KR101461221B1 - Method for Producing Nanobubble Liquid of Organ Preservation - Google Patents

Abstract

(A) a long-term storage liquid feedstock supply unit (A, B, and C); (b) a gas supply part (F) for supplying gas to the first reaction tank (9), the second reaction tank (26) and the third reaction tank (29); (b) an ultrapure water supply part (D); (c) a base water supply unit G; (d) a long-term storage liquid manufacturing section (E); And a cleaning liquid supply unit (H) for producing a nano bubble ozonated water and supplying it to the metering tank (a) and performing a CIP (Clean-in-place) And a cleaning-in-place (CIP) step of cleaning residual materials during the process using the same, and a method for manufacturing a long-term preservation liquid for implantation containing nano bubbles
Since a device for manufacturing a long-term preservation liquid for implants according to the present invention is equipped with a CIP (Clean-In-Place) device, the device is cleaned at the same time as the manufacturing process and the process operation is convenient. Reperfusion injury < / RTI > effect that has been previously clinically used for the preservation of transplantation organs, is very useful in the medical industry.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for producing a nanobubble-

The present invention relates to a method and an apparatus for producing a long-term preserved liquid containing nano bubbles.

The organ transplants are intended to provide long-term survival to patients who have not survived about a year or more due to end-stage malfunctions of various organs of the human body, by replacing organs from organs donated by organ donors, including brain dead or deceased. For example.

Such organ transplantation is becoming more and more common due to recent technological advances in surgery and the development of drugs such as immunosuppressants. However, even though the number of patients awaiting organ transplantation is large, there is an absolute shortage of organ donors and current limitations of long-term preservation techniques, It is a fact that it can not support.

There is an ischemia time during the resumption of blood perfusion due to the connection of organs to the blood vessels of the recipient since the organs of the donor are inevitably extracted from the donor organ during the organ transplantation, To preserve the organs for transplantation, a method of reperfusion after storage at a low temperature that inhibits metabolism occurring within the organ of 20 degrees or less, generally 4 degrees Celsius, rather than providing a physiological environment similar to that in vivo And these methods are inevitable not only for damage caused by oxidative stress by ischemia-reperfusion in organ transplantation but also for microcirculatory disturbance due to cell contraction and endothelial cell morphology, (eg, inflammation damage due to migration) It is known

Cellular changes during ischemic time are primarily caused by the production of metabolites in oxygenated and oxygenated states of loss of oxygen and nutrients, and intracellular ATP is rapidly reduced. ATP is crucial for maintaining electrolyte balance in the cell membrane. When ATP is lost, K ⁺ and Mg ^{2 +} ions are extracellularly intracellularly, while Na ⁺ and Ca ^{2 +} ions move into cells, resulting in edema. As a result of anaerobic metabolism, lactic acid is formed and pH is decreased in the cells. This destabilizes the cell membrane of the lysosome containing proteolytic enzymes in the cell and liberates iron or copper ions attached to the protein. After blood flow is resumed, oxygen is supplied to the cells again. Increased Ca ^{2 +} in the cell activates the cytoplasmic protease, forms xanthine, and superoxide and H ₂ O ₂ Free radicals are generated. This causes structural damage by oxidizing the lipid component of the cell membrane. In particular, damage of the endothelial cell causes blood flow to the organ of the transplant.

During the ischemic time, the cell damage of the organs that are transplanted is inevitable though there is a difference in degree, and development of a method for minimizing cell damage and development of transplantation long term storage is essential. To inhibit ischemia and reperfusion injury, long term storage contains various substances. In order to prevent cell edema, it contains mamnitol, sucrose, hydroxyethyl starch, lactobionace, raffinose and gluconate which do not penetrate the cell membrane. In order to supply the energy into the cell, adenosine, xanthine oxydase inhibitor allopurinol , Glutathione to treat free radicals, insulin and steroids to stabilize the cell membrane, and buffer substances to buffer the cell's pH reduction. In addition, a high concentration of K ⁺ ions may be added to inhibit the release of K ⁺ ions contained in cells and prevent cell edema.

Currently, various preservative solutions have been developed and used clinically to minimize organ damage due to ischemia - reperfusion in organ transplantation. Although Euro-Collin's was initially used as such a preservative solution, UW (Univ. Wisconsin, Wahlberg, JA, et al ., Transplantation , 43: 5, 1987) is the most widely used fluid. However, although the UW liquid has an osmolarity regulating effect due to the use of lactobionic acid and hydroxyethyl starch, it has antioxidant properties such as reduced glutathione (GSH) and allopurinol Although the ability has been added, it is known that it has a problem that it can not effectively prevent high price, high K ⁺ content and long-term damage due to ischemia-reperfusion.

