JP3495106B2 - Blood component adhesion preventing agent, blood test container and blood component adhesion preventing carrier - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the field of clinical examination using a blood sample, more specifically, a blood component anti-adhesion agent, a blood test container, and a blood test container used in tests such as hematology, blood chemistry, and immunoserology. The present invention relates to a carrier for preventing blood component adhesion.

[0002]

2. Description of the Related Art In recent years, with the progress of examination techniques, hematology,
Blood tests such as blood chemistry and immunoserologic tests are remarkably mechanized, and it is possible to obtain test results in a short time if a sample that has undergone appropriate pretreatment can be prepared.For example, on the spot when an outpatient visits a hospital. It is now possible to obtain a doctor's examination based on the test results, which greatly contributes to speeding up the diagnosis of diseases. Of these, the pretreatment in a hematology test using blood itself as a sample is only required to sufficiently mix blood and an anticoagulant, which does not require much time and can be immediately set in the measuring device. In chemical or immunoserologic tests, serum in blood is used as a sample, so in order to obtain the necessary serum, it is necessary to first coagulate blood and then separate the serum components by centrifugation or the like. It takes time, not a little. Therefore, it is not enough to mechanize the analysis process in order to shorten the time required for all the steps of the test including pretreatment of blood to obtain a sample and obtaining a measurement result. , The serum collection time should also be shortened.

By the way, the blood used for the test is stored in
Conventionally, glass containers were often used as blood test containers used for coagulation and separation of serum by centrifugation, but blood samples that were vulnerable to mechanical shock and leaked or scattered when damaged. Due to the fear of infection and the increased burden on patients due to re-collection of blood, the proportion of plastic containers has increased recently.

However, the plastic container has a poor activation power for blood coagulation factor XII, factor XI and the like, and as it is, it requires a significantly longer time for blood coagulation than glass products and is not practical. In addition, blood components such as platelets, various proteins in blood, and fibrin produced in the final stage of blood coagulation tend to adhere very strongly to the plastic surface.

Therefore, in order to shorten the blood coagulation time, fine powders of mineral substances such as glass, kaolin, bentonite, silica and celite, or organic substances such as ellagic acid having a blood coagulation accelerating property are previously used for blood tests. It may be applied on the inner wall surface of the container, or may be stored in the container by carrying the above-mentioned fine powder on a carrier such as a non-woven fabric or a plastic sheet which is substantially insoluble in blood and chemically inert. However, when the coagulation is forcibly promoted by such a method, it often happens that the adhesion of the aforementioned blood components is rather enhanced. The blood component attached to the inner wall surface or the carrier surface of the test container is not easily peeled off under normal centrifugation conditions of about 1000 to 1800 G for 5 minutes. Therefore, the inner wall surface of the container and the blood clot coagulated. At the interface with and, platelets and erythrocytes are destroyed due to the strong shearing force due to centrifugal force, and these contents leak out, which adversely affects various tests.

Therefore, when a blood coagulation-promoting substance is contained in a test container, a nonionic surfactant should be simultaneously present on the inner wall surface of the container or the surface of the carrier for the purpose of preventing adhesion of blood components. (Japanese Patent Laid-Open Nos. 58-105063 and 58-105064).

However, in recent years, with the progress of the technology for increasing the sensitivity of immunoserologic tests, there have been more and more occasions when the related substances of nonionic surfactants are used as sensitizers, and these containers have been used for test containers from the beginning. If substances are used, they may contaminate the serum, and in some immunoserologic test items, false-positive reactions that may be caused by oversensitization may occur, and measures should be taken. ing.

On the other hand, in a hematological test using an anticoagulant such as ethylenediaminetetraacetate or citrate, the frequency of blood components is low, but the adhesion of platelets to the plastic surface may be a problem. When platelets adhere to the inner wall surface of the testing container, the count value of platelets may show an abnormally low value, or the blood coagulation function test may be adversely affected. In addition, heparin salt, which is a type of anticoagulant, is used when a chemical test must be performed urgently, though it is not common. Enzymes may leak into plasma, causing problems such as abnormally high values for these measurement items. In the past, the frequency was smaller than when blood was coagulated, so it was not a problem so much.However, in today's world where there is an increasing demand for improvement in the accuracy of hematology, blood chemistry, and immunoserologic testing. Therefore, a better anti-adhesion agent for blood components that can be universally used is desired.

