Test fluid for evaluating protein recovery performance of body cavity fluid concentrator and its manufacturing method

Abstract

The present invention relates to a test solution for evaluating the protein recovery performance of a body cavity fluid concentrator with excellent reliability and a method for producing the same. The test solution has a total protein concentration of 0.5 to 2.4 g / dL, an albumin to globulin ratio (A / G ratio) of 0.8 to 1.5, and a blood cell count of 1×10 2 less than 1 μg / μL.

Description

Technical Field

[0001] The present invention belongs to the field of medical devices and relates to a test fluid used in the performance evaluation of medical devices and a manufacturing method thereof, and in particular to a test fluid used in the performance evaluation of a device or component for processing human body cavity fluid and a manufacturing method thereof. Background Art

[0002] As a treatment for refractory ascites, there is a cell-free and concentrated ascites reinfusion therapy: ascites is removed from the patient, pathogenic substances such as bacteria and cancer cells are removed from the ascites, the ascites is dehydrated while retaining useful components such as albumin, and the concentrated solution after dehydration is reinfused into the body.

[0003] Patients with ascites can be broadly divided into hepatic ascites (caused by diseases such as cirrhosis) and cancerous ascites (caused by cancers such as stomach cancer, ovarian cancer, and colorectal cancer). Previously, the above treatments were primarily used for hepatic ascites, but in recent years, their effectiveness in treating cancerous ascites has been increasingly recognized, leading to an increase in the number of treatments available for this condition.

[0004] In the above-mentioned treatment methods, an ascites treatment device is generally used. In this ascites treatment device, an ascites bag, a filter, a concentrator, and an ascites concentrated ascites bag are connected in series in this order, and the ascites is filtered and concentrated by flowing the ascites using a drop or a pump.

[0005] For example, for patients with liver cirrhosis and other conditions prone to accumulation of ascites and pleural effusion (hereinafter collectively referred to as ascites), in order to utilize the protein in the ascites to increase the patient's blood protein concentration, the following ascites filtration, concentration, and intravenous injection method is performed: by using two filters with hollow fiber membranes, the ascites, which is discharged from the body after a needle is inserted into the effusion, is filtered and concentrated to obtain a concentrated protein solution, which is then injected into the patient intravenously. The first of the two filters is used to remove cellular components such as cancer cells and blood cell components contained in the ascites. A membrane with a pore size that does not allow cellular components to pass but allows solute components such as water and protein to pass can be used. On the other hand, the other filter is a concentration filter used to remove water from ascites with a dilute protein concentration and concentrate the protein. A membrane that substantially does not pass protein components but allows water, electrolytes, etc. to pass can be used. Generally, from the perspective of convenience, a method can be adopted in which the cellular components are filtered and separated using a filter and the filtered ascites is concentrated using a concentrator. An apparatus that allows these two processes to be performed continuously can be used.

[0006] Furthermore, as such an ascites treatment apparatus, Reference 1 discloses an ascites treatment apparatus equipped with a system for automatically switching from filtration to recirculation.

[0007] References:

[0008] Reference 1: Japanese Patent Application Laid-Open No. 2013-188427 Summary of the Invention

[0009] Problems to be solved by the invention

[0010] Various practical devices and apparatuses have been developed for treating body cavity fluids. These devices and apparatuses are subject to extremely high safety and reliability requirements in practical use. As previously mentioned, concentrators are used to concentrate nutrients in body cavity fluids that are beneficial to the human body. Typically, these concentrated body cavity fluids are considered free of viruses and excess water, and therefore, are often intended to be returned directly to the human body as quickly as possible.

[0011] When these concentrators are actually used to concentrate ascites, the concentrators are usually controlled by various control means provided by the equipment or devices. However, the inventors of the present invention have found that:

[0012] On the one hand, although the structure of the equipment or device itself, or the data of the main components, can provide some reference for the concentration capacity and effect, in actual operation, adjusting the concentrator to achieve satisfactory parameters may be difficult in some situations, and problems may also arise in terms of stable operation.

[0013] On the other hand, in order to obtain satisfactory results during the use of the concentrator, in addition to being regulated by various control means, the material of the concentrator itself (usually a porous membrane material with a certain pore size) has an important influence on the body cavity fluid concentration effect. However, the inventors of the present invention have also found that the selection of these materials currently relies solely on the parameters provided by the membrane material manufacturers, and the parameters given by the manufacturers of these membrane materials are usually only tested and marked based on the testing means (i.e., physical and chemical tests) in non-medical fields. Under normal circumstances, after the selection of the membrane material, it is assumed that these membrane materials themselves are qualified for processing human body cavity fluid. Therefore, the safety and reliability of the use of the concentrator are mainly improved by adjusting the concentrator working state / parameters as described above, or adjusting the concentrator structure.

[0014] Furthermore, concentrators face a variety of complexities during actual use. In some cases, even qualified concentrators may still struggle to achieve reliable concentration results, potentially leading to increased concentration time or excessive loss of nutrients. In other cases, during ascites treatment, the body fluids treated by concentrators are often those filtered by upstream filters. These fluids vary not only due to different formation mechanisms, such as those caused by cirrhosis or cancer, but also due to differences in composition between upstream filters and their filtration conditions. Therefore, even if a concentrator or batch of concentrators achieves satisfactory treatment results for one specific patient or when used in combination with a specific upstream filter, it may pose risks when used with another specific patient or in combination with another specific upstream filter, posing a challenge to concentrator suitability. Consequently, testing methods for evaluating concentrator suitability are urgently needed.

[0015] Because the body cavity fluid treatment equipment containing the concentrator is a closed, one-to-one device, if the concentrator experiences an abnormality in its treatment of body cavity fluid (such as an abnormal decrease in concentrating capacity due to a blockage or other reasons), it is generally not possible to simply repair or replace the device. Due to the lack of a viable solution, the entire equipment may need to be replaced, and the economic costs and actual losses of such a solution are often unbearable. Similarly, due to the lack of a viable solution, such problems are often attributed to unavoidable systemic risks.

