Novel process for extracting D-glucosamine hydrochloride

Abstract

The invention relates to the technical field of bioengineering, in particular to a novel process for extracting D-glucosamine hydrochloride, and through the synergistic effect of composite enzymolysis and microwave-assisted treatment, the degradation efficiency of chitin is remarkably improved; microwave enhanced acid hydrolysis is combined with ion exchange resin neutralization, so that the reaction time is effectively shortened, and the acid consumption and the energy consumption are reduced; by utilizing precise concentration of a nanofiltration membrane and a crystallization control technology based on a solubility prediction model, high-efficiency purification and crystallization process optimization of the product are realized, so that the purity and yield of the product are improved, the environmental pollution is reduced, and the production cost is reduced. The method solves the technical problems of low efficiency, high energy consumption, difficult product purity and quality guarantee and easy environmental pollution in the extraction process of D-glucosamine hydrochloride in the prior art.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of bioengineering, and particularly relates to a new process for extracting D-glucosamine hydrochloride. BACKGROUND

[0002] D-glucosamine hydrochloride is an important bioactive substance and has wide application value in many fields such as medicine, food, cosmetics and the like. In the field of medicine, it is an important raw material for synthesizing joint cartilage components and can be used for treating and preventing joint diseases such as osteoarthritis, and can effectively relieve joint pain and improve joint function; in the food industry, it can be added to various foods as a nutritional fortifier to supplement necessary nutrients for the human body; in the field of cosmetics, it has the effects of moisturizing and anti-aging and can improve the gloss and elasticity of the skin.

[0003] However, the existing D-glucosamine hydrochloride extraction process has the problems of low efficiency, high energy consumption, difficult guarantee of product purity and quality, and easy pollution of the environment. SUMMARY

[0004] The present application provides a new process for extracting D-glucosamine hydrochloride, which solves the technical problems of low efficiency, high energy consumption, difficult guarantee of product purity and quality, and easy pollution of the environment in the existing D-glucosamine hydrochloride extraction process.

[0005] To achieve the above-mentioned purpose, the present application provides a new process for extracting D-glucosamine hydrochloride, which comprises the following steps: The chitin raw material is crushed to 40-60 mesh, and then is placed in a composite enzyme solution, and is subjected to enzymolysis at 45-55 DEG C and pH 5.5-6.5 for 2-3 hours; after the enzymolysis, microwave-assisted treatment is performed, the microwave power is 400-600 W, and the treatment time is 10-15 minutes; The raw material after the microwave-assisted treatment is mixed with a hydrochloric acid solution with a concentration of 2-4 mol / L, and is placed in a microwave reactor, and is subjected to hydrolysis reaction at 80-95 DEG C for 40-60 minutes; the power of the microwave reactor is 500-800 W, and the frequency is 2450 MHz; After the hydrolysis reaction is completed, the residue is removed by filtration to obtain a hydrolysis liquid; the hydrolysis liquid is cooled to 30-40 DEG C, and is subjected to neutralization through a resin column filled with a strong basic anion exchange resin, the flow rate is controlled so that the contact time of the hydrolysis liquid with the resin is 10-20 minutes, and the effluent is collected until the pH value is stabilized at 6.5-7.5; The effluent after the neutralization is first concentrated through a nanofiltration membrane, the nanofiltration membrane has a molecular weight cut-off of 200-300 Da, and the operating pressure is 1.5-2.5 MPa to obtain a concentrated liquid; The concentrate is introduced into the crystallization vessel, and the crystallization optimization control program based on the solubility prediction model is started to dynamically control the ethanol addition rate and the cooling program of the crystallization vessel. The crystallization is allowed to stand at 0-4℃ for 12-24 hours. The crystallized solid is separated and dried using a vacuum freeze-drying method. The pre-freezing temperature is -45℃ to -35℃, the vacuum degree is less than 30 Pa, and the product is dried until the moisture content is less than 2.0%. This yields the D-glucosamine hydrochloride product.

