CN111647097A - Method for extracting chitin from waste shrimp shells - Google Patents
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Abstract
The invention relates to a method for extracting chitin from waste shrimp shells, firstly, carrying out deproteinization treatment on shrimp shell powder by using protease to obtain protein enzymatic hydrolysate, then carrying out decalcification treatment on the deproteinized shrimp shell powder by using dilute hydrochloric acid to obtain crude chitin, then adding a sodium acetate solution into the solution obtained after the decalcification treatment to obtain solid calcium acetate and a sodium chloride solution, concentrating the sodium chloride solution to a saturated solution, salting out the protein enzymatic hydrolysate to obtain solid protein, and finally carrying out decoloration treatment on the crude chitin by using ethanol and distillation to obtain the chitin with higher purity; the method has simple process, avoids generating acid-base waste liquid, and solves the problem of waste water pollution puzzling the production of chitin; and the recycling of inorganic calcium, protein and astaxanthin is realized, the resource waste is reduced, and the utilization value of the waste shrimp shells is improved.
Description
Technical Field
The invention relates to the technical field of chitin extraction, in particular to a method for extracting chitin from waste shrimp shells.
Background
Chitin, also known as chitin, was discovered in 1811 by the french scholars in blakenor, and was extracted from crustacean shells in 1823 by orgel, and was light beige to white, soluble in concentrated hydrochloric acid/phosphoric acid/sulfuric acid/acetic acid, insoluble in alkali and other organic solvents, and insoluble in water. Chitosan which is a deacetylation derivative of chitin is insoluble in water and soluble in partial dilute acid, can be added into food as a preservative, has the functions of preserving and inhibiting bacteria, does not harm human bodies, and does not influence the flavor of the food.
The existing chitin extraction process adopts a large amount of acid and alkali to remove inorganic calcium and protein in shrimp shells or crab shells so as to obtain chitin, a large amount of high-concentration acid and alkali waste liquid is generated in the process, and the inorganic calcium and the protein in the waste liquid are difficult to extract and utilize, so that the resource waste is caused; meanwhile, the high-concentration acid-base waste liquid is very difficult to treat and has high cost, so that the yield of the chitin is very low, the price is high, the yield of the chitosan is low, and the chitosan cannot be popularized and applied.
Therefore, there is a need to provide a new technical solution to overcome the above-mentioned drawbacks.
Disclosure of Invention
The invention aims to provide a method for extracting chitin from waste shrimp shells, which can effectively solve the technical problems.
In order to achieve the purpose of the invention, the following technical scheme is adopted:
a method for extracting chitin from waste shrimp shells comprises the following steps:
firstly, soaking and cleaning collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 1-2 hours after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 4-6 hr, filtering, washing with water, and mixing filtrates to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding dilute hydrochloric acid with the concentration of 1mol/L into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 5-6, carrying out filtration and washing after reacting for 3-5 hours to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 1-4 hours, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with the protein enzymolysis solution, stirring for 2-6 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding 95% ethanol solution into the dried crude chitin, soaking for 1-1.5 hours, filtering, and washing with water to obtain filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate at 60-75 deg.C to extract astaxanthin.
Preferably, the protease in the protease hydrolysate is papain, and the addition amount of the papain is 500-600U/g.
Preferably, the protein enzymolysis liquid prepared in the second step is heated for 0.4 to 0.8 hour and then mixed with a saturated solution of sodium chloride, and the heating temperature is 60 ℃.
Preferably, the temperature of enzymolysis in the second step is 30-35 ℃.
Preferably, the volume ratio of the calcium chloride solution to the sodium acetate solution in the fourth step is 1: 3-7.
Preferably, the amount of the ethanol solution added in the sixth step is 30-50 ml/g.