Studies have been actively conducted on various organ preservatives that can more effectively prevent organs damage by ischemia-reperfusion, and have been actively studied for the preparation of crystalline cardioplegia solution containing histidine, tryptophan and ketoglutarate as main components (Histidine-Tryptophan-Ketoglutarate solution), a long-term preservation solution containing polyamines such as polythymine, putrescine, spermidine, and spermine and a carrier (Korean Patent No. 304594), polyphenol (Korean Patent Application Laid-Open No. 2001-0029692), a long-term storage liquid containing perfluorocarbon (Korean Patent Application Laid-Open No. 2001-0029692), a long-term preservation including cystamine (Korean Patent No. 665976) and the like have been developed and reported, but they have not yet been widely used in preserving organs for transplantation Effective preservative is a situation that is not known.

Nano - bubbles are ultrafine bubbles which can not be seen by eyes of 1 ㎛ or less. They have 1 / 2,000 size of normal bubbles. By instantly inducing ultra - high temperature and ultra - high pressure state in bubble generation and extinction process, they generate heat ultrasonic waves, It also emits.

Nano bubbles generate negative ions through the ultrasonic wave to remove nutrients and oxygen from the outside of the skin as well as skin keratin and pores. It also promotes blood circulation, facilitates metabolism, relieves fatigue and muscle elasticity It is also effective for pimples, bacteria, sebum, and keratin which are parasitic on the pores, thus helping the skin to regenerate. In addition, according to the results of experiments conducted by Korea Far Infrared Application Evaluation Research Institute, it is known that 99% high sterilizing power against Escherichia coli, Pseudomonas aeruginosa, Salmonella and Vibrio bacteria is known.

Therefore, the inventors of the present invention have developed a long-term storage liquid manufacturing apparatus capable of forming a nano-bubble inclusion liquid as a base water and having various compositions in order to develop a technique for utilizing a solution containing nanobubbles, Clean-In-Place (NBB) device equipped with a nano bubble can be more conveniently obtained.

An object of the present invention is to provide an apparatus for manufacturing a long-term preservation liquid for implantation using a base water in which a nano bubble is mixed in a mixed solution of ultrapure water and sodium containing a cleaning liquid supply unit for CIP (clean-in-place) And to provide a method for producing a long-term preservation liquid.

In order to achieve the above object, the present invention provides a process for producing a long-term storage liquid feedstock comprising: (a) a long-term storage liquid feedstock supply unit (A, B, and C); (b) a gas supply part (F) for supplying gas to the first reaction tank (9), the second reaction tank (26) and the third reaction tank (29); (b) an ultrapure water supply part (D); (c) a base water supply unit G; (d) a long-term storage liquid manufacturing section (E); And a cleaning liquid supply unit (H) for producing a nano bubble ozonated water and supplying it to the metering tank (a) and performing a CIP (Clean-in-place) Thereby providing a storage liquid producing apparatus.
(A) preparing ultrapure water through three-step sequential filtration of raw water and storing it in ultrapure water storage tank 5; (b) producing a base water using ultrapure water; (c) transferring the organ preservation liquid raw material and the base water to the second reaction tank 26 and mixing them; (d) a first CIP (Clean-in-Place) step for the tanks and valves of the long term storage liquid feedstock (A, B, C); (e) preparing a long-term preservation liquid; And a second clean-in-place (CIP) step of cleaning the tanks and valves of the organically preserved stock feed, and (f) the residual feedstock in the first reservoir 26, And a method for producing a long-term preservation liquid for implantation.

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Since a device for manufacturing a long-term preservation liquid for implants according to the present invention is equipped with a CIP (Clean-In-Place) device, the device is cleaned at the same time as the manufacturing process and the process operation is convenient. Reperfusion injury < / RTI > effect that has been previously clinically used for the preservation of transplantation organs, is very useful in the medical industry.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a configuration diagram of a long-term storage liquid preparation device for implantation.
Fig. 2 is a structural view of the apparatus for preparing a long-term storage liquid for transplantation.

Hereinafter, the present invention will be described in detail.