[0006] The complex enzyme is composed of protease and chitinase in a mass ratio of 1:1.5-2, and the total amount of enzyme added is 0.5%-1.0% of the raw material mass. The protease is Bacillus subtilis protease, and the chitinase is derived from Aspergillus fermentation extraction. The enzyme activity of Bacillus subtilis protease is ≥10000 U / g, and the enzyme activity of chitinase is ≥5000 U / g. Furthermore, the enzyme activity retention rate of the compound enzyme is ≥90% under pH conditions of 5.5-6.5.

[0007] The specific implementation method of the crystallization optimization control program based on the solubility prediction model is as follows: The solubility prediction model is a support vector machine regression model. This model is trained using historical crystallization data. The input variables include the temperature, conductivity and viscosity of the concentrate, and the output is the instantaneous solubility. The control system adjusts the crystallization vessel temperature by gradient reduction based on the relationship between predicted solubility and target supersaturation, and controls the addition rate of ethanol peristaltic pump according to proportional feedback, so that the crystallization process is always maintained within the optimal supersaturation range, and the nucleation density is controlled at 10^3-10^4 nuclei / mL.

[0008] Specifically, in the step "mixing the microwave-treated raw material with a hydrochloric acid solution of concentration 2-4 mol / L, placing it in a microwave reactor, and hydrolyzing it at 80-95℃ for 40-60 minutes; the power of the microwave reactor is 500-800W, and the frequency is 2450MHz", The process is monitored using an endpoint prediction system based on near-infrared spectroscopy; Specifically, the spectral data of the hydrolysate is acquired in real time by an online near-infrared fiber optic probe, and the spectral data is processed by a pre-established partial least squares regression model to predict the concentration of D-glucosamine hydrochloride in the hydrolysate in real time; when the predicted concentration change rate is less than the set threshold, the hydrolysis endpoint is automatically determined and the reaction is terminated. The threshold is set at a concentration change rate of <0.5% per minute.

[0009] The partial least squares regression model was trained using a large number of historical samples covering different hydrolysis times, temperatures, and acid concentrations, along with their corresponding high-performance liquid chromatography (HPLC) measurements. After training, the model underwent cross-validation, with the root mean square error (RMSE) of the cross-validation being ≤0.05.

[0010] The strongly basic anion exchange resin is a 201×7 type resin, and the regeneration of the resin is achieved through the following methods: Record the cumulative volume of hydrolysate flowing through the resin and the pH value of the effluent. When the pH value drops below 6.0 or the cumulative volume reaches 90%-95% of the resin's theoretical exchange capacity, it indicates that regeneration should be performed. Regeneration is performed using a 5-10% sodium hydroxide solution at a flow rate of 2-4 BV / h.

[0011] The resin exchange capacity recovery rate after regeneration is ≥95%. If it is less than 90%, it indicates that the resin should be replaced.

[0012] The microwave-assisted treatment and microwave hydrolysis reaction are carried out continuously in the same multifunctional reactor. The reactor is lined with polytetrafluoroethylene and integrates a temperature-pressure feedback system, which automatically releases pressure when the pressure exceeds 1.5 MPa.

[0013] When the microwave-treated raw material is mixed with a hydrochloric acid solution with a concentration of 2-4 mol / L, the ratio of the microwave-treated raw material to the hydrochloric acid solution is 1:8-1:12 (g / mL).

[0014] After extraction is complete, wastewater is recycled, and the specific methods are as follows: The crystallization mother liquor and washing wastewater are collected and treated using a reverse osmosis-multi-effect evaporation combined process. The reverse osmosis operating pressure is 1.5-3.0 MPa, and the multi-effect evaporation temperature is 60-90℃. 85% of the water is recovered for raw material cleaning or acid preparation. The salt obtained from evaporation is centrally treated, and the COD of the recovered water is ≤50 mg / L.

