Medicinal and edible plant chromatographic separation and extraction process and equipment based on enzymolysis method

Abstract

The invention relates to the technical field of chromatographic separation and extraction of medicinal and edible plants, in particular to a process and equipment for chromatographic separation and extraction of medicinal and edible plants based on an enzymolysis method, which are characterized in that a regenerated macroporous resin chromatographic column can be quickly replaced to quickly enter a chromatographic filtration state, so that the advantage of convenience in regeneration is preliminarily achieved; through mutual clamping of a butt joint seat and a clamping head, a second motor is used for driving the butt joint seat to rotate to drive the clamping head and a plurality of stirring pipes in the macroporous resin chromatographic column to rotate, and the movement frequency of macroporous resin in the chromatographic column is improved through a stirring mode, so that the macroporous resin cleaning efficiency is further improved; the cleaning liquid is finally discharged from the orifices of the plurality of stirring pipes and directly acts on the surface of the macroporous resin in the chromatographic column, so that the cleaning efficiency of the macroporous resin is further improved, the macroporous resin does not need to be taken out from the chromatographic column in the whole regeneration process, and the device has the advantages of few operation steps and convenience in regeneration.

Description

Technical Field

[0001] The invention relates to the technical field of chromatographic separation and extraction of edible and medicinal plants, and in particular to a chromatographic separation and extraction process and equipment for edible and medicinal plants based on an enzymatic method. Background Art

[0002] The chromatographic separation and extraction process of medicinal and edible plants based on enzymatic hydrolysis is an advanced process that combines enzymatic hydrolysis technology and chromatographic separation technology, and is designed to efficiently extract and separate active ingredients from medicinal and edible plants.

[0003] Chromatographic separation technology is a method that achieves separation based on the different distribution coefficients of components in a mixture between the stationary phase and the mobile phase. In the extraction process of medicinal and edible plants, chromatographic separation technology is used to further separate and purify the active ingredients.

[0004] The chromatography separation and extraction method taking puerarin as an example is roughly as follows: a crude extract or puerarin powder containing puerarin is obtained as a starting material, and a suitable solvent is selected for dissolution and elution according to the solubility of puerarin and the properties of the chromatography medium. Common solvents include water, ethanol, etc., and a suitable chromatography medium is selected, such as polyamide, macroporous resin, silica gel, etc., for pretreatment and column loading. The dissolved puerarin crude extract or pueraria powder solution is loaded onto the chromatography column and eluted with an appropriate solvent. According to the adsorption and desorption properties of puerarin on the chromatography medium, suitable elution conditions and flow rates are selected. Common elution solvents include water, ethanol, methanol, etc., and sometimes the pH value of the solution needs to be adjusted. During the elution process, the eluate is collected in sections and tested and analyzed. The puerarin content in the eluate is usually determined by ultraviolet spectrophotometry, high performance liquid chromatography, etc. If the purity of puerarin needs to be further improved, an extraction method can be used for further separation.

[0005] However, when chromatographic columns such as macroporous resins are used for a certain period of time, their adsorption capacity decreases or they are seriously contaminated and need to be regenerated. Traditional regeneration methods usually include soaking, rinsing and column passing with acid and alkali solutions to restore the adsorption capacity of the resin. However, traditional regeneration methods require the chromatographic column to be disassembled and cleaned in multiple steps. After cleaning, it needs to be reinstalled and reset, which results in inconvenience in regeneration, thereby affecting the smooth progress of the chromatographic separation process. Summary of the invention