As a result of intensive efforts to develop a technology capable of exploiting the usefulness of nano bubbles, the present inventors have developed a device capable of producing a long-term preservation liquid based on a nano bubble mixed liquid, It is possible to obtain a long-term preservative solution for transplantation and a cleaning solution for surgery, and completed the present invention.

In one aspect of the present invention, there is provided a method for purifying ultrapure water, comprising the steps of: (a) mixing sodium chloride with ultrapure water purified through a three-stage filtration process of an ultrafilter, an ion filter, and deionization (EDI) (b) generating nano bubbles at a high pressure by adding oxygen to the mixed solution; (c) passing the nano bubble mixed solution through an ultrafilter and an ion filter to remove ionic substances and sodium ions to produce a base water containing nano bubbles; (d) mixing a long-term stock solution and a base water; (e) adding a base water to the mixture of (d); And (f) adding oxygen and carbon dioxide to the mixture of (e) and allowing the nano bubbles to mix at a high pressure.

In the method for producing a long-term preservation liquid for implantation of the present invention, 10 g / L to 15 g / L of sodium chloride is added before the step (b) in which nanobubbles are introduced, more preferably 13.56 g / L .

The sodium chloride is ionized in a solution to shield the bubble at the interface between the bubble and the solution due to the electrostatic adsorption capability, so that the gas can not be dispersed, thereby stabilizing the bubble.

In the method for producing a long-term preservation liquid for implantation of the present invention, the raw water may be tap water, but is not limited thereto.

In the manufacturing method of the present invention, the oxygen and the carbon dioxide in the step (f) are supplied as a mixed gas to produce a nano bubble, and the two gases are mixed at a ratio of 2: 1. The reaction time is 15 minutes To 30 minutes.

In the manufacturing method of the present invention, the mixture in step (f) is mixed in step (d) with 5% of the raw material for long term preservation liquid and 40% 15%, 20% and 20% of the additional base water 45% are sequentially introduced.

The additional base water is used to clean the residual components in piping and valves disposed between the weighing tank and the second reaction vessel after supplying the organ preservation liquid raw material to the second reaction vessel 26 of the apparatus, Since three metering tanks are provided in the system, they are sequentially injected three times.

In the method for producing a long-term preservation liquid for transplantation of the present invention, the long-term preservation liquid raw material contains an osmotic control agent as an active ingredient.

In the present invention, the osmotic conditioning agent induces hyperosmolarity and inhibits cell oedema, which is frequently found in long-term refrigerated storage. The osmotic modulating agent may be glucose, lactobionate, But is not limited to, at least one selected from the group consisting of raffinose, hydroxyethyl starch, sucrose, gluconate, and mannitol.

The osmotic agent may be composed of Collins liquid, Euro-Collins liquid, UW liquid, Histidine-Tryptophan-Ketoglutarate solution and the like. Recently, (Korean Patent No. 304594), technology using polyphenol (Korean Patent Application Laid-Open No. 2001-0029692), technology using perfluorocarbon (Korea Patent No. 304594), technology using phthalocyanine, (Patent Application Publication No. 2001-0029692) and a technique using cystamine (Korean Patent No. 665976) can be used.

In the preparation method of the present invention, the amount of the osmotic modulating agent is not particularly limited, and if hydroxyethyl starch is used, 20-70 g / l, when lactobionate is used, 60-120 mM, raffinose is used 10 mM to 50 mM is preferable.

As used herein, the term "organ" is only a specific representation of the subject to which the composition of the present invention is applied, and the composition of the present invention is not limited to a narrow organs but also to a part of organs in a narrow sense, . Thus, the term "organ" used in connection with the long-term preservation solution for transplantation of the present invention is interpreted in its broadest sense to include the organs and parts thereof, tissues and parts of tissues in the narrow sense. The tissue is not particularly limited and includes, for example, an epithelial tissue, a connective tissue, a muscle tissue, a nerve tissue, and the like. In addition, the organ in the narrow sense is not particularly limited and includes, for example, skin, blood vessel, cornea, kidney, heart, liver, umbilical cord, intestine, nerve, lung, placenta, .

The long-term storage liquid for transplantation prepared by the method of the present invention may contain additional raw materials as long as the organ is stored outside the body, a substance having an effect of preventing damage due to various stresses applied to organs, and a combination thereof.

The additional raw materials contained in the organ transplantation liquid prepared by the method of the present invention are subjected to a step of transplantation after being stored in vitro in a surgical operation procedure for transplanting various organs of an animal or a human, (Adenosine triphosphate) source, which prevents depletion of adenosine 5'-triphosphate (ATP), which is one of the important factors causing organ damage, as well as an osmotic agent, An antioxidant that suppresses oxidative stress by ischemia-reperfusion, and an electrolyte ion, or the like, alone or in combination.

