CN111560063A - Raw material medicine purification device for animal pancreas-derived insulin and use method - Google Patents
Raw material medicine purification device for animal pancreas-derived insulin and use method Download PDFInfo
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- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 title claims abstract description 186
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- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/62—Insulins
- C07K14/625—Extraction from natural sources
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Endocrinology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a raw material medicine purification device of animal pancreas-derived insulin, comprising: a high-pressure electrophoresis separation system, a peristaltic pump, a one-way valve and a detector; the high-voltage electrophoresis separation system is provided with at least two electrode chambers, and the electrode chambers comprise a positive electrode chamber and a negative electrode chamber; wherein the positive electrode chamber is connected with the detector, and the negative electrode chamber is connected with the peristaltic pump; the one-way valve is connected with the detector. The device for purifying the insulin raw material medicine provided by the invention adopts a multi-stage high-pressure electrophoresis separation system for the first time, is used for separating and purifying the insulin, has less impurity types and contents obtained by separation and purification, has high insulin purity, improves the yield of the insulin, and effectively reduces the inactivation rate of the insulin in the separation process.
Description
Technical Field
The invention relates to the technical field of biological substance separation and purification devices, in particular to a raw material drug purification device for animal pancreas-derived insulin and a using method thereof.
Background
Insulin is a protein hormone secreted by the beta cells of the islets of langerhans in the pancreas stimulated by endogenous or exogenous substances such as glucose, lactose, ribose, arginine, glucagon, etc. Insulin is the only hormone in the body which reduces blood sugar and promotes the synthesis of glycogen, fat and protein. Exogenous insulin is mainly used for treating diabetes and consumptive diseases. Promote glucose in blood circulation to enter liver cells, muscle cells, fat cells and other tissue cells to synthesize glycogen, reduce blood sugar, and promote synthesis of fat and protein. The function of the insulin of mammals (human, cattle, sheep, pigs, and the like) of different ethnic groups is roughly the same, and the components are slightly different, so that the extraction of medicinal insulin from the pancreas of the animals such as the pigs, the cattle, the sheep, and the like has wider application value and the same medicinal effect.
The insulin can be dissolved in diluted alcohol and diluted propanol with the concentration of less than 80%, acid and alkali, and is not dissolved in ethanol and other organic solvents with the concentration of more than 90%, is stable when the pH value is 2-4, is easy to damage when meeting protease, strong acid and strong base, is easy to change by heating, has the isoelectric point of 5.35-5.45 (shown in table 1), and is stored: shading, sealing, and storing at below-15 deg.C. The crystalline insulin has typical protein properties, is easily soluble in dilute acid or dilute alkali solution, is stable in subacidity (pH value of 2.5-3.5), is extremely easy to change in alkalinity, is more easily damaged at high temperature, can be dissolved in 90% phenol and dilute alcohol, and has levorotatory property. Adding alkali into the insulin water solution to adjust the pH value to 5.1-5.3, separating out a precipitate, adding acid to enable the pH value to 3.3-3.8, dissolving the precipitate completely, adding alkali to enable the pH value to 8.0-8.5, generating slight turbidity, and enabling the activity of insulin to be damaged by reducing agents such as hydrogen sulfide, formaldehyde, formic acid, vitamin C, cysteine, sodium thiosulfate and the like.
Insulin related physicochemical information
TABLE 1
Electrophoresis is a technique in which charged particles are moved toward an electrode of an opposite polarity to the charged particles by an electric field and are separated by using the difference in the moving speed of the charged particles in the electric field, and is generally used for measuring the purity and content of proteins.
The traditional production process is to purify insulin from animals (pigs), extract and separate the insulin by acid-alcohol solution, separate and purify the insulin after salting out, and aims to remove impurity proteins, anions, cations, pyrogen substances and the like. At present, the pig pancreas is taken as a material, and the insulin is prepared by an acid-alcohol extraction reduced-pressure concentration method, a direct zinc salt precipitation method and an ion exchange resin method which are commonly used, wherein the acid-alcohol extraction reduced-pressure concentration method has the defects of the most complex method and less obtained insulin, and the direct zinc salt precipitation method has the defects of low product yield, more operation steps, long production period and high auxiliary material consumption.
Therefore, aiming at the defects in the prior art, the problem to be solved by those skilled in the art is to provide a device for purifying raw material drug of insulin from animal pancreas.
