JP2006088046A - Concentration instrument and concentration method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a system and a method for concentrating an object to be concentrated while keeping a completely closed system. <P>SOLUTION: A concentration instrument includes at least one vessel which is used for housing the object to be concentrated containing a component to be concentrated and which has at least one of a liquid outlet and a liquid inlet, wherein at least a part of the vessel is composed of a non-porous hydrophilic film or membrane. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a concentration device such as a solution and a concentration method. More specifically, the present invention relates to the concentration of various liquids including protein solutions such as collagen, polysaccharide solutions such as hyaluronic acid, physiologically active substance solutions, polymer solutions, and plant cells / tissues, animal cells / tissues. The present invention relates to a concentration device and a concentration method that can be suitably used for concentration of a mixture of a solid and a liquid.

  Conventional methods for removing and concentrating water from fruit juices, protein solutions, etc., forcibly evaporating water by applying heat (heating evaporation method), and forcibly evaporating water under vacuum (vacuum) Evaporation method), after freezing, a method of sublimating ice under vacuum (lyophilization method), a membrane concentration method using a reverse osmosis membrane or an ultrafiltration membrane, and the like have been performed.

  Among these technologies, in the heat evaporation method, components are altered by heat, and because they are not closed systems, there is a high risk of contamination such as bacteria, viruses, and foreign substances. There are many problems, such as the fact that the energy consumption is large because the phase change of water due to heating is easy, and the concentration of trace components is particularly difficult.

  In addition, since the vacuum evaporation method is not a closed system, there is a high risk of contamination with bacteria, viruses, foreign substances, etc., and it is necessary to apply mechanical stimulation such as stirring to prevent bumping. Since phase change is used, there are problems such as large energy consumption.

  Furthermore, the freeze-drying method is not a closed system as in the above method, so there is a high risk of contamination with bacteria, viruses, foreign substances, etc., there is a risk that the components will be altered by freezing, and contact with air. Thus, there are problems such as high energy costs because the components are susceptible to oxidation reactions and are accompanied by phase changes such as freezing and sublimation.

  On the other hand, with respect to the reverse osmosis membrane method and ultrafiltration membrane method using a membrane, alteration of components due to heating, freezing, etc. is suppressed, and energy consumption due to phase conversion accompanying heating, decompression, freezing, sublimation, etc. is reduced. There is a big advantage that there are few. However, the membrane method has the following problems.

  That is, the reverse osmosis membrane method and the ultrafiltration membrane method are not completely occluded systems, and there is a risk of contamination with bacteria, viruses, foreign substances, and the like. In addition, since a pressurizing step is essential in the membrane method, the membrane cannot be used as it is, and it is necessary to make a module such as a hollow module or a tubular module, which is expensive.

  Furthermore, there are problems such as high sterilization operation, clogging cleaning operation, and high operating costs. In addition, the membrane method is originally used for the purpose of separating the components according to the size using the pores of the membrane, and the pore diameter of the reverse osmosis membrane is several angstroms to several tens of angstroms. In the case of a filtration membrane, it is several tens of μm to several μm. Therefore, since components smaller than the pore diameter pass through the membrane, they are not suitable as a concentration step for removing only moisture. In particular, the ultrafiltration method is used for the purpose of removing components having a specific size or less, and a reverse osmosis membrane is used for removing only water.

  However, even in the reverse osmosis membrane method, there is a problem that if a membrane having a high water removal capability is used, necessary components such as salt are permeated. As mentioned above, there are several important problems with conventional methods for removing and concentrating moisture. In particular, in the conventional method, it is difficult to concentrate in a completely closed system and there is a risk of contamination with bacteria, viruses, and foreign matters, and it is necessary to install a sterilization or sterilization step or a foreign matter removal step after the concentration step. Further, contact with air could not be completely blocked during the concentration process, and it was difficult to prevent denaturation at the solid-liquid interface, which is particularly problematic with proteins and the like, at the same time as being susceptible to oxidation reactions.

  An object of the present invention is to provide a concentrating device and a concentrating method which have solved the above-mentioned drawbacks of the prior art.

  Another object of the present invention is to eliminate the difficulty of concentrating while maintaining a completely closed system, which is a drawback of the prior art, and to concentrate the system while maintaining a substantially completely closed system. And to provide a method.

