Fluid type dynamic ultrasonic integrated temperature control circulating equipment and method for extracting biomass by fluid type dynamic ultrasonic integrated temperature control circulating equipment

Abstract

The invention discloses fluid type dynamic ultrasonic integrated temperature control circulating equipment and a method for extracting biomass by using the fluid type dynamic ultrasonic integrated temperature control circulating equipment. The equipment provided by the invention comprises the liquid storage tank, the ultrasonic generator, the gas-liquid separator, the circulating device and the collector, the biological activity can be better retained and the yield can be improved by performing circulating reflux extraction through the equipment, the equipment is suitable for extracting various biomasses, and the dissolving capacity of a solvent to a target substance can be fully exerted. Meanwhile, the equipment is suitable for extracting different eutectic solvents or gas response switchable solvents, hydrophobic / hydrophilic conversion can be realized, separation of a target object and recovery of the solvents are facilitated, and the equipment has higher economic benefits. Meanwhile, the invention further provides a method for extracting biomass by adopting the equipment, extraction is performed in the specific equipment, the extraction environment condition can be accurately controlled, the problems of component damage, oxidation, pollution and the like caused by high temperature can be better avoided, and the content of active components in the biomass and the effect of the active components in the biomass can be effectively improved.

Description

Technical Field

[0001] This invention relates to the field of biomass extraction technology, and in particular to a fluid-powered ultrasonic integrated temperature-controlled circulation device and a method for extracting biomass. Background Technology

[0002] Extraction technology plays a crucial role in the development and utilization of biomass resources. Traditional biomass extraction methods often suffer from numerous problems. For example, many methods require the use of large amounts of organic solvents, which not only increases production costs but may also pollute the environment. Furthermore, some extraction processes are energy-intensive and have limited extraction efficiency, making it difficult to fully extract the target components from the biomass. For some easily oxidized and deactivated biomass, traditional extraction methods struggle to avoid degradation and denaturation during the extraction process, resulting in a significant reduction in the quality and activity of the extract.

[0003] In recent years, with the continuous advancement of science and technology, ultrasonic extraction technology has gradually attracted attention. Ultrasound waves can generate cavitation effects in liquids, creating a localized high-temperature and high-pressure environment, thereby accelerating the penetration and dissolution of biomass by the solvent and improving extraction efficiency. However, existing ultrasonic extraction equipment and methods still need further improvement in areas such as solvent recycling, target analyte separation, and precise control of the extraction process.

[0004] For example, a biomass extraction ultrasonic purification device disclosed in existing research uses an ultrasonic extraction tank for extraction. This tank is equipped with a heating mechanism, a stirring mechanism, and an injection component, enabling continuous, controllable, and uniform heating and extraction. However, because some biomass active components are easily oxidized and volatilized under high temperatures, and prolonged ultrasonic extraction can lead to excessively high extraction temperatures and inactivation of the target analyte, this ultrasonic purification device, which relies on heating extraction, is difficult to apply to the extraction of target analytes that are easily inactivated under high-temperature conditions. Therefore, existing ultrasonic extraction equipment still suffers from problems such as: ultrasonic heat generation in a closed environment leading to easy oxidation and inactivation of the extract; difficulty in accurately controlling the separation and extraction of the target analyte; insufficient extraction; insufficient recycling of the ultrasonic extraction solvent; and waste due to the large amount of organic solvent used.

[0005] With the development of more novel green solvents, many solvents can switch their properties according to extraction conditions to extract different biomass. However, current extraction equipment struggles to utilize these solvents. Currently, there are relatively few devices and methods that combine eutectic solvents with ultrasonic extraction technology to achieve integrated solvent recycling. Furthermore, the application of gas-responsive switchable solvents, and how to achieve hydrophobic / hydrophilic conversion through extraction methods to facilitate target separation and solvent recovery, are also issues that require further exploration and resolution in the field of biomass extraction. Therefore, to extract biomass more efficiently and conveniently, it is necessary to develop more methods and equipment that can precisely control extraction conditions, perform integrated recycling extraction, and simultaneously apply to different extraction solvents. Summary of the Invention

[0006] The technical problem to be solved by this invention is to overcome the above-mentioned defects and deficiencies of existing ultrasonic extraction equipment or methods, and to provide a fluid-powered ultrasonic integrated temperature-controlled circulation device and its method for extracting biomass, which can achieve efficient, green and precise extraction of biomass, while improving solvent utilization and economic benefits, and providing a more advantageous technical solution for the development and utilization of biomass.

[0007] The purpose of this invention is to provide a fluid-powered, ultrasonically integrated, temperature-controlled, circulating biomass extraction device.

[0008] Another objective of this invention is to provide a method for extracting biomass using a fluid-driven, ultrasonically integrated temperature-controlled circulation device.

[0009] The above-mentioned objective of this invention is achieved through the following technical solution: This invention provides a fluid-powered ultrasonic integrated temperature-controlled circulation extraction device for biomass, comprising a storage tank, an ultrasonic generator, a gas-liquid separator, an air pump, a collector, and a circulation device; The ultrasonic generator includes: a sealed acoustic energy processing tank, a whistle, a filter, and a temperature controller A; the whistle is placed at one end inside the acoustic energy processing tank, and the filter is placed at the other end inside the acoustic energy processing tank away from the whistle; a liquid storage tank is connected to the end of the ultrasonic generator near the whistle via a pipe, and a gas-liquid separator is connected to the rear end near the filter via a pipe; a collector is connected to the rear end of the filter via a pipe; the temperature controller A is installed on the outside of the ultrasonic generator; and the air pump is connected to the whistle in the ultrasonic generator via a pipe. The circulation device includes: a circulation pipe and a liquid pump; the liquid pump is connected to the circulation pipe; one end of the circulation pipe is connected to the air whistle in the ultrasonic generator, and the other end is connected to the end of the ultrasonic generator near the filter; a liquid pump C is provided between the ultrasonic generator and the collector.