The above-mentioned problems relating to the adhesion of blood components are not limited to the plastic inspection container, and the frequency thereof is low, but the glass inspection container is not completely free. In fact, the adverse effect on the test value, which is thought to be caused by the adhesion of platelets, may occur, and improvement is required as with plastic products.

[0010]

SUMMARY OF THE INVENTION The present invention is intended to solve the above-mentioned problems, and its object is to provide an inner wall surface of a container for testing blood components without giving false positive results to immunoserologic tests. A blood component adhesion preventive agent capable of effectively preventing the adhesion of blood components, a blood test container having the prevention agent present on the inner wall surface, and a blood component adhesion preventing carrier having the prevention agent present on the surface are provided. Especially.

[0011]

As a result of intensive studies, the present inventors have found a substance that can effectively solve the above-mentioned problems, and completed the present invention. The blood component adhesion preventive agent of the present invention is
A random copolymer comprising a monomer component (a) in which the homopolymer is water-soluble and a monomer component (b) in which the homopolymer is water-insoluble, the monomer component (b) Content of 1
It is composed of 0 to 90 mol% of the copolymer.

As the monomer component (a) in which the homopolymer is water-soluble, vinyl alcohol, vinylpyrrolidone, ethylene oxide, acrylate, styrenesulfonate, vinylphosphonate, allylamine salt, hydroxymethyl ( Examples thereof include meth) acrylate, glycosylethyl (meth) acrylate, sugars such as glucose, and amino acids such as glutamic acid, and these may be used in combination of two or more kinds.

The monomer component (b) in which the homopolymer is water-insoluble includes ethylene, propylene, propylene oxide, vinyl acetate, vinyl chloride and alkyl (meth).
Examples thereof include acrylate, styrene, acrylonitrile, acrolein, and the like, and these may be used in combination of two or more kinds.

A random copolymer of the monomer component (a) and the monomer component (b) can be obtained by a known addition polymerization, condensation polymerization or the like. As the monomer component (a), vinylpyrrolidone or A random copolymer comprising vinyl alcohol and vinyl acetate as the monomer component (b) is easily available and suitable. As a commercially available product, as a random copolymer composed of vinylpyrrolidone and vinyl acetate, BASF Corporation, trade name “Rubiscol VA” described in Table 1 below,
Grade number VA73, VA64, VA55, VA3
7, VA28, etc. Further, as a random copolymer composed of vinyl alcohol and vinyl acetate, Table 1 described below is used.
Made by Unitika Ltd., trade name "Unitika Poval", grade number E-180, UMR-10M, UMR-3
0L, UMR-150L and the like.

In the random copolymer used for the blood component adhesion preventive agent of the present invention, when the amount of the monomer component (b) in which the homopolymer is water-insoluble is small, the monomer component (a) is substantially contained. ), The adsorption rate to the inner wall surface or carrier surface of the test container is small, and its solubility in blood is too large. Since it is washed out from the surface of the carrier, the effect as an anti-adhesion agent for blood components cannot be achieved, and false positive immunoserologic tests are likely to occur.

On the contrary, when the amount of the monomer component (b) in which the homopolymer is water-insoluble is large, it is substantially the same as the homopolymer of the monomer component (b) into the blood. Since the solubility of the blood component is substantially lost, the effect of the blood component as an anti-adhesion agent is also reduced, and further, the blood component anti-adhesion agent is used in combination with a blood coagulation promoting substance or an anticoagulant. When applied to the inner wall surface or carrier surface of the test container and dried, a film insoluble in blood is formed on the surface of the coagulation promoting substance or anticoagulant, and blood coagulation factors XII and XI And the like cannot bind to the surface of the coagulation-promoting substance, resulting in the inconvenience that blood coagulation is difficult to be promoted and the anticoagulation is not sufficiently performed because the solubility of the anticoagulant is impaired. It will be easier. For the above reasons, the content of the monomer component (b) in the random copolymer used in the present invention is limited to 10 to 90 mol%. In view of the above, in order to prevent the adhesion of blood components, it is considered necessary to have both adsorptivity to the inner wall surface of the test container and carrier surface and weak solubility in blood.