  Still another object of the present invention is to provide a concentrating device and a concentrating method capable of concentrating a target component while avoiding heating, freezing, and contact with air as much as possible, which are the causes of alteration of the component, which is a drawback of the prior art. It is to provide.

  Still another object of the present invention is to provide a concentration device and a concentration method capable of avoiding as much as possible external operations such as heat, pressure reduction, freezing, sublimation, pressurization, and stirring required for concentration, which are disadvantages of the prior art. It is in.

  As a result of diligent research, the present inventor is extremely effective in solving the above problems by using a film or membrane that does not transmit water but transmits water vapor (that is, has moisture permeability) in combination with a container having a predetermined shape. I found out.

  The concentration device of the present invention is based on the above knowledge, and more specifically, a concentration device including at least a container having at least one doorway and containing an object to be concentrated containing a component to be concentrated. And at least one part of this container is comprised from a nonporous hydrophilic film or film | membrane, It is characterized by the above-mentioned.

  According to the present invention, there is further provided a container for containing an object to be concentrated containing one or more outlets and containing a component to be concentrated, and a sealable gas impermeability disposed outside the container. There is provided a concentrating device comprising at least a film or a membrane having at least a part of the container made of a nonporous hydrophilic film or membrane.

According to the present invention, it further comprises at least a container for containing a substance to be concentrated containing one or more outlets and containing a component to be concentrated, and at least a part of the container is nonporous and hydrophilic. There is provided a concentration method comprising concentrating an object to be concentrated by contacting the object to be concentrated with the hydrophilic film or membrane using an apparatus for concentration composed of a film or a membrane.
When the concentration device of the present invention having the above-described configuration is used, the object to be concentrated is brought into contact with a nonporous hydrophilic film or membrane that forms part of the concentration device of the present invention. Moisture in the body is absorbed into the film or membrane. As a result, the object to be concentrated can be concentrated by being evaporated (scattered) from the surface of the film or membrane to the outside of the concentration device.

As described above, according to the present invention, since the external operation such as heating, freezing, and decompression is not essential as in the conventional method by using the concentration device of the present invention, it is possible to effectively prevent alteration of the concentrated components. it can.
Furthermore, in the concentration device of the present invention, since the contact of the concentrated component with air is suppressed, it is easy to suppress the denaturation of the concentrated component due to the oxidation reaction and the solid-liquid interface.
In addition, when the concentration device of the present invention is used, concentration is performed in a substantially closed system, so that contamination with bacteria, viruses, foreign substances, etc. can be easily prevented as compared with the conventional method.
Further, by using the concentrating device of the present invention, it is easy to control the concentration factor of the object to be concentrated, and at the same time, the concentrating device of the present invention itself can also serve as a container for the concentrated material.

  Hereinafter, the present invention will be described more specifically with reference to the drawings as necessary. In the following description, “parts” and “%” representing the quantity ratio are based on mass unless otherwise specified.

(Equipment for concentration)
The concentration device of the present invention includes at least a container for containing an object to be concentrated containing a component to be concentrated. This container has one or more inlets and outlets for allowing the concentrator and the like to enter and exit, and at least a part of the container is composed of a nonporous hydrophilic film or membrane. As long as the efficiency of concentration as a “concentration device” as described below is shown, it contacts the mass, volume or outside air of the nonporous hydrophilic film or membrane in the concentration device of the present invention (or the container which is a component thereof). The ratio of the surface area to be used is not particularly limited. Similarly, as long as the concentration efficiency as the “concentration device” is shown, the shape, size, thickness, material, etc. of the concentration device of the present invention (and the nonporous hydrophilic film or membrane constituting the device) are particularly Not limited.
The concentration device of the present invention may be substantially composed of the above-mentioned “container” itself, and, if necessary, other parts (for example, holding of the container, reinforcement, objects to be concentrated, etc.) It may have a part having an entry / exit assist function.

(Film or film having moisture permeability)
There are two known moisture permeable films that do not transmit water but transmit water vapor: (1) a porous type and (2) a non-porous type. As the former porous type, there is a hydrophobic polymer film provided with a large number of micropores. In this type, water vapor permeates through the micropores and is hydrophobic. Cannot penetrate into the micropores and the film does not penetrate water. On the other hand, in the case of the latter non-porous type, since the film has hydrophilicity, water migrates into the film, and the water evaporates (scatters) from the surface of the film. The property that it does not permeate water and permeates only water vapor is manifested.

