JPH09143081A - Plasma separation filter, separation of plasma using the filter and plasma separation apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To efficiently, easily and quickly provide plasma free from hemocyte and hemoglobin. SOLUTION: The objective plasma separation filter, plasma separation method and plasma separation apparatus enables the easy, quick and safe separation of plasma having a protein concentration same as that of blood even from a small amount of blood without damaging the hemocyte component of the blood. The plasma separation filter is produced by filling an ultrafine fiber bundle in a vessel having an inlet and an outlet in such a manner as to get an average hydraulic radius of 0.5-3.0μm and an L/D ratio of 0.15-6 wherein D is the flow diameter and L is the flow length of the blood component. The invention also relates to a plasma separation method and apparatus using the separation filter. The filter considerably contributes to the automation, speedup and safety-improvement of clinical examination.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a plasma separation filter, a blood separation method using the filter, and a plasma separation device equipped with the filter. More specifically, a small amount of plasma necessary for blood tests and the like, which does not substantially contain blood cells and / or hemoglobin and has almost the same protein and electrolyte composition as in blood, can be simply, quickly and safely collected. The present invention relates to a plasma separation filter, a blood separation method using the filter, and a plasma separation device equipped with the filter.

[0002]

2. Description of the Related Art So-called biochemical tests for measuring components in blood are widely used for diagnosis and follow-up of various diseases,
It has an important position as a clinical test. The analytical technique has made remarkable progress in recent years. For example, the development of various automatic analysis machines has made it possible to analyze a large number of specimens accurately and quickly.

However, in the field of biochemical tests, plasma or serum separated from blood in advance is used because contamination of red blood cells and the like interferes with the analysis of the target substance. Plasma or serum is obtained by collecting blood from a patient or a subject in advance, then once coagulating blood cell components and centrifuging. Coagulation and centrifugation take time to operate,
Not only does it hinder the reduction of clinical examination time, but it also requires a large centrifuge. Therefore, except for large hospitals, clinical tests are currently outsourced to external contractors. As a result, it takes several days to obtain the test results.

As described above, in spite of the fact that many clinical examination processes are automated, the present situation is that the separation of plasma still depends on human labor. Therefore, the plasma separation operation is
Not only is the inefficiency of the clinical examination, but there is also the problem that the worker is exposed to the risk of contact with blood and infection.

As a means for solving the above problems, a technique generally called dry chemistry is known.
This is because, when a small amount of blood is dropped on a small plate composed of a serum separation layer composed of an ultrafine fiber filter such as glass fiber and a reaction layer located below the serum, the serum is separated by the serum separation layer and the lower layer is separated. Reaction and color development occur in the reaction layer, which is measured with a spectrophotometer. This dry chemistry is a simple method that does not use a liquid coloring reagent and does not require cumbersome serum collection by centrifugation.However, the measurable items are compared to general biochemical analysis and immunoassay using liquid reagents. The number is limited, one inspection item is used for one plate, so a large number of plates must be used for inspecting a plurality of items. Therefore, it has not been widely used.

Membrane separation methods are available as means for rapidly obtaining plasma. Japanese Patent Application Laid-Open No. 53-72691 discloses a fine tube-shaped filter element (pore diameter: 0.
A method for separating plasma from blood using a filter medium (05-0.5 μm) is disclosed. However, in this method, blood cells adhere to the surface of the filter element, so that it takes a very long time to filter plasma, and the permeability of components such as proteins contained in plasma is poor. On the other hand, when the filtration pressure is increased to increase the filtration rate, there is a problem such as hemolysis (the red blood cell membrane is broken and hemoglobin inside is released). Further, JP-A-60-111166 discloses a filtration cartridge using a hollow fiber bundle (pore size 0.05 to
1 μm) has been proposed to separate plasma from blood. However, in this method, it is necessary to perform priming (wetting the hollow fiber membrane with physiological saline etc.) as preparation for plasma separation work. However, the plasma is diluted with physiological saline or the like, which causes a problem that accurate analysis data cannot be obtained.

Since the above-mentioned separation method using a membrane is a separation method based on molecular size, the permeability of substances having a relatively large molecular weight such as proteins in blood is low. Therefore, there arises a problem that the composition of proteins contained in plasma does not accurately reflect the composition of proteins before separation. Further, if the pore size of the membrane is too large, there is a problem of hemolysis due to clogging of red blood cells.

In addition to the above, various techniques for separating serum or plasma for clinical examination using a fibrous filter have been proposed. Japanese Patent Application Laid-Open No. 61-38608 discloses a solid-liquid separation device made of fibrous material using a volume filtration effect. This solid-liquid separation device pressurizes blood to flow into the fibrous material,
Plasma can be obtained. However, since the pressure loss is large and the resistance of the filter medium is large, it takes several minutes to obtain plasma. In addition, the protein in blood is adsorbed on the fiber, and there is a problem that the protein concentration of plasma obtained in the initial stage is lowered, so that it has not yet been put into practical use.

Japanese Unexamined Patent Publication Nos. 4-208856 and 5-
196620 includes a separation filter having a glass fiber containing a polyacrylic ester derivative and polyethylene glycol and a lectin impregnated layer, a method for separating and collecting serum or plasma components using the filter, and serum or plasma using the separation filter. A separation device is disclosed. Although these methods and instruments can collect serum or plasma for clinical examination without using centrifugation, the amount of serum or plasma obtained is as small as about 100 μl, and the time required for separation is about 2 minutes. Therefore, it cannot be said that the time is sufficiently shortened as compared with centrifugation. Furthermore, since these techniques use glass fibers as a separating material, the concentration of electrolytes, phosphorus, and lipids in the obtained plasma is increased due to elution of metal ions from the glass fibers and adsorption of blood components to the fibers. Has a drawback that it is significantly different from the concentration in blood before separation. Therefore, these technologies have not been widely spread.

[0010]

As described above, as a clinical test application, a small amount of blood can be used efficiently in a short time.
At present, there is no filter having sufficient performance as a filter for safely separating plasma / serum components.

[0011]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a plasma or serum separation filter which can be separated from blood simply, quickly and safely and which is excellent in assemblability. According to the present invention, plasma or serum components having the same composition as blood can be obtained without damaging blood cells in blood. The present invention provides for separating blood components using an assembly of ultrafine fibers. The mechanism of separation of plasma or serum by the assembly of ultrafine fibers of the present invention is to cause a difference in the migration rate of red blood cells and plasma or serum that move in the assembly of ultrafine fibers. This difference in the moving speed is due to factors such as the material of the ultrafine fibers, the average fine diameter and the size of the fiber gap, the length of the blood cell separation layer, the morphology of the ultrafine fibers, the flow direction of blood, and the surface condition of the fibers. It is achieved by optimizing. As a result, plasma or serum in blood is separated and collected from blood cell components such as red blood cells. Further, since the filter of the present invention has a low pressure loss, the blood is rapidly processed, and the concentration of the electrolyte or protein of the obtained plasma or serum is substantially the same as the concentration before separating the blood. Therefore, according to the present invention, it is possible to obtain plasma or serum equivalent to plasma or serum obtained by ordinary centrifugation. Hereinafter, the present invention will be described in detail.

The present invention relates to a plasma separation filter. The filter of the present invention is a filter in which an assembly of ultrafine fibers is attached to a container having an inlet and an outlet, and the filter has the following characteristics: (1) An assembly of the attached ultrafine fibers The ratio (L / D) of the blood flow channel diameter (D) to the blood flow channel length (L) is 0.
15-6; and (2) the average hydrodynamic radius of the attached ultrafine fiber aggregate is 0.5-3.0 μm.

In a preferred embodiment, the plasma separation filter of the present invention has a red blood cell concentration of 6 to 8 × 10 ⁹ cells /
Fresh cow blood (ml) was separated at a pressure of 0.2 to 0.4 Kg / cm ² , and from the start of separation, an amount of plasma equivalent to 10% of the void volume of the attached ultrafine fiber aggregate was recovered. At times, (a) the concentration of red blood cells in plasma is 0.1% or less with respect to the concentration of red blood cells in blood before separation; and (b) substantially red blood cells are not hemolyzed.

Further, in the filter of the present invention, when plasma is collected under the above conditions, the electrolyte concentration in the separated plasma is 90% or more as compared with the electrolyte concentration in plasma obtained by centrifugation. It has a characteristic that the protein concentration in the retained or separated plasma is 90% or more as compared with the protein concentration in the plasma obtained by centrifugation.

In a preferred embodiment, the ultrafine fibers are polyester, polypropylene, polyamide, or polyethylene.

Furthermore, in a preferred embodiment, the aggregate of the ultrafine fibers is a non-woven fabric.

In a preferred embodiment, the non-woven fabric is attached to the container alone or in a laminated form, and the face of the non-woven fabric of the attached ultrafine fiber aggregate or the flat face of the laminated non-woven fabric is attached. On the other hand, the blood is configured to flow substantially in parallel.

[0018] In a further preferred embodiment, the container and the non-woven fabric are disc-shaped, the inlet is configured so as to be able to supply blood over the entire circumferential side surface of the disc-shaped non-woven fabric, and the outlet is It is configured such that plasma flows out from the center of the disc-shaped nonwoven fabric.

The present invention also relates to a filter in which a hydrophilizing agent is fixed to ultrafine fibers.

In a preferred embodiment, the hydrophilizing agent is fixed on the surface of the ultrafine fibers.

[0021] In a further preferred embodiment, the hydrophilizing agent is polyvinylpyrrolidone. The filter has the following characteristics: (1) The ratio (L / D) of the blood flow channel diameter (D) and the blood flow channel length (L) of the attached ultrafine fiber assembly is 0.
15 to 6; and (2) the aggregate of the attached ultrafine fibers has an average hydraulic radius of 0.5 to 3.0 μm.

In a preferred embodiment, the filter in which the hydrophilizing agent is fixed to the ultrafine fibers has the following characteristics: 0.2 to 0 of fresh bovine blood having a red blood cell concentration of 6 to 8 × 10 ⁹ cells / ml. When separated at a pressure of 4 kg / cm ² and recovering an amount of plasma equivalent to 10% of the void volume of the attached ultrafine fiber aggregate from the start of separation, (a) the concentration of red blood cells in plasma was It has characteristics of 0.1% or less with respect to the concentration of red blood cells in blood before separation; and (b) substantially no red blood cells are hemolyzed.

Further, in the filter of the present invention in which the hydrophilizing agent is fixed to the ultrafine fibers, when the plasma is collected under the above conditions, the electrolyte concentration in the separated plasma is plasma obtained by centrifugation. 90% or more of the concentration of electrolytes in the blood, or 90% or more of the protein concentration in the separated plasma compared to the concentration of protein in the centrifuged plasma Have.

[0024] In a preferred embodiment, the ultrafine fibers are polyester, polypropylene, polyamide, or polyethylene.

Furthermore, in a preferred embodiment, the aggregate of the ultrafine fibers is a non-woven fabric.

In a preferred embodiment, the non-woven fabric is attached to the container alone or in a laminated form, and the face of the non-woven fabric of the attached ultrafine fiber aggregate or the plane of the laminated non-woven fabric. On the other hand, the blood is configured to flow substantially in parallel.

[0027] In a further preferred embodiment, the container and the non-woven fabric are disc-shaped, the inlet is configured so as to be able to supply blood over the entire circumferential side surface of the disc-shaped non-woven fabric, and the outlet is It is configured such that plasma flows out from the center of the disc-shaped nonwoven fabric.

The present invention also relates to a method for separating plasma using a plasma separation filter having the above characteristics or a plasma separation filter in which ultrafine fibers are fixed with a hydrophilizing agent.

[0029] In a preferred embodiment, a filter in which a nonwoven fabric is used alone or in a laminated form and attached to the container is used.

In a further preferred embodiment, the linear velocity of blood to be treated is 0.05 to 50 cm / min.

Further, the present invention relates to a plasma separation device having a plasma separation filter having the above characteristics or a plasma separation filter having ultrafine fibers fixed with a hydrophilizing agent.

In a preferred embodiment, the apparatus of the present invention further comprises a blood supply means for supplying blood to the filter, and a blood supply means for separating plasma from the supplied blood. It has a pressurizing means for applying pressure and a plasma collecting means for collecting the separated plasma.

In a further preferred embodiment, the device of the present invention separates blood cells and / or hemoglobin detecting means for detecting blood cells and / or hemoglobin in separated plasma, and plasma contaminated with blood cells and / or hemoglobin. And a blood cell and / or hemoglobin-contaminated plasma collection means for collecting the separated blood cells and / or plasma mixed with hemoglobin.

In a preferred embodiment, in the device of the present invention, the filter is detachably attached between the blood supply means and the plasma recovery means.

In a preferred embodiment, the apparatus of the present invention has a blood supply means for supplying a fixed amount of blood and / or a plasma recovery means for recovering a fixed amount of plasma, or both of them.

[0036]

BEST MODE FOR CARRYING OUT THE INVENTION The blood used in the present invention generally contains blood cells, plasma and other components. Blood is
It can be derived from humans, cows, goats, dogs, rabbits, etc. Blood can be used as it is, or blood added with an additive such as an anticoagulant or a hemagglutination agent can also be used. Furthermore, when the blood is left without additives, or when a coagulant is added, the fibrinogen in the blood changes to fibrin, and blood coagulation proceeds, but these coagulable blood also remain unchanged. Can be used. Furthermore, blood that has been chemically treated after being subjected to a treatment such as centrifugation may also be used.

The plasma referred to in the present invention means plasma that does not substantially contain blood cell components. Therefore, it is not strictly limited to plasma that does not contain any blood cell components. further,
When the solid content in the blood is separated and removed after the blood is coagulated, serum containing no fibrinogen is obtained.
In the present invention, the term "plasma" includes serum unless otherwise used.

The plasma separation filter, the plasma separation method and the plasma separation device will be described below in this order.