The blood test container of the present invention is characterized in that the blood component adhesion-preventing agent of the present invention comprising a random copolymer is present on its inner wall surface. When the amount of the random copolymer existing on the inner wall surface of the inspection container decreases, it becomes impossible to obtain a sufficient adhesion preventing effect of blood components,
When the amount is large, the possibility of adversely affecting various inspection values becomes strong, so 1 × 10 ⁻¹⁰ to 1 × 10 ⁻² g / cm ² is preferable.

As the material for the above-mentioned blood test container, one made of a known material may be used, and various plastics, thermoplastic elastomers and elastomers which are substantially insoluble in blood and chemically inactive. Chemically modified cellulose, or hard, soft, borosilicate, quartz glass, or the like is used.

To manufacture the blood test container of the present invention,
The blood component adhesion preventive agent of the present invention may be present on the inner wall surface of the test container by various known methods. As this method, for example, a method of kneading a blood component adhesion preventive agent in advance into a plastic material constituting a container for inspection and molding the container by injection molding or blow molding, purified blood component or alcohol etc. There is a method of spraying or dipping on the inner wall surface of the container and then drying.

In the blood test container of the present invention, in addition to the anti-adhesion agent for blood components comprising the random copolymer of the present invention being present on the inner wall surface thereof, in addition, in the container, a thixotropic agent is added. A material capable of providing a partition wall between the blood serum or plasma layer and the solid component layer of blood, such as a blood plasma separating agent or a separating member, may be contained. As the serum or plasma separating agent, for example, liquid acrylic resin, chlorinated polybutene or liquid dicyclopentadiene (DCPD)
Examples thereof include a resin as a main component, and an inorganic powder such as finely powdered silica, alumina, or glass added for the purpose of adjusting specific gravity and imparting thixotropy. The presence of such a septum-forming substance allows the serum or plasma to be stored for a long time without affecting the test value.

The blood component anti-adhesion carrier of the present invention is characterized in that the blood component anti-adhesion agent of the present invention comprising a random copolymer is present on the surface of the carrier.

The blood component adhesion preventive carrier of the present invention is used by being housed inside a blood test container.

Known carriers are used as the above carrier, and various shapes such as pellets, sheets, non-woven fabrics and woven fabrics can be used.

As the material of the above carrier, as in the case of the blood test container, one made of a known material may be used, and various plastics which are substantially insoluble in blood and chemically inert, and heat. Plastic elastomers, elastomers, chemically modified celluloses, etc., or hard, soft, borosilicate, quartz glass, etc. are used.

When the amount of the random copolymer existing on the surface of the carrier is small, the effect of preventing the adhesion of blood components cannot be sufficiently obtained, and when the amount is large, there is a strong possibility that various test values are adversely affected. Therefore, 1 × 10 ^-10 to 1
× 10 ^-2 g / cm ² is preferable.

In order to produce the blood component anti-adhesion carrier of the present invention, the blood component anti-adhesion agent of the present invention may be present on the surface of the carrier by various known methods. As this method, for example, a blood component adhesion preventive agent is kneaded in advance in a plastic material constituting the carrier, and the carrier is molded by injection molding or blow molding, or the blood component adhesion preventive agent is dissolved in purified water or alcohol. After that, there is a method in which the surface of the carrier is sprayed or dip-coated and dried.

The blood component adhesion preventive agent of the present invention may be used alone as described above, but it is a mineral substance such as glass, kaolin, bentonite, silica and celite, or an organic blood coagulation promoting agent such as ellagic acid. It may be used in combination with a substance having, or may be used in combination with an anticoagulant such as ethylenediaminetetraacetate, citrate, heparin salt, oxalate, or a glycolysis inhibitor such as a fluoride salt or mannose. is there.

As the blood component adhesion preventing agent, a vinylpyrrolidone-vinyl acetate copolymer is used, which is composed of one or a mixture of two or more selected from the group consisting of glass, kaolin, bentonite, silica and celite. When used in combination with an adsorptive inorganic substance, the vinylpyrrolidone-vinyl acetate copolymer preferably has a vinylpyrrolidone content of 10 to 70 mol% and a vinyl acetate content of 30.
~ 90 mol% is preferred. When the amount of vinylpyrrolidone is small, the blood clot adheres to the inner wall surface of the tube and loses the clot-separating action. When the amount of vinylpyrrolidone is large, it is dissolved in blood and does not remain on the inner wall surface of the tube, so that the clot-separating action is lost.