Therefore, in the method of the present invention for producing a long-term preservation liquid for transplantation, the organ preservation liquid raw material is also at least one selected from the group consisting of an adenosine triphosphate source, an antioxidant, an antibiotic, and an electrolyte ion do.

In the production method of the present invention, the source of the adenosine triphosphate is at least one selected from the group consisting of glucose, adenosine, and alpha-ketoglutarate. However, it is not limited thereto.

The antioxidant may be at least one selected from the group consisting of flavonoids, glutathione, catechin, polyphenol, ascorbic acid and allopurinol. But is not limited thereto.

In the method of the present invention for producing a transplantable organ preservation solution, the antibiotic is selected from the group consisting of Penicillin, Streptomycin, Kanamycin, Gentamycin, Neomycin, and Apramycin. And the electrolyte ion is at least one selected from the group consisting of sodium ion (Na ⁺ ), potassium ion (K ⁺ ), hydrogen phosphate ion (H ₂ PO ^4- ), hydrogen phosphate ion (HPO ₄ ^2- ) Mg ²⁺ ), sulfate ion (SO ₄ ^2- ), chloride ion (Cl-), and carbonate ion (H ₂ CO ^3- ).

In the method of the present invention for preparing a long-term preservation liquid for transplantation, the long-term preservation liquid raw material is at least one further selected from the group consisting of hormones, growth factors, enzymes, and natural extracts, But is not limited thereto.

The term "nano bubble" as used herein is characterized by having a size of less than 1 [micro] m including an active gas such as oxygen, ozone, carbon dioxide or nitrogen monoxide.

In the manufacturing method of the present invention, since the base water is ultrapure water containing oxygen water or oxygen nano bubbles, sodium is added to stabilize the bubbles in step (b), and the step of removing sodium hydroxide is performed in step (c) have.

Generally, when nano bubbles are produced only with pure water, bubbles disappear within 5 minutes, but when sodium is added as a catalyst, the bubbles can be stabilized by bonding with nano bubbles.

In step (d) of the present invention, 10% of the organosoluble raw material mixture is added to the base water in an amount of 5% based on 40% of the final reaction weight, and in step (e) 15% to 20% of base water is further added.

The reason for adding the base water in the step (e) is that it may remain in the piping and the valve of the apparatus in the process of adding it, since there is more than one raw material of the organically preserved liquid, Therefore, after adding one kind of raw material, all of the remaining raw materials are put into the reaction tank, and the effect of cleaning the device is obtained.

In another aspect, the present invention provides a method of treating a raw water introduced through a raw water inflow pipe (1) in three stages (ultrafilter (2), ion filter (3), and deionization An ultrapure water supply unit including a filtration unit and an ultrapure water storage tank (5); (b) a nano bubble generator 8 connected to a gas suction device 7 and a first reaction tank 9, a sodium chloride supply device 6 and a storage tank 10 each of which is connected to an upper layer of the ultrapure water inflow pipe from the ultrapure water supply unit, A base water producing unit comprising: (c) a long term storage liquid raw material supply section including raw material supply tanks 11 to 20, weighing tanks 21 to 23, and a supply pipe; (d) a second reaction vessel (26) having an upper portion mounted with a nano bubble generator (25) to which the base water inflow portion and the raw material supply pipe from each of the weighing tanks are fastened and a gas inhalation device A long - term storage liquid manufacturing section; And (e) a long-term storage liquid storage unit.

The apparatus for producing a long-term storage liquid for implants according to the present invention includes a third reaction tank 29 on which a base water inlet and a gas generator 27 are connected and a nano bubble generator 28 connected thereto, And a cleaning liquid supply unit for manufacturing and supplying cleaning nano-ozone water.

In the apparatus of the present invention, the gas suction apparatuses 7 and 24 receive oxygen, ozone, or carbon dioxide from the gas generator 27 of the gas supply unit and transfer the same to the generator, and the gas suction apparatus is connected to the first reaction vessel 9 ), And a second reaction tank (26), and supplies a gas mixture of oxygen and oxygen and carbon dioxide to the nano bubble generator installed in each reaction tank through the gas supply pipe, and the third reaction tank (29) And the ozone is supplied directly from the device 27. [

The gas inhalation device (7, 24) supplies gas to a generator of each reaction tank. The gas inhalation device is composed of a pump suction venturi pipe and line mixing. The sucked gas is circulated into the generator and discharged into the reservoir Create a nano bubble.