[0006] In view of the above-mentioned shortcomings of the prior art, the present invention provides a chromatographic separation and extraction process and equipment for medicinal and edible plants based on enzymatic hydrolysis, which can effectively solve the problems raised in the background technology.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions: The present invention provides a chromatographic separation and extraction process of medicinal and edible plants based on an enzymatic method, comprising the following steps: S1, raw material pretreatment, select fresh, dry, non-mildewed Pueraria root as raw material, then peel and wash the Pueraria root, grind it with a grinder and filter it, select a 10-mesh sieve to obtain Pueraria root powder; S2, enzymatic hydrolysis, selecting cellulase as an enzymatic hydrolyzing agent, and adding kudzu root powder and cellulase into a beaker in equal proportions, respectively, and enzymatically hydrolyzing the kudzu root powder at an appropriate temperature and pH value to release puerarin; S3, extraction and filtration, extracting puerarin from kudzu root powder by hot water extraction, filtering the solution after the extraction to remove undissolved kudzu root powder and other impurities, and collecting the filtrate after filtration to obtain the extract containing puerarin; S4, chromatographic separation, collecting the eluate through a chromatographic separation device, the specific method is to select a suitable chromatographic medium, such as a macroporous resin, perform pretreatment and column loading, load the dissolved puerarin crude extract or pueraria root powder solution onto the chromatographic column, use a suitable solvent, such as ethanol, for elution, pass the eluate through the chromatographic column at a suitable flow rate, collect the eluate, select suitable elution conditions and flow rate according to the adsorption and desorption properties of puerarin on the chromatographic medium, commonly used elution solvents include water, ethanol, methanol, etc., and the pH value of the solution needs to be adjusted during the process; S5. Extraction and purification: concentrate the ethanol eluate to recover the ethanol, then add n-butanol for extraction to further purify puerarin. Crystallize the puerarin solution obtained by extraction, filter out the crystals and dry them to obtain puerarin concentrate. A chromatographic separation and extraction device for medicinal and edible plants based on an enzymatic hydrolysis method, the chromatographic separation device comprising a U-shaped bracket, a discharge pipe being rotatably mounted on the U-shaped bracket, a rotary connector being rotatably mounted between two opposite inner side walls of the U-shaped bracket and being connected to an inner cavity of the discharge pipe, two chromatographic separation cylinders being symmetrically mounted on the top and bottom of the rotary connector respectively, the two chromatographic separation cylinders being connected to the same discharge pipe via the rotary connector, solenoid valves being arranged at the connection points between the two chromatographic separation cylinders and the rotary connector to maintain unidirectional flow, two fine-mesh filters being symmetrically distributed up and down being mounted on the inner cylinder walls of the two chromatographic separation cylinders, and macroporous resin being filled between the two fine-mesh filters in the same chromatographic separation cylinder; The two feed stirring assemblies are respectively installed in the corresponding chromatographic separation cylinders, and both include a clamping joint rotatably installed at the end of the corresponding chromatographic separation cylinder, and a rotating tube rotatably installed inside the chromatographic separation cylinder and located between two fine-mesh filters. The outer tube walls of the two rotating tubes are installed with a plurality of stirring tubes connected to the corresponding clamping joints. The docking seat is lifted and slid in the U-shaped bracket, and rises to the highest point to dock with the corresponding clamping joint. The docking seat can rotate to drive the clamping joint and the corresponding rotating tube and stirring tube to rotate. The cleaning pump is installed on the outer side wall of the U-shaped bracket, and the output end is connected to the inner cavity of the chromatographic separation cylinder through the docking seat and the clamping joint.

[0008] Furthermore, a first motor is vertically fixedly installed on the outer wall of the U-shaped bracket, the output end of the first motor is connected to the rotating rod, the other end of the rotating rod passes through the U-shaped bracket and is vertically fixedly connected to the outer ring wall of the rotating connector, and a limit ring is fixedly sleeved on the outer tube wall of the discharge pipe, and the limit ring is rotatably sleeved in the U-shaped bracket.

[0009] Furthermore, the top and bottom of the rotary connector are provided with sockets for convenient insertion of the chromatography separation cartridge, the inner walls of the two sockets are provided with inner grooves, and two one-way drain valves are installed at the connection between the rotary connector and the discharge pipe.

[0010] Furthermore, one end of the chromatographic separation cylinder is equipped with a solenoid valve respectively inserted into the corresponding inner slot, and the outer cylinder wall of the other end of the chromatographic separation cylinder is equipped with an exhaust pipe, which is connected to the inner cylinder cavity of the chromatographic separation cylinder, and an exhaust valve is installed in the exhaust pipe.

[0011] Furthermore, the feed stirring assembly includes a rotating seat rotatably installed at the port of the corresponding chromatographic separation cylinder, the rotating seat is penetrated by a through hole, the clamping joints are respectively fixedly installed on the corresponding rotating seats, the rotating tube is installed at the bottom of the rotating seat and is connected to the inner cavity of the clamping joint through the through hole, and several of the stirring tubes are obliquely installed on the outer wall of the corresponding rotating tube.

[0012] Furthermore, a docking hole adapted to the card joint is provided at the top of the docking seat, a rotating cavity and a square hole are provided at the bottom of the docking seat, the square hole is connected to the docking hole, a square block is adapted to be inserted into the square hole, a connecting round seat is installed at the bottom of the square block, a connecting round tube is connected between the connecting round seat and the square block, the connecting round seat is connected to the docking hole through the connecting round tube, the square block and the square hole, and a plurality of connecting small holes are provided on the outer ring wall of the connecting round seat through the inner cavity.