[0016] In response to the aforementioned issues identified by the present invention, the inventors believe that the previous reliance on isolated empirical evidence regarding material parameters, control methods, and equipment structure to determine concentrator performance may have contributed to the aforementioned systemic risks. Further research has revealed that such systemic risks are not unavoidable. Therefore, the present invention aims to provide a testing tool (test solution) for evaluating concentrators to accurately reflect their concentrating performance.

[0017] Solutions for solving problems

[0018] According to long-term research by the inventors of the present invention, it is found that the above technical problems can be solved by implementing the following technical solutions:

[0019] The present invention provides a test solution for evaluating the protein recovery performance of a body cavity fluid concentrator, wherein the total protein concentration in the test solution is 0.5 to 2.4 g / dL, the albumin to globulin ratio (A / G ratio) is 0.8 to 1.5, and the blood cell count is 1×10 2 less than 1 μg / μL.

[0020] In some specific embodiments, the test fluid further contains an anticoagulant.

[0021] In some specific embodiments, the anticoagulant comprises at least one substance selected from heparin and its salts, ethylenediaminetetraacetic acid salts, citrates, oxalates, and hirudin.

[0022] In some specific embodiments, the anticoagulant comprises more than 2 units / mL of heparin and / or its salt.

[0023] In some specific embodiments, the total protein concentration of the test fluid is 0.5-0.9 g / dL.

[0024] In some specific embodiments, the total protein concentration of the test fluid is 2.0-2.4 g / dL.

[0025] In some specific embodiments, the test fluid is used to evaluate the protein recovery performance of a body cavity fluid concentrator.

[0026] The present invention also provides a method for producing a test fluid for evaluating the protein recovery performance of a body cavity fluid concentrator, which is characterized by comprising the following steps:

[0027] a filtration step of filtering the raw material liquid using a filter membrane having an average pore size of less than 0.2 μm;

[0028] The concentration adjustment step is to adjust the total protein and / or albumin concentration in the filtered filtrate.

[0029] In some specific embodiments, the filter membrane has a pore size distribution in which the ratio of the number of pores with a size of 0.08 to 0.12 μm relative to the total number of pores is 60% or more.

[0030] In some specific embodiments, the feedstock solution comprises non-human animal plasma.

[0031] In some specific embodiments, the non-human animal plasma is bovine plasma.

[0032] In some specific embodiments, in the concentration adjustment step, the total protein concentration is adjusted to 0.5-2.4 g / dL.

[0033] In some specific embodiments, in the concentration adjustment step, the total protein concentration is adjusted to 0.5-0.9 g / dL.

[0034] In some specific embodiments, in the concentration adjustment step, the total protein concentration is adjusted to 2.0-2.4 g / dL.

[0035] In some specific embodiments, in the concentration adjustment step, the ratio of albumin to globulin (A / G ratio) is adjusted to 0.8 to 1.5.

[0036] In some specific embodiments, the manufacturing method further comprises:

[0037] freeze-thaw process, and

[0038] Fibrin removal process.

[0039] In some specific embodiments, the fibrin removal process comprises filtering through gauze to remove the fibrin after the freeze-thaw process.

[0040] In some specific embodiments, the freeze-thawing step and the fibrin removal step are performed before the filtration step, or after the filtration step and before the concentration adjustment step.

[0041] The present invention also provides a kit for evaluating the protein recovery performance of a body cavity fluid concentrator, characterized in that the kit includes a containing portion and a test liquid placed in the containing portion, and the test liquid includes the above-mentioned test liquid of the present invention or the test liquid obtained according to the above-mentioned manufacturing method of the present invention.

[0042] In some specific embodiments, the kit is used to evaluate the protein recovery performance of a body cavity fluid concentrator.

[0043] Effects of the Invention

[0044] The present invention provides a test fluid for evaluating the protein recovery performance of a body cavity fluid concentrator, a method for manufacturing the test fluid, a kit containing the test fluid or the test fluid manufactured by the manufacturing method, and applications of the test fluid and the kit. Implementation of the above technical solution can achieve the following technical effects:

[0045] (1) By using the test liquid provided by the present invention, the protein recovery performance of the body cavity fluid concentrator can be evaluated in a holistic and systematic manner. The test liquid and its manufacturing method have not been reported in the literature disclosed in the field, and provide a reliable evaluation reagent for the concentrators used in the field.

[0046] (2) The test fluid and the like provided by the present invention can truly reflect the treatment effect and treatment capacity of the concentrator when concentrating human body cavity fluid, and thus can accurately evaluate whether the concentrator of that type or batch can reliably and safely provide medical services to patients, and can avoid deviations or accidents that may occur during the use of the concentrator as much as possible, thereby reducing the medical burden and medical risks of patients.

[0047] (3) The test solution etc. provided by the present invention has a wide range of applicability, that is, it is applicable not only to the evaluation of concentrators for ascites caused by liver cirrhosis but also to the evaluation of concentrators for ascites caused by cancer.

[0048] (4) By using the test solution provided by the present invention, a convenient and operable evaluation method can be implemented for the concentrator, thereby enabling rapid detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 : A schematic diagram of the structure of a body cavity fluid concentrator as an evaluation object of the test fluid of the present invention

[0050] Figure 2 Schematic diagram of the apparatus used in the test for evaluating the protein recovery of a body cavity fluid concentrator using a test fluid according to a specific embodiment of the present invention

[0051] Figure 3 : Transmembrane pressure (TMP) data of each concentrator in the embodiment of the present invention and the reference example during operation

[0052] Description of Reference Numerals

[0053] 1: Concentrator body 2: Concentrate outlet (1st output port)

[0054] 3: Concentrator input port 4: Filtrate outlet (second output port)

[0055] 5: Test liquid storage 6: Concentrate storage 7: Filtrate storage

[0056] a / b / c: pipeline DETAILED DESCRIPTION

[0057] The following is a detailed description of the present invention. The following description of the technical features is based on representative embodiments and specific examples of the present invention, but the present invention is not limited to these embodiments and specific examples. It should be noted that:

[0058] In this specification, a numerical range expressed using "a numerical value A - a numerical value B" means a range including the endpoints A and B.