Compared with the prior art, the invention has the following beneficial effects:
according to the method, firstly, protease is used for deproteinizing shrimp shell powder to obtain protein enzymatic hydrolysate, then dilute hydrochloric acid is used for decalcifying the deproteinized shrimp shell powder to obtain crude chitin, and then sodium acetate solution is added into the solution obtained after decalcification, so that on one hand, the sodium acetate solution can neutralize residual dilute hydrochloric acid in the solution to avoid acid waste liquid, on the other hand, the sodium acetate solution reacts with calcium chloride in the solution to obtain solid calcium acetate and sodium chloride solution, and the calcium acetate is organic calcium with high nutritional value and can be absorbed and utilized by a human body, so that the recycling of inorganic calcium is realized; the sodium chloride solution reacts with the protein enzymolysis solution after being concentrated to saturation, and can salt out the protein in the protein enzymolysis solution, so that solid protein is obtained, the recycling of the protein is realized, and the treatment cost of waste liquid generated after the reaction is low; in addition, the crude chitin is soaked in ethanol and distilled to be decolorized, so that the process is simple, and the astaxanthin is recycled.
In conclusion, the method has simple process, not only avoids generating acid-base waste liquid, but also solves the problem of waste water pollution puzzling the production of chitin; the recycling of inorganic calcium, protein and astaxanthin is realized, the resource waste is reduced, and the utilization value of the waste shrimp shells is improved; meanwhile, the purity and the extraction rate of the chitin are improved, the large-scale production is realized, and the yield of the chitin and the chitosan is improved.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the following will clearly and completely describe the technical solutions in the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments.
Example 1
The invention provides a method for extracting chitin from waste shrimp shells, which comprises the following steps:
firstly, soaking and cleaning collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 1 hour after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 4 hours, filtering and washing the shrimp shell powder after stirring is finished, and combining filtrate to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding 1mol/L dilute hydrochloric acid into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 5, reacting for 3 hours, filtering and washing to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 1 hour, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with a protein enzymolysis solution, stirring for 2 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding the dried crude chitin into a 95% ethanol solution, soaking for 1 hour, filtering and washing to obtain a filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate at 60 deg.C to extract astaxanthin.
Specifically, the protease in the protease hydrolysate is papain, and the adding amount of the papain is 500U/g.
Specifically, the protein enzymolysis liquid prepared in the step two is heated for 0.4 hour and then mixed with a saturated solution of sodium chloride, and the heating temperature is 60 ℃.
Specifically, the temperature of enzymolysis in the second step is 30 ℃.
Specifically, the volume ratio of the calcium chloride solution to the sodium acetate solution in the fourth step is 1: 3.
Specifically, the amount of the ethanol solution added in the sixth step is 30 ml/g.
Example 2
The invention provides a method for extracting chitin from waste shrimp shells, which comprises the following steps:
firstly, soaking and cleaning the collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 1.3 hours after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 4.5 hr, filtering, washing with water, and mixing filtrates to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding dilute hydrochloric acid with the concentration of 1mol/L into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 5.3, performing filtration and washing after reacting for 3.5 hours to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 2 hours, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with a protein enzymolysis solution, stirring for 3 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding the dried crude chitin into a 95% ethanol solution, soaking for 1.2 hours, and then filtering and washing to obtain a filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate at 60 deg.C to extract astaxanthin.
Specifically, the protease in the protease hydrolysate is papain, and the addition amount of the papain is 530U/g.
Specifically, the protein enzymolysis liquid prepared in the step two is heated for 0.5 hour and then mixed with a saturated solution of sodium chloride, and the heating temperature is 60 ℃.
Specifically, the temperature of enzymolysis in the second step is 32 ℃.
Specifically, the volume ratio of the calcium chloride solution to the sodium acetate solution in the fourth step is 1: 4.
Specifically, the amount of the ethanol solution added in the sixth step is 35 ml/g.
Example 3
The invention provides a method for extracting chitin from waste shrimp shells, which comprises the following steps:
firstly, soaking and cleaning the collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 1.5 hours after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 5 hours, filtering and washing the shrimp shell powder after stirring is finished, and combining filtrate to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding dilute hydrochloric acid with the concentration of 1mol/L into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 5.5, performing filtration and washing after reacting for 4 hours to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 2.5 hours, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with a protein enzymolysis solution, stirring for 4 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding the dried crude chitin into a 95% ethanol solution, soaking for 1.3 hours, and then filtering and washing to obtain a filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate at 60 deg.C to extract astaxanthin.