[0015] This invention discloses a novel process for extracting D-glucosamine hydrochloride. Through the synergistic effect of combined enzymatic hydrolysis and microwave-assisted treatment, the degradation efficiency of chitin is significantly improved. Microwave-enhanced acid hydrolysis combined with ion exchange resin neutralization effectively shortens reaction time and reduces acid and energy consumption. Utilizing nanofiltration membrane for precise concentration and crystallization control technology based on a solubility prediction model, efficient purification of the product and optimization of the crystallization process are achieved. This improves product purity and yield while reducing environmental pollution. This approach solves the technical problems of low efficiency, high energy consumption, difficulty in ensuring product purity and quality, and easy environmental pollution inherent in existing D-glucosamine hydrochloride extraction processes. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.

[0017] Figure 1 This is a flowchart of the novel process for extracting D-glucosamine hydrochloride according to the present invention. Detailed Implementation

[0018] The embodiments of the present invention are described in detail below. Examples of the embodiments are shown in the accompanying drawings. The embodiments described below with reference to the accompanying drawings are exemplary and intended to explain the present invention, but should not be construed as limiting the present invention.

[0019] Please see Figure 1 , Figure 1 This is a flowchart of the novel process for extracting D-glucosamine hydrochloride according to the present invention.

[0020] This invention provides a novel process for extracting D-glucosamine hydrochloride, comprising the following: S1. Crush the chitin raw material to 40-60 mesh, then place it in a compound enzyme solution and enzymatically hydrolyze it for 2-3 hours at 45-55℃ and pH 5.5-6.5. After enzymatic hydrolysis, perform microwave-assisted treatment with a microwave power of 400-600W for 10-15 minutes. In this specific embodiment, the composite enzyme is composed of protease and chitinase in a mass ratio of 1:1.5-2, and the total amount of enzyme added is 0.5%-1.0% of the raw material mass. The protease is Bacillus subtilis protease, and the chitinase is derived from Aspergillus fermentation extraction. The enzyme activity of Bacillus subtilis protease is ≥10000 U / g, and the enzyme activity of chitinase is ≥5000 U / g. Furthermore, the enzyme activity retention rate of the compound enzyme is ≥90% under pH conditions of 5.5-6.5.

[0021] S2. Mix the microwave-assisted treated raw material with a hydrochloric acid solution with a concentration of 2-4 mol / L, place it in a microwave reactor, and hydrolyze it at 80-95℃ for 40-60 minutes; the power of the microwave reactor is 500-800W, and the frequency is 2450MHz. In this specific embodiment, when the microwave-treated raw material is mixed with a hydrochloric acid solution with a concentration of 2-4 mol / L, the ratio of the microwave-treated raw material to the hydrochloric acid solution is 1:8-1:12 (g / mL).

[0022] The process is monitored using an endpoint prediction system based on near-infrared spectroscopy; Specifically, the spectral data of the hydrolysate is acquired in real time by an online near-infrared fiber optic probe, and the spectral data is processed by a pre-established partial least squares regression model to predict the concentration of D-glucosamine hydrochloride in the hydrolysate in real time; when the predicted concentration change rate is less than the set threshold, the hydrolysis endpoint is automatically determined and the reaction is terminated. The threshold is set at a concentration change rate of <0.5% per minute.

[0023] The partial least squares regression model was trained using a large number of historical samples covering different hydrolysis times, temperatures, and acid concentrations, along with their corresponding high-performance liquid chromatography (HPLC) measurements. After training, the model underwent cross-validation, with the root mean square error (RMSE) of the cross-validation being ≤0.05.