(Preferable non-porous hydrophilic film or membrane)
The “non-porous hydrophilic film or membrane” that can be suitably used in the present invention is a film or membrane exhibiting the above-mentioned “osmotic evaporation (scattering)” phenomenon (in addition, according to the study by the present inventors). Non-hydrophilic films or membranes do not show the “osmotic evaporation” phenomenon).
More specifically, the “nonporous hydrophilic film or membrane” that can be suitably used in the present invention has a transpiration rate exhibited by the film or membrane when in contact with water as Rc (g / m ^2). 24hrs), and when the transpiration rate of the film or membrane when not in contact with water is Rn (g / m ² · 24hrs), the value of these ratios Rr = Rc / Rn is 1.5 The above is preferable. The value of this ratio Rr = Rc / Rn is further preferably 2 or more, and particularly preferably 3 or more (particularly 4 or more). Such values of Rc and Rn can be suitably measured under the conditions of Example 1 described later.

Example 1 shows the transpiration rate (g / m ² · 24 hrs) of water from the surface of a nonporous hydrophilic film and a microporous hydrophobic film. In the case of a microporous hydrophobic film, The transpiration rate was almost the same (approximately 730 g / m ² · 24 hrs) whether the film was in direct contact with water or not, whereas in the case of a nonporous hydrophilic film, the water was In a non-contact state, it is about 380 g / m ² · 24 hrs, but when it comes into direct contact with water, it is greatly improved to about 2,200 g / m ² · 24 hrs. That is, the phenomenon of osmotic transpiration appears only when a nonporous hydrophilic film is in direct contact with water.

  In order to achieve the object of the present invention, (2) a non-porous type film or membrane is suitable. The reason why the hydrophobic film having microporosity of (1) is unsuitable for achieving the object of the present invention is as follows. 1) In the case of a microporous hydrophobic film as described in Example 1, the film surface Even if it is in direct contact with water, the transpiration rate of water is significantly lower than the osmotic transpiration of a nonporous hydrophilic film when in direct contact with water, and 2) the concentration of the present invention. In an amphiphilic system such as protein as a target component or a mixed system of a hydrophilic component and a hydrophobic component, the component is adsorbed on the surface of the micropore and the wettability to water is increased. The problem is that water penetrates and water itself permeates the membrane.

(Nonporous type film)
Polyvinyl alcohol (PVA), cellophane, cellulose acetate, cellulose nitrate, ethyl cellulose, hydrophilic polyester, etc. can be used as the material of the nonporous type film. The thickness of the film is not particularly limited, but is usually 500 μm or less, more preferably about 300 to 5 μm, and particularly preferably about 200 to 30 μm.
(Systematization)
At least a part of the container for storing the component to be concentrated is formed by the non-porous hydrophilic film or membrane having the above-described pervaporation (scattering) property. As described in Example 1, the transpiration rate of water from the surface of the nonporous hydrophilic film is directly increased to about 6 times when it is in contact with water as compared with non-contact. Furthermore, the shape of the container is preferably such that the contact area between the concentrated component and the film or membrane is maximized.

  In the container, usually, an inlet and outlet for concentrated components are installed. However, the inlet and the outlet may be the same. A feature of the system is that it has both a function of concentrating the components and a function of storing the concentrated components and storing them while maintaining the concentration ratio. An embodiment of the system is shown in FIG. 1, but is not limited thereto.

(Concentrate)
As long as it is an object that can be concentrated by the concentration container or concentration method of the present invention, the substance to be concentrated is not particularly limited. From the viewpoint of easy entry / exit operation with respect to the container of the present invention, it is preferable that the material to be concentrated has a certain degree of fluidity at room temperature (25 ° C.), but is not limited to this (that is, almost nearly solid) In some cases, the present invention may be applicable).

  More specifically, the concentrate in the present invention has a concentration factor 48 hours after the start of measurement under the conditions of Example 6 described later (however, the temperature is about 25 ° C. and the relative humidity is about 50%). And those showing 1.2 or more are preferred. The concentration ratio is preferably 1.3 or more (particularly 1.4 or more).