[0059] In this specification, "%" refers to weight or mass percentage unless otherwise stated.

[0060] In this specification, the use of "may" includes both the meaning of performing a certain process and the meaning of not performing a certain process.

[0061] In this specification, the use of "optional" or "optional" indicates that certain substances, components, execution steps, application conditions and other factors are used or not used.

[0062] In this manual, the unit names used are all international standard unit names.

[0063] In this specification, unless otherwise stated, "a plurality of" means two or more.

[0064] In this specification, references to "some specific / preferred embodiments," "other specific / preferred embodiments," "embodiments," etc., mean that the specific elements (e.g., features, structures, properties, and / or characteristics) described in connection with the embodiments are included in at least one embodiment described herein, and may or may not be present in other embodiments. In addition, it should be understood that the elements may be combined in various embodiments in any suitable manner.

[0065] The present invention provides a test solution for evaluating the protein recovery performance of a body cavity fluid concentrator, wherein the total protein concentration in the test solution is 0.5 to 2.4 g / dL, the albumin to globulin ratio (A / G ratio) is 0.8 to 1.5, and the blood cell count is 1×10 2 less than 1 μg / μL.

[0066] <Body cavity fluid concentrator>

[0067] The body cavity fluid concentrator (sometimes simply referred to as a concentrator) applicable to the present invention can be any concentrator in the art for concentrating body cavity fluid.

[0068] Figure 1 The typical structure of the concentrator of the present invention is shown in FIG. 1 , wherein 1 represents the concentrator body and 3 represents the inlet of the concentrator. When the patient is actually treated, the filter ( Figure 1 The body cavity fluid (not shown) treated by the concentrator enters the concentrator body 1 from the concentrator input port 3. In contrast, when the test fluid of the present invention is used to evaluate the performance of the concentrator, the test fluid enters the concentrator body 1 from the input port 3. Figure 1 Reference numeral 2 indicates the concentrated liquid outlet (first outlet) of the concentrator. In actual use, the concentrated liquid processed by the concentrator is discharged from the concentrated liquid outlet 2. Figure 1 Reference numeral 4 indicates the filtrate outlet (second output port), which is used to discharge the filtrate generated by the concentrator during the concentration process.

[0069] In addition, in some other specific embodiments, the concentrator of the present invention further has a discharge port for residual liquid ( Figure 1 (not shown) for discharging the body cavity fluid remaining in the concentrator body 1.

[0070] The concentrator body 1 is provided with a concentrating unit. In some specific embodiments of the present invention, the concentrating unit is selected from a porous membrane. There is no particular limitation on the porous membrane, and a common ultrafiltration membrane in the art can be used. Generally speaking, the pore size of the porous membrane used in the concentrator is smaller than the membrane used in the filter upstream thereof. The porous membrane is mainly used to concentrate proteins in low-concentration protein solutions and separate an aqueous filtrate from which the protein has been substantially removed on the other side of the membrane.

[0071] In some preferred embodiments of the present invention, the ratio of the number of pores having a diameter of 0.08 μm or more and 0.12 μm or less relative to the total number of pores in the porous membrane suitable for the present invention is 60% or more, preferably 70% or more. In other preferred embodiments, the ultrafiltration performance of the concentrator of the present invention is 85 mL / min / 200 mmHg to 150 mL / min / 200 mmHg. When the ultrafiltration performance is below this range, the discharge of the filtrate will decrease during the concentration process, and a fully concentrated protein solution cannot be obtained. In addition, when the ultrafiltration performance is above 85 mL / min / 200 mmHg, the possibility of clogging is lower, and therefore it is preferably, more preferably, above 95 mL / min / 200 mmHg. When the ultrafiltration performance is above 150 mL / min / 200 mmHg, protein sometimes leaks into the filtrate, and a sufficient protein concentration may not be obtained. The above-mentioned preferred porous membranes are widely used due to their excellent performance. When the test solution of the present invention is used to evaluate the performance of a concentrator equipped with such a porous membrane, the effects of the present invention can be further exerted.

[0072] There is no particular limitation on the type of porous membrane. In some preferred embodiments, a fiber membrane with a hollow structure can be used from the perspective of concentration efficiency. The hollow fiber membrane mentioned here is not particularly limited in shape and size, as long as it has the above-mentioned ultrafiltration performance. Regarding the material, polysulfone raw materials are preferably used because the pore size is easy to control during membrane formation and the chemical stability is excellent. Alternatively, it can be sulfones or celluloses, which also include polyethersulfone. Polysulfone polymers are aromatic compounds, so they have particularly excellent radiation resistance, and are also very heat-resistant and chemically resistant, and have excellent safety. Therefore, various membrane forming conditions can be selected, and radiation sterilization is possible, making them particularly preferred as membrane materials for concentrators. In addition, "~ type" not only refers to homopolymers, but also includes copolymers composed of other monomers and chemically modified derivatives.

[0073] The polysulfone polymers (hereinafter sometimes referred to as PSf) mentioned here are a general term for polymer compounds having a sulfone bond, and there is no particular limitation. For example, polysulfone polymers whose repeating units are represented by the following formula (1), formula (2), formula (3), formula (4) and formula (5) can be listed. It can also be a modified polymer in which a substituent is introduced into a part of the above aromatic ring. Based on the viewpoint of easy industrial availability, aromatic polysulfone polymers whose repeating units are represented by formula (1), formula (2) and formula (3) are preferred, among which polysulfones having a chemical structure represented by formula (1) are particularly preferred. The bisphenol-type polysulfone resin is sold, for example, by Solvay Advanced Polymers LLC under the trade name of "Udel (registered trademark)". There are many types of bisphenol-type polysulfone resins according to the degree of polymerization, but they are not particularly limited.