[0024] S3. After the hydrolysis reaction is completed, filter to remove the residue and obtain the hydrolysate; cool the hydrolysate to 30-40℃ and pass it through a resin column packed with strong base anion exchange resin for neutralization. Control the flow rate so that the contact time between the hydrolysate and the resin is 10-20 minutes. Collect the effluent until the pH value stabilizes at 6.5-7.5. In this specific embodiment, the strongly basic anion exchange resin is a 201×7 type resin, and the regeneration timing of the resin is achieved in the following manner: Record the cumulative volume of hydrolysate flowing through the resin and the pH value of the effluent. When the pH value drops below 6.0 or the cumulative volume reaches 90%-95% of the resin's theoretical exchange capacity, it indicates that regeneration should be performed. Regeneration is performed using a 5-10% sodium hydroxide solution at a flow rate of 2-4 BV / h.

[0025] The resin exchange capacity recovery rate after regeneration is ≥95%. If it is less than 90%, it indicates that the resin should be replaced.

[0026] S4. The neutralized effluent is first concentrated by nanofiltration membrane. The nanofiltration membrane has a molecular weight cutoff of 200-300 Da and an operating pressure of 1.5-2.5 MPa to obtain the concentrate. S5. Introduce the concentrate into the crystallization vessel and start the crystallization optimization control program based on the solubility prediction model to dynamically control the ethanol addition rate and the cooling program of the crystallization vessel. Allow the crystallization to stand at 0-4℃ for 12-24 hours. For this specific implementation method, the crystallization optimization control program based on the solubility prediction model is implemented as follows: The solubility prediction model is a support vector machine regression model. This model is trained using historical crystallization data. The input variables include the temperature, conductivity and viscosity of the concentrate, and the output is the instantaneous solubility. The control system adjusts the crystallization vessel temperature by gradient reduction based on the relationship between predicted solubility and target supersaturation, and controls the addition rate of ethanol peristaltic pump according to proportional feedback, so that the crystallization process is always maintained within the optimal supersaturation range, and the nucleation density is controlled at 10^3-10^4 nuclei / mL.

[0027] S6. Separate the crystallized solid and dry it using a vacuum freeze-drying method. The pre-freezing temperature is -45℃ to -35℃, the vacuum degree is less than 30 Pa, and the moisture content is less than 2.0% to obtain the D-glucosamine hydrochloride product.

[0028] In this specific embodiment, microwave-assisted treatment and microwave hydrolysis reaction are carried out continuously in the same multifunctional reactor. The reactor lining is made of polytetrafluoroethylene and integrates a temperature-pressure feedback system, which automatically releases pressure when the pressure exceeds 1.5 MPa.

[0029] After extraction is complete, wastewater is recycled, and the specific methods are as follows: The crystallization mother liquor and washing wastewater are collected and treated using a reverse osmosis-multi-effect evaporation combined process. The reverse osmosis operating pressure is 1.5-3.0 MPa, and the multi-effect evaporation temperature is 60-90℃. 85% of the water is recovered for raw material cleaning or acid preparation. The salt obtained from evaporation is centrally treated, and the COD of the recovered water is ≤50 mg / L.

[0030] Furthermore, during the extraction process, key process parameters are monitored and recorded in real time. The key process parameters, the purity and yield data of the corresponding final product are associated and stored, and blockchain technology is used for evidence storage to ensure the integrity and immutability of the data chain. Based on the stored historical data of multiple batches, an optimization model between key process parameters and product purity and yield is established through regression analysis or machine learning algorithms. Based on the optimization model, the key process parameter settings for the next batch of production are dynamically adjusted.

[0031] The key process parameters include at least the alkali concentration, temperature, and time during deacetylation detection, as well as the acid concentration, temperature, and pressure during the hydrolysis process.

[0032] Furthermore, after "filtering to remove residue and obtaining hydrolysate", a rapid quality assessment of the hydrolysate is performed: Take a small amount of hydrolysate and use a portable refractometer to determine its soluble solids content (Brix value) and a pH meter to determine its acidity. When the Brix value is in the range of 10-15% and the pH value is below 1.5, the hydrolysate is deemed to be of qualified quality and can proceed to the next neutralization process. This rapid evaluation method serves as an auxiliary verification method for the online near-infrared endpoint prediction system.