[0010] The fluid-driven ultrasonic integrated temperature-controlled circulation device provided by this invention combines a fluid-driven ultrasonic generator with temperature control, air pump, gas-liquid separation, and circulation devices. This integrated device performs cyclic temperature-controlled extraction, fully controlling extraction conditions and enabling intermittent cyclic reflux extraction. This better avoids oxidation and loss of biomass, preserves biological activity, and improves yield and extraction efficiency. Furthermore, through flexible gas-liquid adjustment and control, it can achieve multiple cyclic extractions with various solvents, fully utilizing the solvent's dissolving power for the target substance. It is suitable for gas-responsive solvent switching and can recover solvents.

[0011] The operating principle of this equipment is as follows: the storage tank and collector are used to store or collect the sample to be extracted or the extracted substance; the ultrasonic generator mainly performs the ultrasonic extraction reaction, and can also introduce gas, adjust temperature parameters, and filter through its internal filter; the gas-liquid separator is used to discharge gas and balance gas pressure; the circulation device circulates the extraction solution or solvent back to the ultrasonic generator for further extraction; in the ultrasonic generator, the acoustic energy processing tank performs ultrasonic extraction, and its external temperature controller effectively prevents the target substance from being deactivated due to excessively high temperature caused by prolonged ultrasonic time; inert gases such as nitrogen are introduced into the acoustic energy processing tank through a gas whistle by an air pump, which not only balances the gas pressure imbalance caused by temperature changes in the tank, but also further avoids oxidation during the extraction process, which would lead to the deactivation of biomass active ingredients; in addition, the flexible adjustment of the gas introduced by the air pump allows this hydrodynamic ultrasonic fluid generator to be used in conjunction with gas-responsive polarity switchable solvents, responding to the concept of green and sustainable chemistry. The circulating device enables the extraction solution to be circulated and refluxed for extraction, fully utilizing the solvent's extraction capacity. Intermittent extraction through reflux also protects the product's activity. Furthermore, after extraction, the circulating device can recover solvents with switchable polarity. When extraction is not performed through reflux, extraction can be carried out directly in the acoustic energy treatment tank, which can better avoid oxidation and contamination and improve extraction efficiency.

[0012] The purpose of the integrated circulating biomass extraction device provided by this invention is to achieve the following: (1) For solvents with switchable polarity in gas response, taking carbon dioxide response as an example, carbon dioxide can be introduced to make the solvent hydrophobic and dissolve hydrophobic substances. Then, by heating and introducing nitrogen, carbon dioxide can be removed, making the solvent hydrophilic and separating it from the previously extracted hydrophobic substances. The solvent can be reused by separating the liquid into the collector. (2) For ordinary solvents, new plant materials can be continuously added and circulated in the same solvent to make the solvent reach the upper limit of its extraction capacity and reach the most saturated state of the solvent for the target substance. (3) Another purpose of the circulation is intermittent extraction, that is, heating is only performed in the acoustic energy treatment tank. Combined with the reflux extraction of the circulation device, it is beneficial to protect the product activity and improve the yield and extraction efficiency.

[0013] Preferably, valve A is provided between the liquid storage tank and the ultrasonic generator.

[0014] Preferably, valve B is provided between the gas-liquid separator and the ultrasonic generator.

[0015] Preferably, the circulation pipeline is a double-layered pipeline, with the inner layer carrying the circulated extractable liquid and the outer layer carrying the external cooling liquid. The reflux design of the circulation pipeline is to prevent the active substances from oxidizing and becoming inactive due to temperature changes during prolonged ultrasound extraction. Intermittent extraction protects the product's activity, thereby improving yield and extraction efficiency.

[0016] Preferably, two liquid pumps are connected to the circulation pipeline; liquid pump A controls the circulation of liquid flowing through the inner pipeline, and liquid pump B controls the circulation of liquid flowing through the outer pipeline.

[0017] Preferably, the gas-liquid separator is covered with a temperature controller B.

[0018] Preferably, the device further includes a central control unit connected to valve A, acoustic energy processing tank, air pump, temperature controller A, gas-liquid separator, valve B, collector, liquid pump A, liquid pump B, liquid pump C, temperature controller B, and valve C. The external central control unit can simultaneously and precisely regulate various parameter values ​​of the device, as well as valves and switches.

[0019] Preferably, a centrifuge device is connected to the collector.

[0020] Meanwhile, this invention also provides a method for extracting biomass using a fluid-driven, ultrasonically integrated, temperature-controlled, circulating biomass extraction device. The extraction process involves adding the sample to be extracted and solvent to a storage tank in a specific ratio, adjusting the parameters of the acoustic energy processing tank and temperature controller A, turning on the air pump to introduce gas, and starting the acoustic energy processing tank for extraction. After extraction, the extract is pumped through a filter into a circulation pipeline by a liquid pump, and then returned to the acoustic energy processing tank for further cyclic extraction. After extraction, the gas-liquid separator is turned on to remove gas, and the extracted sample is pumped into a collector by a liquid pump C. After the extracted sample separates into layers, biomass is obtained.