It is preferable to use the adsorptive inorganic material having a particle size of 50 μm or less and an average particle size of 30 μm or less. The adsorptive inorganic substance that is particularly effective for shortening the blood coagulation time is silica, and particularly porous silica containing 20% by weight or more of an amorphous component exerts an excellent effect. Such an adsorptive inorganic substance has an action of promoting activation of a blood coagulation factor when it comes into contact with blood and also promoting platelet aggregation.

In this case, it is not present on the inner wall surface of the blood test container.
The amount of vinylpyrrolidone-vinyl acetate copolymer to be added is small.
When it disappears, the serum separation effect is lost.
1 x 10 because it affects the value^-Ten~ 1 x 10^-2g /
cm²Is preferred. It also exists on the inner wall surface of the blood test container.
When the amount of adsorbing inorganic substances is reduced, the effect of promoting coagulation is increased.
Lost and increased, blood test values will be affected,
1 x 10^-6~ 1 x 10 ^-2g / cm²Is preferred. Also,
The total amount of each component is 1 x 10^-2g / cm²The following
And is desirable.

As described above, according to the blood test container in which the vinylpyrrolidone-vinyl acetate copolymer and the adsorptive inorganic substance are used in combination, the blood coagulation factor is rapidly activated and the time required for blood coagulation is significantly shortened. In addition, blood clots resulting from blood coagulation do not adhere to the inner wall surface of the container, and serum and blood clots are easily separated, and the clot component remaining in the separated and collected serum is mixed. Is also eliminated, and the serum yield is significantly increased.

When the above-mentioned blood component adhesion preventive agent is a vinylpyrrolidone-vinyl acetate copolymer and is used in combination with an anticoagulant such as ethylenediaminetetraacetate, citrate, heparin salt or oxalate, The vinyl acetate content in the vinylpyrrolidone-vinyl acetate copolymer is 30 to
90 mol% is preferable.

As the above-mentioned ethylenediaminetetraacetate compound, those conventionally used as blood anticoagulants can be used, and for example, disodium ethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate, tripotassium ethylenediaminetetraacetate. Etc.

As the heparin compound, those conventionally used as blood anticoagulants can be used, and examples include heparin sodium and heparin lithium.

As the citric acid compound, those conventionally used as blood anticoagulants can be used, and examples thereof include trisodium citrate.

As the oxalic acid compound, those conventionally used as blood anticoagulants can be used, and examples thereof include sodium oxalate and potassium oxalate.

In order to allow these blood coagulation-promoting substances, anticoagulants and glycolysis inhibitors to be present on the inner wall surface of the test container or on the surface of the carrier to be contained in the container, it is known in the same manner as described above. Various methods may be used. For example, after the blood component adhesion preventing agent is present on the inner wall surface or the carrier surface in advance, the blood coagulation promoting substance or the anticoagulant may be sprayed or impregnated on the inner wall surface or the carrier surface in a separate step.
Alternatively, all components may be simultaneously dissolved or suspended in an appropriate medium, and the solution or suspension may be sprayed or dip-coated and dried.

[0038]

The blood component adhesion preventive agent of the present invention is a random copolymer composed of a monomer component (a) in which the homopolymer is water-soluble and a monomer component (b) in which the homopolymer is water-insoluble. The polymer is characterized by comprising the copolymer having a content of the monomer component (b) of 10 to 90 mol%. The conventional blood component anti-adhesion agents, so-called nonionic surfactants, consist of block copolymers of hydrophilic monomer components and hydrophobic monomer components, or graft polymerization of blocks. It has been conventionally considered that such a chemical structure is an extremely important index for obtaining an excellent anti-adhesion agent for blood components.

However, the nonionic surfactant is
As it is often known to have properties as an excellent sensitizer for immunoserologic tests, it has become widely used in the field. The blood component anti-adhesion agent of the present invention has a chemical structure that is difficult to predict from conventional recognition,
In addition to having excellent anti-adhesion properties, it does not exhibit properties as a sensitizer for immunoserologic tests, so it does not cause oversensitization in immunoserologic tests and does not cause false-positive reactions. .