(Concentration method)
For example, a solution containing a component to be concentrated (or an object to be concentrated such as a mixture of a solid and a liquid) is introduced into the container from the inlet of the container partially composed of a nonporous hydrophilic film. . At this time, the mixing of air into the container is prevented as much as possible. When the concentrate is sterilized or sterilized, the filling of the concentrate into the container is extremely preferably performed aseptically.

  The inlet is sealed after the container is filled with the concentrator. The condensate comes into contact with the nonporous hydrophilic film or membrane constituting the container, and water and low molecular weight substances (electrolyte, sugar, amino acid, etc.) in the solution are absorbed into the film or membrane, Only moisture is released out of the container by osmotic evaporation. In this process, moisture is removed from the concentrate, and the concentrate components are concentrated without any substantial loss. This concentration process is characterized by the fact that there is no need for external operations such as heating, pressurization, decompression, etc., and that there is no problem of denaturation or deactivation of the concentrated components.

  Further, in this concentration process, the concentrated component is prevented from coming into direct contact with air by the film or membrane, so that the concentrated component comes into contact with air as in the conventional method, resulting in denaturation, deactivation or oxidation reaction, etc. Not receive. Furthermore, since this concentration process is carried out in a completely closed system, it is possible to effectively prevent external contamination with bacteria, viruses, foreign substances and the like.

  Example 7 to be described later shows hemoglobin permeability of the film or membrane of the present invention, but hemoglobin was impermeable. The size of hemoglobin is about 100 mm (for example, New Bio-Separation Vol. 2 Application P80, issued by CMC, 1988).

  Therefore, it is considered that the permeability limit of the film or membrane of the present invention is 100 mm or less. On the other hand, the size of the virus is several hundred Å and the size of the bacteria is several thousand Å to several μm (for example, New Bioseparation Vol. 2 Application P80, published by CMC Co., 1988). Therefore, it is considered that there is no risk of contamination with bacteria, viruses, or foreign substances from outside the container during the concentration process or storage process.

(Hemoglobin impermeability)
The concentration device of the present invention preferably has hemoglobin impermeability. This hemoglobin impermeability is below the limit of quantification when the hemoglobin in the phosphate buffer in the beaker is measured at an absorption wavelength of 409 nm using a spectrophotometer (Hitachi, U-2001) under the conditions of Example 7 described later. (OD: 0.01 or less).
(Myoglobin concentration rate / recoverability)
The concentration device of the present invention preferably exhibits a myoglobin concentration ratio of 1.2 or more after standing for 5 hours under the conditions of Example 8 described later (however, the temperature is about 25 ° C. and the relative humidity is about 50%). This myoglobin concentration rate is preferably 1.5 or more (particularly 2.0 or more).
Similarly, the concentration device of the present invention exhibits a myoglobin recovery rate of 80% or more after standing for 5 hours under the conditions of Example 8 described later (however, the temperature is about 25 ° C. and the relative humidity is about 50%). Is preferred. The myoglobin recovery rate is preferably 90% or more (particularly 95% or more).

(Concentration efficiency)
On the other hand, in order to increase the concentration efficiency in the concentration process of the present invention, it is preferable to improve the permeation evaporation (scattering). For example, 1) increase the water absorption of the film or membrane of the present invention, that is, increase the water content, 2) reduce the thickness of the film or membrane as shown in Example 2, and 3) the object to be concentrated. Increase the contact area (s) with the film with respect to the volume (v) of the container, that is, design the container shape so that s / v increases. For example, temperature, reduced pressure, air flow rate on the film surface, and the like are controlled.

  In particular, in order to increase the efficiency of the concentration process while preventing deterioration of the concentrated components, a container that maximizes the s / v is used, and the humidity as an external condition of the container is reduced below room temperature without heating. In addition, a method such as increasing the air flow rate is optimal.

On the other hand, another feature of the present concentration method is that, when a predetermined concentration rate is achieved, the container is included in a second container (the second container including a film or a film having a gas barrier property as a constituent element if necessary). The container may be encapsulated in substantially the whole of a container or a film or a film having a gas barrier property.
As described above, the film is further covered or sealed (sealed) with a film or a film having a gas barrier property on the outside of the concentration apparatus of the present invention, which includes at least a container for containing a substance to be concentrated. In this embodiment, there is an advantage that storage can be performed while suppressing further osmotic evaporation (dispersion) of the object to be concentrated (that is, while fixing the concentration rate to a predetermined value) (FIG. 2).