[0013] Furthermore, a shell is provided below the docking seat and is sleeved around the connecting round seat. A movable cavity is provided in the shell. A first rotating circular hole is provided at the top of the shell and passes through the movable cavity to facilitate the rotating installation of the connecting round tube. A second rotating circular hole is provided at the bottom of the shell and passes through the movable cavity.

[0014] Furthermore, a second motor is installed at the bottom of the inner cavity of the U-shaped bracket, and a driving rod is connected to the output end of the second motor. The top of the driving rod passes through the second rotating circular hole and is fixedly connected to the lower bottom of the connecting round seat. A lifting cylinder is fixedly installed on the other outer side wall of the U-shaped bracket, and an L-shaped lifting rod is installed at the output end of the lifting cylinder. The L-shaped lifting rod passes through the U-shaped bracket and is fixedly connected to the outer ring wall of the outer shell.

[0015] Furthermore, the output end of the cleaning pump is connected to a cleaning pipe, the end of the cleaning pipe passes through the U-shaped bracket and is connected to the outer wall of the shell, and the cleaning pipe is connected to the active cavity.

[0016] Compared with the known prior art, the technical solution provided by the present invention has the following beneficial effects: 1. The end parts of two macroporous resin chromatography columns are installed head to head in a linear manner, and the regenerated macroporous resin chromatography column can be quickly replaced by driving the two macroporous resin chromatography columns to rotate at the same time, so that the regenerated macroporous resin chromatography column can be quickly put into the chromatography filtration state, reducing the steps of disassembly and assembly required for regenerating the macroporous resin chromatography column, thereby initially having the advantage of convenient regeneration.

[0017] 2. A liquid inlet stirring assembly is provided in the macroporous resin chromatography column. When the macroporous resin chromatography column rotates to a position where the clamp is aligned with the docking seat, the docking seat and the clamp are engaged with each other, and a second motor is used to drive the docking seat to rotate, thereby driving the clamp and a plurality of stirring tubes in the macroporous resin chromatography column to rotate. The movement frequency of the macroporous resin in the chromatography column is increased by stirring, thereby further improving the cleaning efficiency of the macroporous resin. Cleaning liquid is injected into the docking seat accompanied by the stirring of the plurality of stirring tubes, and the cleaning liquid is finally discharged from the nozzles of the plurality of stirring tubes to directly act on the surface of the macroporous resin in the chromatography column, thereby further improving the cleaning efficiency of the macroporous resin. The macroporous resin does not need to be taken out of the chromatography column during the entire regeneration process, which has the advantages of fewer operation steps and convenient regeneration. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 It is a schematic diagram of the overall structure of the present invention.

[0020] Figure 2 It is a schematic diagram of the installation structure of the chromatography separation cartridge of the present invention.

[0021] Figure 3 For the present invention Figure 2 Enlarged structural diagram at A in the middle.

[0022] Figure 4 This is a schematic diagram of the installation structure of the feed stirring assembly of the present invention.

[0023] Figure 5 It is a schematic diagram of the structure of the feed stirring assembly of the present invention.

[0024] Figure 6 It is a schematic diagram of the installation structure of the docking socket of the present invention.

[0025] Figure 7 It is a schematic diagram of the installation structure of the connecting round seat and the docking seat of the present invention.

[0026] Figure 8 It is a schematic diagram of the shell structure of the present invention.

[0027] The numbers in the figure represent: 1. U-shaped bracket; 11. first motor; 12. rotating rod; 13. discharge pipe; 14. limiting ring; 2. Rotating connector; 21. Socket; 22. Internal slot; 23. One-way drain valve; 3. Chromatographic separation cylinder; 31. Solenoid valve; 32. Fine-mesh filter; 33. Exhaust pipe; 41. Rotating seat; 42. Card joint; 43. Rotating tube; 44. Stirring tube; 5. docking seat; 51. docking hole; 52. rotating cavity; 53. square block; 54. connecting round seat; 55. connecting small hole; 6. housing; 61. active cavity; 7. Second motor; 71. Driving rod; 8. Lifting cylinder; 81. L-shaped lifting rod; 9. Clean the pump; 91. Clean the pipe. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] The present invention will be further described below in conjunction with the embodiments.