[0021] The method provided by this invention not only effectively preserves the active components of various biomass and avoids oxidative deactivation during ultrasonic extraction, but also improves extraction yield. It is applicable to various solvent extraction methods and solvent recycling. When using a gas-responsive switchable solvent, hydrophobic / hydrophilic conversion can be achieved, facilitating the separation of target substances and solvent recovery, resulting in higher economic benefits. Furthermore, the entire extraction process is carried out in a closed loop within the device, effectively avoiding oxidation and contamination problems. The extraction temperature can be adjusted at any time, effectively preventing component damage caused by high temperatures. It is suitable for the extraction of various biomass, providing more effective methods and pathways for extracting easily oxidized and deactivated biomass. It solves the problems of existing extraction methods and equipment, such as high organic solvent consumption, low extraction efficiency, insufficient recycling of ultrasonic extraction solvents, non-closed extraction environment, and easy oxidation and deactivation of extracts caused by ultrasound. The extraction method and equipment provided by this invention not only improve extraction efficiency but also better preserve biological activity, increase biomass yield, and recycle solvents, avoiding pollution and waste.

[0022] Preferably, the sample to be extracted is selected from one or more of plants, animals, and microorganisms.

[0023] Preferably, the solvent is selected from one or more of the following: gas-responsive switchable solvents, eutectic solvents, organic solvents, and water.

[0024] More preferably, the gas-responsive switchable solvent includes, but is not limited to, one or more of the following types: amines and their derivatives, organic base solvents, guanidine solvents, ionic liquid gas-responsive solvents, amphiphilic molecular solvents, and redox-responsive solvents.

[0025] When a gas-responsive switchable solvent is used, the hydrophilic-hydrophobic conversion of the solvent is simply divided into hydrophilic and lipophilic. For example, if the substance to be extracted is hydrophilic, the switchable solvent is set to process the raw material in a hydrophilic state, allowing the hydrophilic components of the raw material to dissolve in the solvent. Subsequently, the switchable solvent is changed to lipophilic. The hydrophilic target extract then has a different polarity than the current lipophilic solvent, resulting in stratification and separation of the solvent and target substance, allowing for solvent recovery.

[0026] As a preferred implementation method, when extracting biomass from plants using the above-mentioned equipment, such as extracting Polygonatum polysaccharides, the specific method conditions are as follows: Polygonatum polysaccharides are extracted using a gas-responsive switchable solvent, N,N-dimethylbutylamine, and water. N,N-dimethylbutylamine and water are mixed evenly at a volume ratio of 1:1 to prepare the gas-responsive switchable solvent for later use. Then, dried Polygonatum powder and the gas-responsive switchable solvent are placed in storage tank 1 at a solid-liquid ratio of 1:20 g / g. Valve A is opened to inject the Polygonatum powder and the gas-responsive switchable solvent into the acoustic energy treatment tank. The temperature of temperature controller A is set to 50℃. Cooling water is introduced into the outer pipe of circulation pipe 61. CO2 is introduced into air pump 4. Liquid pumps A and B are turned on to start the circulation device. The acoustic energy treatment tank 21 is started with a power of 500 W and an extraction time of 60 minutes for ultrasonic extraction. Simultaneously, new plant material can be continuously added for cyclic extraction, allowing the solvent to reach its maximum extraction capacity and achieve maximum saturation of the target substance. After extraction, the gas pump was switched to nitrogen gas for 1 hour at a flow rate of 100 mL / min to remove CO2. The solution was then injected into collector 5. After settling, the solution separated into an upper oil phase and a lower aqueous phase. The lower aqueous phase is the extract rich in Polygonatum polysaccharides.

[0027] As a preferred implementation method, when extracting biomass from animals using the above-mentioned equipment, such as extracting chondroitin sulfate, the specific method and parameters are as follows: Chondroitin sulfate is extracted from bovine laryngeal cartilage using betaine and urea. Betaine and urea are prepared at a molar ratio of 1:2 to a solvent with a water content of 40%, and set aside. Valve B is kept open; nitrogen gas is introduced by an air pump at a flow rate of 100 mL / min. The solvent is mixed evenly with 60-mesh dried bovine laryngeal cartilage powder at a solid-liquid ratio of 30:1 mL / g, and injected into the acoustic energy treatment tank from the storage tank. The extraction conditions in the acoustic energy treatment tank are adjusted to 200W, temperature controller A is turned on to maintain the extraction environment at 110℃, and the device is started for extraction for 2 hours. After extraction, liquid pumps A and B are turned on to pump the extract through a filter into a circulation device for recirculation back to the acoustic energy treatment tank for intermittent extraction. After extraction is complete, the temperature of temperature controller A is set to 20℃, and after the solution cools, it is pumped into a collector to obtain the crude extract.

[0028] Further, the crude extract was centrifuged at 5000 rpm for 30 min, and the supernatant was filtered. Ethanol was added to bring the ethanol volume fraction of the system to 75%, and the mixture was allowed to stand overnight at 4°C. The mixture was then centrifuged (10000 rpm, 15 min, 4°C), and the above steps were repeated. The precipitate was resuspended in ultrapure water, placed in a 10 kDa dialysis bag, and dialyzed in ultrapure water at 4°C for 2 days. The dialysate was collected and lyophilized to obtain chondroitin sulfate.