Disclosure of Invention
In view of the above, the present invention provides a purification device for raw material drugs of insulin from animal pancreas, the purification device uses a multi-stage high-voltage electrophoresis separation system for the first time to separate and purify insulin, the insulin is amphoteric molecule, when the pH of an external solution is greater than the isoelectric point of the insulin, the insulin releases protons with negative charges, the insulin moves to a positive electrode chamber under high-voltage direct-current frequency conversion electrophoresis, a sample adopts U-shaped multi-stage flow, multi-stage separation, and the finally separated and purified insulin has few impurities and high purity, the yield of the insulin is improved, and the inactivation rate of the insulin in the separation process is effectively reduced.
In order to achieve the purpose, the invention adopts the following technical scheme:
an animal pancreas derived insulin raw material medicine purification device, comprising: a high-pressure electrophoresis separation system, a peristaltic pump, a one-way valve and a detector;
the high-voltage electrophoresis separation system is provided with at least two electrode chambers, and the electrode chambers comprise a positive electrode chamber and a negative electrode chamber;
wherein the positive electrode chamber is connected with the detector, and the negative electrode chamber is connected with the peristaltic pump; the one-way valve is connected with the detector.
By adopting the technical scheme, the separation and purification of the insulin by using the high-voltage electrophoresis separation system are realized, the sample adopts U-shaped multi-stage flow direction and multi-stage separation, and finally, the types and the contents of the impurity proteins of the insulin obtained by extraction and purification are less and the purity is high.
Furthermore, the high-voltage electrophoresis separation system is also provided with a high-voltage direct-current variable-frequency power supply, and the high-voltage direct-current variable-frequency power supply comprises a voltage-stabilizing filtering module, a variable-frequency control module, a voltage boosting and transforming module, a high-voltage rectifying module, an error analysis module, a setting synthesis module and an output module;
the electric signal passes through the voltage-stabilizing filtering module, the frequency-converting control module, the voltage-boosting and voltage-transforming module, the high-voltage rectifying module, the error analysis module and the output module in sequence, and the electric signal of the error analysis module passes through the setting synthesis module and is fed back to the frequency-converting control module.
Furthermore, the error analysis module comprises a 0-30kV DC filtering, voltage stabilizing and constant current error molecule module.
Furthermore, the setting and synthesizing module comprises a control signal input and feedback signal output setting and synthesizing module.
Further, the high-voltage electrophoresis separation system is also provided with an electrophoresis column at least comprising two separation sections, and the electrophoresis column is matched with the electrode chamber;
the electrophoresis column is filled with a separation medium, and the detector is arranged between the separation sections.
Furthermore, the electrophoresis column is made of transparent hard glass, and the separation medium is linear polyacrylamide buffer solution. By adopting the technical scheme, the linear polyacrylamide buffer solution used in the invention has low viscosity, thereby being beneficial to the separation of insulin and impurities.
Further, still include the spiral line condenser, the separation pipeline is wrapped up in the spiral line condenser.
By adopting the technical scheme, the high-pressure electrophoresis separation system is used for separation and purification at low temperature, the temperature is kept below 25 ℃, the special high-efficiency spiral pipeline condenser is high in condensation efficiency, and the thermal denaturation of insulin can be effectively prevented.
The invention also provides a using method of the animal pancreas derived insulin raw material medicine purification device, which comprises the following steps:
s1, raw material pretreatment: the animal pancreas is frozen in deep immediately after being separated from the body, the pancreas is frozen and crushed, then acidic ethanol is added for stirring and extraction, and centrifugation is carried out to obtain the supernatant; wherein the residue is extracted again by repeating the above operations, and the extracting solutions are combined to obtain crude insulin;
s2, separating and purifying a sample: the condenser is started first, and the temperature display reaches the required temperature; then turning on the ultraviolet detector, and setting the detection wavelength; then the peristaltic pump and the one-way valve are opened, the flow rate is adjusted, and the flow rate is stabilized; then turning on a high-voltage power supply to set various parameters, and inputting crude insulin for separation and purification after instrument equipment is stable;
s3, detecting insulin: and (5) detecting on line in real time by using a detector, continuously separating and purifying if the insulin is unqualified until the insulin is qualified, and adjusting a one-way valve to obtain the separated and purified insulin.
Further, sources of insulin raw materials include, but are not limited to, human, bovine, ovine, porcine pancreas, and other extracts that yield insulin that require further purification.