[0074]

[0075] The polysulfone hollow fiber membrane in the present invention preferably has hydrophilicity by utilizing a hydrophilic polymer. This is because, with only polysulfone polymers, the surface of the hollow fiber membrane is hydrophobic, and proteins are easily adsorbed on this surface, which becomes the reason for reducing the recovery performance of the protein. As hydrophilic polymers, polyvinyl pyrrolidone (hereinafter sometimes referred to as PVP), polyethylene glycol, polyvinyl alcohol, polypropylene glycol, etc. can be used. Among them, PVP is preferred from the perspective of hydrophilization effect and safety. Regarding PVP, there are several types depending on the molecular weight, for example, as commercially available products, there are PVP K-15, K-30, K-90 (all produced by International Specialty Chemicals (ISP) Company), etc. The molecular weight (viscosity average molecular weight) of PVP used in the present invention is 10,000 to 2 million, preferably 50,000 to 1.5 million. The content of hydrophilic polymer in the membrane is 3% to 20% of the total weight of the polymer, preferably 3% to 10%. When the content is 3% or less, the effect as a hydrophilizing agent is weakened, and when the content exceeds 20%, the viscosity of the membrane-forming stock solution becomes too high, which is not preferable in terms of production.

[0076] The method for producing hydrophilized polysulfone hollow fiber membranes can utilize known dry and wet membrane-forming techniques. First, a polysulfone polymer and a hydrophilic polymer such as polyvinyl pyrrolidone are dissolved in a common solvent for both to prepare a uniform spinning dope. When the hydrophilic polymer is polyvinyl pyrrolidone, the common solvent can be, for example, dimethylacetamide (hereinafter referred to as DMAC), dimethyl sulfoxide, N-methyl-2-pyrrolidone, dimethylformamide, sulfolane, dioxane, or a mixture of two or more of the above solvents. Furthermore, additives such as water can be added to the spinning dope to control pore size.

[0077] When making a hollow fiber membrane, a spinneret with a tube in orifice is used, and the spinning dope from the hole of the spinneret and the hollow inner liquid from the tube are ejected into the air at the same time. The hollow inner liquid is a substance used to coagulate the spinning dope, and water or a coagulation liquid with water as the main component can be used. The composition of the hollow inner liquid can be determined based on the target ultrafiltration performance of the hollow fiber membrane. Although it cannot be generalized, a mixed solution of the solvent used for the spinning dope and water can usually be appropriately used. For example, a DMAC aqueous solution of 0% by weight to 65% by weight can be used as the hollow inner liquid. The spinning dope ejected together with the hollow inner liquid from the spinneret travels in the air walk portion, is introduced into a coagulation bath with water as the main component arranged at the bottom of the spinneret, and is immersed in it to complete coagulation. After a cleaning step of the solidified hollow fibers, the wet hollow fiber membranes are wound using a winder to obtain a hollow fiber membrane bundle, which is then dried. Alternatively, a hollow fiber membrane bundle can be obtained by washing the membranes and then drying them in a dryer. The production method is not limited.

[0078] Since the concentrator needs to rely on a certain pressure when working, for the convenience of explanation, the side of the porous membrane with higher pressure in the concentrator is called the primary side of the porous membrane, and the side of the porous membrane with lower pressure is called the secondary side of the porous membrane.

[0079] In this process, the protein component is concentrated on the primary side, and the filtrate (water-containing component) is separated on the secondary side. Figure 1 The concentrated liquid outlet 2 of the intermediate concentrator is communicated with the space where the primary side of the porous membrane is located, and the filtrate outlet 4 is communicated with the space where the secondary side of the porous membrane is located.

[0080] <Test fluid>

[0081] The present invention provides a test fluid to test the concentrating and separation capabilities of a concentrator. Theoretically, to fully reflect the actual concentrating and separation capabilities of the concentrator under test, using patient body cavity fluid for evaluation and testing is the most reliable method. However, such a theory is rarely practical.

[0082] On the one hand, it's nearly impossible to obtain patient body fluid in advance, and even more difficult to obtain filtered body fluid in a standardized manner. On the other hand, using body fluid from a specific patient to evaluate a concentrator only yields results specific to that specific patient at a specific time and without universal applicability. Furthermore, since concentrators themselves are typically expensive and rarely reused after testing, this approach would cost a patient at least double their medical expenses. Furthermore, once a patient's body fluid is used for testing, it cannot be recovered, making it unacceptable from a medical ethical perspective. Therefore, using patient body fluid to evaluate the concentrator's performance clearly lacks practical value.

[0083] Therefore, it is important to provide a reliable test solution for evaluating concentrators. The present inventors believe that, since the most important function of a concentrator is to concentrate and recover protein-containing bodily fluids, it is feasible to provide a test solution containing a certain concentration of protein to evaluate the concentrator's concentrating performance. Furthermore, due to the varying formation of body cavity fluids, the composition of body cavity fluids varies. Consequently, during actual processing, the composition of the body cavity fluids extracted from upstream filters encountered by concentrators cannot be generalized. Therefore, selecting a test solution of appropriate concentration is also crucial.

[0084] The test fluid for evaluating the protein recovery performance of the body cavity fluid concentrator of the present invention has a total protein concentration of 0.5 to 2.4 g / dL, an albumin to globulin ratio (A / G ratio) of 0.8 to 1.5, and a blood cell count of 1×10 2 less than 1 μg / μL.

[0085] In the present invention, the number of blood cells in the test solution exceeds 1×10 2 When the number of blood cells in the test solution is 1×10 / μL, the coagulation cascade is likely to occur (thrombus-like masses are likely to form), and the concentration performance cannot be tested with good reproducibility. From this point of view, the present invention controls the number of blood cells in the test solution to 1×10 2 The number of blood cells is preferably 50 or less, more preferably 10 or less, and even more preferably substantially free of blood cells. From the perspective of process cost, the number of blood cells may be, for example, 5 or more.

[0086] In the present invention, in order to improve the universal applicability and reliability of the test liquid, a solution with a protein content of 0.5 to 2.4 g / dL, especially an aqueous solution, is used as the test liquid. If the protein concentration is too low, the concentration effect cannot be evaluated (when the protein is too dilute, the concentrator actually concentrates the water, and the results of the evaluation test cannot reflect the actual effect of the concentrator in treating human body cavity fluid). At the same time, by controlling the protein content below 2.4 g / dL, the TMP (transmembrane pressure difference experienced by the porous membrane in the concentrator) will not rise during the evaluation test time (30 minutes to 1 hour, the same as the treatment time usually used for patients), thereby ensuring good reproducibility of the evaluation test.