(Plasma Separation Filter) In the plasma separation filter of the present invention, an assembly of ultrafine fibers is attached to a container having an inlet and an outlet, and the average fluid radius of the installed assembly of ultrafine fibers is , Preferably 0.5 μm to 3.
0 μm. The thickness is more preferably 0.5 to 2.5 μm, and particularly preferably 0.5 to 2.0 μm. Here, the average fluid radius is expressed as a concept in place of the diameter when the voids of the attached ultrafine fiber aggregate are non-circular,
It is defined as follows: Mean hydraulic radius = cross-sectional area of conduit / length of circumference of pipe = volume of fluid in pipe / internal surface area of pipe in contact with liquid = void volume of porous body / porosity Surface Area of Voids of the Body In the present invention, the hydraulic radius is calculated by the following formula (1): DH = R × (ρ−rm) / 4rm (1) DH: of the attached ultrafine fiber aggregate Average hydraulic radius (μ
m) R: average fiber diameter of ultrafine fibers (μm) ρ: density of ultrafine fibers (g / cm ³ ) rm: average bulk density of attached ultrafine fiber aggregates (g /
cm ³ ) As shown in the formula (1), the average hydraulic radius DH of the attached ultrafine fiber aggregate is R and R when the ultrafine fibers of the same material are used (that is, when ρ is constant). determined by rm.

If the average fluid radius is more than 3.0 μm, blood cells will easily pass through the fiber gap. in this case,
It only provides the same effect as conventional filters such as leukocyte removal filters or leukocyte separation filters that remove leukocytes from blood. White blood cells (including some platelets)
As described above, only the component having specific adhesiveness is adsorbed and removed by the ultrafine fiber aggregate.

When the average fluid radius is less than 0.5 μm, the fiber gap in the filter, that is, the blood flow path becomes too narrow, and the blood cell component is likely to be clogged. Further, when pressure is applied to the filter in order to increase the amount of liquid passing through, the pressure loss increases, and hemolysis of red blood cells is likely to occur.

The average fluid radius is 0.5 μm to 3.0 μm.
In the range of m, the smaller the average fluid radius is, the smaller the particle size such as the plasma component is, but the permeability is not affected. On the other hand, those having a large particle size such as blood cell components are less likely to pass through the filter. Therefore, the average fluid radius is preferably 0.5 μm to 2.8 μm, particularly preferably 0.5 μm to 2.0 μm.

Furthermore, the fluid dynamic radius of the ultrafine fiber assembly of the present invention can be constant over the axial direction from the blood supply side to the plasma outlet side, and can be varied depending on the portion of the ultrafine fiber assembly. obtain. Also, the hydraulic radius can be gradually reduced from the blood inlet to the passage liquid outlet. As a result, the separation efficiency between the blood cell component and the plasma component near the plasma outlet can be increased.

In the present invention, the term "average fluid radius" refers to an assembly of ultrafine fibers attached to a container having an inlet and an outlet, which can substantially participate in plasma separation. The average hydraulic radius in the aggregate of. Therefore, for example, when an aggregate of ultrafine fibers is used as the base material 17 for filling the voids 16 in FIG. 2 (FIG. 3)
Since the aggregate of the ultrafine fibers of the base material 17 cannot participate in plasma separation, it means the average hydraulic radius of the part excluding this part.

When this is seen from another side, when the aggregate of the ultrafine fibers attached to the filter is viewed as a whole, the average fluid radius may be outside the preferable range above. However, even in this case, the fact that plasma can be separated is nothing but to show that at least a part of the attached ultrafine fiber aggregate has an average fluid radius in the above-mentioned preferred range.

In the present invention, a prefilter for removing foreign matters in blood may be installed in front of the assembly of ultrafine fibers for plasma separation. The average pore diameter and the average hydraulic radius of these prefilters are naturally larger than the average hydraulic radius of the aggregate of the ultrafine fibers, but the average hydraulic radius of the filter as a whole should take the average pore diameter of the prefilter into consideration. Instead, the average hydraulic radius of the main filter section must be used.

In the present invention, the aggregate of ultrafine fibers is
It refers to a state in which ultrafine fibers are randomly gathered. This state is
For example, it can be obtained by, for example, compressing ultrafine fibers in the form of cotton, non-woven fabric, woven fabric, or knit fabric, either alone or in combination. The ultrafine fibers to be attached to the filter are preferably non-woven or cotton-like ultrafine fibers from the viewpoints of moldability, processability, easy handling, and difficulty in channeling or uneven flow after molding. In particular, it is preferably in the form of a non-woven fabric. This is because when the aggregate of ultrafine fibers is filled in the filter case, the uniformity is likely to be maintained, and it is difficult to partially densify and densely flow the blood evenly.

The material of the ultrafine fibers is not particularly limited, and examples thereof include polyester, polypropylene, polyamide or polyethylene. Preferably, a hydrophobic polypropylene or polyester material (for example, polyethylene terephthalate) can be used.

These materials are preferable because they do not adsorb plasma or serum components when they come into contact with blood, and conversely, some of the materials do not elute into plasma or serum. When plasma or serum separation filters that use glass fibers are used as described in the section of the prior art, metal ions are eluted from the glass fibers, and phosphorus and lipids are adsorbed to the glass fibers, so these substances are measured. I couldn't.

In the present invention, the blood cell separation layer has a length of 5 m.
m or more is preferable. The length of the blood cell separation layer means the length from the point where the aggregate of ultrafine fibers and blood come into contact to the point where the blood (plasma or serum) separates from the aggregate of ultrafine fibers. As described above, the present invention separates blood cells and plasma by utilizing the difference in the moving speed of blood components in the aggregate of ultrafine fibers. Blood is moved in the aggregate of ultrafine fibers by applying pressure from the inlet side of the blood cell separation layer, depressurizing from the outlet side, or both at the same time. At this time, the blood cell component repeatedly collides with the fine fibers in the gaps between the fine fibers. Adhesive white blood cells and platelets are adsorbed on ultrafine fibers, and red blood cells do not have adhesive properties, so they move while repeatedly deforming. On the other hand, plasma or serum
Since it is a liquid component, it moves through the ultrafine fibers faster than red blood cells and reaches the outlet earlier. At this time, if the length of the blood cell separation layer is less than 5 mm, a sufficient difference does not occur in the migration distance between blood cells and plasma, and the separation of the two becomes insufficient, which is not preferable. The longer the length of the blood cell separation layer, the higher the separation efficiency of blood cells and plasma, but on the other hand, there arises a problem that the pressure loss becomes large or the amount of necessary ultrafine fibers and blood amount increase. Therefore, the length of the blood separation layer is determined by the required amount of plasma or serum, the amount of blood used, the size limit of the filter, etc., but there is no theoretical upper limit.

When a non-woven fabric is used, it is preferable that blood flow parallel to the plane of the non-woven fabric (the plane of the laminated non-woven fabric). Generally, when a nonwoven fabric is used as a filter, the flow direction of the treatment liquid is perpendicular to the plane of the nonwoven fabric (plane of the laminated nonwoven fabric). However, in the present invention, by allowing blood to flow parallel to the surface of the nonwoven fabric, the efficiency of separating blood cells and plasma components is improved. When blood flows in parallel to the plane direction of the non-woven fabric, when blood flows from the inlet to the outlet, there is no break in the ultrafine fibers over the entire flow path length, and therefore the blood flow uniformity is improved. However, the reason is not limited to this.

The ultrafine fibers to which the hydrophilizing agent is fixed are also
It can be used as the filter of the present invention. The immobilization of the hydrophilizing agent can be performed physically or chemically. By fixing the hydrophilizing agent on the fiber surface, the affinity between the ultrafine fibers and blood is increased. Therefore, when separating blood into blood cells and plasma, the pressure loss is reduced and the separation speed can be increased. The hydrophilizing agent is not particularly limited as long as it does not interfere with the analysis even when mixed with plasma or serum. Polyvinylpyrrolidone is preferred. Polyvinylpyrrolidone has a relatively large molecular weight, so the elution rate is relatively slow, but
Elute in blood. However, it does not affect the analysis of blood components. The method for immobilizing polyvinylpyrrolidone is not particularly limited, and a known method can be used. For example, by dipping an aqueous solution of polyvinylpyrrolidone in an aggregate of ultrafine fibers and then drying the aggregate, it can be easily physically fixed on the fiber surface. Further, polyvinyl pyrrolidone can be easily cross-linked by heat treatment or radiation treatment of an aggregate of ultrafine fibers having such polyvinyl pyrrolidone physically fixed on the surface. By cross-linking, elution of polyvinylpyrrolidone into blood can be suppressed to a lower level.

In the case of heat treatment, the method is not particularly limited. For example, a method of heating under pressure, such as an autoclave treatment, or a method of leaving it in a constant temperature bath may be used. The temperature of the heat treatment is not particularly limited, but is preferably 70 ° C or higher, more preferably 100 ° C or higher. The higher the heating temperature, the higher the efficiency of crosslinking.
The upper limit temperature is not uniquely determined by the properties of the ultrafine fibers used, the heat resistance of polyvinylpyrrolidone itself, etc.,
The temperature is preferably 200 ° C or lower, more preferably 150 ° C or lower. The heating time is preferably long for sufficient crosslinking, but is limited by the properties of the ultrafine fibers used and the modification of polyvinylpyrrolidone. Generally, the time is preferably from 20 minutes to 2 hours. Further, the crosslinking by heating can be performed either when the ultrafine fibers are immersed in the hydrophilic agent aqueous solution (WET state) or when the ultrafine fibers are dried after the immersion (DRY state). In each case,
Polyvinylpyrrolidone can be fixed to the ultrafine fibers. Unreacted polyvinylpyrrolidone can be removed by washing with water.

The method of fixing the hydrophilizing agent using radiation is not particularly limited. For example, γ-ray irradiation, electron beam irradiation, corona discharge, etc. may be mentioned. Irradiation with γ-rays is preferred in terms of the thickness that can be processed and the operability. The irradiation dose is not particularly limited as long as the polyvinyl pyrrolidone is sufficiently crosslinked. As a range that does not cause modification of the ultrafine fiber material and polyvinylpyrrolidone itself by irradiation
It is preferably KGy or more and 50 KGy or less. Further, the radiation irradiation can be performed in the WET state or the DRY state. Unreacted polyvinylpyrrolidone can be removed by washing with water or the like.

Polyvinylpyrrolidone is available in various molecular weights. In order to avoid elution into the blood, it is particularly preferred to use one having a large molecular weight.

A filter is produced using the aggregate of ultrafine fibers to which the hydrophilizing agent is fixed.

The filter can be produced by laminating and compressing cotton-like, non-woven fabric-like, woven fabric-like, and knitting-like ultrafine fibers, alone or in combination.

In the present invention, the shape of the filter container is not particularly limited. Examples include a rectangular parallelepiped shape, a disk shape, a cylindrical shape, a truncated cone shape, and a fan shape. Of these, when a filter having a rectangular parallelepiped shape, a disk shape, or a fan shape is used to flow parallel to the surface of the aggregate of ultrafine fibers, the separation performance is improved. In the case of a rectangular parallelepiped, blood is made to flow from one end surface to the other end surface, or is formed into a disk shape, and blood is made to flow from the peripheral surface toward the center. The filter can be sealed by pressing the non-woven fabric with a container of these shapes. Therefore, it is particularly preferable because it is not necessary to use an adhesive. Among them, the disk shape or the fan shape is more preferable because the cross-sectional area of the blood flow path gradually decreases as the blood moves, and unevenness in the lateral movement of the blood component decreases. In particular, the disc-shaped filter is excellent in operability and is most preferable. In the case of a disk-shaped container,
The attached non-woven fabric also has a disc shape. It is preferable to configure the blood inlet so that blood can be supplied over the entire circumference of the disc-shaped nonwoven fabric. For example, a gap may be provided between the disc-shaped non-woven fabric and the disc-shaped container at the circumference thereof, and one or a plurality of inlets communicating with the gap may be opened on the side surface, the top surface or the bottom surface of the container.

The average fiber diameter of the ultrafine fibers used in the filter of the present invention is preferably 0.5 to 3.5 μm, more preferably 0.5 to 2.8 μm, and particularly preferably 0.5 to It is 2.0 μm.

The ultrafine fibers having the above average fiber diameter are
It can be obtained by a conventional ultrafine fiber spinning method such as melt blowing.

Here, the average fiber diameter of the ultrafine fibers is obtained by measuring the diameter of 50 ultrafine fibers randomly selected from a photograph of an aggregate of the ultrafine fibers taken with an electron microscope of 2000 times with a caliper or a scale loupe. The average value of the values obtained by

When the average fiber diameter of the fibers exceeds 3.5 μm, the length of the fibers per unit volume of the ultrafine fiber assembly is shortened, and as a result, the number of entangled points between the fibers is reduced,
The fiber gap becomes large. As a result, even components with a large particle size, such as blood cells, can easily pass through the aggregate of ultrafine fibers,
Poor separation of blood cells and plasma. Furthermore, the adsorptivity to white blood cells and platelets is reduced.

When the average fiber diameter of the ultrafine fibers is less than 0.5 μm, the length of the fibers per unit volume becomes long, and as a result, the number of entangled points between the fibers becomes large and the fiber gap becomes small. As a result, blood cells are easily clogged in the filter. Furthermore, since the pressure loss of the aggregate of ultrafine fibers is large, hemolysis of red blood cells is likely to occur.

The average bulk density of the attached ultrafine fiber aggregate used in the present invention is preferably 0.15 to 0.6.
0 g / cm ³ , and more preferably 0.18-0.
50 g / cm ³ , and particularly preferably 0.25 to 0.
40 g / cm ³ .

Here, the average bulk density of the aggregate of ultrafine fibers is a value obtained by dividing the weight of the aggregate of ultrafine fibers by the volume of the aggregate of ultrafine fibers.

When the average bulk density is less than 0.15 g / cm ³ , the average bulk density immediately after spinning of the ultrafine fiber aggregate (for example, 0.08 to 0.10.
g / cm ³ ) is small, the compressibility of the ultrafine fiber aggregate is small. Therefore, densification is likely to occur partially in the attached ultrafine fiber assembly, and unevenness may occur in the blood passage speed. Moreover, since the fiber gap is large on average, the separation of blood and plasma becomes insufficient.