 In the apparatus of the present invention, the raw material tanks (11 to 20) store raw materials for a long term preservation liquid for transplantation, and the raw materials include osmotic control agents, adenosine triphosphate source, antioxidants, antibiotics, Factors, enzymes and natural extracts may be used. Therefore, one or more or fifteen or less of them may be used for storing them individually, and those skilled in the art can easily determine and install them according to the kind of raw materials required.

The concentrations of the raw materials stored in the raw material tanks 11 to 20 are as follows: raw material 132 ppm, 3L, raw material 12, raw material 24, 756 ppm, 3L, raw material tank 13, raw material C, raw material 24,756 ppm, Tank (14) D Raw material 49,788 ppm. 3L and raw material tank 15 E raw material 40,458 ppm 3L raw material tank 16 raw material 52,602 ppm 3L raw material tank 17 raw material 68,609 ppm 8L raw material tank 18 raw material 99,365 ppm 8L, Raw material tank (19) I Raw material 68,609 ppm, 63L, and raw material tank (20) J Raw material 12,378 ppm, 3L can be stored respectively. Raw material stored in each raw material tank can be stored for 24 times Adjust and store.

In the apparatus of the present invention, the nano bubble generator 9, 25, 28 may be an impeller type or a screw type, but is not limited thereto.

The HMI (Human Machine Interface) system, which monitors the senses such as the automatic valve, the load cell, and the temperature included in the apparatus of the present invention and controls it automatically and unattended, is characterized by controlling a programmable logic controller (PLC) .

In the method for producing a long-term preservation liquid for use by the apparatus of the present invention, the accuracy of a predetermined amount for stabilizing the raw material and stabilization of the nano bubble are the most important factors. The apparatus of the present invention can reduce the error to less than 0.2% by measuring a plurality of raw materials introduced from the raw material tank by attaching a meter attached to each of the reservoirs in order to manufacture a preservable liquid having a good function.

The apparatus of the present invention further includes a cleaning step during the manufacturing process to minimize contamination between raw materials and to facilitate maintenance of the apparatus.

The cleaning process is characterized in that after the raw materials are put into the second reaction tank 26 in the preservative liquid manufacturing process, the base water is transferred to the raw material weighing tanks 21 to 23 to clean the remaining raw materials. In this case, Since the amount of raw material to be weighed in the tank is different, there is a characteristic that there is a difference in the amount of base water transferred for cleaning.

The weighing tanks included in the apparatus of the present invention are characterized by a capacity of 60 L to 1500 L, but are not limited thereto.

The apparatus of the present invention can be upgraded by incorporating nano bubbles into a long-term preservation liquid that is commercially available or developed by a third party, without newly mixing.

The long-term preservative solution is a long-term preservation solution (Korean Patent No. 304594) comprising a crystalline cardioplegant solution (HTK solution), hydroxyethyl starch, polyamine, putrescine, spermidine, (Korean Patent Application Laid-Open No. 2001-0029692) and polyphenol (Japanese Patent Application Laid-open No. 2001-0029692) containing perfluorocarbon by adding polyphenol to a long-term preserved liquid to improve the preservation period of the organ for transplantation, , And a long-term preservation composition containing cystamine (Korean Patent No. 665976), but is not limited thereto.

The method for producing a long-term preservation liquid for implantation containing nanobubbles using the apparatus of the present invention comprises the steps of: (a) preparing an ultrapure water; (b) a base water producing step in which nano bubbles are mixed; (c) a process for preparing a long-term storage liquid for transplantation by mixing the additional raw materials; And (d) the pipe cleaning process.

The present invention will be described in more detail with reference to the following examples. However, the following examples are provided to aid understanding of the present invention, and the scope of the present invention is not limited by these examples in any sense.

In this case, unless otherwise defined, technical terms and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the following description and the accompanying drawings, A description of known functions and configurations that may unnecessarily obscure the description of the present invention will be omitted.

Example 1 : Method of manufacturing long-term preservation liquid for transplantation by mixing one or more raw materials

The method for producing a long-term preservation liquid for implantation containing nanobubbles using the apparatus of the present invention comprises the steps of: (a) preparing an ultrapure water; (b) a base water producing step in which nano bubbles are mixed; (c) a process for preparing a long-term storage liquid for implantation by mixing raw materials; And (d) the pipe cleaning process.