[0087] The inventors discovered that by setting the protein content in the test fluid within the above range, the evaluation results more closely match the actual performance of the concentrator. Furthermore, adjusting the protein content within this range significantly improves the universality of the test fluid of the present invention, enabling accurate evaluation of the actual performance of the concentrator for concentrating body fluids from liver cirrhosis as well as for concentrating body fluids from various cancers.

[0088] Furthermore, in some preferred embodiments of the present invention, the protein content of the test fluid is controlled within the range of 0.5 to 0.9 g / dL. Such a test fluid provides greater reliability, particularly when evaluating the actual performance of a concentrator for human body cavity fluid resulting from liver cirrhosis.

[0089] In other preferred embodiments of the present invention, the protein content of the test fluid is controlled within a range of 2.0 g to 2.4 g / dL. Such a test fluid provides greater reliability, particularly when evaluating the actual performance of a concentrator for concentrating body cavity fluids derived from various cancers.

[0090] In addition, the present invention can use proteins including albumin and proteins other than albumin. Preferably, the protein other than albumin can be globulin. In addition, to reduce clogging of the concentrator by components in the test solution, preferably, a small amount of or no fibrin is used among the proteins.

[0091] In some specific embodiments of the present invention, the albumin to globulin ratio (A / G ratio) in the test solution is 0.8 to 1.5. The present invention believes that the test solution formed by controlling the albumin to globulin ratio within the above range will provide evaluation results that better reflect the actual performance of the concentrator.

[0092] Furthermore, in some preferred embodiments of the present invention, the albumin to globulin ratio (A / G ratio) of the test fluid is 0.8 to 1.2. Such a test fluid has better reliability, particularly when evaluating the actual performance of a concentrator for human body cavity fluid resulting from liver cirrhosis.

[0093] Furthermore, in some preferred embodiments of the present invention, the albumin to globulin ratio (A / G ratio) of the test fluid is 0.9 to 1.5. Such a test fluid has better reliability when evaluating the actual performance of a concentrator for human body cavity fluids generated by various cancers.

[0094] Furthermore, in actual use, the test fluid is not only expected to exhibit excellent reliability during evaluation and testing, but also to possess uniform and stable composition. This facilitates both practical use and storage of the test fluid. Therefore, in some preferred embodiments of the present invention, a certain amount of anticoagulant is used in the test fluid to improve its stability while maintaining reliability during use.

[0095] There are no particular requirements for the anticoagulant that can be used in the test solution of the present invention, and the anticoagulant can be selected from at least one of heparin and its salts, ethylenediaminetetraacetic acid salts, citrates, oxalates, and hirudin, or a mixture of multiple thereof.

[0096] In some preferred embodiments, the anticoagulant includes at least heparin or a salt thereof. From the perspective of effectively improving the stability of the test fluid, the anticoagulant includes more than 2 units / mL, preferably more than 3 units / mL, and further preferably more than 5 units / mL of heparin and / or a salt thereof.

[0097] The test solution for evaluation of the present invention may contain, in addition to the aforementioned protein and optional anticoagulant, other auxiliary components as needed, as long as the presence of these components does not affect the evaluation effect of the test solution. These auxiliary components may include inorganic salts and pH buffering components.

[0098] <Method for producing test solution>

[0099] The method for producing the test solution of the present invention comprises the following steps:

[0100] a filtration step of filtering the raw material liquid using a filter membrane having an average pore size of less than 0.2 μm;

[0101] The concentration adjustment step is to adjust the total protein and / or albumin concentration in the filtered filtrate.

[0102] In some preferred embodiments of the present invention, the test fluid can be prepared using non-human animal plasma. Because animal plasma also contains proteins and is biologically similar to human body cavity fluid, using animal plasma as the raw material can simplify the test fluid preparation process and improve the test fluid's effectiveness.

[0103] The animal plasma is not particularly limited, and blood from (relatively larger) mammals, such as cattle, sheep, pigs, horses, and deer, can be used. For ease of collection, cattle or sheep plasma is preferred. Furthermore, the plasma can be fresh or refrigerated; for ease of handling, fresh animal plasma is preferred. Optionally, an anticoagulant, as described above, particularly heparin, can be added to the plasma.

[0104] In the filtration process, the filtration membrane is not particularly limited. In some specific embodiments of the present invention, a common porous membrane in the art can be used, typically a hollow fiber membrane.

[0105] In some specific embodiments of the present invention, the filtration membrane can be used in the form of a filter comprising a hollow fiber membrane bundle within a cylindrical container. In the cylindrical container, a raw liquid is passed from the outside of the hollow fiber membranes of the hollow fiber membrane bundle to the inside of the hollow fiber membranes, thereby removing specific substances from the body cavity fluid. The hollow fiber membranes are dispersed so that the filling rate of the hollow fiber membrane bundle is greater than 20% and less than 41% of the internal cross-section of the cylindrical container, and the average distance between the hollow fiber membranes in the hollow fiber membrane bundle is greater than 150 μm.

[0106] In some preferred embodiments, the maximum distance between hollow fiber membranes in the hollow fiber membrane bundle is greater than 300 μm; the effective membrane area of the hollow fiber membrane bundle is 0.7 m 2 Above 3.0m 2 below; the inner diameter of the hollow fiber membrane is 50 μm or more and 500 μm or less.

[0107] In some preferred embodiments, from the perspective of balancing filtration performance, the filtration membrane has a pore size distribution in which the ratio of the number of pores larger than 0.08 μm and smaller than 0.12 μm to the total number of pores is greater than 60%, preferably greater than 70%, and more preferably greater than 80%.

[0108] In some other specific embodiments of the present invention, prior to the filtration step, filtration can be performed using a coarse-pore filter membrane, such as gauze, non-woven fabric, or filter paper (hereinafter referred to as coarse filtration). These coarse-pore filter membranes typically have an average pore size of, for example, 10 μm, 100 μm, or 1 mm, and can be used as a single layer or in a stacked manner.