[0033] The concentrated waste liquid and hydrolysis residue generated by the wastewater recycling system are combined and subjected to anaerobic fermentation. The biogas produced is used to provide part of the heat energy for the multi-effect evaporator. The residue after fermentation can be used as raw material for organic fertilizer after composting, thereby realizing the resource utilization of waste and the green closed loop of the entire process.

[0034] The novel extraction process for D-glucosamine hydrochloride according to this invention significantly improves the degradation efficiency of chitin through the synergistic effect of combined enzymatic hydrolysis and microwave-assisted treatment. Microwave-enhanced acid hydrolysis combined with ion exchange resin neutralization effectively shortens the reaction time and reduces acid and energy consumption. Precise concentration using nanofiltration membranes and crystallization control technology based on a solubility prediction model achieves efficient product purification and optimized crystallization process. This improves product purity and yield while reducing environmental pollution. This approach solves the technical problems of low efficiency, high energy consumption, difficulty in ensuring product purity and quality, and easy environmental pollution in existing D-glucosamine hydrochloride extraction processes.

[0035] The above-disclosed embodiments are merely one or more preferred embodiments of this application and should not be construed as limiting the scope of this application. Those skilled in the art can understand that all or part of the processes for implementing the above embodiments and equivalent changes made in accordance with the claims of this application still fall within the scope of this application.

Claims

1. A novel process for extracting D-glucosamine hydrochloride, characterized in that, Including the following: Chitosan raw material is pulverized to 40-60 mesh, then placed in a compound enzyme solution and enzymatically hydrolyzed for 2-3 hours at 45-55℃ and pH 5.5-6.

5. After enzymatic hydrolysis, it is subjected to microwave-assisted treatment with a microwave power of 400-600W and a treatment time of 10-15 minutes. The microwave-assisted treated raw material is mixed with a 2-4 mol / L hydrochloric acid solution and placed in a microwave reactor. The mixture is then subjected to hydrolysis at 80-95℃ for 40-60 minutes. The microwave reactor has a power of 500-800W and a frequency of 2450MHz. After the hydrolysis reaction is completed, the residue is removed by filtration to obtain the hydrolysate; the hydrolysate is cooled to 30-40℃ and neutralized by passing it through a resin column packed with a strong base anion exchange resin. The flow rate is controlled so that the contact time between the hydrolysate and the resin is 10-20 minutes. The effluent is collected until the pH value stabilizes at 6.5-7.

5. The neutralized effluent is first concentrated by nanofiltration membrane with a molecular weight cutoff of 200-300 Da and an operating pressure of 1.5-2.5 MPa to obtain a concentrated solution. The concentrate is introduced into the crystallization vessel, and the crystallization optimization control program based on the solubility prediction model is started to dynamically control the ethanol addition rate and the cooling program of the crystallization vessel. The crystallization is allowed to stand at 0-4℃ for 12-24 hours. The crystallized solid is separated and dried using a vacuum freeze-drying method. The pre-freezing temperature is -45℃ to -35℃, the vacuum degree is less than 30 Pa, and the product is dried until the moisture content is less than 2.0%. This yields the D-glucosamine hydrochloride product.

2. The novel process for extracting D-glucosamine hydrochloride as described in claim 1, characterized in that, The complex enzyme consists of protease and chitinase in a mass ratio of 1:1.5-2, with the total enzyme content being 0.5%-1.0% of the raw material mass. The protease is Bacillus subtilis protease, and the chitinase is derived from Aspergillus fermentation extraction. The enzyme activity of Bacillus subtilis protease is ≥10000 U / g, and the enzyme activity of chitinase is ≥5000 U / g. Furthermore, the enzyme activity retention rate of the compound enzyme is ≥90% under pH conditions of 5.5-6.

5.