One) Ultrapure water  Manufacture process

When supplied with tap water as raw water to remove the suspended solids in the primary ultra-filter (2), an ion filter 3 bite secondary ionic, bacteria (colony Total Counts) total coliforms (Total Coliforms) Escherichia coli (E. E. coli ) heavy metals and the like, and EDI (electrodeionization) (4) is used as a tertiary filter to produce ultrapure water. The treated water has a dissolved oxygen content (DO) of 0.46 ppb (1 / billionth of a degree) and an organic carbon content (TOC) of 2.18 ppb or more with a resistivity of 18 MΩ.cm or more. The produced ultrapure water is used for producing base water, It is supplied to the long-term preservation liquid manufacturing process.

2) Nano bubble  The incorporated base Water  Manufacture process

The base water manufacturing process includes a sodium chloride storage device 6, a first reaction tank 8, an ultrapure water supply pipe, an instrument, a gas suction device 7, an ultrafilter 30, an ion filter 31 and a base water storage tank 10 .

The sodium chloride storage device 6 includes a reservoir for storing 1 TON of the sodium reagent, and the reagent is transferred to the supply tube and mixed with the base water while the screw rotates at 30 rpm.

In order to manufacture the base water, 350 L of ultra-pure water was transferred to the first reaction tank 8, 9,492 g of sodium chloride was added to the first reaction tank so that the measurement error of the load cell of the meter 31 was within ± 0.5% It receives 350L by car. When the metering is completed, the generator 9 is operated for 180 seconds to mix ultrapure water and sodium chloride.

Next, in order to generate nano bubbles, the gas suction unit 7 of the first reaction tank 8 is operated for 25 minutes so that oxygen is circulated into the generator 9, and when the operation is completed, The filter 30 and the ion filter 31 are passed to produce a base water in which sodium and ionic substances are removed and oxygen nano bubbles are mixed.

The base water is characterized by having nano bubbles of 100 nm mixed in ultrapure water. The base water is stored in the storage tank 10 until it is used in a preservation liquid manufacturing process or a cleaning process.

3) Long-term preservation liquid manufacturing process for transplant

end. Long-term storage liquid raw material

The concentrations of the raw materials stored in the raw material tanks (11 to 20) are as shown in Table 1, and the amount of the raw materials to be mixed is adjusted to 5% of the total reaction weight in accordance with the final concentration of the nine raw materials.

number Raw material Mass (g / mol) Final concentration (mol / L) One Sodium chloride 58.44 0.015 2 Potassium chloride 75.55 0.009 3 Potassium hydrogen 2-ketoglutarate 184.19 0.001 4 Magnesium chloride hexate 203.30 0.004 5 L-histidine hydrochloride 209.23 0.018 6 Histidine 155.18 0.18 7 Tryptophan 204.23 0.002 8 Mannitol 182.17 0.03 9 Calcium chloride dihydrate 147.02 0.000015

I. Base Water  And raw material mixing

The pump is operated from the base water storage tank 10 to transfer the base water 40% to the second reaction tank 26 and the long term storage liquid raw material (s) is added to the reaction tank so as to be 5% The pump is restarted from the reservoir 10 to transfer the base water 15%, 20%, and 20% to the weighing tank 1 (21), weighing tank 2 (22), and weighing tank 3 So that the raw materials remaining in the piping and the valve are introduced into the reaction tank while being transferred to the second reaction tank 26.

When the raw material is more than one, the raw material supply from the raw material tank and the piping and the valve cleaning process by the base water are repeated after each raw material supply process so that the raw material is mixed or not contaminated in the pipe or valve.

Weighing error of each raw material is within 0.2%.

All. Nano bubble  produce

When a plurality of raw materials and base water are fed to the second reaction tank 26, the gas suction device 24 and the generator 25 are operated for 180 seconds to mix the raw materials. At this time, the generator 25 is variablely operated at 1000 rpm at 3000 rpm and circulated at a pump operation 50% turnover rate.

The gas suction device 24 is connected to a gas suction device 24 so that raw materials of the reaction tank are line-mixed while oxygen and carbon dioxide are transferred to a supply pipe At this time, the oxygen is operated at 20 ppm and the carbon dioxide is operated at 15 ppm for 5 to 30 minutes.