[0040]

EXAMPLES The present invention will be described below with reference to Examples using 9 kinds of random copolymers shown in Table 1. In addition,
Examples 2 and 3 are substances that promote blood coagulation, and Examples 4 and 5 are examples in which an anticoagulant is used in combination.
According to the gist of the present invention, the coagulation promoting substance and the anticoagulant do not have any physical or chemical reason for limiting these types to specific ones. Therefore, in Examples 2 and 3, silica was used as the coagulation promoting substance, and in Example 4
In Examples 5 and 5, heparin sodium and heparin lithium are only exemplified as the anticoagulant, but it goes without saying that the constitution of the invention is not subject to any limitation.

[0041]

[Table 1]

Examples 1-1 to 1-27 As a blood component adhesion preventive agent, a random copolymer composed of vinylpyrrolidone and vinyl acetate shown in Table 2 (in Table 2, only grade numbers are shown. ), A purified aqueous solution having a predetermined concentration (% by weight) shown in Table 2 was prepared. In addition, a purified aqueous solution of a random copolymer composed of vinyl alcohol and vinyl acetate shown in Table 3 (shown only by grade number in Table 3) at a predetermined concentration (% by weight) shown in Table 3 was prepared. Prepared. However, only UMR-150L was a methanol solution.

As shown in Table 2 or Table 3, 1% of each solution was used.
About 50 μl of a polyethylene terephthalate blood collection tube having a volume of 0 ml was sprayed and dried with a blow dryer at 60 ° C. to obtain a blood test container. 3 ml of fresh rabbit blood was collected in this blood test container and left standing at room temperature of 23 to 25 ° C. After 4 hours, it was confirmed that the blood was completely coagulated, and the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed, and none was found to have adhered. Then 2
Centrifugation was performed under the conditions of 5 ° C., 1300 G, and 5 minutes, and the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed, and the yield of the separated serum was measured. As a result, none of the blood clots adhered to the inner wall surface of the blood test container. The serum yield was as shown in Tables 2 and 3.

[0044]

[Table 2]

[0045]

[Table 3]

(Comparative Example 1-1) As a blood component adhesion preventive agent, a vinylpyrrolidone homopolymer (manufactured by BASF) was used instead of a 0.05% by weight aqueous solution of a random copolymer of vinylpyrrolidone and vinyl acetate. A test was conducted in the same manner as in Example 1-1, except that a 0.05% by weight aqueous solution of trade name "Rubiscol K-30" having a weight average molecular weight of 30,000) was used. As a result, it was confirmed that the blood was completely coagulated 4 hours after the blood was collected, and the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed to find that the blood spots were scattered. The presence or absence of blood clots adhered to the inner wall surface of the blood test container after centrifugation was found to be scattered and strongly hemolyzed. The yield of serum after centrifugation was 1.4 ml.

(Comparative Example 1-2) As a blood component adhesion preventive agent, instead of a 0.05% by weight aqueous solution of a random copolymer composed of vinylpyrrolidone and vinyl acetate, a homopolymer of vinyl alcohol (manufactured by Unitika Ltd., A test was performed in the same manner as in Example 1-1, except that a 0.05% by weight aqueous solution of a trade name "Unitika Poval UF200G") was used. As a result, it was confirmed that the blood was completely coagulated 4 hours after the blood was collected, and the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed to find that the blood spots were scattered. Regarding the presence or absence of adherence of blood clots to the inner wall surface of the blood test container after centrifugation, the adhesion was scattered and weak hemolysis was observed.
The yield of serum after centrifugation was 1.4 ml.

(Comparative Example 1-3) As a blood component adhesion preventive agent, instead of a 0.05% by weight aqueous solution of a random copolymer of vinylpyrrolidone and vinyl acetate, a copolymer of vinyl acetate and ethylene (Hoechst) was used. A test was performed in the same manner as in Example 1-1, except that a 0.05% by weight methanol solution of Synthetic Co., Ltd., trade name "Movinyl E45") was used. As a result, it was confirmed that the blood was completely coagulated 4 hours after blood collection, and the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed to find that they were significantly attached. The presence or absence of blood clots on the inner wall surface of the blood test container after centrifugation was significantly attached. Therefore, the serum yield after centrifugation was 0.0 ml.