Specifically, the protease in the protease hydrolysate is papain, and the addition amount of the papain is 550U/g.
Specifically, the protein enzymolysis liquid prepared in the step two is heated for 0.6 hour and then mixed with a saturated solution of sodium chloride, and the heating temperature is 60 ℃.
Specifically, the temperature of enzymolysis in the second step is 33 ℃.
Specifically, the volume ratio of the calcium chloride solution to the sodium acetate solution in the fourth step is 1: 5.
Specifically, the amount of the ethanol solution added in the sixth step is 40 ml/g.
Example 4
The invention provides a method for extracting chitin from waste shrimp shells, which comprises the following steps:
firstly, soaking and cleaning the collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 1.7 hours after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 5.5 hr, filtering, washing with water, and mixing filtrates to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding dilute hydrochloric acid with the concentration of 1mol/L into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 5.7, filtering and washing after reacting for 5 hours to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 3 hours, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with the protein enzymolysis solution, stirring for 5 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding the dried crude chitin into a 95% ethanol solution, soaking for 1.4 hours, and then filtering and washing to obtain a filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate at 60 deg.C to extract astaxanthin.
Specifically, the protease in the protease hydrolysate is papain, and the addition amount of the papain is 570U/g.
Specifically, the protein enzymolysis liquid prepared in the step two is heated for 0.7 hour and then mixed with a saturated solution of sodium chloride, and the heating temperature is 60 ℃.
Specifically, the temperature of enzymolysis in the second step is 34 ℃.
Specifically, the volume ratio of the calcium chloride solution to the sodium acetate solution in the fourth step is 1: 6.
Specifically, the amount of the ethanol solution added in the sixth step is 45 ml/g.
Example 5
The invention provides a method for extracting chitin from waste shrimp shells, which comprises the following steps:
firstly, soaking and cleaning collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 2 hours after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 6 hours, filtering and washing the shrimp shell powder after stirring is finished, and combining filtrate to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding 1mol/L dilute hydrochloric acid into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 6, filtering and washing after reacting for 5 hours to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 4 hours, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with a protein enzymolysis solution, stirring for 6 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding the dried crude chitin into a 95% ethanol solution, soaking for 1.5 hours, and then filtering and washing to obtain a filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate to extract astaxanthin, wherein the distillation temperature is 75 ℃.
Specifically, the protease in the protease hydrolysate is papain, and the addition amount of the papain is 600U/g.
Specifically, the protein enzymolysis liquid prepared in the step two is heated for 0.8 hour and then mixed with a saturated solution of sodium chloride, and the heating temperature is 60 ℃.
Specifically, the temperature of enzymolysis in the second step is 35 ℃.
Specifically, the volume ratio of the calcium chloride solution to the sodium acetate solution in the fourth step is 1: 7.
Specifically, the amount of the ethanol solution added in the sixth step is 50 ml/g.
The control group comprises chitin extracted by acid-base method
The raw materials of shrimp and crab shell are cleaned and dried in the sun in time to keep the quality;
pickling, wherein each hundred jin of crab shells are processed with 30 jin of industrial hydrochloric acid (30 degrees of Pomex) and 200 jin of water in a water tank. The crab shells should be turned over frequently at first and then every 4 hours. Generally, pickling needs 30-40 hours, if some raw materials are not softened after pickling, and no bubbles are generated in the acid liquor, which indicates that the acid quantity is insufficient, some concentrated acid should be added. Pickling, and washing crab shell with water to neutral pH 6-7;
alkali liquor cooking: after pickling, the main components of the soft shell are protein and a certain amount of grease, and in order to remove the protein and the grease, the soft shell needs to be boiled with alkali for 40 minutes (16-18 ℃ of vermex) and then washed with water to be neutral;
secondary pickling, wherein in order to further remove calcium, secondary pickling is needed, the concentration of the acid liquor is lower than that of the first pickling, and the acidity is reduced; soaking for 10-20 hours at about 5 ℃ of permafrost, turning over frequently, pickling, washing with water, and sun drying (or pickling once for labor saving but thorough);
secondary alkali cooking, namely, in order to further degrease, performing alkali cooking for 30 minutes (8-10 ℃ of Veronica);
drying, namely cleaning and drying in the sun to obtain a semi-finished product.
