CN113527413B - Antibacterial peptide crude liquid separation and purification device and method - Google Patents
Antibacterial peptide crude liquid separation and purification device and method Download PDFInfo
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- CN113527413B CN113527413B CN202110860483.2A CN202110860483A CN113527413B CN 113527413 B CN113527413 B CN 113527413B CN 202110860483 A CN202110860483 A CN 202110860483A CN 113527413 B CN113527413 B CN 113527413B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D36/00—Filter circuits or combinations of filters with other separating devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D36/00—Filter circuits or combinations of filters with other separating devices
- B01D36/04—Combinations of filters with settling tanks
- B01D36/045—Combination of filters with centrifugal separation devices
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
- C07K1/306—Extraction; Separation; Purification by precipitation by crystallization
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
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Abstract
The invention discloses an antibacterial peptide crude liquid separation and purification device, which relates to the technical field of antibacterial peptide filtration and comprises a storage barrel for storing antibacterial peptide crude liquid, wherein a first liquid outlet pipe and a liquid inlet pipe are arranged at the upper end of the storage barrel, the lower end of the storage barrel is connected with a separation box through a second liquid outlet pipe, the outer wall of the storage barrel is sleeved with a heat exchange pipe, one end of the heat exchange pipe is connected with the liquid inlet end of a refrigerator, the other end of the heat exchange pipe is connected with the liquid outlet end of the refrigerator, and the outer wall of the storage barrel is provided with temperature monitoring equipment. In the invention, when the temperature is too high, the solution in the detection tube group begins to melt, and the metal balls in the detection tube group can be correspondingly changed, so that the detection tube group is convenient for staff to observe; the temperature inside the storage barrel is led out through the temperature guide assembly, so that the temperature inside the storage barrel can be conveniently detected by workers.
Description
Technical Field
The invention relates to the technical field of antibacterial peptide filtration, in particular to an antibacterial peptide crude liquid separation and purification device and method.
Background
In the process of long-term storage of the antibacterial peptide crude liquid in a low-temperature freezing environment, inorganic salts which are originally remained in the centrifugal sedimentation wet substance are gradually separated out and stay at the bottom of the container.
The general storage barrel can be at internal installation temperature sensor detects temperature, but temperature sensor can damage in long-time use, and sensor has damaged in the storage barrel, if open the storage barrel at this moment and carry out the temperature measurement, can lead to the inside temperature of storage barrel to change, leads to the unable better crystallization of thick liquid, and simultaneously can lead to dust particle to get into in the air at the open storage barrel detection temperature, leads to the thick liquid to receive the pollution.
Disclosure of Invention
The invention aims at: in order to solve the problems, a device and a method for separating and purifying the antibacterial peptide crude liquid are provided.
In order to achieve the above purpose, the antibacterial peptide crude liquid separation and purification device adopts the following technical scheme: the antibacterial peptide crude liquid storage device comprises a storage barrel for storing antibacterial peptide crude liquid, wherein a first liquid outlet pipe and a liquid inlet pipe are arranged at the upper end of the storage barrel, a separation box is connected to the lower end of the storage barrel through a second liquid outlet pipe, a heat exchange pipe is sleeved on the outer wall of the storage barrel, one end of the heat exchange pipe is connected with the liquid inlet end of a refrigerator, the other end of the heat exchange pipe is connected with the liquid outlet end of the refrigerator, and a temperature monitoring device is arranged on the outer wall of the storage barrel;
the temperature monitoring equipment comprises a first protective cover, a second protective cover is arranged in the first protective cover, vacuum is arranged between the first protective cover and the second protective cover, heat-insulating cotton is sleeved on the outer wall of the first protective cover, the first protective cover is rotationally connected with the heat-insulating cotton, a mounting ring is fixed on the outer wall of the heat-insulating cotton, the heat-insulating cotton and the mounting ring are both fixed on the outer wall of a storage barrel, a detection pipe group is arranged in the second protective cover, and a heat-conducting component is arranged at the position of the storage barrel, which is located in the temperature monitoring equipment;
the detection tube group is composed of a first sealing tube, a second sealing tube and a third sealing tube, wherein the first sealing tube is internally filled with saturated brine, the second sealing tube is internally filled with sodium chloride solution with concentration of 10%, the third sealing tube is internally filled with water, metal balls are respectively arranged in the first sealing tube, the second sealing tube and the third sealing tube, and the solutions in the first sealing tube, the second sealing tube and the third sealing tube are only half filled.