If the average bulk density of the attached ultrafine fiber aggregate is larger than 0.60 g / cm ³ , a special heat compression process or the like is required for the production of the ultrafine fiber aggregate, and the compression process is complicated. become. Furthermore, since the fiber gaps of the attached ultrafine fiber aggregates are small, the blood cell components are likely to cause clogging of the filter, and the pressure loss of the ultrafine fiber aggregates is increased, resulting in hemolysis of red blood cells. It will be easier.

The average bulk density is 0.15 to 0.60 g /
By increasing the average bulk density in the range of cm ³ , the homogeneity of the aggregate of ultrafine fibers is further improved. On the other hand, since the workability is lowered, it is preferably 0.18 to 0.
50 g / cm ³ , and particularly preferably 0.25 to 0.
40 g / cm ³ .

The bulk density of the aggregate of ultrafine fibers mounted on the filter of the present invention can be partially changed. For example,
The bulk density of the attached ultrafine fiber assembly can be gradually increased from the inlet to the outlet of the filter container. In this way, the separability between blood cells and plasma improves as the blood component moves toward the outlet.

The ratio (L / D) of the flow path length (L) to the flow path diameter (D) of the blood component of the ultrafine fiber assembly mounted on the filter of the device of the present invention is 0.15 to 6. . It is preferably 0.25 to 4, particularly preferably 0.5 to
2.

Here, the flow path length (L) of the blood component means the linear distance (generally, the ultrafine fiber) inside the ultrafine fiber assembly from the contact of blood with the ultrafine fiber until the plasma leaves the ultrafine fiber. The length of the aggregate of fibers). The flow path diameter (D) of the blood component means the equivalent circle diameter of the cross-sectional area of the surface of the aggregate of ultrafine fibers at the blood inlet portion which is perpendicular to the flow path length direction.

The equivalent circle diameter is obtained from the sectional area (A) by the following equation (2).

D = 2 (A / π) ^1/2 (2) The surface of the aggregate of ultrafine fibers has small irregularities due to the bending of the ultrafine fibers in the strict sense. Ignore and calculate as a plane. In addition, in addition to the unevenness due to the bending of the ultrafine fibers, when there is a large unevenness formed by the surface processing of the aggregate of the ultrafine fibers, the cross-sectional area is calculated as a flat surface that averages the unevenness.

When L / D is smaller than 0.15, the flow path is too short and the moving distance of each component in blood becomes short. Further, since the cross-sectional area is large, the moving speed of each component in blood is uneven in the lateral direction. Therefore, the separation of the blood cell component and the plasma component becomes insufficient.

When L / D exceeds 6, the separation efficiency is improved, but since the moving distance is long, the pressure loss is high and hemolysis of red blood cells may occur.

The filter of the present invention has the following characteristics.

Fresh bovine blood having a red blood cell concentration of 6 to 8 × 10 ⁹ cells / ml was separated at a pressure of 0.2 to 0.4 kg / cm ² , and from the start of the separation, the assembly of the attached ultrafine fibers was separated. When the amount of plasma equivalent to 10% of the void volume was collected,
(a) The concentration of red blood cells in plasma is 0.1% or less of the concentration of red blood cells in blood before separation; and (b) the red blood cells are not substantially hemolyzed.

Furthermore, the electrolyte concentration in the separated plasma is maintained at 90% or more as compared with the electrolyte concentration in the plasma obtained by centrifugation.

In the present invention, the electrolyte concentration in plasma separated by the plasma separation filter is preferably maintained at 90% or more as compared with the electrolyte concentration in plasma obtained by ordinary centrifugation. . In other words, the decrease in electrolyte concentration after filter separation is 1
It is preferably within 0%. It is more preferably within 5%. When the decrease in the electrolyte concentration in plasma exceeds 10%, the reliability of biochemical diagnosis becomes low, which is not preferable. Judging from the measurement accuracy of the biochemical test, if the difference between the two is within 10%, there is virtually no problem, and within 5%, there is almost no problem. Here, plasma refers to a supernatant obtained by adding an anticoagulant to blood obtained by collecting blood and then centrifuging the blood. The centrifugation is performed at 1000 G for 10 minutes.

In the present invention, it is preferable that the plasma concentration of the plasma separated by the plasma separation filter is maintained at 90% or more at the beginning and end of the collection.
That is, it is preferable that the decrease in the protein concentration after the filter separation is within 10% as compared with the case of the centrifugal separation. It is more preferably within 5%. When the decrease in the protein concentration in plasma exceeds 10%, the reliability of biochemical diagnosis is lowered, and the values at the beginning and the end of collection may be different and accurate diagnosis may not be possible. Further, if the difference in protein concentration exceeds 10%, the composition of plasma proteins may fluctuate significantly, and it may not be used for diagnosis of a medical condition, which is not preferable. Usually, if the difference is within 10%, there is no major problem in clinical diagnosis, and if the difference is within 5%, it is within the error range of the measurement, which is preferable.

The material of the filter container of the present invention is not particularly limited, and examples thereof include metal, glass, plastics,
For example polyethylene, polypropylene, nylon, polycarbonate, polystyrene, polyester, ABS
Resins. When observing the internal state, a transparent or semitransparent material may be selected. Plastics are preferable from the viewpoints of workability, resistance to cracking and light weight.

The filter used in the plasma separation apparatus of the present invention will be described below with reference to the drawings. The filter of the present invention is most simply shown in FIG. FIG.
FIG. 3 is a view showing a filter 11 in which a cylindrical flat ultrafine fiber assembly 14 is attached to a container 15 having an inlet 12 and an outlet 13. In this filter 11,
The ultrafine fibers are compressed and mounted so that an appropriate space 16 is provided between the inner peripheral surface of the container 15 and the outer peripheral surface of the ultrafine fiber assembly 14. The outer periphery of the attached ultrafine fiber assembly 14 may be made of, for example, a plastic plate having appropriate holes through which blood can pass. When blood is supplied from the inlet 12 of the container 15 at the upper part of the outer peripheral portion of the ultrafine fiber assembly 14 mounted in a cylindrical shape, the void 1
6, the blood is uniformly supplied to the ultrafine fiber aggregate 14 to which the blood is attached, the blood flows from the outer peripheral portion of the ultrafine fiber aggregate 14 toward the central portion, and is separated into plasma and blood cells in the meantime. Plasma is collected from the outlet 13 at the center of the container 15. Hereinafter, the aggregate 14 of ultrafine fibers simply means an aggregate of ultrafine fibers mounted on the filter.

As another embodiment, the filter 1 of FIG.
There is one. The filter 11 has a cylindrical ultrafine fiber assembly 14 mounted in a cylindrical container 15. Blood is supplied from the inlet 12 of the container 15, and the plasma separated by the ultrafine fiber aggregate 14 is collected from the outlet 13 of the container 15. The inlet 12 and the outlet 13 can be arranged at arbitrary positions.

Similar to the filter of FIG. 1, the cylindrical ultrafine fiber assembly 14 of FIG. 2 may have a void 16 in the vicinity of the inlet 12. The voids 16 can be arranged when supplying blood and pressurizing uniformly.

The filter 11 of FIG. 3 is a filter in which an appropriate base material 17 is arranged in the space 16 of the filter 11 of FIG. The base material 17 to be arranged is a base material having a fiber gap larger than the fiber gap of the aggregate 14 of the ultrafine fibers, and any one can be used as long as it can pass all components in blood. Can be done. The base material may be, for example, a filter medium having a plate shape, a paper shape, a fiber shape, or the like.
Also in the filter of FIG. 1, the base material 17 is provided in the gap 16.
Can be placed.

The filter 11 shown in FIG. 4 is a filter in which an assembly 14 of ultrafine fibers is attached to a container 15 having an opening 18 and an outlet 13.

The filter 11 can be removably attached to the blood supply means and / or plasma recovery means described below. As another embodiment, FIG.
And as illustrated in FIG. 5B, the container 15 has the inlet 1
2 or a portion having an opening 18 (15a in FIG. 5),
It is composed of three parts, a part having the ultrafine fiber aggregate 14 and a part having the outlet 13 (15b in FIG. 5), and each part can be detachably configured. With this configuration, only the part of the assembly of the ultrafine fibers to be mounted can be replaced. The exchangeable microfiber assembly portion may be coated with a blood permeable membrane or the like and may be in the form of a removably matable cassette. The detachable mounting means may be a fitting means, a screwing means, or a magnet means. These attachments and detachments can be performed automatically.

As described above, when the filter of the present invention is manufactured by laminating the ultrafine fibers, the direction of blood flow is such that the filter is almost perpendicular to the laminating surface, or almost the laminating surface. It can be parallel. In the former case, a columnar shape or a conical shape is suitable. For example, when flowing blood from the upper part to the lower part of a cylindrical filter (Figs. 2 to 4), or using a truncated cone-shaped filter with the apex portion cut into a conical shape, the blood is directed from the bottom of the cone toward the apex direction. This is the case when the water is drained. In the latter case, a disc-shaped filter (for example, Fig. 1) is used, and the filter is laminated parallel to the lamination surface from the periphery to the center, or from the side surface of one side of the flat plate-shaped filter. This is the case when blood is shed. The frustoconical or disk-shaped filter gradually reduces the cross-sectional area of the blood flow path as the blood moves, which increases the moving speed of the blood components, causing uneven movement of the blood components in the lateral direction. Is smaller, the blood separation efficiency is improved.

A plurality of filters of the present invention can be connected. Separation improves when connected in series. Connecting in parallel increases the amount of blood to be processed.

Further, the filter and the blood sampling container may be combined and used. The filter combined with such a sampling container can be used in an automatic analyzer having a means for collecting a certain amount of blood and supplying it to the filter and a means for automatically changing the used filter. (Plasma separation method) In the method for separating plasma, the pressure loss at the outlet is 0.03 to 5 kg / cm ² . This value depends on the shape of the filter and the processing speed of blood.

When the pressure loss is less than 0.03 kg / cm ^{2, the} load on the blood in the aggregate of the ultrafine fibers is small. Therefore, when ultrafine fibers having high hydrophobicity are used, blood cannot be sent to the aggregate of ultrafine fibers,
Alternatively, there is a problem that the processing takes too long.
Further, there is no difference in the moving speed between the blood cell component and the plasma component in the aggregate of the ultrafine fibers, and the separation of plasma becomes insufficient.

If the pressure loss is larger than 5 kg / cm ² , the blood flow rate is too high. Therefore, the plasma may not be separated due to a small difference in transit time between the plasma and blood cells. In addition, the pressure may cause hemolysis of red blood cells and damage to the filter or device.

If the pressure is increased within the above range, the amount of plasma obtained increases and the time required to obtain plasma is shortened. On the other hand, there is a possibility of hemolysis, so 0.05 to 3 kg / c
A range of m ² is preferred. The range of 0.1 to 1 kg / cm ² is more preferable.

In the plasma separation method of the present invention, the ratio of the treated blood volume to the total area of the ultrafine fibers is not particularly limited, but is preferably 0.1 to ³ m ² per 1 ml of blood.
Ratio of treated blood volume to total area of ultrafine fibers When the blood volume is less than 0.1 m ² per 1 ml of blood, the separation efficiency of plasma becomes poor. The ratio of the treated blood volume to the total area of ultrafine fibers is blood 1
When it is larger than 3 m ² per ml, the blood volume is too small, which makes it difficult to send blood and collect plasma. Further, as a result of the plasma protein being adsorbed on the ultrafine fibers, the protein component in the plasma component may not accurately reflect the protein component in the original blood. Furthermore, pressure loss is also likely to occur.

When the surface area of the ultrafine fibers is increased within the above range, the amount of plasma obtained increases. On the other hand, the amount of protein adsorbed on the fibers also increases. Therefore, the above ratio is 0.2
The range of ˜2 m ² is preferred. The range of 0.3 to 1.5 m ² is more preferable.

In the plasma separation method of the present invention, the linear velocity of blood treatment is not particularly limited,
It is about 0.05 to 50 cm / min. The linear velocity is 0.
If it is less than 05 cm / min, the time for the blood component to pass through the aggregate of ultrafine fibers becomes long. As a result, the separated plasma and blood cells diffuse during migration, resulting in insufficient separation.

When the processing linear velocity is higher than 50 cm / min,
The pressure loss increases and red blood cells are hemolyzed.

When liquid is sent from the outer peripheral part of a disc shape (for example, FIG. 1) toward the center part, or when a conical container is used to send liquid toward the apex of the cone, Depending on the linear velocity. The linear velocity in such a case refers to the average processing linear velocity from the time when blood comes into contact with the microfiber until it leaves.

In the plasma separation method of the present invention, the blood volume to be treated (B) and the void volume (A) of the aggregate of ultrafine fibers.
It is preferable that the ratio (B / A) to is 0.2 or more. Ratio is less than 0.2 (volume of blood is 2 of interstitial volume)
(Less than 0%), the blood volume is too small, which makes it difficult to deliver blood and collect plasma. Further, as a result of the plasma protein being adsorbed on the ultrafine fibers, the protein component in the plasma component may not accurately reflect the protein component in the original blood. The ratio is preferably 0.5 or more, particularly preferably 0.7 or more.

In the plasma separation method of the present invention, a piston, pressurized air or the like can be used as a method for supplying and pressurizing blood. There is also a method of extruding with a liquid that does not react with blood components, for example, a liquid with high viscosity to prevent mixing, paraffin, glycerin, or the like. The method using pressurized air or paraffin is suitable when the volume of blood to be treated is smaller than the interstitial volume of the aggregate of ultrafine fibers. However, in the case of pressurized air, the blood in the ultrafine fiber aggregate may not be able to flow due to the surface tension generated in the gap in the ultrafine fiber aggregate. On the other hand, the method using paraffin or glycerin is preferable because the problem of surface tension does not occur.