[0030] Example 1 Reference Figure 1-8 , which is the first embodiment of the present invention, a chromatographic separation and extraction device for medicinal and edible plants based on an enzymatic hydrolysis method, comprises a U-shaped bracket 1, a discharge pipe 13 is rotatably mounted on the U-shaped bracket 1, a rotary connector 2 is rotatably mounted between two opposite inner side walls of the U-shaped bracket 1, and is connected to the inner cavity of the discharge pipe 13, two chromatographic separation cylinders 3 are symmetrically mounted on the top and bottom of the rotary connector 2, respectively, the two chromatographic separation cylinders 3 are connected to the same discharge pipe 13 through the rotary connector 2, solenoid valves 31 are arranged at the connection points between the two chromatographic separation cylinders 3 and the rotary connector 2, and one-way flow is maintained, two fine mesh filters 32 symmetrically distributed up and down are installed on the inner cylinder walls of the two chromatographic separation cylinders 3, macroporous resin is filled between the two fine mesh filters 32 in the same chromatographic separation cylinder 3, and the macroporous resin is always located between the two fine mesh filters 32 in the chromatographic separation cylinder 3, and the macroporous resin is prevented from being poured out and loaded while ensuring that the macroporous resin is thoroughly cleaned and regenerated, and the operation steps are reduced while preventing the macroporous resin from being secondary contamination.

[0031] The two feed stirring assemblies are respectively installed in the corresponding chromatographic separation cylinders 3, and both include a clamping joint 42 rotatably installed at the end of the corresponding chromatographic separation cylinder 3, and a rotating tube 43 rotatably installed inside the chromatographic separation cylinder 3 and located between the two fine-mesh filters 32. The rotating tube 43 penetrates one of the fine-mesh filters 32 and extends between the two fine-mesh filters 32. The rotating connection between the rotating tube 43 and the penetrated fine-mesh filter 32 is sealed. The outer tube walls of the two rotating tubes 43 are each installed with a plurality of stirring tubes 44 connected to the corresponding clamping joint 42. The ends of the plurality of stirring tubes 44 away from the corresponding rotating tubes 43 are all open. The docking seat 5 is lifted and slid in the U-shaped bracket 1, and rises to the highest point to dock with the corresponding clamping joint 42. The docking seat 5 can rotate to drive the clamping joint 42 and the corresponding rotating tube 43 and stirring tube 44 to rotate. The cleaning pump 9 is installed on the outer side wall of the U-shaped bracket 1, and the output end is connected to the inner cavity of the chromatographic separation cylinder 3 through the docking seat 5 and the clamping joint 42. Example 2

[0032] Reference Figure 1-7 , which is the second embodiment of the present invention. This embodiment is different from the first embodiment in that: a first motor 11 is vertically fixedly installed on the outer wall of the U-shaped bracket 1, and the output end of the first motor 11 is connected to the rotating rod 12. The other end of the rotating rod 12 penetrates the U-shaped bracket 1 and is vertically fixedly connected to the outer ring wall of the rotating connector 2. A limit ring 14 is fixedly sleeved on the outer tube wall of the discharge pipe 13, and the limit ring 14 is rotatably sleeved in the U-shaped bracket 1. The top and bottom of the rotating connector 2 are provided with sockets 21 for convenient insertion of the chromatography separation cylinder 3. The inner hole walls of the two sockets 21 are provided with inner card grooves 22. Two one-way discharge valves 23 are installed at the connection between the rotating connector 2 and the discharge pipe 13. The one-way discharge valves 23 control the two The liquid path of the chromatographic separation cylinder 3 is opened and closed, so that it can be quickly switched according to the current working state of the chromatographic separation cylinder 3. The two chromatographic separation cylinders 3 share a discharge pipe 13, but the two do not interfere with each other through their own independent one-way discharge valves 23. One end of the chromatographic separation cylinder 3 is equipped with a solenoid valve 31 that is respectively inserted into the corresponding inner slot 22. An exhaust pipe 33 is installed on the outer cylinder wall of the other end of the chromatographic separation cylinder 3. The exhaust pipe 33 is connected to the inner cylinder cavity of the chromatographic separation cylinder 3, and an exhaust valve is installed in the exhaust pipe 33. The exhaust pipe 33 is designed to facilitate the discharge of excess air in the chromatographic separation cylinder 3 when the kudzu root powder solution is introduced into the chromatographic separation cylinder 3, and at the same time, the waste liquid generated by cleaning is discharged during the regeneration process of the macroporous resin.