Further, in step S2, the high-voltage dc variable frequency power supply is a 0-30kV dc variable frequency high-voltage power supply, and the variable frequency mode of the high-voltage dc variable frequency power supply is spike pulse, positive ripple pulse, and square wave pulse.
By adopting the technical scheme, the high-voltage direct-current variable-frequency power supply is low in frequency, high in separation efficiency and large in heat effect. The square wave pulse (conventional) is beneficial to intermittent separation, the heat dissipation is good, but the radial diffusion is severe; sine wave pulses (unconventional) are voltage changes in a sine curve form and are suitable for separation of heat-sensitive substances; spike pulses (unconventional) are suitable for the separation of other specific species.
Further, in step S2, the pH of the high-pressure electrophoresis is 3.5 to 4.8; the temperature of the high-pressure electrophoresis is 0-25 ℃.
The beneficial effect of adopting above-mentioned technical scheme is: on the premise of no deterioration of the sample, the separation efficiency is improved.
Further, in step S2, the electric field intensity of the high-voltage electrophoresis is 0.09k V/cm-2.82kV/cm
By adopting the technical scheme, the voltage is high in electric driving force, the swimming speed is high, but a large heat effect is generated, and the sample is easy to deteriorate.
Further, in step S3, the detector is: ultraviolet-visible detector, UV detection wavelength: 276 nm.
According to the invention, when the pH value of the electrophoresis separation medium is greater than the isoelectric point of insulin, the proton released by the insulin carries negative charges, the insulin moves to the positive electrode chamber under high-voltage direct-current frequency conversion electrophoresis, and the sample adopts U-shaped multi-stage flow direction and multi-stage separation, so that the separation efficiency is improved.
Through the technical scheme, compared with the prior art, the beneficial effects of the invention comprise the following points:
(1) the animal pancreas-derived insulin raw material medicine purification device provided by the invention adopts a multi-stage high-pressure electrophoresis separation system for the first time, is used for separating and purifying insulin, and has the advantages of low impurity types and contents in the separated and purified insulin and high product purity.
(2) The raw material medicine purification device for the animal pancreas-derived insulin is provided with the one-way valve, so that the backflow of fluid reaching a certain separation effect can be effectively prevented, and the separation efficiency is further improved.
(3) The detector provided by the invention can monitor the whole separation process in real time.
(4) The sample obtained by the method for purifying the raw material drug of the animal pancreas-derived insulin has unchanged protein higher structure and low inactivation rate of biological activity.
(5) The device for purifying the raw material drug of the animal pancreas-derived insulin is suitable for industrial and large-scale production.
(6) The method for purifying the raw material drug of the animal pancreas-derived insulin has high separation efficiency of the insulin, and can efficiently separate unstable complex components.
(7) The raw material medicine purification device for the animal pancreas-derived insulin can also expand the separation range by replacing different fluid media, such as traditional Chinese medicine active ingredients with complex ingredients and the like.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a schematic view of an insulin raw material drug purification apparatus provided by the present invention.
FIG. 2 is a structural diagram of a 0-30kV high-voltage direct-current variable-frequency power supply.
Fig. 3 is a diagram of a voltage pulse square waveform.
In the figure, 1 is a peristaltic pump, 2 is a detector, 3 is a one-way valve, 4 is a negative electrode chamber, 5 is a positive electrode chamber, and 6 is a spiral line condenser.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment discloses a raw material drug purification device of animal pancreas-derived insulin, as shown in fig. 1, comprising: the electrophoresis separation device comprises a high-pressure electrophoresis separation system, a peristaltic pump 1, a one-way valve 2 and a detector 3; the high-voltage electrophoresis separation system is provided with four electrode chambers, the electrode chambers comprise a positive electrode chamber 4 and a negative electrode chamber 5, the positive electrode chamber 4 is connected with the detector 3, the negative electrode chamber 5 is connected with the peristaltic pump 1, and the one-way valve 2 is connected with the detector 3.
In order to further optimize the technical scheme, the high-voltage electrophoresis separation system is further provided with a high-voltage direct-current variable-frequency power supply, and the high-voltage direct-current variable-frequency power supply comprises a 220V AC voltage-stabilizing filtering module, a variable-frequency control module, a 0-30kV voltage-boosting and voltage-transforming module, a high-voltage rectifying module, an error analysis module, a setting synthesis module and a three-phase DC 0-30kV output module. The error analysis module comprises a 0-30kV DC filtering, voltage stabilizing and constant current error molecule module, and the setting synthesis module comprises a control signal input and feedback signal output setting synthesis module. The electric signal passes through the voltage-stabilizing filtering module, the frequency conversion control module, the voltage boosting and transforming module, the high-voltage rectifying module, the error analysis module and the output module in sequence, and the electric signal of the error analysis module is fed back to the frequency conversion control module through the setting synthesis module.