(Plasma Separation Apparatus) The plasma separation apparatus of the present invention includes a plasma separation filter, preferably blood supply means for supplying blood to the filter, pressurizing means for pressurizing the supplied blood, and separated plasma. A plasma collection means for collecting plasma is included. Further, a detection means for detecting blood cells and / or hemoglobin in the separated plasma, a switching means for separating plasma mixed with blood cells and / or hemoglobin, a blood cell for collecting plasma mixed with blood cells and / or hemoglobin, and It may include a hemoglobin-containing plasma recovery means, a plasma recovery means for recovering blood cells and / or plasma confirmed to be free of hemoglobin. Further, the blood supply means may be a blood supply means that supplies a fixed amount of blood, and the plasma recovery means may be a plasma recovery means that recovers a fixed amount of plasma. Each of the above means will be described below.

<Blood Supply Means> In the apparatus of the present invention, known means can be used as a means for supplying blood to the filter. For example, means for supplying blood using a pump, pressurizing a container containing blood, means for supplying blood using the pressure, depressurizing the inside of the filter, and utilizing the pressure difference from the blood storage container. Means for supplying blood (for example, a method using a vacuum pump, or FIG.
(B), a method of moving the piston upward to reduce the pressure inside the cylinder and guiding blood from the inlet 12) and the like. Of course, means for manually supplying blood such as a pipette may also be included. The means for supplying blood to the filter can be connected to the inlet of the filter. As a connection method, any method can be used as long as blood does not leak. For example, the blood outlet of the blood supply means and the filter inlet can be connected by fitting, joining, or screwing together.

Further, the outlet of the blood supply means may be closed, and the pierceable means may be arranged at the inlet of the filter. On the contrary, the inlet of the filter may be closed and the blood supply means may be provided with a pierceable means. Examples of the pierceable means include hollow and tubular means such as an injection needle. In either case, the perforations allow blood to be supplied to the filter. Such a pierceable connecting means is particularly effective in automating blood supply and avoiding contact with blood. When plasma is separated from blood collected by a vacuum blood collection tube which has been used in recent years, a vacuum blood collection tube containing blood is directly drilled in a closed inlet of a filter. By reducing the pressure in the container of the filter, blood is supplied to the ultrafine fiber assembly attached to the filter. It is also possible to use a device in which when blood is supplied, the vacuum blood collection tube is pulled out and pressurized from the inlet side of the filter.

The blood supply means may be a supply means for supplying a fixed amount of blood. For example, a certain amount of blood supply device such as a roller pump or a cylinder pump may be used. The supply means may be stopped when the blood is supplied at a constant flow rate. For example, a sensor may be provided in the blood storage device, the amount of decrease in blood in the blood storage tank may be measured, and the supply means may be stopped.

The volume of the blood component processed by the device of the present invention (the amount of blood supplied by the supply means) is preferably 20% or more of the interstitial volume of the aggregate of ultrafine fibers mounted on the filter, It is more preferably 50% or more, and particularly preferably 70% or more. If it is less than 20%, it may be difficult to pump blood into the attached ultrafine fiber assembly, and a part of the plasma component may remain in the gap and may not be collected. Here, the void volume of the attached ultrafine fiber aggregate is defined as the volume of the attached ultrafine fiber aggregate- (total weight of the attached ultrafine fiber aggregate / density of the attached ultrafine fiber). To be done.

<Pressurizing Means> For blood held or supplied to the attached ultrafine fiber assembly, pressurize the blood side and / or depressurize the permeate side of the attached ultrafine fiber assembly. As a result, it moves to the permeate side in the assembly of the attached ultrafine fibers, and is separated into plasma and blood cells by utilizing the difference in moving speed between plasma and blood cells. At this time, white blood cells and blood platelets, which are blood cell components, are adsorbed by the attached ultrafine fiber aggregates. The method of pressurizing and depressurizing is not particularly limited. Pressurization by pump, pressurization by gas such as compressed air, liquid that does not react with and incompatible with blood components (for example,
Paraffin, glycerin, etc.) may be pressurized and extruded, or a vacuum pump or a cylinder may be used for suction. The method of extruding with paraffin which is incompatible with blood components is particularly effective when the amount of blood is small and does not cause the problem of fluidity due to the surface tension of blood components, which is preferable.

FIG. 6 shows an example of a separation device having a pressurizing means.
It shows in (a) and FIG.6 (b). In the device of FIG. 6A, blood supplied from the opening 18 is inserted from the opening 18 of the filter by inserting the slidable pressurizing means 24 having the contacting means 22 in airtight contact with the inner wall 13 of the container 15. Pressurize. The pressurized blood is separated while passing through the ultrafine fiber assembly 14, and plasma is collected from the outlet 13.

In the apparatus shown in FIG. 6 (b), a pressurizing means 24 having a contact means 22 for airtightly contacting the inner wall 19 of the container 15 of the filter 11 is slidably attached. The device has a valve 20 at the inlet 12. With the upward movement of the pressurizing means 24, the inside pressure is reduced, the valve 20 is opened, and blood is supplied from the inlet 12. Further, as the pressurizing means 24 moves downward, the valve 20 at the inlet 12 is closed and at the same time blood is pressurized and plasma is separated. By repeating this procedure, plasma can be continuously separated. In this case, a known means can be used as a means for pressurizing the piston.

In the device of the present invention, the blood supply means can also be used as the pressurizing means. FIG. 7 is a diagram showing an example. In FIG. 7, the inlet 12 of the filter 11 and
The device is connected to the blood supply means 25. The blood supply means 25 is provided with a member 24 having an inner wall 23 and an intimate contact means 22 in airtight contact with each other, and blood 26 is stored therein. When the member 24 is pushed down, the blood 26 is supplied to the filter 11 and is pressurized to separate the plasma. Therefore, the blood supply means 25
At the same time, it has a pressurizing means 24 and can be used as a pressurizing means. For example, a syringe for injection is a specific example.

When the pressurizing or depressurizing means is used, a pressure adjusting means for adjusting the degree of pressurizing and depressurizing may be provided in order to adjust the moving speed of plasma and blood cells. Pumps may be placed before and after the attached ultrafine fiber assembly to adjust the pressure so that hemolysis does not occur. In this case, in particular, the pressure of the blood component before passing through the attached ultrafine fiber assembly and the pressure of the plasma component after passing can be independently adjusted. At this time, the degree of pressurization and depressurization can be automatically controlled by a computer or the like.

The pressure loss (pressure difference between the inlet and the outlet of the filter) due to the pressurization and / or depressurization depends on the shape of the aggregate of ultrafine fibers and the blood treatment rate, but is preferably 0.03 to 5 kg / cm ^2. And more preferably 0.05 to 3 kg
/ cm ² , and particularly preferably 0.1 to 1 kg / cm ² . If the pressure difference is less than 0.03 kg / cm ² , processing time will be longer,
Separation of plasma may be inadequate. At this time, if ultrafine fibers having high hydrophobicity are used, it may be difficult to send blood into the aggregate of ultrafine fibers. Differential pressure is 5kg
When it exceeds / cm ² , the time between passing through the assembly of ultrafine fibers where plasma is attached and the time when passing through the assembly of ultrafine fibers where blood cells are attached are short, so blood cells are mixed, Collection of plasma can be difficult. Furthermore, red blood cells may be hemolyzed, and the device and the attached ultrafine fiber aggregate may be damaged.

<Recovery Means> The plasma separated by the aggregate of ultrafine fibers and moved to the permeate side is recovered for use in inspection and the like. A known means can be used as a means for collecting. For example, the separated blood plasma may be collected in the sample container by using the differential pressure applied to the blood in the filter as it is, or by sucking the permeated liquid with a pump or sending out the permeated liquid. The separated plasma can be collected in a sample container just below the outlet of the filter. Alternatively, a hollow tube can be connected to the outlet and introduced into the sample container. When connecting the outlet of the filter and the collecting means, the same means as the connecting means of the blood supplying means and the filter can be used.

Plasma stored in a sample container or the like is used as a test sample. Examples of the sample container include a tube and a vial, and any material can be used as long as it does not affect storage of the obtained plasma. For example, the same material as the container used for the filter can be used.

During the separation of plasma by the assembly of ultrafine fibers, blood cells permeate with a delay with the passage of time.
Blood cells may be mixed in the separated plasma. Further, depending on the operating conditions, erythrocytes may be destroyed in the aggregate of the ultrafine fibers to which the red blood cells are attached, and the hemoglobin thus eluted may be mixed in the permeate. When separating plasma for clinical examinations, it is necessary to avoid mixing blood cells and hemoglobin in the obtained plasma. When blood cells and hemoglobin are mixed in the permeate plasma in a certain amount or more, the obtained plasma cannot be used as a clinical test sample.

To solve the above problems, the initial permeate (plasma) containing no blood cells can be collected in a preselected amount. For example, under the conditions described herein, the interstitial volume of the loaded microfiber aggregates
By recovering a volume of plasma corresponding to 10%, a substantially blood cell-free plasma can be obtained. Such a quantification can be performed, for example, by disposing a liquid meter in the plasma recovery container, as in the measurement of the blood supply amount. The liquid volume can be measured, for example, by detecting with a sensor. After a certain amount of plasma has been collected, the plasma collection container can be moved to separate the same plasma sample into several fractions. This movement can be done manually or automatically. In this case, means may be used in which the plasma is once pooled and aliquoted. Furthermore, when a certain amount of plasma is obtained, blood supply means,
The device may be configured to turn off the pressurizing means. These controls can be performed using a computer. Or,
After collecting a fixed amount, the permeating plasma can be disposed so as to be discarded or collected in another container by the switching means described below. The blood cells and / or
Alternatively, when the hemoglobin detection means is provided, the plasma quantification device is preferably arranged near the outlet to the container for collecting the blood cell and / or hemoglobin-containing sample or in the plasma sample container.

<Means for Detecting Blood Cells and / or Hemoglobin> When blood cells are mixed with plasma as described above, they cannot be used as test samples. Therefore, the apparatus of the present invention can preferably be provided with a blood cell and / or hemoglobin detection means for detecting blood cells and / or hemoglobin on the permeate side. When blood cells and / or hemoglobin are detected in the permeate, pressurization and blood supply may be stopped, and the line may be switched to a collection means for collecting or discarding plasma contaminated with blood cells and / or hemoglobin. This ensures that plasma useful as a clinical test sample can be obtained without contamination with blood cells and hemoglobin. In addition,
The blood plasma in which blood cells are detected, or the blood cell components that have permeated late can be returned to the living body or the subject.

Examples of means for detecting blood cells and / or hemoglobin (blood cells and / or hemoglobin) in the plasma permeate include optical means and color developing means by chemical reaction. Conveniently, blood cells and / or hemoglobin can be detected directly by optical means. For example, when blood cells are mixed in plasma, the light transmittance is lowered, or when hemoglobin is mixed, the plasma becomes red. Either case can be detected by optical means. For example, a blood leak sensor that detects blood leak above a certain value by an optical method may be used. The blood leak sensor is a sensor that detects blood cells and hemoglobin in a solution, and, for example, the absorbance or the transmittance of the solution at a certain wavelength is measured by a spectrophotometer. Specifically, leakage of blood cells and / or hemoglobin in the solution is measured by measuring the turbidity of the blood cells in the solution (eg, the absorbance at a wavelength of 630 nm) or the absorbance of hemoglobin (eg, the absorbance at a wavelength of 430 nm). Can be detected.

As a coloring means by a chemical reaction, there is a method of utilizing coloring by a chemical reaction with blood cells and / or hemoglobin. This color development can be detected by optical means. For example, a method utilizing the peroxidase-like action of hemoglobin can be mentioned. By choosing the appropriate peroxide and chromogen, the sensitivity can be adjusted.

In view of the object of the present invention, any means other than optical means may be used as long as it is possible to detect blood cells and / or hemoglobin. For example, blood cells can be detected by a cell counter which is an electric cell counter.

Of the blood cell and / or hemoglobin detecting means, the optical means may be arranged between the outlet which is the flow path of plasma and the collecting means. For example, a sensor may be inserted externally and contacted directly with plasma to detect blood cells and / or hemoglobin. Alternatively, the blood cells and / or hemoglobin can be detected by arranging them so as to sandwich a pipe, which is a flow path of plasma, and causing light, laser, or the like to emit light from one side and receiving the emitted light on the other side. In this case, the pipe is preferably transparent so that light can pass therethrough. When a pipe having poor light transmittance is used, the light transmittance of only the pipe may be measured in advance. Further, the pipe may be provided with a detection means by fine sampling, and plasma may be collected to detect the mixing of blood cells and / or hemoglobin with the above-mentioned coloring reagent, for example, a test paper. When the blood cells and / or hemoglobin are detected, the blood cell and / or hemoglobin-containing plasma may be discarded or collected by the switching means. further,
This blood cell and / or hemoglobin detection means can be connected to a blood supply means, a pressurization means, or a switching means described below, and can be linked to processing such as stopping the supply or pressurization, discarding or collecting by the switching means.

[0121] Plasma generally contains some hemoglobin even in the absence of hemolysis, and its concentration varies among individuals. Further, in blood collection using a vacuum blood collection tube which has been widely used in recent years, it is known that some hemolysis occurs during blood collection. However, it has been found that the presence of hemoglobin in plasma at such a low concentration does not significantly affect clinical laboratory data. Therefore, the concentration of blood cells or hemoglobin contained in plasma has an allowable range, and there is no particular problem even if blood cells and / or hemoglobin are contained within the range. For example, the concentration of red blood cells in the obtained plasma is 0.1% with respect to the concentration of red blood cells in blood before separation.
The following is acceptable.

Therefore, in the device of the present invention, when the blood cell and / or hemoglobin detection means detects a blood cell and / or hemoglobin concentration exceeding a certain concentration, the switching means is operated to activate the blood cell and / or hemoglobin. It can be connected to discard hemoglobin-enriched plasma or to stop the supply or pressurization of blood.