Further, the inside of the separation box is provided with a polyethersulfone organic membrane for filtration.
Further, the first drain pipe runs through the top of the storage barrel and extends to the inside of the storage barrel, a gap is reserved between the bottom end of the first drain pipe and the bottom end inside the storage barrel, and the lower end of the first drain pipe is sleeved with a polyether sulfone organic film.
Further, the temperature guide assembly comprises temperature guide plate and temperature guide wire, the outer wall in temperature guide plate both sides is installed to the temperature guide wire, the temperature guide wire is provided with the multiunit, the one end that the temperature guide wire is located the storage barrel is loudspeaker form distribution, the through-hole of installation temperature guide plate has been seted up to the storage barrel outer wall, storage barrel and temperature guide plate are fixed connection and sealed.
Further, the first sealing tube, the second sealing tube and the third sealing tube are made of glass with different colors, and scale marks are arranged on the outer walls of the first sealing tube, the second sealing tube and the third sealing tube.
Further, the first protective cover and the second protective cover are made of transparent glass.
Further, the method for separating and purifying the antibacterial peptide crude liquid comprises the following steps:
step S1: filtering fermentation liquor obtained after the bacillus amyloliquefaciens is fermented to prepare antibacterial peptide through a plate frame of a 100-mesh filter cloth, pre-coating diatomite on the filter cloth, wherein the coating amount of the diatomite is 1.5kg per hundred liters of fermentation liquor, the average filtering flux is 40 liters/hour per square meter, and removing thalli to prepare clarified liquor;
step S2: adding dilute hydrochloric acid into the clarified liquid prepared in the step S1 until the pH value is 2.5 under the stirring condition, adding 0.6mol of hydrochloric acid into each liter of clarified liquid per minute, standing at room temperature for more than 16 hours, discarding the supernatant, centrifuging, and removing the supernatant to obtain slurry fluid with the solid wet weight of 20 g/L;
step S3: adding food-grade absolute ethyl alcohol into the pasty fluid prepared in the step S2, wherein the addition amount of the food-grade absolute ethyl alcohol is 5% of the volume of the fermentation liquor in the step S1, stirring uniformly, extracting for 4 hours at 10 ℃, centrifuging, and carrying out microfiltration through a 0.22-micrometer polyether sulfone organic membrane, wherein the filtration speed is 51L/(square.min), so as to obtain filtrate;
step S4: and (3) standing the filtrate obtained in the step (S3) at the temperature of minus 20 ℃ for 7 days for crystallization, and filtering out the crystallization by a 0.22-micrometer polyether sulfone organic membrane microfiltration to obtain the antibacterial peptide solution.
Compared with the prior art, the invention has the following beneficial effects:
in the invention, when the temperature is too high, the solution in the detection tube group begins to melt, and the metal balls in the detection tube group can be correspondingly changed, so that the detection tube group is convenient for staff to observe; the temperature inside the storage barrel is led out through the temperature guide assembly, so that the temperature inside the storage barrel can be conveniently detected by workers.
Drawings
FIG. 1 is a front view of a crude antimicrobial peptide liquid separation and purification device according to the present invention;
FIG. 2 is a schematic diagram of the inside of a storage barrel of the antibacterial peptide crude liquid separation and purification device provided by the invention;
FIG. 3 is a side view of a temperature monitoring device of the antibacterial peptide crude liquid separation and purification device provided by the invention;
FIG. 4 is a front view of a temperature monitoring device of the antibacterial peptide crude liquid separation and purification device provided by the invention;
fig. 5 is a schematic diagram of the filling amounts of the liquid medicine in the first sealing tube, the second sealing tube and the third sealing tube of the antibacterial peptide crude liquid separation and purification device.
In the figure: 1. a storage tub; 2. a temperature monitoring device; 201. a first protective cover; 202. a second protective cover; 203. a detection tube group; 2031. a first sealing tube; 2032. a second sealing tube; 2033. a third sealing tube; 204. thermal insulation cotton; 206. a metal ball; 3. a temperature guiding component; 301. a temperature guide plate; 302. a temperature guiding wire; 4. a heat exchange tube; 5. a first liquid outlet pipe; 6. a separation box; 7. a liquid inlet pipe.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments.
In the description of the present invention, it should be understood that the terms "upper," "lower," "front," "rear," "left," "right," "top," "bottom," "inner," "outer," and the like indicate or are based on the orientation or positional relationship shown in the drawings, merely to facilitate description of the present invention and to simplify the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention.