[0033] The feed stirring assembly includes a rotating seat 41 rotatably mounted at the port of the corresponding chromatographic separation cylinder 3. The port of the chromatographic separation cylinder 3 is also provided with a rotating groove for facilitating the rotating seat 41 to be rotatably mounted. A through hole is formed through the rotating seat 41. The clamping joints 42 are respectively fixedly mounted on the corresponding rotating seat 41. The rotating tube 43 is mounted at the bottom of the rotating seat 41 and communicates with the inner cavity of the clamping joint 42 through the through hole. A plurality of stirring tubes 44 are obliquely mounted on the outer tube wall of the corresponding rotating tube 43. In the process of cleaning the macroporous resin, the gravity of the macroporous resin can be reversely overcome, and the macroporous resin can be prevented from being completely deposited and stacked by the oblique upward centrifugal force. The docking seat 5 is stacked together, thereby improving the efficiency of stirring the macroporous resin and reducing the pressure of stirring the macroporous resin. A docking hole 51 adapted to the card joint 42 is provided at the top of the docking seat 5, and a rotating cavity 52 and a square hole are provided at the bottom of the docking seat 5. The square hole is connected to the docking hole 51, and a square block 53 is inserted into the square hole. A connecting round seat 54 is installed at the bottom of the square block 53. A connecting round tube is connected between the connecting round seat 54 and the square block 53. The connecting round seat 54 is connected to the docking hole 51 through the connecting round tube, the square block 53, and the square hole. The outer ring wall of the connecting round seat 54 passes through the inner cavity to open a plurality of connecting small holes 55.

[0034] The remaining structures are the same as those of Example 1. Example 3

[0035] Reference Figure 1 , 6 -8 is the third embodiment of the present invention, which is different from the second embodiment in that: a shell 6 is provided below the docking seat 5 and is sleeved around the connecting round seat 54, and a movable cavity 61 is provided in the shell 6. The inner diameter of the movable cavity 61 is larger than the outer diameter of the connecting round seat 54, which reduces contact and wear while ensuring that the cleaning liquid entering the movable cavity 61 can directly enter the connecting round seat 54 through the connecting small hole 55. A first rotating circular hole is provided at the top of the shell 6 through the movable cavity 61 to facilitate the rotating installation of the connecting round tube. The inner diameter of the first rotating circular hole is consistent with the outer diameter of the connecting round tube to ensure that the seal does not leak. The bottom of the shell 6 A second rotating circular hole is opened through the active cavity 61, and a second motor 7 is installed at the bottom of the inner cavity of the U-shaped bracket 1. The output end of the second motor 7 is connected to a driving rod 71, and the top of the driving rod 71 passes through the second rotating circular hole and is fixedly connected to the lower bottom of the connecting round seat 54. A lifting cylinder 8 is fixedly installed on the other outer side wall of the U-shaped bracket 1, and an L-shaped lifting rod 81 is installed at the output end of the lifting cylinder 8. The L-shaped lifting rod 81 passes through the U-shaped bracket 1 and is fixedly connected to the outer ring wall of the outer shell 6. The output end of the cleaning pump 9 is connected to a cleaning pipe 91, and the end of the cleaning pipe 91 passes through the U-shaped bracket 1 and is connected to the outer wall of the outer shell 6, and the cleaning pipe 91 is connected to the active cavity 61.

[0036] The remaining structure is the same as that of Example 2. Example 4

[0037] The fourth embodiment of the present invention provides: a chromatographic separation and extraction process of medicinal and edible plants based on enzymatic hydrolysis, characterized in that it comprises the following steps: S1. Raw material pretreatment: fresh, dry, and mildew-free kudzu root is selected as the raw material, the kudzu root is peeled and cleaned, crushed by a grinder, and then filtered. A 10-mesh sieve is selected to obtain kudzu root powder, so as to facilitate the dissolution of the effective ingredients in the subsequent extraction process and improve the efficiency of subsequent enzymatic hydrolysis and extraction.

[0038] S2. Enzymatic hydrolysis: cellulase is selected as the enzymatic agent because cellulose is one of the main components of the cell wall of kudzu root. Pueraria powder and cellulase are added to the beaker in equal proportions. The kudzu root powder is enzymatically hydrolyzed at an appropriate temperature and pH value to destroy the cell wall structure and release puerarin.

[0039] S3, extraction and filtration, extracting puerarin from kudzu root powder by hot water extraction method, the specific process is to place the beaker containing the mixture of puerarin and kudzu root powder after enzymatic hydrolysis in a constant temperature water bath, set a suitable temperature for extraction, the extraction temperature of puerarin is usually within a certain range, and needs to be selected and adjusted according to actual conditions, in the constant temperature water bath process, use a stirrer to stir the solution to promote the dissolution and extraction of puerarin, after the extraction is completed, the solution is filtered to remove undissolved kudzu root powder and other impurities, and the filtrate after filtration is collected to obtain the extract containing puerarin.