<Switching Means> The switching means for separating plasma containing blood cells and / or hemoglobin is arranged after the outlet. Examples of the switching means include a switching cock and a switching valve. The switching means may be either automatic or manual. In the case of automatic operation, the blood cell and / or hemoglobin detection means is arranged at a position before plasma passes through the switching means. The switching means is connected to the blood cell and / or hemoglobin detection means. Plasma in which blood cells and / or hemoglobin is mixed with blood cells and / or hemoglobin from a line for collecting plasma when the blood cells and / or hemoglobin exceed a preset value, and the signal from the blood cell and / or hemoglobin detection means activates the switching means. Switch to a line for collection or disposal. Recovery,
As the discarding means, the above plasma collecting means can be used.

(Function) The separation mechanism of blood cells and plasma in the present invention mainly utilizes the difference in the moving speed in the aggregate of the ultrafine fibers of both components. The migration velocity of the ultrafine fibers in the aggregate is higher in the plasma component than in the blood cell component. Therefore, when blood is supplied to the aggregate of ultrafine fibers and pressure is applied, the plasma moves ahead of the blood cells, and after moving a certain distance, they are completely separated. Therefore, first the plasma flows out, then the blood cells flow out,
Plasma can be collected by utilizing this time difference.

As described above, the present invention is fundamentally different from the centrifugal separation method utilizing the difference in specific gravity of blood components or the membrane separation method utilizing the difference in size of blood components. Furthermore, it is also different from the method for separating white blood cells, which utilizes only the adsorptivity of blood components to ultrafine fibers.

[0126]

The plasma separation device of the present invention utilizes the difference in the moving speed of plasma and blood cells. Therefore, the obtained plasma component does not undergo dilution, does not change the component, and is no different from the plasma obtained from centrifugation. Furthermore, by providing a blood cell and / or hemoglobin detection device on the permeate side and a cock interlocked with this, it is possible to prevent blood cells and / or hemoglobin from being mixed in the plasma for sampling. As described above, according to the device of the present invention, a blood plasma sample free of blood cells and / or hemoglobin can be efficiently, conveniently, rapidly and safely obtained. Therefore, the device of the present invention is useful in a field such as a clinical test where it is required to separate a small amount of blood into plasma or serum in a short time. Further, the plasma separation device of the present invention enables automation of collection of plasma samples for clinical tests, which has hitherto relied on human hands, and can greatly contribute to automation, speedup, and improvement of safety of clinical tests.

Hereinafter, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

[0128]

EXAMPLES In the examples, as an assembly of ultrafine fibers, ultrafine fibers made of polyethylene terephthalate spun by an ordinary melt blow method (density: 1.38 g / cm 2).
^A non-woven fabric consisting of ³ ) was used. As bovine blood, fresh bovine blood within 8 hours after blood collection was used, but an anticoagulant (ACD: dextrose citrate) was added immediately after blood collection to prevent blood coagulation. The red blood cell concentration in this bovine blood was 7.2 × 10 ⁹ cells / ml, the white blood cell concentration was 6.8 × 10 ⁶ cells / ml, and the platelet concentration was 2.1 × 10 ⁸ cells / ml.

The performance of the plasma separation filter is determined by measuring the concentration of blood cells (erythrocytes, white blood cells, platelets) in plasma, the presence or absence of hemolysis of red blood cells, and blood components (serum amylase, total cholesterol, neutral fat, urea nitrogen, blood glucose, Total protein concentration,
Albumin) was analyzed and evaluated. The plasma obtained by centrifuging the above bovine blood was used for comparison.

The concentration of contaminated blood cells in plasma was measured by the Coulter counter method. The detection limit of each blood cell is 2 × 10 ⁵ cells / ml in red blood cell concentration and 1 × 10 ⁵ cells / m in white blood cell concentration.
1 and the platelet concentration was 1 × 10 ⁶ cells / ml.

Hemolysis was detected by measuring the hemoglobin concentration in plasma using the O-toluidine method.

The analysis of each blood component was performed by the following method.

(I) Serum amylase: G5CNP method (ii) Total cholesterol: cholesterol oxidase
Peroxidase coloring method (iii) Neutral fat: LPL / glycerol triphosphate oxidase / peroxidase coloring method (iv) Urea nitrogen: urease / indophenol method (v) Blood glucose: glucose oxidase / peroxidase coloring method (vi) Total protein concentration: Buuret Method (vii) Albumin: BCG (Bromcresol Green) Method

Example 1 A plasma separation filter as shown in FIG. 1 was prepared. 1.0 mm diameter hole at the top edge of the container as an inlet and 1.0 m diameter at the center of the bottom surface as an outlet
m hole, the inside diameter of the container is 52.0 mm, and the thickness is 2.
A 0 mm acrylic disk-shaped container was prepared. In this container, a non-woven fabric made of polyethylene terephthalate ultrafine fibers having an average fiber diameter of 1.8 μm as an aggregate of ultrafine fibers (Basis weight 50 g / m ² , thickness about 2 mm) 1.4 g (14
The sheets were laminated so that the diameter was 50.0 mm and the thickness was 2.0 mm. At the same time, a gap having a width of 1.0 mm was provided between the outer peripheral surface of the ultrafine fiber aggregate and the inner peripheral surface of the container. Fabricated filter, the volume of the aggregate of ultrafine fibers: about 3.9 cm ^3, gap volume: of about 0.32 cm ^3. The total surface area of the ultrafine fibers, the average bulk density of the aggregate of the ultrafine fibers, the average hydraulic radius, the pore volume and L / D in this plasma separation filter are as described in Table 1. Each value was calculated by the following formula. The density of polyethylene terephthalate is 1.38 g as described above.
/ Cm ³ .

Total surface area = 4 × weight of aggregate of ultrafine fibers / (density of ultrafine fibers × average fiber diameter) = 4 × 1.4 g / (1.38 g / cm ³ × 1.8 μm) = 2.3 m ² Average bulk density = weight of aggregate of ultrafine fibers / volume of aggregate of ultrafine fibers = 1.4 g / 3.9 cm ³ = 0.36 g / cm ³ average hydraulic radius = average fiber diameter × (density of ultrafine fibers− average bulk density) / (4 × average bulk density) = 1.8μm × (1.38g / cm 3 -0.36g / cm 3) /(4×0.36g/cm 3) = 1.28μm pore volume = Volume of ultrafine fiber aggregate- (weight of ultrafine fiber aggregate / density of ultrafine fiber) = 3.9 cm ^3- (1.4 g ÷ 1.38 g / cm ³ ) = 2.9 cm ³ L / D ( flow path length of the blood component / channel diameter of a blood component) = L / 2 (the cross-sectional area of the aggregate surface of the ultrafine fibers of the blood inlet / π) ^0.5 = (2 5cm) / 2 ((5.0cm × π × 0,2cm) ÷ π) 0.5 = 1.25 above the plasma separation filter bovine blood 4ml injected from the inlet of the container, the outer peripheral portion surface of the aggregate of ultrafine fibers After filling in the gap on the inner peripheral surface of the container, a pressure of 0.2 kg / cm ² was applied to push out bovine blood. Bovine blood enters from the outer peripheral surface of the ultrafine fiber aggregate, moves horizontally from the outer peripheral portion toward the center of the circle of the ultrafine fiber aggregate, and after pressurization 2
After 0 seconds, plasma started to flow out from the outlet of the container, and 4 seconds after that, red blood cells began to be mixed in the effluent. Therefore, it started to flow out and collected plasma for about 2 seconds,
0.29 ml was obtained. The average linear processing speed was 75 mm / min (= 25 mm / 20 seconds).

The concentration of red blood cells, white blood cells and platelets in 0.29 ml of plasma obtained above was below the detection limit. Therefore, the red blood cell concentration in the obtained plasma (hereinafter referred to as the red blood cell contamination ratio) was 0.003 with respect to the red blood cell concentration in the bovine blood before separation.
% (= (2 × 10 ⁵ ÷ 7.2 × 10 ⁹ ) × 100) or less. The hemoglobin concentration of the obtained plasma was 5 mg.
/ Dl, and the analysis results of plasma components are shown in Table 1.

Example 2 A plasma separation filter was prepared in the same manner as in Example 1. However, the filling amount of non-woven fabric is 0.8
g (8 layers).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average fluid radius, the pore volume and L / D are as shown in Table 1, respectively.

Plasma was separated in the same manner as in Example 1 using the above plasma separation filter. Plasma started to flow out 15 seconds after the pressurization, and 2 seconds after that, the red blood cells started to be mixed. Therefore, plasma was collected for about 1.5 seconds after starting to flow out, and 0.33 ml was obtained. The average linear processing speed was 100 mm / min.

The white blood cell and platelet concentrations in 0.33 ml of plasma obtained above were below the detection limit, and the red blood cell concentration was 3 × 10 ⁵ cells / ml. Therefore, the red blood cell contamination rate was 0.004%. The hemoglobin concentration of the obtained plasma was below the detection limit, and the analysis results of the plasma components are shown in Table 1.

Example 3 A plasma separation filter as shown in FIG. 3 was prepared. 1.0 mm diameter hole at the top edge of the container as an inlet and 1.0 m diameter at the center of the bottom surface as an outlet
with a diameter of 29.0 mm and a thickness of 6.
A columnar container made of 5 mm polypropylene was prepared.
In this container, an average fiber diameter of 0.
Nonwoven fabric made of ultrafine fibers made of polyethylene terephthalate of 8 μm (Basis weight 50 g / m ² , thickness about 3 mm) 1.2
g (12 layers), diameter 29.0 mm, thickness 6.0
so that the gap becomes 0.5 mm, a gap of 0.5 mm in thickness is provided between the upper surface of the aggregate of ultrafine fibers and the inner surface of the container,
A non-woven fabric made of ultrafine fibers made of polyethylene terephthalate having an average fiber diameter of 10 μm (weight per unit area: 50 g /
m ^2, a thickness of about 1 mm) 0.33 g (two-layer) to prepare a plasma separation filter filled with (collection of the ultrafine fiber volume: about 4.0 cm ^3, gap volume: about 0.33c
m ³ ).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average fluid radius, the pore volume and L / D are as shown in Table 1, respectively.

4 ml of bovine blood in the above plasma separation filter
Was injected from the inlet of the container to fill the ultrafine fibers in the gap between the upper surface of the aggregate of ultrafine fibers and the upper part of the inner surface of the container, and
Bovine blood was extruded by applying a pressure of kg / cm ² . Bovine blood enters from the upper surface of the ultrafine fiber aggregate, moves downward inside the ultrafine fiber aggregate, and 30 seconds after pressurization, plasma begins to flow out from the outlet of the container, and further after that. 5
Erythrocytes began to be mixed in the effluent at the time of elapse of seconds. Therefore, the plasma was collected for about 3 seconds after it started to flow out,
ml were obtained. The average linear processing speed was 12 mm / min.

The blood cell concentrations in 0.31 ml of plasma obtained above were all below the detection limit. Therefore, the red blood cell contamination rate was 0.003% or less. The hemoglobin intensity of the obtained plasma was 8 mg / dl, and the analysis results of the plasma components are shown in Table 1.

Example 4 A plasma separation filter as shown in FIG. 1 was prepared. 1.0 mm diameter hole at the top edge of the container as an inlet and 1.0 m diameter at the center of the bottom surface as an outlet
Has a hole of m, the inside diameter of the container is 134 mm, and the thickness is 0.3.
A 0 mm acrylic disk-shaped container was prepared. The vessel nonwoven (basis weight 50 g / m ^2, thickness 2 mm) made of the average fiber diameter 3.0μm polyethylene terephthalate of the ultrafine fibers as a collection of ultrafine fibers 2.0g of (3 sheets of layer), diameter 130mm The layers were laminated so that the thickness was 0.30 mm. At the same time, a gap having a width of 2.0 mm was provided between the outer peripheral portion of the ultrafine fiber aggregate and the inner peripheral portion of the container.
A plasma separation filter as shown in (1) was prepared (volume of ultrafine fiber aggregate: about 4.0 cm ³ , gap volume: about 0.
25 cm ³ ).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average fluid radius, the pore volume and L / D are as shown in Table 1, respectively.

Plasma was separated in the same manner as in Example 1 using the above plasma separation filter. However, 0.4 kg / c
Bovine blood was extruded by applying a pressure of m ² . The plasma started to flow 120 seconds after the pressurization, and 18 seconds after that, the red blood cells started to be mixed. Therefore, plasma was collected for 12 seconds from the start of outflow to obtain 0.26 ml. The average linear processing speed was 32.5 mm / min.

The white blood cell and platelet concentrations in 0.26 ml of the plasma obtained above were below the detection limit, and the red blood cell concentration was 2.9 × 10 ⁸ cells / ml. Therefore, the red blood cell contamination rate was 0.04%. The hemoglobin concentration of the obtained plasma was 4 mg / dl, and the analysis results of the plasma components are shown in Table 1.

Example 5 A plasma separation filter as shown in FIG. 2 was prepared. 1.0 mm diameter hole at the top edge of the container as an inlet and 1.0 m diameter at the center of the bottom surface as an outlet
Has a hole of m, the inside diameter of the container is 15,2 mm, and the thickness is 24.
A polypropylene cylindrical container of mm was prepared. In this container, an average fiber diameter of 1.8μ as an aggregate of ultrafine fibers
m of polyethylene terephthalate ultrafine fiber 1.4 g of cotton with a thickness of 2.0 mm on the upper surface of the aggregate of ultrafine fibers and the upper surface of the inner surface of the container to have a diameter of 15.2 mm and a thickness of 22 mm. A plasma separation filter filled with a gap was produced (volume of ultrafine fiber aggregate: about 4.
0 cm ³ , gap volume: about 0.36 cm ³ ).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average hydraulic radius, the pore volume and L / D are as shown in Table 1, respectively.