In order to further optimize the technical scheme, the high-voltage electrophoresis separation system is also provided with four separation sections of electrophoresis columns, separation media are filled in the electrophoresis columns, detectors 3 are arranged between the separation sections, the electrophoresis columns are matched with the electrode chambers and are made of transparent hard glass, and the separation media are linear polyacrylamide buffer solutions.
In order to further optimize the technical scheme, the biological activity sample heat treatment device further comprises a spiral pipeline condenser 6, a specially-made high-efficiency spiral pipeline condenser is high in condensation efficiency, and the biological activity sample heat denaturation can be effectively prevented.
The setting range of each parameter of the multi-stage high-voltage electrophoresis separation system is as follows:
1. initial switching time: 0.1 second to 70000 second
2. Final switching time: 0.1 second to 70000 second
3. Separation time: 0.1 to 999 hours
4. Electric field intensity (Volts/cm): 0.09k V/cm-2.82 kV/cm;
the calculation formula of the electric field strength is as follows:
in the formula, D is the electrode distance, and the design is 10 cm; dj is the thickness of the insulating material, here designed to be 0.8cm,jdielectric constant of the insulating material, here 3.6;rdielectric constant of the buffer solution system, here 5; u is input voltage and takes a value rangeThe voltage is 1-30 kV. The input voltage-corresponding electric field strength is as follows.
Electric field strength corresponding to different input voltages:
| input voltage/kV | 1 | 2 | 4 | 6 | 8 | 10 | 15 | 20 | 25 | 30 |
| Electric field strength/kV/cm | 0.09 | 0.19 | 0.38 | 0.56 | 0.75 | 0.94 | 1.41 | 1.88 | 2.35 | 2.82 |
5. Frequency conversion mode: square wave pulses, sine wave pulses, spikes, wherein the voltage pulses have a square wave shape, as shown in fig. 3.
6. Voltage range: the 0-30kV direct current variable frequency high-voltage power supply has strong voltage electric driving force and high swimming speed, but has larger heat effect, and the sample is easy to deteriorate.
7. Electrophoresis pipeline pressure: 0-5MPa, high pipeline pressure and large resistance, and is favorable for quick separation.
8. Flow rate of the electrophoretic medium: 0-1000mm/min, high flow rate and fast flow, which is not favorable for effective separation, but favorable for reducing radial diffusion.
The embodiment discloses a use method of the insulin raw material medicine purification device, which comprises the following steps:
s1, raw material pretreatment: the porcine pancreas is immediately frozen after being separated from the body, the pancreas is frozen and crushed, then acidic ethanol is added for stirring and extraction, and the mixture is centrifuged to obtain supernatant; wherein the residue is extracted again by repeating the above operations, and the extractive solutions are combined to obtain crude insulin.
S2, separating and purifying a sample: the condenser is started first, and the temperature display reaches the required temperature; then turning on the ultraviolet detector, and setting the detection wavelength; then the peristaltic pump and the one-way valve are opened, the flow rate is adjusted, and the flow rate is stabilized; then, the high-voltage power supply is turned on to set various parameters, and the specific parameters comprise: initial switching time: 5 seconds, final switching time: 20 seconds, separation time: 2 hours, the electric field intensity is 1.0kV/cm, and the frequency conversion mode is as follows: square wave pulse, voltage range: 20kV direct current frequency conversion high voltage power supply, electrophoresis pipeline pressure: 3MPa, electrophoresis medium flow rate: 50ml/min, pH of high pressure electrophoresis is 4.0; the temperature of high-pressure electrophoresis is 10 ℃, and crude insulin is input for separation after instrument equipment is stabilized.
S3, detecting insulin: and (5) detecting on line in real time by using a detector, continuously separating and purifying if the insulin is unqualified until the insulin is qualified, and adjusting a one-way valve to obtain the separated and purified insulin.