[0040] S4, chromatographic separation, collecting the eluent through a chromatographic separation device, the specific method is to select a suitable chromatographic medium, such as a macroporous resin, for pretreatment and column loading, wherein the pretreatment is carried out at room temperature, the macroporous resin is soaked in 95% ethanol by volume for 24 hours, stirred at regular intervals to ensure that the resin is fully immersed in ethanol, and the ethanol solution is replaced every 8 hours to remove impurities and oil stains on the surface of the resin. After the soaking is completed, the resin is washed with deionized water until there is no ethanol smell, and then the resin is soaked in 5% hydrogen chloride solution for 4 hours. The hydrogen chloride solution can neutralize the alkaline substances on the surface of the resin and remove some insoluble impurities. After the soaking is completed, the resin is washed with deionized water until the pH value of the washing solution reaches 7, and then the resin is soaked in 5% sodium hydroxide solution for 4 hours. The sodium hydroxide solution can further remove the acidic substances and residues on the surface of the resin. After the soaking is completed, the resin is also washed with deionized water until the pH value of the washing solution reaches 7. After the alkali treatment, the resin is soaked in distilled water for 24 hours. The purpose of this step is to remove the residual acid and alkali solution and impurities in the resin. The soaked resin is dried. Appropriate temperature and time should be selected during drying to avoid damage to the resin structure. Common drying methods include natural drying and hot air drying. The dissolved puerarin crude extract or pueraria powder solution is loaded onto the chromatography column and eluted with an appropriate solvent, such as ethanol. The eluent is passed through the chromatography column at an appropriate flow rate and the eluent is collected. During this process, the eluent can be replaced multiple times to improve the elution efficiency of puerarin. According to the adsorption and desorption properties of puerarin on the chromatographic medium, appropriate elution conditions and flow rates are selected. Common elution solvents include water, ethanol, methanol, etc. The pH value of the solution needs to be adjusted during the process. During the elution process, the eluent is collected in sections and tested and analyzed. The puerarin content in the eluent is usually determined by ultraviolet spectrophotometry, high performance liquid chromatography, and other methods.

[0041] The kudzu root powder solution is poured into the rotating tube 43 from the clamping joint 42 on the top of the upper chromatographic separation cylinder 3, and is naturally discharged from each stirring tube 44 by the gravity of the solution and evenly dispersed in the macroporous resin, which changes the traditional method of pouring the solution completely into the top of the macroporous resin and impregnating the entire chromatographic separation cylinder 3 by sinking and infiltration. It has the advantages of reducing waiting time and improving reaction efficiency. Then, the eluent is injected through the clamping joint 42. After the addition, the clamping joint 42 is driven to rotate by an external motor or manual rotation, thereby driving the rotating tube 43 and the stirring tube 44 to rotate, thereby improving the elution efficiency of the eluent. Finally, the corresponding solenoid valve is opened and closed regularly, and the discharged eluent is collected in batches through the discharge pipe 13.

[0042] While the upper chromatographic separation cylinder 3 is performing chromatographic separation, the macroporous resin in the lower chromatographic separation cylinder 3 can be cleaned to ensure that the cleaned chromatographic separation cylinder 3 can be used directly when the position is changed next time, without waiting time, thus reducing waiting time and improving operation efficiency. The specific cleaning steps are as follows: The lifting cylinder 8 is started, and the housing 6 and the docking seat 5 are raised by the L-shaped lifting rod 81 until the clamping joint 42 at the end of the chromatographic separation cylinder 3 below is just inserted into the docking hole 51 of the docking seat 5, and then the second motor 7 is started, and the connecting round seat 54, the connecting round tube, and the square block 53 are driven by the driving rod 71 to drive the docking seat 5 to rotate, and then the clamping joint 42 is driven to rotate, and the rotating seat 41, the rotating tube 43 and the stirring tube 44 are driven to rotate. At the same time, the cleaning pump 9 is started, and clean water is injected into the active cavity 61 through the cleaning tube 91, and enters into the connecting round block 54 through a plurality of connecting holes 55, and then enters into the docking hole 51 through the connecting round tube and the square block 53, and finally Water is discharged from the plurality of stirring tubes 44 through the clamping joint 42, the through hole and the rotating tube 43. During this process, the control valve on the exhaust pipe 33 is opened to discharge the sewage generated by flushing with clean water and cleaning the surface of the macroporous resin. Then, the macroporous resin regeneration liquid with a good ratio is extracted through the cleaning pipe 91 in the above-mentioned manner. The regeneration liquid includes acid and alkali solution, etc. The perforated resin regeneration liquid is evenly distributed in the chromatographic separation cylinder 3 to regenerate the macroporous resin. After waiting for a period of time, it is discharged through the exhaust pipe 33. Finally, the residual macroporous resin regeneration liquid can be cleaned with clean water. Finally, the surface drying of the macroporous resin in the chromatographic separation cylinder 3 can be accelerated by stirring it separately, and the excess water is discharged from the exhaust pipe 33.