4 ml of bovine blood in the above plasma separation filter
Was injected from the inlet of the container to fill the gap between the upper surface of the ultrafine fiber aggregate and the upper part of the inner surface of the container, and then 0.4 kg / cm
A pressure of ² was applied to extrude bovine blood. Raw blood enters from the upper surface of the ultrafine fiber aggregate, moves downward inside the ultrafine fiber aggregate, and plasma starts to flow out 300 seconds after pressurization, and 70 seconds after that. After that, red blood cells started to mix. Therefore, plasma was collected for 30 seconds from the beginning of the outflow to obtain 0.30 ml. The average linear processing speed was 4.4 mm / min.

The blood cell concentration in 0.30 ml of plasma obtained above was all below the detection limit. Therefore, the red blood cell contamination rate was 0.003% or less. The hemoglobin concentration of the obtained plasma was 6 mg / dl, and the analysis results of the plasma components were as shown in Table 1.

(Example 6) A plasma submerged filter was prepared in the same manner as in Example 3. However, as the aggregate of the ultrafine fibers, cotton made of polyethylene terephthalate ultrafine fibers having an average fiber diameter of 0.8 μm was used as the aggregate of the ultrafine fibers.

The total surface area of the ultrafine fibers, the average bulk density of the aggregate of the ultrafine fibers, the average hydraulic radius, the pore volume and the L / D in the above plasma separation filter are the same as in Example 3 and are shown in Table 1. As stated.

Plasma was separated in the same manner as in Example 3 using the above plasma separation filter. The plasma started to flow out 880 seconds after pressurization, and the red blood cells started to mix after a further 115 seconds. Therefore, 90
The second blood plasma was collected to obtain 0.31 ml. The average processing linear velocity was 0.41 mm / min.

The blood cell concentration in 0.31 ml of plasma obtained above was all below the detection limit. Therefore, the red blood cell contamination rate was 0.003% or less. The hemoglobin concentration of the obtained plasma was 10 mg / dl, and the analysis results of the plasma components are shown in Table 1.

Example 7 A plasma separation filter was prepared in the same manner as in Example 5. However, the diameter inside the container is 10.0
mm using a container of 55.0 mm in thickness, 2.0 g of cotton made of ultra-fine fibers having an average fiber diameter of 3.2 μm as an aggregate of ultra-fine fibers to have a diameter of 10.0 mm and a thickness of 51.0 mm. A gap having a thickness of 4.0 mm was provided between the upper surface of the ultrafine fiber aggregate and the upper surface of the container (volume of the ultrafine fiber aggregate: about 4.0 cm ³ , volume of the gap: about 0.31 c).
m ³ ).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average hydraulic radius, the pore volume and L / D are as shown in Table 1, respectively.

Plasma was separated in the same manner as in Example 5 using the above plasma separation filter. Effluent started 264 seconds after pressurization of the plasma, and erythrocytes began to be mixed after 25 seconds thereafter. Therefore, plasma was collected for 22 seconds after starting to flow out, and 0.26 ml was obtained. The average linear processing speed was 11.6 mm / min.

The white blood cell and platelet concentrations in 0.26 ml of plasma obtained above were below the detection limit, and the red blood cell concentration was 4.3 × 10 ⁸ cells / ml. Therefore, the red blood cell contamination rate was 0.06%. The hemoglobin concentration of the obtained plasma was below the detection limit, and the analysis results of the plasma components are shown in Table 1.

Example 8 A plasma separation filter was prepared in the same manner as in Example 5. However, the filling amount of cotton composed of ultrafine fibers was set to 0.8 g. The total surface area of the ultrafine fibers of this filter, the average bulk density of the aggregate of the ultrafine fibers, the average hydraulic radius, the void volume and L / D are shown in Table 1, respectively.

Example 5 using this plasma separation filter
Plasma separation was performed in the same manner as in. The plasma started to flow out about 150 seconds after the pressurization, and after a further 20 seconds, the red blood cells started to be mixed. Therefore, 1
Plasma was collected for 7 seconds to obtain 0.34 ml. The average processing linear velocity was 8.8 mm / min.

The concentration of white blood cells and platelets in 0.34 ml of plasma obtained above was below the detection limit, and the concentration of red blood cells was 2.2 × 10 ⁸ cells / ml. Therefore, the red blood cell contamination rate was 0.03%. The hemoglobin concentration of the obtained plasma was below the detection limit, and the analysis results of the plasma components are shown in Table 1.

[0164]

[Table 1]

Comparative Example 1 A plasma separation filter was prepared in the same manner as in Example 1. However, as an aggregate of ultrafine fibers, a non-woven fabric made of ultrafine fibers having an average fiber diameter of 2.5 μm (Basis weight 50 g / m ² , thickness 2 mm) 0.8 g (8 layers are stacked)
Was charged. The total surface area of the ultrafine fibers in this filter,
The average bulk density, average hydraulic radius, pore volume, and L / D of the ultrafine fiber aggregate are as shown in Table 2.

Plasma was separated in the same manner as in Example 1 using the above plasma separation filter. The effluent began to appear 10 seconds after the pressurization, but blood cells were mixed in the effluent from the beginning. Collect the effluent for about 1.0 seconds after it begins to flow out,
0.33 ml was obtained.

The white blood cell and platelet concentrations in the 0.33 ml effluent obtained above were below the detection limit, but the red blood cell concentration was 3.8 × 10 ⁸ cells / ml. Therefore, the erythrocyte contamination rate was 5.3%. The hemoglobin concentration in plasma of the obtained effluent was below the detection limit.
Analysis of plasma components was omitted.

Comparative Example 2 A plasma separation filter was produced in the same manner as in Example 1. However, as an aggregate of ultrafine fibers, a non-woven fabric made of ultrafine fibers having an average fiber diameter of 1.0 mm (Basis weight: 50 g / m ² , thickness: 2 mm) 2.0 g (20 layers are stacked)
Was charged. The total surface area of the ultrafine fibers in this plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average fluid radius, the pore volume and L / D are as described in Table 2.

Plasma was separated in the same manner as in Example 1 using the above plasma separation filter. However, 0.4 kg / cm
Pressed at ² . The effluent started to appear 1365 seconds after pressurization, but the effluent was red from the beginning. The effluent for 1600 seconds after starting to flow out was collected to obtain 0.26 ml.

The blood cell concentration in the 0.26 ml effluent obtained above was all below the detection limit, but the hemoglobin concentration in plasma of the effluent was 95 mg / dl. Analysis of plasma components was omitted.

Comparative Example 3 A plasma separation filter was prepared in the same manner as in Example 3. However, the diameter inside the container is 50.0m
m, 2.3 mm thick acrylic columnar container, non-woven fabric made of polyethylene terephthalate ultrafine fibers with an average fiber diameter of 1.8 μm as aggregate of ultrafine fibers (Basis weight 50 g / m ² , thickness about 1 mm) 1.4g (14 layers are stacked) with a diameter of 50.0 mm and a thickness of 2.0 mm, with a gap of 0.3 mm between the upper surface of the ultrafine fiber aggregate and the upper inner surface of the container The non-woven fabric made of ultrafine fibers made of polyethylene terephthalate having an average fiber diameter of 10 μm (weight per unit area: 50 g / m ² , thickness: about 1 m).
m) A plasma separation filter filled with 0.17 g (1 sheet) was prepared (volume of ultrafine fiber aggregate: 3.9 cm ³ ,
Gap volume: 0.59 cm ³ ).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average hydraulic radius, the pore volume and L / D are as shown in Table 2, respectively.

Plasma was separated in the same manner as in Example 3 using the above plasma separation filter. The effluent began to appear 15 seconds after pressurization, but blood cells were mixed in the effluent from the beginning. Collect the effluent about 1.5 seconds after it started to flow out,
30 ml was obtained.

The concentration of white blood cells and platelets in 0.30 ml of the effluent obtained above was below the detection limit, but the concentration of red blood cells was 6.9 × 10 ⁹ cells / ml. Therefore, the red blood cell contamination rate was 55%. The hemoglobin concentration in plasma of the obtained effluent was below the detection limit. The analysis of blood components was omitted.

(Comparative Example 4) A plasma separation filter was prepared in the same manner as in Example 5. However, the diameter inside the container is 8.0m
m, a polypropylene container having a thickness of 86.0 mm, and the aggregate of ultrafine fibers has a diameter of 8.0 mm and a thickness of 80.0 mm. And filled with a gap of 6.0 mm (the volume of the ultrafine fiber aggregate: 4.0 cm ³ , the gap volume: 0.30 c).
m ³ ). The total surface area of the ultrafine fibers, the average bulk density of the aggregate of the ultrafine fibers, the average fluid radius, the void volume and L / D in this plasma separation filter are as described in Table 2.

Plasma was separated in the same manner as in Example 5 using the above plasma separation filter. The effluent is pressurized after 1920
It started to appear after 2 seconds, but the effluent was red from the beginning. It started to flow out and got effluent for 3600 seconds,
Only 0.25 ml was obtained.

The blood cell concentration in the 0.25 ml effluent obtained above was all below the detection limit, but the hemoglobin concentration in the effluent plasma was 75 mg / dl. Analysis of plasma components was omitted.

(Comparative Example 5) The same plasma separation filter as in Example 1 was prepared, and plasma was separated in the same manner as in Example 1.
However, the amount of bovine blood used was 0.5 ml.

A permeate could not be obtained even if all the blood was fed into the ultrafine fibers. Therefore, pressurized air was further sent from the container inlet, but the pressurized air passed through the ultrafine fibers faster than blood, and only bubbled blood containing air was obtained at the outlet. It was considered that this was because the amount of blood to be treated was too small and the blood could not be uniformly fed, as compared with the void volume of 3.0 ml of the aggregate of ultrafine fibers.

Comparative Example 6 A plasma separation filter was prepared in the same manner as in Example 3. However, the diameter inside the container is 35.0
mm, 4.6 mm thick polypropylene container, a non-woven fabric made of polyethylene terephthalate ultrafine fibers having an average fiber diameter of 1.8 μm (aggregate weight 50 g / m ² , thickness 1 mm) as an aggregate of ultrafine fibers 0.87 g ( 18 layers) are filled with a gap of 0.4 mm in thickness between the upper surface of the aggregate of ultrafine fibers and the upper surface of the container so that the diameter is 35.0 mm and the thickness is 4.2 mm. A non-woven fabric made of ultrafine fibers made of polyethylene terephthalate having an average fiber diameter of 10 μm (weight per unit area: 50 g / m ² , thickness: 1 mm) in the gap.
87 g (one layer was overlaid) was filled (volume of ultrafine fiber aggregate: 4.0 cm ³ , gap volume: 0.38 cm ³ ).

The total surface area of the ultrafine fibers in the plasma separation filter, the average bulk density of the aggregate of the ultrafine fibers, the average fluid radius, the pore volume and L / D are as shown in Table 2.

Plasma separation was carried out in the same manner as in Example 3 using the above plasma separation filter. The effluent began to emerge 15 seconds after pressurization, or the effluent was contaminated with red blood cells from the beginning. Collect the effluent for about 1.5 seconds after it begins to flow out,
0.34 ml was obtained. Average processing linear velocity is 16.8 mm /
Minutes.

The white blood cell and platelet concentrations in the 0.34 ml effluent obtained above were below the detection limit, but the red blood cell concentration was 1.1 × 10 ⁹ cells / ml. Therefore, the residual amount of red blood cells was 15%. The hemoglobin concentration in plasma of the obtained effluent was below the detection limit. Analysis of plasma components was omitted.

[0184]

[Table 2]

Example 9 In the same disc-shaped container as in Example 1, 2 g of polyethylene terephthalate ultrafine fiber nonwoven fabric having an average fiber diameter of 1.5 μm is 50 mm in diameter and 2.0 m in thickness.
It was cut into m and filled with a gap of 1.0 mm between the inner peripheral surface of the container and the container. The average hydraulic radius at this time is 1. 0
4 μm.

ACD solution was added to this as an anticoagulant,
Bovine blood having a hematocrit of 45% was fed from the outer peripheral portion of the container to the inner portion at a pressure of 0.5 kg / cm ² . Blood passed through the assembly of ultrafine fibers in the container in the horizontal direction with respect to the non-woven fabric surface, and after about 30 seconds, plasma had permeated from the outlet. The permeation of plasma lasted for about 45 seconds, after which blood cells began to mix in the permeate. Therefore, plasma without blood cell contamination could be collected for 30 to 45 seconds after pressurization, and the total volume was 0.7 ml. The hemoglobin concentration of the obtained plasma was 3 mg / d.
Hemolysis was not observed below l. In addition, red blood cells, white blood cells, and platelets measured by a blood cell analyzer were all below the detection limit. The biochemical analysis values of the obtained plasma are shown in Table 3. The permeate at the beginning of collection, the permeate at the end of collection, a sample of 0.25 ml corresponding to 10% of the interstitial volume of the aggregate of ultrafine fibers, and plasma obtained by centrifugation of the same blood were compared. There was no significant difference between them. When the amount of fibrinogen in the obtained plasma was quantified by cellulose acetate electrophoresis, it was reduced to about 30% of the concentration in the plasma obtained by centrifugation, but it was not completely removed. No precipitation of fibrin was observed when left standing overnight.

(Example 10) The same filter as that used in Example 9 was used. 0.5 kg of hematocrit 47% human blood immediately after blood collection without adding anticoagulant
The solution was sent at a pressure of / cm ² . Blood passes through the assembly of ultrafine fibers in the container in the horizontal direction with respect to the surface of the non-woven fabric,
A permeate was obtained from the outlet after 5 seconds. About 55 permeate
Blood cells started to mix in seconds. The permeated liquid free from blood cells can be collected from 35 seconds to 20 seconds after pressurization, and the total amount is 1.
It was 0 ml. The hemoglobin concentration of the obtained permeate was 3 mg / dL or less, and no hemolysis was observed. In addition, red blood cells, white blood cells, and platelets measured by a blood cell analyzer were all below the detection limit. Table 3 shows the biochemical analysis values of the obtained permeated liquid. The permeate at the beginning of collection, the permeate at the end of collection, a sample in which 0.25 ml corresponding to 10% of the interstitial volume of the ultrafine fiber aggregate was collected, and the serum obtained by centrifugation of the same blood after centrifugation were compared. However, there was no significant difference between them.