Comparative Example 1-4 As a blood test container,
The test was performed in the same manner as in Example 1-1, except that a blood component adhesion-preventing agent was not used (that is, an untreated 10 ml-volume polyethylene terephthalate blood collection tube). As a result, it was confirmed that the blood was completely coagulated 4 hours after the blood was collected, and the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed. Regarding the presence or absence of blood clots adhered to the inner wall surface of the blood test container after centrifugation, there was strong adhesion and hemolysis was observed. The yield of serum after centrifugation was 0.5 ml or less.

From the results of Example 1 and Comparative Example 1, the blood component adhesion preventive agent of the present invention was applied to a polyethylene terephthalate blood collection tube (Comparative Example 1-4), which originally has a property of strongly adhering blood components. It can be seen that a random copolymer has an excellent anti-adhesion effect. The serum yield of Example 1 was about 50% of the whole blood volume, and almost the whole amount was recovered. In Comparative Examples 1-1 and 1-2, when the ratio of the monomer component in which the homopolymer is water-insoluble is 0 mol% among the monomer components constituting the random copolymer used in the present invention. In Comparative Example 1-3, the ratio of the monomer component is 10
It represents the case of 0 mol%. Comparative Examples 1-1 and 1-2
Was not so bad as to adversely affect the serum yield, but the adherence of blood clots was scattered on the inner wall surface, the anti-adhesion property was clearly inferior, and hemolysis occurred. On the other hand, in Comparative Example 1-3, there was strong blood clot adhesion, and serum could not be collected. In all of these comparative examples, the ratio of the monomer component in which the homopolymer is water-insoluble is 10 to 90 mol%, which is outside the requirement of the present invention, and the effect of preventing the adhesion of blood components is not recognized. It was

Example 2-1 A vinylpyrrolidone-vinyl acetate random copolymer (B
ASF Co., Rubiscol VA73, vinyl acetate content of about 36 mol%), fine powder silica (average particle diameter 4.0 μm, Japanese Industrial Standard K-5101) which is an adsorbing inorganic substance as a blood coagulation promoting substance. Linseed oil absorption amount measured by 30ml / 100g, BET specific surface area value 12000c
m ² / g, specific resistance value which is the reciprocal of electric conductivity 2.6 × 1
0 ⁴ Ω · cm) was used to prepare a methyl alcohol dispersion liquid such that the concentrations of these components were 0.1% by weight and 1.0% by weight, respectively. A blood test container was obtained by spray coating on the inner wall surface of the blood vessel and air drying. The amount of each component adhered to the inner wall surface of the container per unit area was 2 × 10 ⁻⁶ g / cm ^{2 of} vinylpyrrolidone-vinyl acetate copolymer, 2 × 1 of fine powder silica.
It was 0 ⁻⁵ g / cm ² .

After pouring 8 ml of human fresh blood into the blood test container and leaving it at 20 ° C., the time required until the whole blood does not completely flow is measured as the blood coagulation time to determine the blood coagulation property. evaluated. Immediately after the coagulation was confirmed, centrifugation was performed at 3000 rpm for 5 minutes to observe the serum separation state, and the supernatant serum was pipetted, and the amount was used as the serum yield. The results are shown in Table 4.

(Example 2-2) A vinylpyrrolidone-vinyl acetate random copolymer (B
Lubiscol VA28 manufactured by ASF, vinyl acetate content of about 84 mol%), Example 2 as blood coagulation promoting substance
The same fine powder silica as in -1 was used to prepare a methyl alcohol dispersion liquid such that the concentrations of these components were 0.02% by weight and 1.0% by weight, respectively.
A blood test tube was obtained by spray-applying the inner wall surface of a polypropylene blood collection tube of sufficient capacity and air-drying. The amount of each component adhering to the inner wall surface of the container per unit area was 5 × 10 ⁻⁷ g / cm ^{2 of} vinylpyrrolidone-vinyl acetate copolymer and 3 × 10 ⁻⁵ g / cm ² of fine powder silica. Example 2
Blood coagulability, serum separation state, and serum yield were evaluated in the same manner as in -1. The results are shown in Table 4.