In order to further explain the technical effect of the device for purifying the raw material insulin from the animal pancreas, the invention detects the separated and purified insulin, adopts an insulin detection method of the national drug Community and national drug Community standard detection method, utilizes an ultraviolet visible detector, and has the UV detection wavelength as follows: 276nm, wherein the content of impurity protein in the insulin obtained by using the animal pancreas-derived insulin raw material medicine purification device disclosed by the invention is only 0.98%, while the content of impurity protein in the insulin obtained by using the animal pancreas-derived insulin raw material medicine purification device disclosed by the invention is only 0.98%, so that the animal pancreas-derived insulin raw material medicine purification device provided by the invention adopts a multistage high-pressure electrophoresis separation system for the first time for the separation and purification of insulin, and the types and the contents of impurities in the insulin obtained by separation and purification are lower.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (9)
1. An animal pancreas source insulin raw material medicine purification device which is characterized by comprising: a high-pressure electrophoresis separation system, a peristaltic pump, a one-way valve and a detector;
the high-voltage electrophoresis separation system is provided with at least two electrode chambers, and the electrode chambers comprise a positive electrode chamber and a negative electrode chamber;
wherein the positive electrode chamber is connected with the detector, and the negative electrode chamber is connected with the peristaltic pump; the one-way valve is connected with the detector.
2. The apparatus for purifying raw material drug of insulin from animal pancreas according to claim 1, wherein the high voltage electrophoresis separation system is further provided with a high voltage direct current variable frequency power supply, and the high voltage direct current variable frequency power supply comprises a voltage stabilizing and filtering module, a variable frequency control module, a voltage boosting and transforming module, a high voltage rectifying module, an error analyzing module, a setting and synthesizing module, and an output module;
the electric signal passes through the voltage-stabilizing filtering module, the frequency-converting control module, the voltage-boosting and voltage-transforming module, the high-voltage rectifying module, the error analysis module and the output module in sequence, and the electric signal of the error analysis module passes through the setting synthesis module and is fed back to the frequency-converting control module.
3. The apparatus for purifying raw insulin drug derived from animal pancreas as claimed in claim 2, wherein the high pressure electrophoresis separation system further comprises an electrophoresis column comprising at least two separation sections, the electrophoresis column is adapted to the electrode chamber;
the electrophoresis column is filled with a separation medium, and the detector is arranged between the separation sections.
4. The apparatus for purifying raw insulin drug derived from animal pancreas as claimed in claim 3, wherein the electrophoresis column is made of transparent hard glass material, and the separation medium is linear polyacrylamide buffer solution.
5. The apparatus for purifying raw material insulin from animal pancreas as claimed in claim 1, further comprising a spiral line condenser, wherein the spiral line condenser is wrapped around the separation tube.
6. A method for using the insulin raw material purification device according to any one of claims 1 to 5, comprising the following steps:
s1, raw material pretreatment: the animal pancreas is frozen in deep immediately after being separated from the body, the pancreas is frozen and crushed, then acidic ethanol is added for stirring and extraction, and centrifugation is carried out to obtain the supernatant; wherein the residue is extracted again by repeating the above operations, and the extracting solutions are combined to obtain crude insulin;
s2, separating and purifying a sample: the condenser is started first, and the temperature display reaches the required temperature; then turning on the ultraviolet detector, and setting the detection wavelength; then the peristaltic pump and the one-way valve are opened, the flow rate is adjusted, and the flow rate is stabilized; then turning on a high-voltage power supply to set various parameters, and inputting crude insulin for separation and purification after instrument equipment is stable;
s3, detecting insulin: and (5) detecting on line in real time by using a detector, continuously separating and purifying if the insulin is unqualified until the insulin is qualified, and adjusting a one-way valve to obtain the separated and purified insulin.
7. The use method of the apparatus for purifying a crude drug of insulin derived from animal pancreas according to claim 6, wherein in step S2, the high voltage DC variable frequency power source is a 0-30kV DC variable frequency high voltage power source, and the variable frequency mode of the high voltage DC variable frequency power source is any one of spike pulse, positive sine wave pulse and square wave pulse.
8. The use method of an apparatus for purifying a crude drug of insulin derived from animal pancreas as claimed in claim 6, wherein in step S2, the pH of the high pressure electrophoresis is 3.5-4.8; the temperature of the high-pressure electrophoresis is 0-25 ℃.
9. The use method of the apparatus for purifying a crude drug of insulin derived from animal pancreas according to claim 6, wherein in step S2, the electric field intensity of the high voltage electrophoresis is 0.09k V/cm-2.82 kV/cm.
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