In other words, according to such an embodiment, by concentrating the concentrate to a desired concentration ratio and maintaining the concentration ratio by preventing excessive progress of the concentration process of the concentrate. Can do.
(Film or film having gas barrier properties)
The material of the film or film having gas barrier properties is not particularly limited. From the viewpoint of gas barrier properties, polyvinylidene chloride, polyvinyl chloride, nylon, polyester, polyethylene, polypropylene and the like can be suitably used.

  One of the practical merits of this concentration method is as follows. For sterilization of proteins, polysaccharides, physiologically active substances and the like to which autoclave sterilization, EO gas sterilization, radiation sterilization and the like cannot be applied, filtration sterilization is usually performed. Proteins such as collagen and polysaccharides such as hyaluronic acid are high molecular weight substances and have a very high viscosity even at low concentrations and are difficult to sterilize by filtration. When the concentration method of the present invention is used, filter sterilization is facilitated by adding water to the above-mentioned solution having a high viscosity to reduce the solution viscosity. After filter sterilization, the solution is aseptically filled into the concentration container of the present invention. It becomes possible to concentrate and store at a predetermined concentration while maintaining the sterilized state.

  Hereinafter, the present invention will be described more specifically with reference to examples.

Example 1
An osmotic transpiration test was performed on the nonporous hydrophilic film and the microporous hydrophobic film. As the nonporous hydrophilic film, a polyvinyl alcohol (PVA) film (manufactured by Aicello Chemical Co., Ltd., film thickness 40 μm) and a hydrophilic polyester film (manufactured by DuPont, film thickness 40 μm) were used. As the porous hydrophobic film, a microporous polypropylene film (manufactured by Tokuyama Corporation, thickness 35 μm) was used.
Measurement of the transpiration rate of water from various film surfaces was performed by the following method. 100 mL of tap water was placed in a methacrylic resin cup (top diameter 9.5 cm, bottom diameter 7 cm, height 9 cm), and the film (20 cm × 20 cm) was stretched on the top of the cup and fixed with a rubber band. Two pieces were prepared for each film, one with the cup's mouth facing up on the table, and the other with the cup's mouth facing down, so that there was a gap between the film surface and the table. In the former case, the air in the cup is in contact with the film, and in the latter case, the water in the cup is in direct contact with the film. The cup with the film stretched was placed in a room at a temperature of 24 ° C. and a humidity of 61%, the change in the cup weight with time was measured, converted into the water evaporation rate, and the results are shown in Table 1.
From Table 1, when the nonporous hydrophilic film, the film is 2050~2321g / m ² · cases the film to 378~395g / m ² · 24hrs case in contact with the air is in direct contact with the water Water permeation is about 6 times as large as 24 hrs. On the other hand, if the microporous hydrophobic film, there is no change 734g / m ² · 24hrs of direct contact with the case of water film to 729g / m ² · 24hrs case where the film is in contact with air. It is shown that more water permeates through the film when the nonporous hydrophilic film is in direct contact with the water.

Example 2
Example 1 shows osmotic transpiration of water having a thickness of 25 μm, 40 μm, and 65 μm of a polyvinyl alcohol (PVA) film (manufactured by Aicello Chemical Co., Ltd.), which is a nonporous hydrophilic film used in Example 1. And measured in the same manner as shown in Table 2.
As can be seen from Table 2, the osmotic transpiration of water decreased with increasing film thickness in both cases where the film surface was in contact with air and in water, but the decrease was not significant.

Example 3
Commercially available Meviol (registered trademark) gel (manufacturer: Meviol Co., Ltd., distributor: Ikeda Rika, MB-10, freeze-dried product, sterilized) 0.67 g was dissolved in 16.7 g of distilled water, % Meviol® gel aqueous solution was prepared.
As a container for concentration of the present invention, a polyvinyl alcohol (PVA) film (produced by Aicello Chemical Co., Ltd., film thickness 40 μm) used in Example 1 is heat-sealed to form a 12 cm × 9 cm bag (a container with an inlet) ) Was produced. The aqueous meviol® gel solution was poured into the container without being mixed with air and sealed. As a comparative experiment, 17.5 g of distilled water was similarly sealed in a container similar to the container described above.