[0029] As a preferred implementation method, when extracting biomass from microorganisms using the above-mentioned equipment, such as extracting morel polysaccharides, the specific method conditions are as follows: Morel polysaccharides are extracted using the solvents choline chloride and citric acid. A eutectic solvent with a water content of 44% is prepared by mixing choline chloride and citric acid at a molar ratio of 1:2 and set aside. The morel fruiting bodies are vacuum dried at 60℃ for 12 hours, then ultra-finely pulverized. An appropriate amount of powder is placed in a beaker, and 95% anhydrous ethanol is added at a material-to-liquid ratio of 1:20 g / mL. The mixture is then placed in a constant temperature water bath and stirred (70℃, 30 min), followed by rotary reflux for 2 hours (75℃) to remove some pigments, lipids, and other alcohol-soluble impurities. After standing until complete separation, the upper layer of ethanol is discarded, and the mixture is dried (50℃) and ground into a 40-mesh powder for later use. The eutectic solvent and morel powder are mixed evenly at a material-to-liquid ratio of 1:42 g / mL and injected into the acoustic energy treatment tank through a storage tank. Adjust the extraction conditions of the acoustic energy treatment vessel to 300W, turn on temperature controller A to maintain the extraction environment at 62℃, and start the device for extraction. After 65 minutes of extraction, turn off the ultrasound and adjust temperature controller A to lower the solution temperature to room temperature. Pump the solution into a collector using liquid pump C to obtain the crude extract. Add 4 times the volume of 95% ethanol to the crude extract, incubate overnight at 4℃ for alcohol precipitation, and then centrifuge (10610 rpm, 5 min) to obtain the crude polysaccharide extract.

[0030] The present invention has the following beneficial effects: This invention provides a fluid-powered, ultrasonically integrated, temperature-controlled, and circulating device and a method for extracting biomass. The device includes a storage tank, an ultrasonic generator, a gas-liquid separator, a circulation device, and a collector. Extraction using this integrated device allows for more complete extraction of biomass, increasing the extraction yield and effectively preserving the active ingredients in natural substances, thereby enhancing the activity effect. When extracting biomass using this device: intermittent ultrasonic extraction via the ultrasonic generator and circulation device better preserves active ingredients; a constant extraction temperature is maintained by a temperature controller, preventing component damage due to high temperatures; gas is introduced via a gas pump to balance the gas pressure and prevent oxidation; simultaneously, the device can be used with a gas-responsive, switchable solvent to achieve hydrophobic / hydrophilic conversion, facilitating the separation of the target substance and solvent recovery, resulting in higher economic efficiency; the circulation device allows for multiple extraction cycles, ensuring consistent extraction environmental conditions, fully utilizing the solvent's dissolving power for the target substance, better preserving product activity, and applicability to different extraction solvents and raw materials. The entire extraction process is controllable, minimizing oxidation, pollution, and solvent waste, providing more effective methods and pathways for extracting easily oxidized and deactivated biomass. Attached Figure Description

[0031] Figure 1 This is a schematic diagram of the fluid-driven ultrasonic integrated temperature-controlled circulating extraction device of Example 1.

[0032] Reference numerals in the attached drawings: 1-storage tank, 2-ultrasonic generator, 21-sound energy processing tank, 22-air whistle, 23-filter, 24-temperature controller A, 3-gas-liquid separator, 31-temperature controller B, 4-air pump, 5-collector, 6-circulation device, 61-circulation pipeline, 62-liquid pump, 7-liquid pump C, 8-valve A, 9-valve B. Detailed Implementation

[0033] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0034] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0035] Example 1: A fluid-driven ultrasonic generator and an integrated temperature-controlled circulating extraction device for multiple solvents This embodiment provides a fluid dynamic ultrasonic integrated circulation extraction device for biomass, including a liquid storage tank 1, an ultrasonic generator 2, a gas-liquid separator 3, an air pump 4, a collector 5, and a circulation device 6.

[0036] The ultrasonic generator 2 includes: a sealed acoustic energy processing tank 21, a whistle 22, a filter 23, and a temperature controller A 24; the whistle 22 is placed at one end inside the acoustic energy processing tank 21, and the filter 23 is placed at the other end inside the acoustic energy processing tank 21 away from the whistle 22; the end of the ultrasonic generator 2 near the whistle 22 is connected to a liquid storage tank 1 through a pipe, and the rear end near the filter 23 is connected to a gas-liquid separator 3 through a pipe; a collector 5 is connected to the rear end of the filter 23 through a pipe; a temperature controller A 24 is installed on the outside of the ultrasonic generator 2; an air pump 4 is connected to the whistle 22 in the ultrasonic generator 2 through a pipe; and a liquid pump C 7 is provided between the ultrasonic generator 2 and the collector 5.

[0037] The ultrasonic generator 2 is used for ultrasonic reactions, the gas whistle 22 is used to introduce gas or circulate liquid, and the filter screen 23 is installed before the gas-liquid separator 3 and the collector 5 to filter raw materials or impurities. The extract after ultrasonic treatment in the acoustic energy treatment tank 21 passes through the filter screen and enters the collector 5 or the circulation device 6. The gas-liquid separator 3 is used to balance the gas pressure and discharge gas. Its exterior is covered with a temperature controller B 31 to regulate the temperature, which helps to remove gases such as carbon dioxide from the solvent to adjust the polarity.