[0109] For ease of filtration and cost considerations, the present invention preferably uses a coarse-pore filter membrane, such as the gauze, for the coarse filtration step. This coarse filtration step removes unwanted components, primarily tissue fragments, fibrin, and some blood cells, thereby improving the efficiency of the filtration step. To ensure effective filtration, multiple filtration steps may be performed.

[0110] In some preferred embodiments, to prevent the degradation of test results due to components such as fibrin, the raw material solution may be subjected to a freeze-thaw process before the filtration process, and the fibrin may then be removed by, for example, the coarse filtration process described above. Of course, such a freeze-thaw process and fibrin removal process may also be performed after the filtration process and before the concentration adjustment process.

[0111] The filtrate obtained after the filtration step is further subjected to a concentration adjustment step so that it finally meets the desired composition, preferably the composition of the test solution of the present invention.

[0112] The concentration adjustment step includes adjusting the protein concentration. Typically, the protein concentration can be adjusted to the range defined by the test solution of the present invention by adding protein.

[0113] Furthermore, the test solution obtained according to the above steps is placed in a container for use. There is no particular limitation on the storage conditions of the test solution, and typically, the test solution can be stored in a refrigerator.

[0114] For ease of use, in a preferred embodiment of the present invention, the test solution obtained by the above process can be stored in a test kit. The test kit includes a container and the test solution placed in the container. The test kit can have a flexible or rigid housing, and in some specific embodiments, has at least one outlet or interface that can communicate with the outside world. Such an outlet or interface is sealed before use. In other specific embodiments, the test kit can be evacuated, that is, the interior of the test kit is evacuated before use.

[0115] <Evaluation Method>

[0116] In the present invention, the concentrator was evaluated using the above-mentioned test solution.

[0117] In some embodiments, evaluation of the concentrator is performed using at least a concentrating step and a calculating step.

[0118] Concentration step

[0119] In the present invention, the protein-containing test solution is sent to a concentrator through the concentration step and separated into a concentrate and a filtrate.

[0120] See attached Figure 2 The apparatus shown in FIG. 1 shows an apparatus for performing evaluation tests in accordance with a specific embodiment of the present invention, wherein reference numeral 3 represents a concentrator input port, reference numeral 5 represents a container for storing a test liquid (a test liquid reservoir), reference numeral 6 represents a container for storing a concentrated liquid obtained by treatment with the concentrator (a concentrated liquid reservoir), and reference numeral 7 represents a container for storing a filtrate discharged from the concentrator (a filtrate reservoir). In this case, the concentration step involves introducing the test liquid into the concentrator via a pipeline through the concentrator input port 3 and separating the test liquid into a concentrate and a filtrate via a porous membrane.

[0121] In addition, in some other specific embodiments, the system used for evaluation testing includes Figure 2 In addition to the device or structure shown, it may optionally include: a recycling device that recirculates the concentrated liquid in the concentrated liquid storage 6 to the test liquid storage; a control device that stops the filtration action when the amount of concentrated body cavity fluid stored in the concentrated liquid storage reaches a first specified amount, and uses the recycling device to recirculate the concentrated liquid stored in the concentrated liquid storage 6 to the test liquid storage 5, and then enters the concentrator again and is concentrated.

[0122] In addition, the transfer of materials between the various devices of the present invention can be carried out by a power device. The type of the power device is not particularly limited and can be a transfer pump.

[0123] The evaluation test of the present invention can be carried out at 0-50°C, preferably at 15-35°C. If the temperature is too low, the fluidity of the test liquid will deteriorate, and it will also be detrimental to the concentration process of the concentrator; if the temperature is too high, there is a concern that the protein components may be denatured.

[0124] When conducting the evaluation test of the present invention, the test liquid is transported from pipeline a to the concentrator body 1, and the test liquid is concentrated and the filtrate (mainly water-containing components) is separated by pressurizing the primary side of the porous membrane in the concentrator. The concentrated liquid further enters the concentrated liquid storage 6 through pipeline b, and the filtrate is transported to the filtrate storage 7 through pipeline c.

[0125] In addition, the concentrated liquid in the concentrated liquid storage 6 can be returned to the test liquid storage 5 through a recycling device, thereby performing cyclic concentration and separation.

[0126] The concentrate in the concentrate storage 6 and / or the filtrate in the filtrate storage 7 after the concentration process is tested for components to evaluate the performance of the concentrator.

[0127] Calculation steps

[0128] In the present invention, the components in the test fluid, the concentrated fluid and / or the filtrate are measured through the calculation step, and the protein recovery performance of the body cavity fluid concentrator is calculated.

[0129] In some specific embodiments, the protein recovery performance of a concentrator is evaluated based on the albumin recovery rate of the concentrator.

[0130] An automatic detector (BioMajesty, manufactured by JEOL Ltd.) was used. TM The amount of albumin in the concentrate was detected by a JCA-BM6050, and the albumin recovery rate was calculated according to the following formula (1) based on the amount of albumin originally in the test solution:

[0131] Formula (1) Albumin recovery rate (%) = (amount of albumin in the concentrate / amount of albumin in the test solution)×100.

[0132] In some other specific embodiments, the albumin recovery rate can also be calculated as follows:

[0133] Formula (2) Albumin recovery rate (%) = {(amount of concentrate×albumin concentration in concentrate) / (amount of test solution×albumin concentration in test solution)}×100.

[0134] Furthermore, in some other specific embodiments, the evaluation method of the concentrator also takes into account the residual liquid remaining in the concentrator, especially when the volume of the concentrator is large or the residual liquid is more obvious.

[0135] In this case, the evaluation test of the present invention further includes a residual liquid recovery step to recover the residual test liquid remaining in the body cavity fluid concentrator after the concentration step.

[0136] Therefore, in some specific embodiments, the protein recovery performance is an albumin recovery rate. In the calculation step, the albumin in the test fluid, the concentrated fluid, and the residual test fluid is measured to calculate the protein recovery performance of the body cavity fluid concentrator.