3. The novel process for extracting D-glucosamine hydrochloride as described in claim 2, characterized in that, The specific implementation method of the crystallization optimization control program based on the solubility prediction model is as follows: The solubility prediction model is a support vector machine regression model. This model is trained using historical crystallization data. The input variables include the temperature, conductivity and viscosity of the concentrate, and the output is the instantaneous solubility. The control system adjusts the crystallization vessel temperature by gradient reduction based on the relationship between predicted solubility and target supersaturation, and controls the addition rate of ethanol peristaltic pump according to proportional feedback, so that the crystallization process is always maintained within the optimal supersaturation range, and the nucleation density is controlled at 10^3-10^4 nuclei / mL.

4. The novel process for extracting D-glucosamine hydrochloride as described in claim 3, characterized in that, In the step "mixing the microwave-treated raw material with a 2-4 mol / L hydrochloric acid solution as a feed-liquid mixture, placing it in a microwave reactor, and hydrolyzing it at 80-95℃ for 40-60 minutes; the microwave reactor has a power of 500-800W and a frequency of 2450MHz", The process is monitored using an endpoint prediction system based on near-infrared spectroscopy; Specifically, the spectral data of the hydrolysate is acquired in real time by an online near-infrared fiber optic probe, and the spectral data is processed by a pre-established partial least squares regression model to predict the concentration of D-glucosamine hydrochloride in the hydrolysate in real time; when the predicted concentration change rate is less than the set threshold, the hydrolysis endpoint is automatically determined and the reaction is terminated. The threshold is set at a concentration change rate of <0.5% per minute.

5. The novel process for extracting D-glucosamine hydrochloride as described in claim 4, characterized in that, The partial least squares regression model was trained using a large number of historical samples covering different hydrolysis times, temperatures, and acid concentrations, along with their corresponding high-performance liquid chromatography (HPLC) measurements. After training, the model underwent cross-validation, and the root mean square error (RMSE) of the cross-validation was ≤0.

05.

6. The novel process for extracting D-glucosamine hydrochloride as described in claim 5, characterized in that, The strongly basic anion exchange resin is a 201×7 type resin. The regeneration of the resin is achieved through the following methods: Record the cumulative volume of hydrolysate flowing through the resin and the pH value of the effluent. When the pH value drops to When the volume drops below 6.0 or the cumulative volume reaches 90%-95% of the resin's theoretical exchange capacity, regeneration is indicated. Regeneration should be performed using a 5-10% sodium hydroxide solution at a flow rate of 2-4 BV / h. The resin exchange capacity recovery rate after regeneration is ≥95%. If it is less than 90%, it indicates that the resin should be replaced.

7. The novel process for extracting D-glucosamine hydrochloride as described in claim 6, characterized in that, Microwave-assisted treatment and microwave hydrolysis reaction are carried out continuously in the same multifunctional reactor. The reactor is lined with polytetrafluoroethylene and integrates a temperature-pressure feedback system, which automatically depressurizes when the pressure exceeds 1.5 MPa.

8. The novel process for extracting D-glucosamine hydrochloride as described in claim 1, characterized in that, When mixing the microwave-treated raw material with a hydrochloric acid solution of concentration of 2-4 mol / L, the ratio of the microwave-treated raw material to the hydrochloric acid solution is 1:8-1:12 (g / mL).

9. The novel process for extracting D-glucosamine hydrochloride as described in claim 1, characterized in that, After extraction is complete, wastewater is recycled, and the specific methods are as follows: The crystallization mother liquor and washing wastewater are collected and treated using a reverse osmosis-multi-effect evaporation combined process. The reverse osmosis operating pressure is 1.5-3.0 MPa, and the multi-effect evaporation temperature is 60-90℃. 85% of the water is recovered for raw material cleaning or acid preparation. The salt obtained from evaporation is centrally treated, and the COD of the recovered water is ≤50 mg / L.

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