Each of the above gases is introduced into the generator 25 mounted in the second reaction vessel 26 by the piping. When the gas is supplied, the generator 25 is operated at 5,000 rpm for 300 seconds; 10,000 rpm, 300 seconds; 20,000 rpm, 300 seconds; And 30,000 rpm for 15 minutes, respectively, to prepare a long-term preservative solution for implantation containing nanobubbles, and the prepared preservative solution was transferred to a storage tank.

The pump adjusts the operating rate of the pump to the vacuum sensor according to the set value in the inverter.

Example 2 : Method for manufacturing long-term preservation solution for transplant using single raw material

The long term preservation liquid can be upgraded by incorporating nano bubbles into the long-term preservation liquid commercialized using the apparatus of the present invention, the long term preservation liquid can be introduced into the second reaction vessel 26, the gas suction apparatus 24 and the generator 25 can be operated Then, as described in Example 1, the gas was supplied for 15 minutes so that oxygen and carbon dioxide were 20 ppm and 5 ppm, respectively, so that nano bubbles were generated. The upgraded organ preservation solution thus prepared is transferred to the storage tank 32 for storage.

Example  3: Piping cleaning process

CIP (Clean-in-Place) is essential during the process because raw materials of raw materials may remain in piping and contamination may occur during weighing. In the CIP process, a single or a plurality of raw materials are first metered into the second reaction vessel 26, and 40% of the base water is supplied to the reactor, 15% to the metering tank 1 (21) (22), and 35% is supplied to the metering tank 3 (23), and then flows into the reactor so that the pipe and the valve are cleaned.

In the general CIP method, ultrapure water introduced into the third reaction tank 29 included in the cleaning liquid supply unit and ozone introduced by the generator 28 through the gas generating unit 27 are mixed with each other to form nano bubble ozone water And the ozone water is supplied to the metering tank 1 (21), the metering tank 2 (22), and the metering tank 3 (23) to clean the tank and the pipe. The ozonated water used for cleaning is reused in the recovery tank after the ultrafilter and ion filter process.

1: raw water inlet pipe 2, 30: ultra filter
3, 31: ion filter 4: Electrodeionization (EDI)
5: Ultrapure water storage tank 6: Sodium storage tank
7, 24: gas suction device 8: first reaction tank
9, 25, 28: Generator 10: Base water reservoir
11 to 20: raw material tanks 21 to 23: weighing tank
26: second reaction tank 27: gas generating device
29: third reaction tank 32: measuring instrument
33: gas tank 34: flow meter
35: load cell 36: valve
37: Pump
A, B, C: raw material supply part D: ultrapure water supply part
E: long-term coarse solution producing part F:
G: base water supply part H: cleaning liquid supply part

Claims (14)