[0137] In this case, the albumin recovery rate is calculated according to formula (3):

[0138] Formula (3) Albumin recovery rate (%) = {(amount of albumin in the concentrate + amount of albumin in the residual test solution) / amount of albumin in the test solution} × 100.

[0139] Furthermore, the albumin recovery rate can also be calculated according to formula (4):

[0140] Formula (4) Albumin recovery rate (%) = {(concentrate volume × albumin concentration in concentrate + residual test solution volume × albumin concentration in residual test solution) / (test solution volume × albumin concentration in test solution)} × 100.

[0141] In other specific embodiments of the present invention, the filtrate is recovered after the concentration step. Proteins that have passed through the porous membrane may also be present in the filtrate. Therefore, in this case, the protein recovery performance can also be represented by the albumin permeability. Specifically, in the calculation step, the albumin content of the test solution and the filtrate is measured to calculate the albumin permeability.

[0142] In some specific embodiments, the albumin permeability can be calculated as follows:

[0143] Formula (5) Albumin permeability (%) = (amount of albumin in the filtrate / amount of albumin in the test solution) × 100.

[0144] In some specific embodiments, the albumin permeability can also be calculated as shown in formula (6):

[0145] Formula (6) Albumin permeability (%) = {(filtrate volume×albumin concentration in the filtrate) / (test solution volume×albumin concentration in the test solution)}×100.

[0146] In addition to using albumin concentration to evaluate the actual performance of the concentrator, the recovery rate of α1-MG (α1 microglobulin) can also be used for evaluation.

[0147] In the calculation step, the α1-MG in the test solution and the concentrated solution is measured to calculate the α1-MG recovery rate of the concentrator.

[0148] In some specific embodiments, the α1-MG recovery rate is calculated as follows:

[0149] Formula (7) ···α1-MG recovery rate (%) = (α1-MG amount in the concentrate / α1-MG amount in the test solution)×100.

[0150] In some specific embodiments, the α1-MG recovery rate can also be calculated as shown in formula (8):

[0151] Formula (8) ···α1-MG recovery rate (%) = {(amount of concentrated solution×amount of α1-MG in the concentrated solution) / (amount of test solution×amount of α1-MG in the test solution)}×100.

[0152] Similarly, when the residual liquid is taken into account or cannot be ignored, in the calculation step, the α1-MG in the test liquid, the concentrated liquid, and the residual test liquid is measured to calculate the α1-MG recovery rate of the concentrator.

[0153] In some specific embodiments, the α1-MG recovery rate is calculated as follows:

[0154] Formula (9) ···α1-MG recovery rate (%) = {(α1-MG amount in the concentrate + α1-MG amount in the residual test solution) / α1-MG amount in the test solution}×100.

[0155] In some specific embodiments, the α1-MG recovery rate is calculated as follows:

[0156] Formula (10)···α1-MG recovery rate (%) = {(concentrate volume × α1-MG concentration in concentrate + residual test solution volume × α1-MG concentration in residual test solution) / (test solution volume × α1-MG concentration in test solution)}×100.

[0157] Similarly, the protein recovery performance may be evaluated by the permeability of α1-MG. In this case, in the calculation step, the α1-MG in the test solution and the filtrate is measured to calculate the α1-MG permeability.

[0158] In some specific embodiments, the α1-MG transmittance is calculated as follows:

[0159] Formula (11) ···α1-MG transmittance (%) = (α1-MG amount in filtrate / α1-MG amount in test solution)×100.

[0160] In some specific embodiments, the α1-MG transmittance is calculated as follows:

[0161] Formula (12)···α1-MG permeability (%)={(filtrate volume×α1-MG concentration in the filtrate) / (test solution volume×α1-MG concentration in the test solution)}×100.

[0162] Data processing

[0163] In the present invention, the actual performance of the concentrator is evaluated by the recovery rate of albumin, the permeability of albumin, the recovery rate of α1-MG, or the permeability of α1-MG as described above.

[0164] In some specific embodiments, data calculated according to any one of the above formulas (1) to (12) can be used as an evaluation criterion. In addition, optionally, these data can also be weighted according to existing mathematical methods to obtain a more comprehensive evaluation index. The present invention has no particular limitation on the specific mathematical processing method.

[0165] Example

[0166] Hereinafter, the present invention will be described through specific examples.

[0167] Example

[0168] Bovine plasma was supplemented with heparin, frozen and thawed, and then coarsely filtered through gauze. The plasma was then filtered through a hollow fiber membrane with an average pore size of 0.2 μm or less. The total protein (hereinafter referred to as "TP") concentration was adjusted to 2 g / dL and an albumin / globulin ratio ("A / G ratio") of 1.2. The resulting liquid was used as a test solution. The heparin concentration in this test solution was 3.2 units / mL, and the blood cell count was 0 / μL.

[0169] Place the test fluid in the test fluid reservoir ( Figure 2 5) and adjust the temperature to 27±1℃, and add the test solution to the concentrator input port ( Figure 2 3) is supplied at a flow rate of 50 mL / min, and at the same time, the first output port of the concentrator ( Figure 2 2) flow rate is 5mL / min (collected in the bag), and the concentrator second output port ( Figure 2 4) Produce filtrate (collected in a bag) at a flow rate of 45 mL / min and deliver the solution for 60 minutes (the test liquid volume is 3 L).

[0170] After the concentration was stopped, the recovery rate of albumin was calculated using the test solution and the concentrated solution according to formula (1). At this time, the liquid remaining in the concentrator was 46 mL.

[0171] Albumin recovery rate (%) = (amount of albumin in the concentrate / amount of albumin in the test solution)×100.

[0172] The results of the analysis showed that the amount of albumin in the concentrate was 23.7 g, and the albumin recovery rate was 69.3%.

[0173] Furthermore, air was introduced from the concentrator outlet to recover the residual test solution remaining in the concentrator. The recovery rate of albumin was calculated according to formula (3) using the test solution, the concentrate, and the residual test solution.