[0043] When the two chromatographic separation cylinders 3 need to be replaced, it is only necessary to separate the docking seat 5 from the corresponding card connector 42, and then start the first motor 11 to drive the two chromatographic separation cylinders 3 to rotate 180° to quickly complete the switching of the working states of the two without any disassembly and assembly operations, thereby greatly improving the operating efficiency.

[0044] S5, extraction and purification, the ethanol eluate is concentrated to recover the ethanol, and then n-butanol is added for extraction to further purify the puerarin, the puerarin solution obtained by extraction is crystallized, the crystals are filtered and dried to obtain the puerarin concentrate.

[0045] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements will not cause the essence of the corresponding technical solutions to deviate from the protection scope of the technical solutions of the embodiments of the present invention.

Claims

1. A chromatographic separation and extraction process of medicinal and edible plants based on enzymatic hydrolysis, characterized in that: The steps include: S1, raw material pretreatment, select fresh, dry, non-mildewed Pueraria root as raw material, then peel and wash the Pueraria root, grind it with a grinder and filter it, select a 10-mesh sieve to obtain Pueraria root powder; S2, enzymatic hydrolysis, selecting cellulase as an enzymatic hydrolyzing agent, and adding kudzu root powder and cellulase into a beaker in equal proportions, respectively, and enzymatically hydrolyzing the kudzu root powder at an appropriate temperature and pH value to release puerarin; S3, extraction and filtration, extracting puerarin from kudzu root powder by hot water extraction, filtering the solution after the extraction to remove undissolved kudzu root powder and other impurities, and collecting the filtrate after filtration to obtain the extract containing puerarin; S4, chromatographic separation, collecting the eluate through a chromatographic separation device, the specific method is to select a suitable chromatographic medium, such as a macroporous resin, perform pretreatment and column loading, load the dissolved puerarin crude extract or pueraria root powder solution onto the chromatographic column, use a suitable solvent, such as ethanol, for elution, pass the eluate through the chromatographic column at a suitable flow rate, collect the eluate, select suitable elution conditions and flow rate according to the adsorption and desorption properties of puerarin on the chromatographic medium, commonly used elution solvents include water, ethanol, methanol, etc., and the pH value of the solution needs to be adjusted during the process; S5, extraction and purification, the ethanol eluate is concentrated to recover the ethanol, and then n-butanol is added for extraction to further purify the puerarin, the puerarin solution obtained by extraction is crystallized, the crystals are filtered and dried to obtain the puerarin concentrate.

2. A chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis as claimed in claim 1, comprising a U-shaped bracket (1), on which a discharge pipe (13) is rotatably mounted, characterized in that ； A swivel connector (2) is rotatably mounted between two opposite inner side walls of the U-shaped bracket (1) and is communicated with the inner cavity of the discharge pipe (13); Two chromatographic separation cylinders (3) are respectively installed symmetrically at the top and bottom of the rotating connector (2), and the two chromatographic separation cylinders (3) are connected to the same discharge pipe (13) through the rotating connector (2). Solenoid valves (31) are provided at the connection points between the two chromatographic separation cylinders (3) and the rotating connector (2) to maintain one-way flow. Two fine-mesh filters (32) are installed on the inner cylinder walls of the two chromatographic separation cylinders (3) and are symmetrically distributed in the upper and lower directions. Macroporous resin is filled between the two fine-mesh filters (32) in the same chromatographic separation cylinder (3); Two feed stirring assemblies are respectively installed in corresponding chromatographic separation cylinders (3), and each comprises a clamping joint (42) rotatably installed at the end of the corresponding chromatographic separation cylinder (3), and a rotating tube (43) rotatably installed inside the chromatographic separation cylinder (3) and located between two fine-mesh filter screens (32); the outer tube walls of the two rotating tubes (43) are each installed with a plurality of stirring tubes (44) connected to the corresponding clamping joints (42); The docking seat (5) is lifted and slid in the U-shaped bracket (1) and rises to the highest point to dock with the corresponding clamping joint (42). The docking seat (5) can rotate to drive the clamping joint (42) and the corresponding rotating tube (43) and stirring tube (44) to rotate; The cleaning pump (9) is mounted on the outer wall of the U-shaped bracket (1), and the output end is connected to the inner cavity of the chromatographic separation cylinder (3) through the docking seat (5) and the clamping joint (42).

3. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 1, characterized in that: A first motor (11) is vertically fixedly mounted on the outer wall of the U-shaped bracket (1); the output end of the first motor (11) is connected to a rotating rod (12); the other end of the rotating rod (12) passes through the U-shaped bracket (1) and is vertically fixedly connected to the outer ring wall of the rotating connector (2); a limit ring (14) is fixedly sleeved on the outer tube wall of the discharge pipe (13); and the limit ring (14) is rotatably sleeved in the U-shaped bracket (1).

4. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 1, characterized in that: The top and bottom of the rotary connector (2) are provided with insertion holes (21) for convenient insertion of the chromatography separation cartridge (3), and the inner walls of the two insertion holes (21) are provided with inner slots (22). Two one-way discharge valves (23) are installed at the connection point between the rotary connector (2) and the discharge pipe (13).

5. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 4, characterized in that: One end of the chromatographic separation cylinder (3) is provided with a solenoid valve (31) which is respectively inserted into the corresponding inner slot (22), and the other end of the chromatographic separation cylinder (3) is provided with an exhaust pipe (33) on the outer cylinder wall, the exhaust pipe (33) is connected to the inner cylinder cavity of the chromatographic separation cylinder (3), and an exhaust valve is provided in the exhaust pipe (33).

6. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 1, characterized in that: The feed stirring assembly comprises a rotating seat (41) rotatably mounted at the port of the corresponding chromatographic separation cylinder (3), the rotating seat (41) having a through hole extending therethrough, the clamping joints (42) being fixedly mounted on the corresponding rotating seats (41), the rotating tube (43) being mounted at the bottom of the rotating seat (41) and communicating with the inner cavity of the clamping joint (42) through the through hole, and a plurality of the stirring tubes (44) being obliquely mounted on the outer tube wall of the corresponding rotating tube (43).

7. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 1, characterized in that: The top of the docking seat (5) is provided with a docking hole (51) adapted to the card joint (42); the bottom of the docking seat (5) is provided with a rotating cavity (52) and a square hole; the square hole is communicated with the docking hole (51); a square block (53) is adapted to be inserted into the square hole; a connecting round seat (54) is installed at the bottom of the square block (53); a connecting round tube is connected between the connecting round seat (54) and the square block (53); the connecting round seat (54) is communicated with the docking hole (51) via the connecting round tube, the square block (53) and the square hole; a plurality of connecting small holes (55) are provided on the outer ring wall of the connecting round seat (54) through the inner cavity.

8. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 7, characterized in that: A shell (6) is provided below the docking seat (5) and is sleeved around the connecting round seat (54). A movable cavity (61) is provided in the shell (6). A first rotating circular hole is provided at the top of the shell (6) and passes through the movable cavity (61) to facilitate the rotating installation of the connecting round tube. A second rotating circular hole is provided at the bottom of the shell (6) and passes through the movable cavity (61).

9. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 8, characterized in that: A second motor (7) is installed at the bottom of the inner cavity of the U-shaped bracket (1); the output end of the second motor (7) is connected to a driving rod (71); the top of the driving rod (71) passes through the second rotating circular hole and is fixedly connected to the bottom of the connecting circular seat (54); a lifting cylinder (8) is fixedly installed on the other outer side wall of the U-shaped bracket (1); an L-shaped lifting rod (81) is installed at the output end of the lifting cylinder (8); the L-shaped lifting rod (81) passes through the U-shaped bracket (1) and is fixedly connected to the outer ring wall of the outer shell (6).

10. The chromatographic separation and extraction device for medicinal and edible plants based on enzymatic hydrolysis according to claim 1, characterized in that: The output end of the cleaning pump (9) is connected to a cleaning pipe (91), the end of the cleaning pipe (91) passes through the U-shaped bracket (1) and is connected to the outer wall of the housing (6), and the cleaning pipe (91) is connected to the active chamber (61).

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