Referring to fig. 1-5, the antibacterial peptide crude liquid separation and purification device comprises a storage barrel 1 for storing antibacterial peptide crude liquid, wherein a first liquid outlet pipe 5 and a liquid inlet pipe 7 are arranged at the upper end of the storage barrel 1, a separation box 6 is connected to the lower end of the storage barrel 1 through a second liquid outlet pipe, a heat exchange pipe 4 is sleeved on the outer wall of the storage barrel 1, one end of the heat exchange pipe 4 is connected with the liquid inlet end of a refrigerator, the other end of the heat exchange pipe 4 is connected with the liquid outlet end of the refrigerator, and a temperature monitoring device 2 is arranged on the outer wall of the storage barrel 1;
the temperature monitoring equipment 2 comprises a first protective cover 201, a second protective cover 202 is arranged in the first protective cover 201, vacuum is arranged between the first protective cover 201 and the second protective cover 202, heat preservation cotton 204 is sleeved on the outer wall of the first protective cover 201, the first protective cover 201 is rotationally connected with the heat preservation cotton 204, a mounting ring is fixed on the outer wall of the heat preservation cotton 204, the heat preservation cotton 204 and the mounting ring are both fixed on the outer wall of the storage barrel 1, a detection tube group 203 is arranged in the second protective cover 202, and a temperature guide assembly 3 is arranged at the position of the storage barrel 1, which is located in the temperature monitoring equipment 2;
the detection tube group 203 consists of a first sealing tube 2031, a second sealing tube 2032 and a third sealing tube 2033, wherein the first sealing tube 2031 is filled with saturated brine, the second sealing tube 2032 is filled with 10% sodium chloride solution, the third sealing tube 2033 is filled with water, metal balls 206 are placed in the first sealing tube 2031, the second sealing tube 2032 and the third sealing tube 2033, and the solutions in the first sealing tube 2031, the second sealing tube 2032 and the third sealing tube 2033 are only half filled;
the freezing point of 10% sodium chloride solution is about minus 10 to minus 15 ℃, the freezing point of saturated brine is minus 21 ℃ and the freezing point of water is 0 ℃, and the metal ball 206 is made of corrosion-resistant material;
in practical application, after the coarse liquid is injected, the temperature of the storage barrel 1 will be reduced to 20 ℃, so that 10% sodium chloride solution and water will freeze, after the first protective cover 201 is completely frozen, the detection tube set 203 in the first protective cover 201 is turned over by half a turn, at this time, the metal balls 206 inside the second sealing tube 2032 and the third sealing tube 2033 will be at the upper ends of the second sealing tube 2032 and the third sealing tube 2033, if the temperature inside the storage barrel 1 is increased, the solution inside the second sealing tube 2033 will start to melt when the temperature is higher than minus 10 degrees, and the solution inside the third sealing tube 2032 will start to melt when the temperature is higher than zero degrees, and the metal balls 206 will start to slide down when the solution starts to melt, so that the staff can observe conveniently.
Further, the inside of the separation box 6 is provided with a polyethersulfone organic membrane for filtering, the first liquid outlet pipe 5 penetrates through the top end of the storage barrel 1 and extends into the storage barrel, a gap is reserved between the bottom end of the first liquid outlet pipe 5 and the bottom end inside the storage barrel 1, and the lower end of the first liquid outlet pipe 5 is sleeved with the polyethersulfone organic membrane;
in practical applications, the position of the first liquid outlet pipe 5 in the storage barrel 1 can be changed according to the actual situation; after the crude liquid is cooled, the liquid medicine at the upper end of the storage barrel 1 is pumped out through the first liquid outlet pipe 5, after the liquid medicine at the upper end is pumped out, the rest liquid medicine is guided to the separation box 6 through the second liquid outlet pipe, and the liquid medicine is crystallized and separated through the polyether sulfone organic membrane in the separation box 6, so that the purification is realized.
Further, the temperature guide assembly 3 is composed of a temperature guide plate 301 and temperature guide wires 302, the temperature guide wires 302 are arranged on the outer walls of the two sides of the temperature guide plate 301, a plurality of groups of temperature guide wires 302 are arranged, one end of each temperature guide wire 302, which is positioned in the storage barrel 1, is distributed in a horn shape, through holes for installing the temperature guide plate 301 are formed in the outer wall of the storage barrel 1, and the storage barrel 1 is fixedly connected with the temperature guide plate 301 and sealed; in practical application, the temperature inside the storage barrel 1 is led out through the temperature guide assembly 3, so that a worker can conveniently detect the temperature inside the storage barrel 1; the temperature wire 302 is increased by tilting the temperature wire 302 up and down.