The filled container was placed on an uneven plate, placed indoors (temperature 20 ° C. to 30 ° C., relative humidity, about 50%), and the weight of the concentrated component solution in the container was measured over time. Here, the weight of the concentrated component solution (concentrated material) was measured using a precision scale (manufactured by Shimadzu Corporation, trade name: upper plate balance BL-620S). Since substantially only water is removed from the concentrate by transpiration, the concentration factor = the ratio of the weight W1 of the concentrate immediately before the concentration operation and the weight W2 of the concentrate after the concentration operation for a predetermined time. It can be calculated as W1 / W2.
The obtained results (change over time in the concentration ratio of Meviol® gel aqueous solution) are shown in Table 3 and FIG. As can be seen from Table 3, in the case of distilled water alone, transpiration was completely completed in about one day. In addition, in the case of Meviol (registered trademark) gel solution, it was found that it was concentrated about 4 times in 15 hours and about 13 times concentrated in one day.

Example 4
By heat-sealing using the PVA film used in Example 1, a 12 cm × 18 cm bag (a container with an inlet) was produced.
Milk (Morinaga Milk Co., Ltd. 4.5 milk, nonfat milk solid content 9.0%, milk fat component 4.5%), about 100 mL and soy milk (Kibun Foods Co., Ltd., soybean solid content 8%) in the container About 100 mL of each was injected, and the mixed air was removed and sealed. The sealed container was placed in the same room as in Example 3 (temperature 20 ° C. to 30 ° C., relative humidity, about 50%), and the weight of milk and soy milk in the container was measured over time. This weight measurement was performed in the same manner as in Example 4.

  Table 4 and FIG. 4 show changes with time in the concentration ratio of milk and soy milk. In the middle of the above milk and soy milk concentration process (concentration for 5.6 hours and 6.7 times when concentrated for 32 hours, respectively), the container is wrapped with a wrap film for food packaging (Saran Wrap (registered trademark), polychlorinated). It was completely sealed with vinylidene, manufactured by Asahi Kasei Corporation. As a result of measuring the weights of milk and soy milk after sealing with the wrap film over time, the concentration ratio was 5.6 times and 6.7 times, respectively, and the concentration process could be terminated.

Example 5
By heat-sealing using the PVA film used in Example 1, a 12 cm × 18 cm bag (a container with an inlet) was produced. Use a food processor to crush the pulp except the peels and seeds of tangerines (Oita Kitsuki) and prunes (Nagano Prefecture), so that trehalose (Trehaose from Hayashibara Co., Ltd.) becomes 21.3% and 21.0%, respectively. The mixture was stirred well and mixed uniformly to prepare 154.6 g and 133.3 g of concentration components, respectively. Each of these components was poured into the container to remove air and seal.
The sealed container was placed in the same room as in Example 3 (temperature 20 ° C. to 30 ° C., relative humidity, about 50%), and the weight of the concentration component in the container was measured over time. This weight measurement was performed in the same manner as in Example 4.

  Table 5 and FIG. 5 show changes with time in the concentration ratios of oranges and prunes. The saturation concentration ratio was about 2.3 times for oranges and about 2.5 times for prune.

Example 6
A 12 cm × 18 cm bag (a container with an inlet) was produced by heat sealing using the PVA film used in Example 1. 15.1 eggs (the content excluding the eggshell), 18.5 g of egg yolk, 70 g of Morinaga lactoferrin (Morinaga Milk Industry Co., Ltd.), and 58.4 g of melted cheese were mixed and mixed in the container. Air was removed and sealed. The sealed container was placed in an environment at a temperature of 22 to 30 ° C., a relative humidity and about 60%, and the weight of the contents in the container was measured over time. This weight measurement was performed in the same manner as in Example 4.
Table 6 shows changes over time in the concentration ratios of egg, egg yolk, lactoferrin, and melted cheese. The content was concentrated 1.6 to 2.9 times in 24 hours, and the surface of the contents was not covered, and the appearance was also kept in its initial state and concentrated.

