MHz, performing filter pressing to collect filtrate, and washing the filter residues until the water is nearly colorless;

c. adding 3-5 times of water into filter residues, adjusting the pH to be = 2-5, performing microwave treatment for 60s at power of 650W and frequency of 2450MHz, performing filter pressing to collect filtrate, and combining the filtrate and washing water for 3 times;

d. adding 0.03% of ZTC-A, dissolving in water at a ratio of 1%, and stirring for 5 min; adding 0.06% of ZTC-B, dissolving in water for 1%, and stirring for 30 min; centrifuging to remove precipitates, cooling the clear liquid to normal temperature, performing ultrafiltration by 10-30 ten thousand D, collecting a permeate, performing nanofiltration by 1-2Kd, concentrating to 1/5, and collecting a concentrated solution and a permeate respectively;

e. adjusting the pH of the concentrated solution to be 2-5, controlling the flow rate of 3BV/h, flowing through an ADS-8 resin chromatographic column, concentrating the column passing solution to 20%, washing with deionized water until the effluent is colorless, and combining the effluent with the column passing solution for later use; controlling the flow rate of 2BV/h, carrying out gradient elution by using 20-80% ethanol, and collecting a procyanidine elution peak and a tannin elution peak in a grading manner;

f. respectively carrying out nanofiltration on a proanthocyanidin elution peak and a tannin elution peak through dealcoholization membranes to recover ethanol, concentrating until the concentration is less than or equal to 10%, adjusting the pH value to be = 2-5, controlling the temperature to be 180/70 ℃, and directly carrying out spray drying to obtain proanthocyanidin dry powder and tannin dry powder respectively; wherein the ethanol is repeatedly reused for elution;

the third step: extraction of grape pigment

a. Mixing the 1-2KD permeate liquid from which the procyanidine is extracted with the column passing liquid, adjusting the pH to be = 2-5, performing reverse osmosis at normal temperature, concentrating to 1/5, and respectively collecting the concentrated liquid and the permeate liquid, wherein the permeate liquid is pure water and is reused for extracting the procyanidine and the tannin;

b. controlling the flow rate of the concentrated solution at 1.5BV/h, flowing through an XAD-7 resin chromatographic column, concentrating to 20%, and washing with deionized water until the effluent is transparent; controlling the flow rate to be 1.5BV/h, carrying out gradient elution by using 30-90% ethanol, and collecting grape pigment elution peaks;

c. carrying out nanofiltration on grape pigment elution peaks through a dealcoholization membrane to recover ethanol, concentrating until the ethanol content is less than or equal to 10%, adjusting the pH of a concentrated solution to be = 2-5, controlling the temperature to be 180/70 ℃, and directly spraying the solution to obtain grape pigment dry powder, wherein the ethanol is reused for elution;

the fourth step: extraction of tartaric acid

a. Mixing the lees and the tartaric acid precipitate, adding 5 times of water, adjusting the pH to be =3, controlling the temperature to be 40-60 ℃, stirring and extracting for 0.5h, centrifuging, collecting filtrate, and combining the filtrate with the chromatography waste liquid for extracting the grape pigment;

b. adjusting the pH of the combined solution to be 2-5, controlling the flow rate to be 4-5 BV/h, allowing the combined solution to flow through a D301R resin chromatographic column, adsorbing the combined solution to the end point, and washing the combined solution with water until the pH is 2-5; controlling the flow rate to be 2-3 BV/h, carrying out gradient elution by using 0.1-1% sodium hydroxide solution, and collecting a tartaric acid elution peak;

c. adjusting the pH of a tartaric acid elution peak to be 2-5, carrying out 200-500D nanofiltration, concentrating to be less than or equal to 10%, controlling the temperature to be 180/70 ℃, and directly spraying to dry to obtain tartaric acid dry powder;

the fifth step: extraction of activated carbon

a. Adding 0.5-1 mol/L sodium hydroxide solution which is 2 times of the grape skin seed stalk residue, controlling the temperature to be 90-95 ℃, stirring for alkaline hydrolysis for 1h, filtering, and washing with water to be neutral; adding 1-5 times of 1-5% acid solution, controlling the temperature to be 40-60 ℃, stirring and modifying for 0.5h, and filtering to collect filter residues;

b. controlling the power of 720W and the frequency of 2450MHz, carbonizing and activating filter residue by microwave equipment for 12min, washing the filter residue with deionized water to be neutral, filtering, drying at 105 ℃ and drying by a dryer to obtain activated carbon dry powder with the particle size of less than or equal to 80 meshes;

a sixth step: extracted feed

Mixing various waste liquids, and performing 200-500D nanofiltration desalination; concentrating to no more than 1/10, adding protein precipitate obtained by ZTC clarifier, adjusting pH to =7, controlling temperature to 200/70 deg.C, and directly spray drying to obtain feed dry powder.