A long-term storage liquid preparation apparatus for implantation containing a nano bubble having the following constitution:
(a) a long-term storage liquid raw material supply unit (A, B, and C) provided with a raw material supply tank for preparing long-term storage liquid, a weighing tank, and a supply pipe;
(b) a gas tank 33 for storing ozone, carbon dioxide or oxygen, and a gas generating device 27, and the gas is supplied to the first reaction tank 9, the second reaction tank 26 and the third reaction tank 29 A gas supply part F for supplying gas;
(b) The raw water introduced through the raw water inlet pipe 1 is processed while sequentially passing through the ultrafilter 2, the ion filter 3 and the deionization (EDI) 4 to produce ultrapure water. An ultrapure water supply part (D) including a filtration device and an ultrapure water storage tank (5);
(c) a gas suction device 7 for supplying oxygen and carbon dioxide from the gas supply part F to the reaction tank is connected to the upper part of the reaction tank through a pipe, a nano bubble generator 8 is installed in the hollow upper part of the reaction tank, A first reaction tank 9 in which a pipe is connected to the upper portion of the ultrapure water supply unit D, a sodium chloride supply unit 6 connected to the upper part of the reaction tank by piping, A base water supply unit (G) including a water tank (30), an ion filter (31), and a reservoir (10)
(d) a gas suction device (24) for supplying the base water supply part (G) and the raw material supply pipe from the metering tank (a) above and supplying oxygen and carbon dioxide from the gas supply part (F) And a second reaction vessel (26) connected to the upper portion of the reaction vessel through a pipe, wherein the nano bubble generator (25) is installed in the hollow of the reaction tank. The nano bubble generator (E); And
(e) a third reaction tank (29) having a pipe for supplying ozone from the gas supply unit (F) and having a nano bubble generator (25) installed in a hollow portion in the reaction tank, A cleaning liquid supply unit H for producing bubble ozonated water and supplying it to the metering tank of (a) to perform CIP (Clean-in-place).
A method for producing a long-term preservation solution for implantation containing nano bubbles by the apparatus of claim 1 comprising the steps of:
(a) preparing ultrapure water through a three-step sequential filtration process including an ultrafilter, an ion filter, and deionization (EDI), and storing the ultrapure water in the ultrapure water storage tank 5;
(b) transferring ultrapure water and sodium chloride from the ultrapure water storage tank 5 and the sodium chloride storage device 6 respectively to the first reaction tank 1 and introducing oxygen and carbon dioxide into the reaction tank by the gas suction device 7 in the reaction tank The nano bubble generator 8 is operated to mix nano bubbles with the ultra pure water and sodium chloride mixed solution while generating nano bubbles. Then, the nano bubble mixed liquid is supplied to the ultrafilter 30 ) And an ion filter (31), and transferring the sodium to the storage tank (10);
(c) transferring the starting material from the long term storage liquid feed materials (A, B, C) and the basal water from the storage tank (10) of step (b) to the second reaction vessel (26) and mixing them;
(d) The base water is further transferred to the weighing tanks 21 to 23 from the reservoir 10 in the step (b) to clean the tanks and the remaining materials in the valve of the organ preservation liquid raw material supply parts A, B and C A first in-process cleaning (CIP) step in which the base water containing the residual raw material is input to the second reaction tank 26;
(e) The oxygen and carbon dioxide are introduced into the second reactor 26 through which the base water and the raw material are introduced through the steps (c) and (d) by the gas sucking device 24, and the nano bubble generator 25 Preparing a long-term preserved liquid for allowing the nano bubbles to be mixed into the base water and the raw material; And
(f) transferring ozone from the gas supply unit F to the third reaction tank 29 into which the base water of step (b) is charged, and operating the nano bubble generator 28 mounted on the reaction tank to produce nano bubble ozonated water Next, a second CIP (Clean-in-Place) step is carried out to further transfer the material to the weighing tanks 21 to 23 to clean the tanks and valves of the long term preserving liquid raw material supply portion and the residual materials in the first reservoir 26 .
The method according to claim 2, wherein the base water in step (c) transfers 40% of the final weight of the organ preservation solution to the second reaction tank (26).
The method according to claim 2, wherein the first in-situ cleaning (CIP) of step (d) is repeated.
The method according to claim 2, wherein the cleaning liquid obtained in step (f) is discharged through an outlet provided in a lower part of the first reservoir (26).
3. The method according to claim 2, wherein 10 g / L to 15 g / L of the sodium chloride in step (b) is added.
3. The method according to claim 2, wherein the organ preservation liquid raw material comprises an osmotic conditioning agent as an active ingredient.
3. The method of claim 2, wherein the organ preservation fluid material further comprises at least one further selected from the group consisting of a source of adenosine triphosphate, an antioxidant, an antibiotic, and an electrolyte ion. A method for producing a long - term preserved liquid.
[8] The osmolysis-controlling agent according to claim 7, wherein the osmotic agent is selected from the group consisting of glucose, lactobionate, raffinose, hydroxyethyl starch, sucrose, gluconate, Wherein the at least one selected from the group consisting of mannitol and mannitol is at least one selected from the group consisting of mannitol and mannitol.
9. The method of claim 8, wherein the source of adenosine triphosphate is at least one selected from the group consisting of glucose, adenosine, and alpha-ketoglutarate. A method for producing a long - term preservation liquid for transplantation.
The antioxidant according to claim 8, wherein the antioxidant is at least one selected from the group consisting of flavonoid, glutathione, catechin, polyphenol, ascorbic acid and allopurinol Wherein the method comprises the steps of:
9. The antibiotic according to claim 8, wherein the antibiotic is at least one selected from the group consisting of Penicillin, Streptomycin, Kanamycin, Gentamycin, Neomycin, and Apramycin Wherein the method comprises the steps of:
The method of claim 8, wherein the electrolyte ion is selected from the group consisting of Na ⁺ , K ⁺ , H ₂ PO 4 ^- , H ₂ PO ₄ ^2- , Mg ^{2 +} ), Sulfate ion (SO ₄ ^2- ), chloride ion (Cl ^- ), and carbonate ion (H ₂ CO ^3- ).
[3] The method according to claim 2, wherein the organ preservation liquid raw material further comprises at least one selected from the group consisting of hormones, growth factors, enzymes, and natural extracts.

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