Further, the first sealing tube 2031, the second sealing tube 2032 and the third sealing tube 2033 are made of glass with different colors, and scale marks are arranged on the outer walls of the first sealing tube 2031, the second sealing tube 2032 and the third sealing tube 2033; in practical application, the first sealing tube 2031, the second sealing tube 2032 and the third sealing tube 2033 are made of glass with different colors, thereby facilitating the distinction of staff.
Further, the first protective cover 201 and the second protective cover 202 are made of transparent glass.
Further, the method for separating and purifying the antibacterial peptide crude liquid comprises the following steps:
step S1: filtering fermentation liquor obtained after the bacillus amyloliquefaciens is fermented to prepare antibacterial peptide through a plate frame of a 100-mesh filter cloth, pre-coating diatomite on the filter cloth, wherein the coating amount of the diatomite is 1.5kg per hundred liters of fermentation liquor, the average filtering flux is 40 liters/(hour square meter), and removing thalli to prepare clarified liquor;
step S2: adding dilute hydrochloric acid into the clarified liquid prepared in the step S1 until the pH value is 2.5 under the stirring condition, adding 0.6mol of hydrochloric acid into each liter of clarified liquid per minute, standing at room temperature for more than 16 hours, discarding the supernatant, centrifuging, and removing the supernatant to obtain slurry fluid with the solid wet weight of 20 g/L;
step S3: adding food-grade absolute ethyl alcohol into the pasty fluid prepared in the step S2, wherein the addition amount of the food-grade absolute ethyl alcohol is 5% of the volume of the fermentation liquor in the step S1, stirring uniformly, extracting for 4 hours at 10 ℃, centrifuging, and carrying out microfiltration through a 0.22-micrometer polyether sulfone (PES) organic membrane, wherein the filtration speed is 51L/(square per minute), so as to obtain filtrate;
step S4: and (3) standing the filtrate obtained in the step (S3) at the temperature of minus 20 ℃ for 7 days for crystallization, and filtering out the crystallization by a 0.22-micrometer polyether sulfone (PES) organic membrane microfiltration to obtain the antibacterial peptide solution.
The present invention is not limited to the above-mentioned embodiments, and any person skilled in the art, based on the technical solution disclosed in the present invention, can make equivalent substitutions or modifications within the scope of the present invention.
Claims (5)
1. The antibacterial peptide crude liquid separation and purification device comprises a storage barrel (1) for storing antibacterial peptide crude liquid, wherein a first liquid outlet pipe (5) and a liquid inlet pipe (7) are arranged at the upper end of the storage barrel (1), a separation box (6) is connected to the lower end of the storage barrel (1) through a second liquid outlet pipe, a polyethersulfone organic membrane used for filtering is arranged in the separation box (6), a heat exchange pipe (4) is sleeved on the outer wall of the storage barrel (1), one end of the heat exchange pipe (4) is connected with the liquid inlet end of a refrigerator, and the other end of the heat exchange pipe (4) is connected with the liquid outlet end of the refrigerator;
the temperature monitoring equipment (2) comprises a first protective cover (201), a second protective cover (202) is arranged in the first protective cover (201), vacuum is arranged between the first protective cover (201) and the second protective cover (202), heat preservation cotton (204) is sleeved on the outer wall of the first protective cover (201), the first protective cover (201) is rotationally connected with the heat preservation cotton (204), a mounting ring is fixed on the outer wall of the heat preservation cotton (204), the heat preservation cotton (204) and the mounting ring are both fixed on the outer wall of the storage barrel (1), a detection tube set (203) is arranged in the second protective cover (202), and a temperature guide component (3) is arranged at the position of the storage barrel (1) in the temperature monitoring equipment (2);
the heat conducting assembly (3) consists of a heat conducting plate (301) and heat conducting wires (302), the heat conducting wires (302) are arranged on the outer walls of the two sides of the heat conducting plate (301), the heat conducting wires (302) are provided with a plurality of groups, one ends of the heat conducting wires (302) located in the storage barrel (1) are distributed in a horn shape, the outer wall of the storage barrel (1) is provided with through holes for arranging the heat conducting plate (301), and the storage barrel (1) is fixedly connected with the heat conducting plate (301) and is sealed
The detection tube group (203) is composed of a first sealing tube (2031), a second sealing tube (2032) and a third sealing tube (2033), wherein the first sealing tube (2031) is internally filled with saline with saturated concentration, the second sealing tube (2032) is internally filled with sodium chloride solution with 10% concentration, the third sealing tube (2033) is internally filled with water, metal balls (206) are respectively arranged in the first sealing tube (2031), the second sealing tube (2032) and the third sealing tube (2033), and the solutions in the first sealing tube (2031), the second sealing tube (2032) and the third sealing tube (2033) are only half filled.