[0174] Albumin recovery rate (%) = {(albumin amount in concentrate + albumin amount in residual test solution) / albumin amount in test solution} × 100

[0175] The test results showed that the amount of albumin in the concentrate and the residual test solution was 27.5 g, and the albumin recovery rate was 80.6%.

[0176] Three concentrators from the same batch were tested using this test solution (n=3). The results showed that the average albumin recovery rate calculated according to formula (3) using the test solution, concentrate, and residual test solution was 79.9%, and the relative standard deviation (RSD) was 1.61%.

[0177] Reference Example 1

[0178] The protein concentration in the test solution in the example was adjusted to 4 g / dL, and the other conditions were the same as those in the example to perform the concentration operation.

[0179] Reference Example 2

[0180] The protein concentration in the test solution in the example was adjusted to 7 g / dL, and the other conditions were the same as those in the example to perform the concentration operation.

[0181] <Evaluation>

[0182] The TMP (transmembrane pressure difference) of the embodiment and the two reference examples during the concentration process was detected. TP2 represents the change of TMP during the concentration of the embodiment, TP4 represents the change of TMP during the concentration of reference example 1, and TP7 represents the change of TMP during the concentration of reference example 2. The results are shown in FIG. Figure 3 .

[0183] Figure 3 The results show that the TMP in the examples was below 150 mmHg within the test period (the typical treatment time required for patients, i.e., 30 to 60 minutes). In contrast, in Reference Example 1, the TMP had risen to approximately 400 mmHg by 30 minutes, and in Reference Example 2, the TMP had risen to approximately 400 mmHg by 10 minutes.

[0184] The above observations indicate that the test fluids provided in the examples can yield highly reproducible test results and accurately conduct performance tests. They can simulate the concentration conditions experienced by a concentrator during normal use of human body cavity fluid, providing a basic understanding of the concentrator's actual performance during normal operation.

[0185] In Reference Examples 1 and 2, however, the TMP (transmembrane pressure) increased rapidly (within 30 minutes). This made it impossible to complete the test within the planned processing time. Furthermore, excessive TMP increases could lead to membrane rupture and vessel damage, making further testing impossible. This situation made it impossible to effectively predict the concentrator's performance during normal operation and did not provide reliable reproducibility.

[0186] It should be noted that, although the technical solutions of the present invention are described with specific examples, those skilled in the art will appreciate that the present disclosure should not be limited thereto.

[0187] While various embodiments of the present disclosure have been described above, the above descriptions are illustrative, non-exhaustive, and not intended to be limiting of the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is selected to best explain the principles of the embodiments, their practical applications, or improvements to existing technologies, or to enable others skilled in the art to understand the embodiments disclosed herein.

[0188] Industrial applicability

[0189] The test liquid and the manufacturing method thereof provided by the present invention can be used in industrial production to evaluate concentrators for human body cavity fluid.

Claims

1. A test solution for evaluating the protein recovery performance of a body cavity fluid concentrator, wherein: The total protein concentration in the test fluid was 0.5-2.4 g / dL, the albumin to globulin ratio (A / G ratio) was 0.8-1.5, and the blood cell count was 1×10 2 less than 1 μg / μL. The test solution according to claim 1 , further comprising an anticoagulant. 3 . The test solution according to claim 2 , wherein the anticoagulant comprises at least one substance selected from the group consisting of heparin and its salts, ethylenediaminetetraacetate, citrate, oxalate, and hirudin. The test solution according to claim 3 , wherein the anticoagulant comprises 2 units / mL or more of heparin and / or its salt.

5. The test solution according to any one of claims 1 to 4, wherein The total protein concentration in the test solution is 0.5-0.9 g / dL.

6. The test solution according to any one of claims 1 to 4, wherein The total protein concentration in the test solution is 2.0-2.4 g / dL.

7. Use of the test solution according to any one of claims 1 to 6 in a test for evaluating the protein recovery performance of a body cavity fluid concentrator.

8. The method for producing a test solution for evaluating protein recovery performance of a body cavity fluid concentrator according to any one of claims 1 to 6, wherein: The process includes the following steps: a filtration step of filtering the raw material liquid using a filter membrane having an average pore size of less than 0.2 μm; The concentration adjustment step is to adjust the total protein and / or albumin concentration in the filtered filtrate.

9. The manufacturing method according to claim 8, wherein: The filtration membrane has a pore size distribution in which the ratio of the number of pores having a size of 0.08 to 0.12 μm relative to the total number of pores is 60% or more.

10. The manufacturing method according to claim 8 or 9, wherein: The raw material liquid includes non-human animal plasma.

11. The manufacturing method according to claim 10, wherein: The non-human animal plasma is bovine plasma.

12. The manufacturing method according to claim 8 or 9, wherein: In the concentration adjustment step, the total protein concentration is adjusted to 0.5 to 2.4 g / dL.

13. The manufacturing method according to claim 8 or 9, wherein: In the concentration adjustment step, the total protein concentration is adjusted to 0.5 to 0.9 g / dL.

14. The manufacturing method according to claim 8 or 9, wherein: In the concentration adjustment step, the total protein concentration is adjusted to 2.0 to 2.4 g / dL.

15. The manufacturing method according to claim 8 or 9, wherein: In the concentration adjustment step, the ratio of albumin to globulin (A / G ratio) is adjusted to 0.8 to 1.

5.

16. The manufacturing method according to claim 8 or 9, further comprising: freeze-thaw process, and Fibrin removal process.

17. The manufacturing method according to claim 16, wherein: The fibrin removal process includes removing the fibrin by filtering with gauze after the freeze-thaw process.

18. The manufacturing method according to claim 16, wherein: The freeze-thawing step and the fibrin removal step are performed before the filtration step, or after the filtration step and before the concentration adjustment step.

19. A kit for evaluating the protein recovery performance of a body cavity fluid concentrator, characterized in that: The reagent kit includes a containing portion and a test liquid placed in the containing portion, wherein the test liquid includes the test liquid according to any one of claims 1 to 6.

20. Use of the kit according to claim 19 in a test for evaluating protein recovery performance of a body cavity fluid concentrator.

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