[0038] The circulation device 6 includes a circulation pipe 61 and a liquid pump 62. The liquid pump 62 is connected to the circulation pipe 61. One end of the circulation pipe 61 is connected to the air whistle 22 in the ultrasonic generator 2, and the other end is connected to the end of the ultrasonic generator 2 near the filter screen 23. The circulation pipe is a double-layer pipe; the inner layer pipe carries the circulated extracted liquid, and the outer layer pipe carries the external coolant. Two liquid pumps are connected to the circulation pipe; liquid pump A controls the circulation of the liquid in the inner layer pipe, and liquid pump B controls the circulation of the liquid in the outer layer pipe.

[0039] A valve A 8 is installed between the liquid storage tank 1 and the ultrasonic generator 2, and a valve B 9 is installed between the gas-liquid separator 3 and the ultrasonic generator 2.

[0040] Equipment structure diagram as follows Figure 1 As shown, the extracted sample and solvent are added to the storage tank 1 in proportion. Valve A8 is opened, and after the material enters the acoustic energy treatment tank 21, valve A8 is closed. The ultrasonic extraction parameters are adjusted, and the temperature parameters of the temperature controller A24 are set to maintain a constant extraction temperature. Gas is introduced through the air pump 4, and cooling water (or liquids of different temperatures can be used depending on the extraction scheme) is introduced through the outer pipe of the circulation pipe 61. The acoustic energy treatment tank 21 is then started for circulation extraction. After extraction, gas is introduced through the air pump 4, and the gas-liquid separator 3 is started to discharge the gas. Subsequently, the extracted solution is injected into the collector 5 through the liquid pump C7. After standing, the solution separates into an upper oil phase and a lower aqueous phase. The oil phase or aqueous phase is collected based on the extracted substances, thus obtaining the extracted biomass.

[0041] Example 2: Extraction of biomass from plants using a gas-responsive switchable solvent. This embodiment utilizes the gas-responsive switchable solvent N,N-dimethylbutylamine and water to extract Polygonatum polysaccharides. The fluid-driven ultrasonic integrated circulation extraction device of Example 1 is used. N,N-dimethylbutylamine and water are mixed uniformly at a 1:1 volume ratio to prepare the gas-responsive switchable solvent for later use. Then, dried Polygonatum powder and the gas-responsive switchable solvent are placed in storage tank 1 at a solid-liquid ratio of 1:20 g / g. Valve A8 is opened to inject the Polygonatum powder and gas-responsive switchable solvent into the acoustic energy treatment tank 21. After injection, valve A8 is closed. The temperature of the temperature controller A24 covering the acoustic energy treatment tank 21 is set to maintain the temperature inside the acoustic energy treatment tank 21 at 50°C. Cooling water is introduced into the outer pipe of the circulation pipe 61. CO2 is introduced into the air pump 4. Liquid pumps A 621 and B 622 are turned on to start the circulation device 6. The acoustic energy treatment tank 21 is started at 500W power for 60 minutes for ultrasonic extraction. At the same time, by continuously adding new plant materials and the same solvent for cyclic extraction, the solvent can reach its upper limit of extraction capacity and achieve the most saturated state of the solvent for the target substance.

[0042] After extraction, gas pump 4 was switched to nitrogen gas for 1 hour at a flow rate of 100 mL / min to remove CO2, and the solution was then injected into collector 5. After standing, the solution separated into an upper oil phase and a lower aqueous phase. The lower aqueous phase is extract 1, which is rich in Polygonatum polysaccharides.

[0043] The upper oil phase is then pumped into the circulation pipeline 61 via liquid pump C7, allowing it to circulate back to the acoustic energy treatment tank 21. Water, with a volume equal to that of the lower aqueous phase, is then injected into the acoustic energy treatment tank 21 via the storage tank 1. Turning on the gas pump 4 introduces CO2, completing the solvent recovery and initiating the next extraction cycle.

[0044] Example 3: Extraction of biomass from animals This embodiment utilizes the fluid-driven ultrasonic integrated circulating extraction equipment for biomass extraction described in Example 1. Betaine and urea are used as solvents to extract chondroitin sulfate from bovine laryngeal cartilage. The specific extraction steps using the equipment are as follows: S1: Prepare solvent 2 with a water content of 40% by mixing betaine and urea in a molar ratio of 1:2.

[0045] S2: Keep valve B9 open; introduce nitrogen gas through air pump 4 at a flow rate of 100 mL / min.

[0046] S3: Mix solvent 2 with 60-mesh dried bovine laryngeal cartilage powder at a solid-liquid ratio of 30:1mL / g, and pour the mixture into the acoustic energy treatment tank 21 from the storage tank 1.

[0047] S4: Adjust the extraction conditions of the acoustic energy treatment tank 21 to 200W, turn on the temperature controller A 24 to keep the extraction environment at 110℃, start the device, and extract for 2 hours; after extraction, turn on the liquid pump A 621 and liquid pump B 622 to pump the extract through the filter into the circulation device 6 for circulation back to the acoustic energy treatment tank 21 for intermittent extraction.

[0048] S5: After extraction is complete, set the temperature of the temperature controller A 24 to 20°C. After the solution cools, pump the solution into the collector 5 to obtain the crude extract 2.

[0049] S6: Centrifuge crude extract 2 at 5000 rpm for 30 min, filter to obtain the supernatant, add ethanol to make the ethanol volume fraction of the system reach 75%, let stand overnight at 4℃, centrifuge (10000 rpm, 15 min, 4℃) to obtain crude extract 3, repeat the above steps, centrifuge again. Resuspend the precipitate in ultrapure water, place it in a 10 kDa dialysis bag, dialyze in ultrapure water at 4℃ for 2 days, collect the dialysate and freeze dry to obtain extract 4.