Example 7
A 3 cm × 3 cm bag (a container with an inlet) was prepared by heat-sealing using the PVA film used in Example 1, and hemoglobin (manufactured by Wako Pure Chemical Industries) was phosphate buffered at a concentration of 0.9 wt%. 1 mL of an aqueous solution dissolved in (1/15 M, pH 7) was poured into the bag and sealed by heat sealing.
Into a 500 mL glass beaker containing 300 mL of phosphate buffer (1/15 M, pH 7), the bag containing the above-mentioned hemoglobin aqueous solution is placed and stirred at room temperature (about 25 ° C.) for 30 minutes (magnetic stirrer; diameter The rotational speed of a magnetic rotor having a length of about 5 mm and a length of about 40 mm was about 40 rpm).

  A 3 mL phosphate buffer solution was collected from a beaker and the absorbance at an absorption wavelength of 409 nm was measured with a spectrophotometer (Hitachi, U-2001) to determine the amount of hemoglobin permeation. : 0.01 or less). From this experimental result, it was found that the PVA film does not transmit hemoglobin.

Comparative Example 1
An experiment similar to that of Example 4 was performed using a nylon membrane filter (manufactured by Millipore) having a pore diameter of 0.22 μm instead of the PVA film of Example 7. As a result, 50% or more of the encapsulated hemoglobin permeated in 3 minutes. It was confirmed from the result of the absorbance measurement.

Example 8
Myoglobin (manufactured by Wako Pure Chemical Industries) was dissolved in distilled water at a concentration of 0.9 wt%. This aqueous solution was collected, diluted 300-fold with a phosphate buffer (1/15 M, pH), and UV spectrum (300 nm to 600 nm) was measured with a spectrophotometer (Hitachi, U-2001). As a container for concentration according to the present invention, a polyvinyl alcohol (PVA) film (manufactured by Aicello Chemical Co., Ltd., film thickness 40 μm) is heat-sealed to produce a 12 cm × 9 cm bag (a container with an inlet). 17.5 g of a 9 wt% myoglobin aqueous solution was injected into the container without being mixed with air and sealed.
The filled container was placed on an uneven plate, placed indoors (temperature 20 ° C. to 30 ° C., relative humidity, about 50%) and left for 5 hours. When the weight was measured after standing for 5 hours, the myoglobin aqueous solution was reduced to 8.7 g (that is, the concentration factor was 17.5 / 8.7 = about 2.01).

  1 mL of this concentrated solution was sampled and diluted 603 times (300 × 17.5 / 8.7) with a phosphate buffer (1/15 M, pH 7), and the UV spectrum (300 nm to 600 nm) was measured. The maximum absorption wavelength and the UV spectrum measured by diluting a 9 wt% myoglobin aqueous solution 300-fold completely coincided, and the protein was not denatured by the concentration of the protein using the concentration container of the present invention. Further, the absorbance was 5% lower than the absorbance of the initial solution, and it was not able to be recovered by adsorption to a part of the film, but it was shown that about 95% of the concentrated protein could be recovered.

It is a model perspective view which shows the example of the basic aspect of the instrument for concentration of this invention. In the concentration instrument of this invention, it is a schematic diagram which shows the basic aspect of this system which controls concentration magnification. 6 is a graph showing the change over time in the concentration ratio of an aqueous gel solution obtained in Example 3. It is a graph which shows the time-dependent change of the concentration ratio of the milk and the soymilk obtained in Example 4. It is a graph which shows the time-dependent change of the concentration ratio of the mandarin orange and prunes obtained in Example 5.

Claims (4)

A concentrating device comprising at least a container for containing an object to be concentrated containing a component to be concentrated, having one or more inlets and outlets,
An apparatus for concentration, wherein at least a part of the container is composed of a nonporous hydrophilic film or membrane. A container for containing a substance to be concentrated containing a component to be concentrated, having one or more inlets and outlets;
A concentrating device comprising at least a sealable gas-impermeable film or membrane disposed on the outside of the container,
At least a part of the container is composed of a nonporous hydrophilic film or membrane. At least a container for containing an object to be concentrated containing a component to be concentrated, having at least one entrance, and at least a part of the container is made of a nonporous hydrophilic film or membrane Using a concentration device,
A concentration method comprising: concentrating a substance to be concentrated by bringing the substance to be concentrated into contact with the hydrophilic film or membrane.   The concentration method according to claim 3, wherein after the concentration is concentrated to a desired concentration ratio, the concentration device is sealed with a gas-impermeable film or membrane.

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