2. The antibacterial peptide crude liquid separation and purification device according to claim 1, wherein the first liquid outlet pipe (5) penetrates through the top end of the storage barrel (1) and extends to the inside of the storage barrel, a gap is reserved between the bottom end of the first liquid outlet pipe (5) and the bottom end inside the storage barrel (1), and a polyethersulfone organic membrane is sleeved at the lower end of the first liquid outlet pipe (5).
3. The antibacterial peptide crude liquid separation and purification device according to claim 1, wherein the first sealing tube (2031), the second sealing tube (2032) and the third sealing tube (2033) are made of glass with different colors, and scale marks are arranged on the outer walls of the first sealing tube (2031), the second sealing tube (2032) and the third sealing tube (2033).
4. The antibacterial peptide crude liquid separation and purification apparatus according to claim 1, wherein the first protective cover (201) and the second protective cover (202) are made of transparent glass.
5. The method for separating and purifying a crude antibacterial peptide liquid according to any one of claims 1 to 4, comprising the steps of:
step S1: filtering fermentation liquor obtained after the bacillus amyloliquefaciens is fermented to prepare antibacterial peptide through a plate frame of a 100-mesh filter cloth, pre-coating diatomite on the filter cloth, wherein the coating amount of the diatomite is 1.5kg per hundred liters of fermentation liquor, the average filtering flux is 40 liters/(hour square meter), and removing thalli to prepare clarified liquor;
step S2: adding dilute hydrochloric acid into the clarified liquid prepared in the step S1 until the pH value is 2.5 under the stirring condition, adding 0.6mol of hydrochloric acid into each liter of clarified liquid per minute, standing at room temperature for more than 16 hours, discarding the supernatant, centrifuging, and removing the supernatant to obtain slurry fluid with the solid wet weight of 20 g/L;
step S3: adding food-grade absolute ethyl alcohol into the pasty fluid prepared in the step S2, wherein the addition amount of the food-grade absolute ethyl alcohol is 5% of the volume of the fermentation liquor in the step S1, stirring uniformly, extracting for 4 hours at 10 ℃, centrifuging, and carrying out microfiltration through a 0.22-micrometer polyether sulfone organic membrane, wherein the filtration speed is 51L/(square.min), so as to obtain filtrate;
step S4: and (3) standing the filtrate obtained in the step (S3) at the temperature of minus 20 ℃ for 7 days for crystallization, and filtering out the crystallization by a 0.22-micrometer polyether sulfone organic membrane microfiltration to obtain the antibacterial peptide solution.
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GB603998A (en) * | 1945-11-20 | 1948-06-25 | Ralph Ambrose Kekwick | Improved process for the fractionation of blood plasma |
GB2485862B (en) * | 2010-11-29 | 2013-09-04 | Guangdong Huanxi Biolog Technology Co Ltd | Preparation process of L-arginine-alpha-ketoglutarate by direct crystallization from fermentation broth |
CN104897299B (en) * | 2014-05-14 | 2017-11-10 | 代明浩 | A kind of temperature quality inspection ball |
CN207079183U (en) * | 2017-06-15 | 2018-03-09 | 天津冠勤医药科技有限公司 | A kind of device crystallized for antibacterial peptide with dissolving |
CN207845208U (en) * | 2018-01-10 | 2018-09-11 | 广东省茂名市质量计量监督检测所 | A kind of platiniferous waste liquid extraction equipment |
CN209470790U (en) * | 2019-01-17 | 2019-10-08 | 铜仁学院 | A kind of hot dry rock crack rock temperature-detecting device |
CN210229283U (en) * | 2019-05-10 | 2020-04-03 | 广东药科大学 | Polypeptide low-temperature high-pressure filter element filter |
CN210237509U (en) * | 2019-07-17 | 2020-04-03 | 周口师范学院 | Dual-purpose antibacterial peptide crystallization and dissolving device |
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