[0050] Example 4: Extraction of biomass from microorganisms This embodiment utilizes the fluid-driven ultrasonic integrated circulating extraction equipment from Example 1 for biomass extraction, employing choline chloride and citric acid as solvents to extract morel polysaccharides. The specific extraction steps are as follows: S1: Prepare a eutectic solvent with a water content of 44% by mixing choline chloride and citric acid in a molar ratio of 1:2, and set aside for later use.

[0051] S2: Vacuum-dry morel fruiting bodies at 60℃ for 12 hours, then pulverize them into ultrafine powder. Place an appropriate amount of powder in a beaker, add 95% anhydrous ethanol at a material-to-liquid ratio of 1:20 g / mL, and stir in a constant temperature water bath (70℃, 30 min). Reflux by rotary evaporation for 2 hours (75℃) to remove some pigments, lipids, and other alcohol-soluble impurities. Allow to stand until complete separation, discard the upper ethanol layer, dry (50℃), and grind into 40-mesh powder for later use.

[0052] S3: Mix the eutectic solvent and morel powder evenly at a material-to-liquid ratio of 1:42 g / mL, and inject the mixture into the acoustic energy treatment tank 21 through the storage tank 1.

[0053] S5: Adjust the extraction conditions of the acoustic energy treatment tank 21 to 300W, turn on the temperature controller A 24 to maintain the extraction environment at 62℃, start the device, and carry out extraction.

[0054] S6: After extraction for 65 minutes, turn off the ultrasound and adjust the temperature controller A24 to lower the solution temperature to room temperature.

[0055] S7: The solution is pumped into collector 5 by liquid pump C7 to obtain crude extract 5.

[0056] S8: Add 4 times the volume of 95% ethanol to crude extract 5, precipitate overnight at 4°C, and then centrifuge (10610r, 5min) to obtain crude polysaccharide extract 6.

[0057] Comparative Example 1: Extraction of biomass from microorganisms without starting the circulation process This comparative example utilizes the fluid-driven ultrasonic integrated circulation extraction device from Example 1 for biomass extraction, employing choline chloride and citric acid as solvents to extract morel polysaccharides. The difference from Example 2 is that the circulation device is not activated during extraction; specifically, liquid pumps A 621 and B 622 are not turned on for reflux extraction. The specific operation is as follows: The fluid-driven ultrasonic integrated circulation extraction device of Example 1 was used. N,N-dimethylbutylamine and water were mixed uniformly at a 1:1 volume ratio to prepare a gas-responsive switchable solvent for later use. Then, dried Polygonatum sibiricum powder and the gas-responsive switchable solvent were placed in storage tank 1 at a solid-liquid ratio of 1:20 g / g. Valve A8 was opened to inject the Polygonatum sibiricum powder and the gas-responsive switchable solvent into the acoustic energy treatment tank 21. After injection, valve A8 was closed. The temperature of the temperature controller A24 covering the acoustic energy treatment tank 21 was set to maintain the temperature inside the acoustic energy treatment tank 21 at 50°C. The acoustic energy treatment tank 21 was started with a power of 500 W and an extraction time of 60 min for ultrasonic extraction. After extraction, gas pump 4 was switched to nitrogen gas for 1 hour at a flow rate of 100 mL / min to remove CO2, and the solution was injected into collector 5. After standing, the solution separated into an upper oil phase and a lower aqueous phase. The lower aqueous phase is the extract 3 rich in Polygonatum sibiricum polysaccharides.

[0058] Test Example 1: Determination and Comparison of Antioxidant Activity of Polygonatum Polysaccharides 1. Comparison of yields Extraction of Polygonatum polysaccharides was performed strictly according to the extraction method described in the existing technology, "Research on Extraction and Separation of Plant Polysaccharides Using Environmentally Responsive Switchable Hydrophilic Solvents," to obtain Polygonatum polysaccharide extract 2 (i.e., extraction without using the equipment of Example 1). After extraction, the yields of Polygonatum polysaccharides extracted by conventional methods, the method of Example 2, and the method of Comparative Example 1 were statistically analyzed. The results are shown in Table 1, indicating that the yield of Polygonatum polysaccharides extracted using the equipment and method provided in this invention is higher than that of existing methods. Furthermore, in Comparative Example 1, extraction without the circulation device also yielded a higher yield than conventional extraction methods, and extraction with the circulation device further improved the yield. This data fully demonstrates that the extraction equipment provided by this invention, compared to traditional extraction methods, can extract plant polysaccharides more efficiently and improve the yield, exhibiting significant technical advantages and practical application value.

[0059] Table 1 Effects of different extraction methods on Polygonatum polysaccharides

[0060] 2. Comparison of activity effects Accurately weigh 4 mg of DPPH and dissolve it in methanol, then dilute to a 100 mL volumetric flask to obtain the DPPH solution. Take different volumes (0.1, 0.3, and 0.5 mL) of the following extracts (to bring the volume up to 0.5 mL with methanol): Polygonatum sibiricum polysaccharide extract 1 from Example 2, Polygonatum sibiricum polysaccharide extract 2 extracted using existing technology, and Polygonatum sibiricum polysaccharide extract 3 from Comparative Example 1, and 2 mL of DPPH solution respectively) and place them in the same test tube, then shake well. Incubate in the dark for 30 min, using the corresponding blank solution as a control, and measure the absorbance at 517 nm. The DPPH free radical inhibition rate is calculated using the following formula: Inhibition rate % = [A0 - (Ab - A*)] / A0 * 100%; Where: A0 is the absorbance after the reaction without the addition of sample inhibitor; Ab represents the absorbance after the reaction with the sample inhibitor; A* represents the absorbance without the addition of inhibitors and DPPH.