Fibrinogen was not detected in the obtained permeate, and the obtained liquid was serum instead of plasma. The fibrinogen concentration in the plasma obtained by centrifuging blood to which heparin was added as an anticoagulant from the same person was 220 mg / dL.

Despite performing the same experiment as in Example 1 except that human blood was used, the difference in the amount of permeate that could be collected was observed because the size of the red blood cells was different between human and bovine. It is thought that it is because it is different. Further, it is considered that the reason why fibrinogen was completely removed was that the anticoagulant was not added and thus the fibrinogen was adsorbed on the filter.

(Example 11) The non-woven fabric of polyethylene terephthalate ultrafine fibers used in Example 1 was dipped in a 0.1% polyvinylpyrrolidone K-90 (Koridone K-90 manufactured by BASF, molecular weight 360,000) solution. , 50KG in wet condition
The y-ray irradiation of y was performed. The non-woven fabric after γ-ray irradiation is washed with pure water to remove the non-crosslinked polyvinylpyrrolidone,
It was dried to obtain a nonwoven fabric having polyvinylpyrrolidone fixed on the surface. This nonwoven fabric was filled in a container in the same manner as in Example 1 and the same experiment as in Example 10 was performed. Blood moved in the container in parallel with the laminated surface of the nonwoven fabric, and after about 15 seconds, plasma came out from the outlet. After about 25 seconds, blood cells started to mix. Plasma free from blood cells could be collected for about 10 seconds. The total volume was 1.2 ml. The hemoglobin concentration in the obtained plasma was 3 mg / dl or less, and hemolysis was not observed. In addition, red blood cells, white blood cells, and platelets measured by a blood cell analyzer were all below the detection limit. The analytical values of the obtained plasma are shown in Table 3. The permeate at the beginning of collection, the permeate at the end of collection, a sample in which 0.25 ml corresponding to 10% of the interstitial volume of the ultrafine fiber aggregate was collected, and the serum obtained by centrifugation of the same blood after centrifugation were compared. However, there was no significant difference between them.

When the amount of fibrinogen in the obtained plasma was measured by cellulose acetate electrophoresis,
It was reduced to about 20% of the plasma concentration obtained by centrifugation, but was not completely eliminated. No precipitation of fibrin was observed when left standing overnight.

By fixing polyvinylpyrrolidone to the ultrafine fibers, the affinity of blood for the ultrafine fibers was improved, the resistance of blood passing through was reduced, and the time until the plasma was separated was shortened.

(Comparative Example 7) Glass fiber filter paper (thread diameter 0.8
~ 2.5 μm) is cut into about 2 mm square, water is added and the mixture is stirred with a mixer, dehydrated, and then filled in a 5 mL syringe container as shown in FIG. 4 so that the filling rate is 0.5 g / cm ^3. did. The average hydraulic radius is 1.45 μm (specific gravity of glass fiber 2.2). An ACD solution was added to this as an anticoagulant, and bovine blood having a hematocrit of 45% was sent from the upper part of the container at a pressure of 0.5 kg / cm ² . Blood passed through the assembly of ultrafine fibers from the upper part to the lower part in the container, and after about 60 seconds, plasma had permeated from the outlet. Permeation of plasma continued until after about 90 seconds, after which blood cells started to be mixed in plasma. Plasma without blood cells can be collected for 60 to 30 seconds after pressurization, and the total volume is 0.2m.
l. The obtained plasma hemoglobin concentration is 3 mg.
No hemolysis was observed below / dL. In addition, red blood cells, white blood cells, and platelets measured by a blood cell analyzer were all below the detection limit. The biochemical analysis values of the obtained plasma are shown in Table 3. Permeate at the beginning of collection, permeate at the end of collection, obtained
A 0.2 ml sample and plasma obtained by centrifugation of the same blood were compared. The obtained 0.2 ml sample was used for 10% of the void volume of the ultrafine fiber aggregate.
This is because a sample in which 0.25 ml was collected could not be obtained.
In the permeate at the early stage of collection, the total protein concentration was significantly lower than the others, and the measured values of electrolytes and lipids were different from those of the centrifuged plasma. It ’s a protein, an electrolyte,
It is considered that the lipid is adsorbed to the glass fiber and a part of the electrolyte is eluted from the glass fiber into plasma. Also,
When the amount of fibrinogen in the obtained plasma was determined by a cellulose acetate electrophoresis method, it was reduced to about 30% of the concentration in the plasma obtained by centrifugation, but it was not completely removed. No precipitation of fibrin was observed when left standing overnight. Table 3 shows the results.

(Comparative Example 8) A polyethylene terephthalate nonwoven fabric having a yarn diameter of 3.5 μm was added to a container similar to that of Example 1.2.
4 g was filled. The average fluid radius at this time is 1.13 μm.
It is. When blood was transferred to this in the same manner as in Example 1, a permeate was obtained after about 45 seconds, but this permeate contained red blood cells from the beginning and plasma could not be separated. It is considered that this is because the fiber diameter is as large as 3.5 μm, and therefore the degree of deformation when red blood cells pass through the fiber gap is small, and a difference in the moving speed of red blood cells and plasma did not occur. Table 3 shows the results.

[0195]

[Table 3]

(Embodiment 12) FIG. 8 is a schematic diagram showing an example of the configuration of the apparatus of the present invention. The container 31 is a container for storing a blood sample. The container 31 is connected to the plasma separation filter 11 via a line (or tube) 32 for supplying blood. Further, the container 31 is connected to a supply line 33 for pressurized air.

The container 31 has a lid 3 which can be hermetically attached and detached.
4 is attached. The container 31 may further have a stirring function or a shaking function. A line for supplying an anticoagulant may be attached to the container 31 or the line 32 (not shown).

The blood 26 in the container 31 is pushed out by the compressed air sent from the compressor 35 through the line 33, and is supplied to the filter 11 through the line 32. The pressure of the compressed air is adjusted by the compressor 35.

The blood supplied to the filter 11 passes through the ultrafine fiber assembly 14 by the pressure of the pressurized air sent to the container 31, and is separated into plasma and blood cells therein.
The plasma flowing out from the permeated liquid side of the filter 11 passes through the three-way cock 36, passes through the line (or tube) 37 for collecting plasma, and is collected in the container 38 for storing the sample plasma.

[0200] On the permeate side of the filter 11, blood cells and / or blood cells and / or hemoglobin for detecting contamination of hemoglobin are detected.
Alternatively, the hemoglobin detection means 40 is attached. In addition to the blood cell and / or hemoglobin detection means 40, a sampling port for detecting contamination of blood cells and / or hemoglobin by a chemical reaction method or a cell counter may be provided. When blood cells and / or hemoglobin are detected in the plasma of the permeate, the three-way cock 36 (which may be, for example, a 3-port valve that can be opened and closed by electromagnetic action) is switched, and the permeate is drained (or collected) line 41. Then, it is sent to the waste liquid (or recovery) tank 42. At the same time, the supply of pressurized air to the container 31 is stopped, and the plasma separation operation ends.

These means can be connected to the blood cell and / or hemoglobin detection means, for example, to automatically terminate the plasma separation operation when blood cells and / or hemoglobin are detected.

Further, to the blood delivery line 32 and the plasma delivery line 37, a pure water supply line 43 for supplying pure water as washing water and a dry air supply line 44 for supplying dry air for drying the apparatus. Can be connected. In addition to the above, as a method of cleaning the apparatus of the present invention, a method of mechanically brushing a glass instrument or the like with a rotating brush, a method of using a jet jet to flow a high-pressure water stream through a nozzle, and an ultrasonic method And so on. These washing methods can be used alone or in combination. By the method described above, the line after the plasma separation operation is quickly washed and dried, and the next blood sample can be continuously processed.

In addition to the above three-way cock 36, various members such as a cock, a chamber, and a pressure sensor for switching the flow direction of the fluid may be arranged in each line constituting the plasma separation device (see FIG. (Not shown).

The filter 11 can be detachably attached to a holder or the like (see, for example, FIG. 5). Retention and release by magnets can be applied. The filter 11 includes an inlet side portion (FIGS. 5 and 15a), an outlet side portion (FIGS. 5 and 15b), and an assembly of ultrafine fibers (FIGS. 5 and 14).
Can be separately configured to be detachable. By detaching the portion containing the aggregate of ultrafine fibers to be attached, it can be made disposable,
Contamination can be prevented. If the portion containing the polymer of the ultrafine fibers thus attached is detachable, the cleaning and drying of the line can be efficient.

The device of the present invention may also have a mechanism for automatically flowing a fixed amount of blood sample into a filter for plasma separation. The metering pump described above can be used for this, for example. Further, it may have a mechanism for automatically changing the filter or each component thereof (such as a portion including an assembly of attached ultrafine fibers). Further, it may have a structure in which the plasma separator (aggregation of ultrafine fibers) portion of the conventional plasma separation device is replaced with a portion or a filter including the aggregate of ultrafine fibers to be mounted. Further, the filters can be connected in series if improved separation performance is desired, and in parallel if more blood volume to be treated is desired.

(Example 13) The apparatus of the present invention can be automated from blood supply to collection, and can separate plasma from a large number of blood samples. For example, FIG. 9 is a schematic diagram showing an example of a configuration for automating the apparatus of the present invention. With automation, many types of blood samples can be processed simultaneously.

In FIG. 9, the blood sample is rack 48.
Are arranged so that the inlet of the blood supply line 32 is almost at the bottom surface of the blood sample 26. Filter 1
1 is fixed to the holder 45. Blood supply line 3
The end of 2 is configured to be airtightly fitted to the inlet of the filter 11, and the tip of the blood supply line 32 is pressed by mechanical pressure to be airtightly fitted to the inlet of the filter. Similarly, the tip of the plasma recovery line 37 is configured to be airtightly fitted to the outlet of the filter 11, and the tip of the plasma recovery line 37 is pressed by mechanical pressure to be airtightly fitted to the outlet of the filter 11. Are combined. Next, at the outlet of the line, the sample container 38 for receiving plasma, which is a means for collecting plasma, is arranged in the rack 50 and automatically arranged.

When the lines are thus connected and the sample container for receiving the plasma is arranged, the blood supply pump 46, which is the blood supply means, operates. When a certain amount of blood flows, the blood supply pump 46 stops. When the blood supply pump 46 is stopped, the valve 47 shuts off the blood supply line 32. When all the blood supply pumps 46 are stopped, the compressor 35 operates and compressed air is supplied from the compressor 35 to the filter 11. Blood passes through the ultrafine fiber assembly 14 by air pressure, and plasma is separated.

At the outlet of the filter 11, blood cells and / or
Alternatively, the hemoglobin detection means 40 is arranged. When blood cells and / or hemoglobin are detected, the blood cell and / or hemoglobin switching means, which is a three-way cock 36, is switched, and the blood cell- and / or hemoglobin-containing plasma recovery line 41 converts blood cells and / or hemoglobin-containing plasma. Lead to. At the same time, the air supply valve 47 is switched and the pressurization is stopped. When blood cells and / or hemoglobin are detected, the sample container is marked and ready for the next sample.

A liquid volume meter 49 is provided at the end of the plasma recovery line 37.
Is arranged, and when a certain amount is collected, the switching means 36 is switched to guide unnecessary plasma to the blood cell and / or hemoglobin-containing plasma recovery line 41. At the same time, the pressurization air supply valve 47 is switched to stop the pressurization. When all the valves are switched, the compressor 35 stops.

The line at the end of the plasma collecting means is pulled out from the sample container 38 from which the plasma has been collected. The sample container 38 is covered and stored.

The end of the blood supply line 32, which is airtightly fitted to the inlet of the filter 11, is separated by mechanical pressure. The tip of the plasma recovery line 37, which is airtightly fitted to the outlet of the filter 11, is also separated by mechanical pressure. The filter 11 thus separated from each means is separated from the holder and discarded.

Then, the line is washed. Instead of the filter 11, a hollow container that can be airtightly fitted to the end of the blood supply line 32 and the tip of the plasma recovery line 37 is arranged. A container for receiving the washing solution is automatically set at the outlet of the line 37 which is the end of the plasma recovery means.
The rack 48 on which the blood sample supply containers 51, which have completed the supply of blood, are arranged moves and the blood sample supply container 51 is discarded. Instead, the container containing the cleaning liquid is the rack 48.
And is set so that the inlet of the blood supply line 32 is almost at the bottom of the washing liquid.

Next, the valve 47 is switched to supply the cleaning liquid, and the blood supply pump 46 is operated to clean the lines 32, 37 and 41. Switching means valve 36
To a plasma recovery line 37 and a blood cell and / or hemoglobin-contaminated plasma recovery line 41, respectively,
Wash.

After the cleaning is completed, the compressor 35 is operated to supply air to dry the line for a certain period of time. afterwards,
A rack in which washing water is arranged and a rack in which containers for washing liquid are arranged are moved, and a rack in which blood samples are arranged 4
8. A rack in which containers for collecting plasma are arranged is arranged at a predetermined position. Further, instead of the hollow container used for cleaning, the filter 11 is arranged by the above method.
Then, the above operation is repeated.

The order of these operations can be arbitrarily changed, and the means for recovering the washing liquid is not limited to the above, and it may not be an individual container. In any case, any modification of such an automated device is also included in the present invention.

(Embodiment 14) The apparatus of the present invention usually has at least (A) a filter, a blood supply means, a blood pressurizing means, and a plasma collecting means. In addition to these means, (a) means for supplying a fixed amount of blood,
(B) means for collecting a certain amount of plasma, (c) means for detecting blood cells and / or hemoglobin,
(D) switching means, (e) means for collecting plasma mixed with blood cells and / or hemoglobin, and the like can be arbitrarily combined. In the device of the present invention, each means may be interlockable with each other. Further, each means may be arranged to be capable of interlocking with means for deactivating them. The combination of these means can be arbitrarily changed depending on the purpose of use of plasma, desired plasma level, scale of device, and the like. For example, a device that combines the following means is typical.
(1) A device having (a) in addition to (A). (2)
A device having (b) in addition to (A). (3) A device having (a) and (b) in addition to (A); in this device, by supplying a preselected amount of blood and collecting plasma by the kind of filter to be used,
Samples can be obtained quickly and easily. (4) A device having (c), (d) and (e) in addition to (A); at this time, (c), (d) and (e) are usually interlockable with each other. (5) A device having all of (b) to (e) in addition to A. (6) A device having all of (a) to (e) in addition to A; this is an example of a device capable of performing the most accurate plasma separation among the devices of the present invention.