The chitin extracted in examples 1-5 and the control group was tested for product form, components and extraction rate, and the test results are shown in table 1:
TABLE 1 detection results of the product form, components and extraction rate of chitin
As can be seen from table 1, the chitin extracted by the process of the present invention has the highest purity and extraction rate in terms of product form, components and extraction, particularly, the chitin extracted by the process of example 4.
The recovery rates of inorganic calcium, protein and astaxanthin and the production of acid-base wastewater in examples 1 to 5 and the comparative group were measured, and the results are shown in table 2:
TABLE 2 recovery rates of inorganic calcium, protein, astaxanthin and results of examination on whether acid-base wastewater was generated
As can be seen from table 2, the extraction process adopted in the present invention can recycle inorganic calcium, protein and astaxanthin while extracting chitin, and simultaneously avoid generating acid-base wastewater, and achieve zero-pollution production, thereby reducing production cost and improving economic benefits of enterprises.
Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention.
Claims (6)
1. A method for extracting chitin from waste shrimp shells is characterized by comprising the following steps: the method comprises the following steps:
firstly, soaking and cleaning collected waste shrimp shells, then drying the waste shrimp shells, putting the waste shrimp shells into a grinder to be ground for 1-2 hours after drying, and grinding the waste shrimp shells into powder;
step two: adding protease hydrolysate into shrimp shell powder, stirring for 4-6 hr, filtering, washing with water, and mixing filtrates to obtain deproteinized shrimp shell powder and protein hydrolysate;
step three: adding dilute hydrochloric acid with the concentration of 1mol/L into the deproteinized shrimp shell powder for decalcification treatment, stopping adding acid when the pH value of a system is reduced to 5-6, carrying out filtration and washing after reacting for 3-5 hours to obtain a calcium chloride solution with the residual dilute hydrochloric acid and crude chitin, and drying the crude chitin;
step four: adding a sodium acetate solution into the calcium chloride solution, stirring for 1-4 hours, filtering and washing to obtain a sodium chloride solution and solid calcium acetate, and drying the solid calcium acetate to obtain calcium acetate crystals;
step five: concentrating a sodium chloride solution to obtain a saturated sodium chloride solution, mixing the saturated sodium chloride solution with the protein enzymolysis solution, stirring for 2-6 hours, centrifuging the mixture, and performing solid-liquid separation to obtain solid protein;
step six: adding 95% ethanol solution into the dried crude chitin, soaking for 1-1.5 hours, filtering, and washing with water to obtain filtrate containing astaxanthin and chitin;
step seven: distilling the astaxanthin-containing filtrate at 60-75 deg.C to extract astaxanthin.
2. The method for extracting chitin from waste shrimp shells as claimed in claim 1, wherein: the protease in the protease hydrolysate is papain, and the adding amount of the papain is 500-600U/g.
3. The method for extracting chitin from waste shrimp shells as claimed in claim 1, wherein: and (3) heating the protein enzymolysis liquid prepared in the step (II) for 0.4-0.8 h, and then mixing the protein enzymolysis liquid with a saturated sodium chloride solution, wherein the heating temperature is 60 ℃.
4. The method for extracting chitin from waste shrimp shells as claimed in claim 1, wherein: the temperature of enzymolysis in the second step is 30-35 ℃.
5. The method for extracting chitin from waste shrimp shells as claimed in claim 1, wherein: in the fourth step, the volume ratio of the calcium chloride solution to the sodium acetate solution is 1: 3-7.
6. The method for extracting chitin from waste shrimp shells as claimed in claim 1, wherein: the amount of the ethanol solution added in the sixth step is 30-50 ml/g.
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