[0061] The test results are shown in Table 2. The results show that the Polygonatum polysaccharide extract 1 obtained by the equipment of Example 1 has better activity. At the same concentration, the Polygonatum polysaccharide extract 1 has a significantly better free radical scavenging effect than Polygonatum polysaccharide extract 2 and Polygonatum polysaccharide extract 3 extracted by existing technology. The results show that the extraction using the equipment of Example 1 can better preserve its active substances. Compared with Polygonatum polysaccharide extract 3 which is not subjected to cyclic extraction, Polygonatum polysaccharide extract 1 has a better free radical scavenging effect at the same dosage.

[0062] Table 2. Scavenging effects of Polygonatum polysaccharide extracts obtained by different extraction methods on free radicals.

[0063] Test Example 2: Comparison of chondroitin sulfate yields obtained from different extraction methods and equipment Chondroitin sulfate was extracted from bovine laryngeal cartilage strictly according to the existing technology: "Efficient Extraction and Structural Characterization of Chondroitin Sulfate by Eutectic Solvent Method". Chondroitin sulfate was obtained (the method conditions and parameters were the same, except that the equipment of Example 1 was not used for extraction). The yield was calculated and compared with the yield of extract 4 obtained by the equipment of Example 3.

[0064] The results are shown in Table 3. The results show that the yield of chondroitin sulfate extracted using the equipment of Example 1 is higher than that of existing extraction methods. Extraction by circulating temperature control in an integrated device can better avoid the oxidation and loss of biomass and improve extraction efficiency.

[0065] Table 3. Yields of chondroitin sulfate extracted by different methods

[0066] Test Example 3: Determination of Microbial Polysaccharide Yield and Comparison of Activity Effects 1. Determination of microbial polysaccharide yield Morel polysaccharides were extracted strictly according to the existing technology: "Optimization of the Extraction Process of Morel Polysaccharides with Eutectic Solvent and Its Probiotic Proliferation Effect". The polysaccharide content was determined according to the method in Example 1 (the method conditions and parameters were the same, the difference was that the equipment of Example 1 was not used for extraction). The crude polysaccharide extract 6 was reconstituted with distilled water and the volume was adjusted to 100 mL. The polysaccharide content was determined by phenol-sulfuric acid method.

[0067] The specific steps are as follows: prepare glucose standard solutions of different concentrations, add phenol solution and concentrated sulfuric acid respectively, and measure the absorbance at a wavelength of 490 nm after color development, and plot the standard curve; similarly, treat the polysaccharide sample solution according to the standard solution method, measure the absorbance, calculate the polysaccharide concentration according to the standard curve, and then determine the polysaccharide content.

[0068] The results are shown in Table 4, which shows that the yield of microbial polysaccharides extracted using the equipment of Example 1 is higher than that of existing extraction methods. Under the same extraction conditions, the equipment provided by this invention can further improve the yield and maximize the extraction effect. Moreover, this equipment is suitable for the extraction of biomass from a variety of raw materials, with a higher yield than existing extraction methods. It can better avoid the oxidation and loss of biomass and better preserve active substances.

[0069] Table 4. Yields of polysaccharides extracted by different extraction methods

[0070] 2. The effect of microbial polysaccharides on acid production by probiotics The crude polysaccharide extract 6 extracted in Example 4 was purified according to the purification method in the existing technology, "Optimization of Extraction Process of Morel Polysaccharides by Eutectic Solvent and Its Probiotic Proliferation Effect". The specific steps included potassium ferrocyanide-zinc acetate method, ion exchange column chromatography, and gel column chromatography to obtain polysaccharide extract 7. At the same time, the purified product D-MSP-3 of Morel polysaccharides extracted by the existing technology was used as a control to determine the effect of different microbial polysaccharides on probiotic acid production.

[0071] Strain activation: Using a sterile pipette, 0.3 mL of liquid culture medium (sterilized at 121℃ for 15 min) was added to a lyophilized strain tube (using *Lactobacillus plantarum* and *Lactobacillus acidophilus* strains). The tube was gently shaken until dissolved. The entire bacterial suspension was then inoculated into liquid culture medium and anaerobically cultured at 37℃ for 48 h. Subsequently, 5% (v / v) of the culture medium was inoculated onto MRS solid culture medium, spread evenly, and anaerobically cultured at 48 h. Single bacteria were picked from the cultured culture and inoculated into MRS liquid culture medium, then anaerobically cultured at 48 h to obtain the third-generation activated strain, which was then refrigerated for later use.

[0072] Liquid culture media containing different concentrations (0, 0.5, 1, 1.5, 2, 3%) of polysaccharide extract 7 or D-MSP-3 as a single carbon source were sterilized (121℃, 15 min, the same below), and then activated bacterial strains were inoculated at a ratio of 5% (v / v). After culturing (37℃, anaerobic, the same below) for 48 h, the pH was measured and the values ​​were statistically analyzed.

[0073] The effects of morel polysaccharide extract 7 and D-MSP-3 on acid production of *Lactobacillus plantarum* and *Lactobacillus acidophilus* are shown in Table 5. Acid production capacity is a measure of the quality of acid-producing bacteria cultivation; higher acid production capacity indicates better polysaccharide activity. This shows that morel polysaccharide extract 7 has a better promoting effect on the acid production capacity of *Lactobacillus plantarum* and *Lactobacillus acidophilus* than D-MSP-3 extracted using existing technologies. Therefore, the integrated extraction equipment provided by this invention helps maintain and preserve the activity of the extract, better retaining bioactive substances while improving yield, resulting in better overall performance.