The average processing linear velocity of blood (average processing linear velocity from the inlet to the outlet of the filter) by the plasma separation device of the present invention is not particularly limited, but is preferably 0.5 to 500 mm / min. If the processing linear velocity is low, it is inconvenient to process a large number of specimens, and if it is too fast, hemolysis may occur due to an increase in differential pressure.

(Example 15: Connection with automatic analysis system) The automatic analysis system is to integrate the operations in each process of blood analysis in order and systemize them.
In other words, it is a system that automates the entire process from sample collection to analysis and data recording. The automatic analysis system is divided into two systems, a flow system and a discrete system, depending on the analysis method. The plasma separation device of the present invention can be connected to these systems. That is, in the flow system, some parts are connected by polyethylene pipes, and the sample to be analyzed is analyzed and recorded while flowing through the connected tubes. Can be connected to the sampler part of this system. On the other hand, the discrete system is a system in which each sample is analyzed by an independent reaction tube, and like the flow system, the device of the present invention can be connected to the sampler part. Alternatively, plasma obtained separately by the device of the present invention may be used as a sample and supplied to these automatic analysis systems. In this case, the conventional automatic analysis system can be used as it is. The sampler part of the automatic analysis system and the plasma separation device of the present invention, for example, the plasma collection line 37 of the device of FIG. 9 can be connected.

(Example 16) A syringe having a blood supply means and a pressurizing means was attached to the plasma separation filter of Example 1 to prepare the plasma separation device of FIG. 7.

After pouring 4 ml of bovine blood from the inlet of the container and filling the space between the outer peripheral surface of the ultrafine fiber assembly attached to the filter and the inner peripheral surface of the container with bovine blood, 0.2
Bovine blood was extruded by applying a pressure of kg / cm ² . Bovine blood entered from the outer peripheral surface of the attached ultrafine fiber assembly, and moved horizontally inside the ultrafine fiber assembly from the outer periphery toward the center of the circle. 20 seconds after pressurization,
Plasma began to flow from the outlet of the container. The separated plasma was continuously sampled. 0.5 ml of plasma free of red blood cells was obtained. Average processing linear velocity is 75mm
/ Minute (= 25 mm / 20 seconds).

The plasma sampled at the beginning of the outflow was 0.4
Collect the cells and analyze the concentration of contaminated blood cells (red blood cells, white blood cells, platelets) in plasma, the presence or absence of hemolysis of red blood cells, and blood components (serum amylase, total cholesterol, neutral fat, urea nitrogen, blood sugar), and analyze the device. evaluated. Table 4 shows the results.

[0223]

[Table 4]

The red blood cell concentration in plasma (hereinafter referred to as the red blood cell contamination ratio) obtained with respect to the red blood cell concentration in bovine blood before separation was 0.003% (= 2 × 10 ⁵ ÷), considering the above detection limit.
It was 7.2 × 10 ⁹ ) × 100) or less. There was no hemolysis of red blood cells, and the hemoglobin concentration in the obtained plasma was 5 mg /
dl, which was within the allowable range. Further, the analysis result of the blood components was not different from that of the conventional centrifugation method, and the effectiveness of the device of the present invention was proved.

[Brief description of the drawings]

FIG. 1 is a diagram showing an example of a filter used in the plasma separation device of the present invention.

FIG. 2 is a diagram showing an example of a filter used in the plasma separation device of the present invention.

FIG. 3 is a filter in which a base material is arranged in a void portion of the filter of FIG.

FIG. 4 is a diagram showing an example of a filter used in the plasma separation device of the present invention.

FIG. 5 is a diagram showing an example of a filter used in the plasma separation device of the present invention.

6 is an apparatus in which a pressurizing means is added to the filter of FIG.

7 is an apparatus in which blood supply and pressurizing means are connected to the filter of FIG.

FIG. 8 is a schematic view showing an example of the plasma separation device of the present invention.

FIG. 9 is a schematic diagram showing an example of a configuration for automating the plasma separation device of the present invention.

[Explanation of symbols]

 11: Filter 12: Inlet 13: Outlet 14: Aggregate of ultrafine fibers 15: Container 16: Void 17: Substrate 18: Opening 19: Inner wall of container 22: Close contact means 24: Pressurizing means 25: Blood supply means 26 : Blood 31: Container for blood supply means 32: Blood supply line 33: Pressurized air supply line 34: Lid 35: Compressor 36: Three-way cock or switching valve 37: Plasma recovery line 38: Sample plasma recovery container 40: Blood cells and / or Or hemoglobin detection means 41: Waste liquid recovery line 42: Waste liquid recovery tank 43: Pure water supply line 44: Dry air supply line 45: Filter holder 46: Blood supply pump 47: Valve 48: Blood sample supply container rack 49: Liquid volume meter 50: Sample plasma recovery container rack 51: Blood sample supply container

────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] August 30, 1996

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0185

[Correction method] Change

[Correction contents]

Example 9 In the same disc-shaped container as in Example 1, 2 g of polyethylene terephthalate ultrafine fiber nonwoven fabric having an average fiber diameter of 1.5 μm is 50 mm in diameter and 2.0 m in thickness.
It was cut into m and filled with a gap of 1.0 mm between the inner peripheral surface of the container and the container. The average hydraulic radius at this time is 0.6
4 μm.

[Procedure amendment 2]

[Document name to be amended] Statement

[Correction target item name]

[Correction method] Change

[Correction contents]

[0195]

[Table 3]

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical display location B01D 39/16 B01D 23/10 A G01N 33/48 29/08 520C 540A (72) Inventor Hitoshi Ohno 2-1-1 Katata, Otsu, Shiga Prefecture, Toyobo Co., Ltd.

Claims (25)

[Claims] 1. A filter in which an assembly of ultrafine fibers is mounted in a container having an inlet and an outlet, the filter having the following characteristics: (1) of the installed assembly of ultrafine fibers The ratio (L / D) of the blood channel diameter (D) and the blood channel length (L) is 0.
15-6; (2) The average fluid radius of the attached ultrafine fiber aggregate is 0.5-3.0 μm; (3) Red blood cell concentration is 6-8 × 10 ⁹ cells / ml. Some fresh bovine blood was separated at a pressure of 0.2 to 0.4 Kg / cm ² , and 10% of the void volume of the attached ultrafine fiber aggregate from the start of separation
(A) The concentration of red blood cells in plasma was 0.1% of the concentration of red blood cells in blood before separation when plasma equivalent to
% Or less; and (b) virtually no red blood cells are hemolyzed. 2. The plasma separation filter according to claim 1, wherein the ultrafine fibers are polyester, polypropylene, polyamide, or polyethylene. 3. The aggregate of the ultrafine fibers is a non-woven fabric,
The plasma separation filter according to claim 1 or 2. 4. The non-woven fabric is attached to the container singly or in a laminated manner, and the blood is almost attached to the face of the non-woven fabric of the attached ultrafine fiber aggregate or the plane of the laminated non-woven fabric. Configured to flow in parallel,
The plasma separation filter according to claim 3. 5. The container and the non-woven fabric are disc-shaped, the inlet is configured to be able to supply blood over the entire circumferential side surface of the disc-shaped non-woven fabric, and the outlet is made of the disc-shaped non-woven fabric. The plasma separation filter according to claim 4, which is configured so that plasma flows out from the central portion. 6. A filter in which an assembly of ultrafine fibers is attached to a container having an inlet and an outlet, and the filter has the following characteristics: (1) of the attached assembly of ultrafine fibers The ratio (L / D) of the blood channel diameter (D) and the blood channel length (L) is 0.
15-6; (2) The average hydrodynamic radius of the mounted assembly of ultrafine fibers is 0.5-3.0 μm; (3) A hydrophilizing agent is fixed to the ultrafine fibers. 7. The plasma separation filter according to claim 6, wherein the hydrophilizing agent is fixed on the surface of the ultrafine fibers. 8. The plasma separation filter according to claim 6, wherein the hydrophilizing agent is polyvinylpyrrolidone. 9. The filter has the following characteristics: 0.2 of fresh bovine blood having a red blood cell concentration of 6 to 8 × 10 ⁹ cells / ml.
When separated at a pressure of 0.4 Kg / cm ² and recovering an amount of plasma equivalent to 10% of the interstitial volume of the attached ultrafine fiber aggregate from the start of separation, (a) red blood cells in plasma 9. The concentration according to claim 6, wherein the concentration is 0.1% or less with respect to the concentration of red blood cells in blood before separation, and (b) the red blood cells are not substantially hemolyzed. Plasma separation filter. 10. The plasma separation filter according to claim 6, wherein the aggregate of the ultrafine fibers is a non-woven fabric. 11. The non-woven fabric is attached to the container individually or in a laminated manner, and blood is configured to flow substantially parallel to a plane of the attached non-woven fabric or a plane of the laminated non-woven fabric. The plasma separation filter according to claim 10, wherein 12. The non-woven fabric, wherein the container and the non-woven fabric are disc-shaped, and the blood inlet is configured so as to be able to supply the blood over the entire circumferential side surface of the non-woven fabric and the outlet is disc-shaped. The plasma separation filter according to claim 11, wherein the plasma separated from the central part of the plasma is discharged. 13. The filter has a red blood cell concentration of 6 to 10.
0.2 ~ 0.4K of fresh bovine blood of 8 × 10 ⁹ cells / ml
When separated at a pressure of g / cm ² and collecting an amount of plasma corresponding to 10% of the void volume of the attached ultrafine fiber aggregate from the start of separation, the electrolyte concentration in the separated plasma was The plasma separation filter according to any one of claims 1 to 12, wherein 90% or more of the electrolyte concentration in the plasma obtained by centrifugation is retained. 14. The filter has a red blood cell concentration of 6 to
0.2 ~ 0.4K of fresh bovine blood of 8 × 10 ⁹ cells / ml
When separated at a pressure of g / cm ² and recovering an amount of plasma corresponding to 10% of the void volume of the attached ultrafine fiber aggregate from the start of separation, the protein concentration in the separated plasma was The filter according to any one of claims 1 to 13, which retains 90% or more of the protein concentration in plasma obtained by centrifugation. 15. A method for separating plasma using a plasma separation filter in which an assembly of ultrafine fibers is attached to a container having an inlet and an outlet, the method comprising supplying blood to the plasma separation filter. And a step of pressurizing the filter, the pressure loss at the outlet is 0.03 to 5
kg / cm ² method: Here, the plasma separation filter has the following characteristics: (1) Blood channel diameter (D) and blood channel length (L) of the attached ultrafine fiber aggregate. The ratio (L / D) is 0.15 to 6; and (2) the average fluid radius of the attached ultrafine fiber aggregate is 0.5 to
It is 3.0 μm. 16. The plasma separation filter separates and separates the following characteristics: Fresh bovine blood having a red blood cell concentration of 6 to 8 × 10 ⁹ cells / ml at a pressure of 0.2 to 0.4 kg / cm ^2. From the beginning, 1 of the void volume of the attached ultrafine fiber aggregate
(A) the concentration of red blood cells in the plasma is 0.1% or less of the concentration of red blood cells in the blood before separation when the amount of plasma equivalent to 0% is collected; and (b) substantially red blood cells. Is not hemolyzed; 16. The plasma separation method according to claim 15, wherein: 17. The plasma separation method according to claim 15, wherein a hydrophilizing agent is fixed to the ultrafine fibers of the plasma separation filter. 18. The linear velocity of blood to be treated is 0.05 to
18. The plasma separation method according to claim 15, which has a flow rate of 50 cm / min. 19. A plasma separation device having a plasma separation filter in which an assembly of ultrafine fibers is attached to a container having an inlet and an outlet, the filter having the following characteristics: (1) The attached ultrafine fibers The ratio (L / D) of the blood flow channel diameter (D) and the blood flow channel length (L) of the aggregate of 1.
15 to 6; and (2) the average fluid radius of the attached ultrafine fiber aggregate is 0.5 to 3.0 μm. 20. The plasma separation filter separates (2) fresh bovine blood having an erythrocyte concentration of 6 to 8 × 10 ⁹ cells / ml at a pressure of 0.2 to 0.4 kg / cm ² , and starts separation. The following characteristics were obtained when the amount of plasma equivalent to 10% of the interstitial volume of the attached ultrafine fiber aggregate was collected from: 20% or less; and (b) the red blood cells are not substantially hemolyzed. 21. The plasma separation apparatus according to claim 19, wherein a hydrophilizing agent is fixed to the ultrafine fibers of the plasma separation filter. 22. Blood supplying means for supplying blood to the plasma separation filter, pressurizing means for pressurizing blood supplied to the filter to separate the supplied blood,
22. The plasma separation apparatus according to any one of claims 19 to 21, further comprising plasma recovery means for recovering the separated plasma. 23. Further, a blood cell and / or hemoglobin detecting means for detecting blood cells and / or hemoglobin in separated plasma, a switching means for separating plasma mixed with blood cells and / or hemoglobin, and 23. The plasma separation apparatus according to claim 22, further comprising blood cell and / or hemoglobin-contaminated plasma collection means for collecting plasma mixed with blood cells and / or hemoglobin. 24. The plasma separation apparatus according to claim 22, wherein the filter is detachably attached between the blood supply means and the plasma recovery means. 25. The plasma separation apparatus according to claim 22, wherein the apparatus has blood supply means for supplying a fixed amount of blood and / or plasma recovery means for recovering a fixed amount of plasma. .