[0074] Table 5. Effects of different polysaccharide extracts on acid production by Lactobacillus plantarum and Lactobacillus acidophilus.

[0075] In summary, the fluid-powered ultrasonic integrated temperature-controlled circulating extraction equipment demonstrates superior extraction efficiency, better preservation of biomass active ingredients, and improved extraction efficiency. This equipment enables integrated circulating extraction within a closed environment, precisely controlling extraction temperature conditions. Combined with a gas-liquid circulation system, intermittent extraction effectively prevents overheating and target inactivation due to prolonged ultrasonic treatment, while also balancing gas pressure to avoid oxidation during extraction. Furthermore, the circulation system allows for multiple solvent cycles, maximizing the solvent's solubility for the target substances. Additionally, flexible gas flow adjustment allows for extraction using gas-responsive, polarity-switching solvents, further enhancing solvent solubility and aligning with green and sustainable chemistry principles. This addresses the problems of existing extraction methods and equipment, such as high organic solvent consumption, low extraction efficiency, insufficient solvent recycling in ultrasonic extraction, unsealed extraction environments, and easy oxidation and inactivation of extracts due to ultrasonic heat generation. The extraction method and equipment provided by this invention not only improve extraction efficiency but also better preserve bioactivity, increase biomass yield, and recycle solvents, avoiding pollution and waste.

[0076] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A device for fluid-driven, ultrasonically integrated, temperature-controlled, circulating extraction of biomass, characterized in that, The equipment includes a liquid storage tank (1), an ultrasonic generator (2), a gas-liquid separator (3), an air pump (4), a collector (5), and a circulation device (6); The ultrasonic generator (2) includes: a sealed acoustic energy processing tank (21), a whistle (22), a filter (23), and a temperature controller A (24); the whistle (22) is placed at one end inside the acoustic energy processing tank (21), and the filter (23) is placed at the other end inside the acoustic energy processing tank (21) away from the whistle (22); the end of the ultrasonic generator (2) near the whistle (22) is connected to the liquid storage tank (1) through a pipe, and the rear end near the filter (23) is connected to the gas-liquid separator (3) through a pipe; a collector (5) is connected to the rear end of the filter (23) through a pipe; a temperature controller A (24) is installed on the outside of the ultrasonic generator (2); the air pump (4) is connected to the whistle (22) in the ultrasonic generator (2) through a pipe; The circulation device (6) includes: a circulation pipe (61) and a liquid pump (62); the liquid pump (62) is connected to the circulation pipe (61); one end of the circulation pipe (61) is connected to the air whistle (22) in the ultrasonic generator (2), and the other end is connected to the end of the ultrasonic generator (2) near the filter (23); A liquid pump C (7) is provided between the ultrasonic generator (2) and the collector (5).

2. The device according to claim 1, characterized in that, A valve A (8) is provided between the liquid storage tank (1) and the ultrasonic generator (2); a valve B (9) is provided between the gas-liquid separator (3) and the ultrasonic generator (2).

3. The device according to claim 2, characterized in that, The circulation pipe (61) is a double-layer pipe, with the inner layer containing the circulated extracted liquid and the outer layer containing the external temperature-controlled liquid.

4. The device according to claim 3, characterized in that, Two liquid pumps are connected to the circulation pipe (61). Liquid pump A (621) controls the circulation of liquid in the inner pipe, and liquid pump B (622) controls the circulation of liquid in the outer pipe.

5. The device according to claim 4, characterized in that, The gas-liquid separator (3) is covered with a temperature controller B (31).

6. The device according to claim 5, characterized in that, The device is also connected to a main control unit, which is connected to the liquid storage tank (1), ultrasonic generator (2), gas-liquid separator (3), air pump (4), collector (5), circulation device (6), valve A (8), valve B (9), liquid pump C (7), and temperature controller B (31) via electrical signals.

7. The device according to claim 1, characterized in that, A centrifuge device is connected to the collector (5).

8. A method for extracting biomass using the equipment described in any one of claims 1 to 7, characterized in that, Extraction is performed using the equipment described in any one of claims 1 to 6. The specific method is as follows: the sample to be extracted and the solvent are added to the storage tank (1) in proportion, the parameters of the acoustic energy processing tank (21) and the temperature controller A (24) are adjusted, the gas pump (4) is turned on to introduce gas, the acoustic energy processing tank (21) is started to extract, after extraction, the extract is pumped into the circulation pipeline (61) by the liquid pump (62) through the filter screen (23), and then returned to the acoustic energy processing tank (21) for circulation extraction; after extraction is completed, the gas-liquid separator (3) is turned on to remove the gas, and the extracted sample is pumped into the collector (5) by the liquid pump C (7). After the extracted sample is separated into layers, biomass is obtained.

9. The method according to claim 8, characterized in that, When the solvent is a gas-responsive switchable solvent, after extraction is completed, the gas pump (4) is turned on to introduce gas to switch the solvent polarity, so that the solvent and the extracted target biomass are separated into layers, thereby separating and recovering the solvent.

10. The method according to claim 9, characterized in that, The switchable gas-response solvent is selected from one of the following: amines and their derivatives, organic base solvents, guanidine solvents, ionic liquid gas-response solvents, amphiphilic molecular solvents, and redox-response solvents.

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