(Comparative Example 2-1) A homopolymer of vinylpyrrolidone as a blood component adhesion preventive agent (manufactured by BASF, Rubiscol K30, weight average molecular weight 30,000), and Example 2-1 as a blood coagulation promoting substance. The same fine powder silica is used, and the concentration of each of these components is 0.1% by weight,
A methyl alcohol dispersion liquid was prepared so as to be 1.0% by weight, and this was spray-coated on the inner wall surface of a polypropylene resin blood collection tube having a volume of 10 ml and air-dried to obtain a blood test container. The amount of each component adhering to the inner wall surface of the container per unit area was 3 × 10 ^-6 g / c of vinylpyrrolidone homopolymer.
m ² and finely divided silica 3 × 10 ⁻⁵ g / cm ² . Then, in the same manner as in Example 2-1, blood coagulability, serum separation state, and serum yield were evaluated. The results are shown in Table 4.

(Comparative Example 2-2) A blood coagulation-promoting substance was prepared in the same manner as in Example 2 except that the blood component adhesion-preventing agent was not used.
Using the same finely divided silica as -1, a methyl alcohol dispersion was prepared so that the concentration of this component would be 1.0% by weight, and spray-applied to the inner wall surface of a polypropylene resin blood collection tube of 10 ml capacity, Air-dried to obtain a blood test container. The amount of this component attached to the inner wall surface of the container per unit area was 3 × 10 ⁻⁵ g / cm ² . Example 2-
Blood coagulability, serum separation state, and serum yield were evaluated in the same manner as in 1. The results are shown in Table 4.

[0056]

[Table 4]

(Example 3-1) Vinylpyrrolidone-vinyl acetate random copolymer (B
Using Rubiscol VA64 manufactured by ASF Co., vinyl acetate content of about 46 mol%), a purified aqueous solution was prepared to a concentration of 1.0 wt%. Further, as the blood coagulation promoting substance, the same fine powder silica as in Example 2-1 was used,
A purified water suspension was prepared so that the concentration was 1.0% by weight. About 50 μl of a purified aqueous solution of vinylpyrrolidone-vinyl acetate random copolymer was sprayed onto a 10 ml capacity polyethylene terephthalate blood collection tube, and dried with a blast dryer at 60 ° C., and a purified water suspension of fine powder silica was about 5 μm.
It was sprayed with 0 μl and dried with a blast dryer at 60 ° C. to obtain a blood test container. 5 ml of fresh rabbit blood was collected in this blood test container and allowed to stand at room temperature of 23 to 25 ° C. The time at which blood loses fluidity and serum starts to exude was measured as blood coagulation time. Then, after 1 hour of blood collection and after centrifugation, the presence or absence of blood clots on the inner wall surface of the blood test container was visually observed in the same manner as in Example 1-1 to measure the serum yield. The results are shown in Table 5.

Next, in order to confirm the influence of the blood component adhesion preventive agent on the immune serum test value, immediately after the above evaluation was completed, the whole amount of serum was immediately taken and transferred to a clean hard glass test tube. I changed it. Using the serum, HBs-Ab test reagent (manufactured by Toa Medical Electronics Co., Ltd.) and F as an immunoserologic test
HB using ree T4 test reagent (Kodak)
When tested for s-Ab and Free T4, the test values were negative and not false positive. The results are shown in Table 5.
It was shown to.

(Example 3-2) In Example 3-1,
Instead of the purified aqueous solution of vinylpyrrolidone-vinyl acetate random copolymer having a concentration of 1.0% by weight, a vinyl alcohol-vinyl acetate random copolymer having a concentration of 1.0% by weight (Unitika Poval UMR-30L, manufactured by Unitika Ltd.). , Vinyl acetate content of about 60 mol%) A blood test container was obtained in the same manner as in Example 3-1 except that a purified aqueous solution was used, and the results were tested in the same manner as in Example 3-1. Indicated.

(Comparative Example 3-1) In Example 3-1,
A blood test container was obtained in the same manner as in Example 3-1, except that the vinylpyrrolidone-vinyl acetate random copolymer was not used (that is, only silica was used). Example 3-1
Tests were conducted in the same manner as in, and the results are shown in Table 5. However, for immunoserologic tests, the blood could not be collected because the blood clot did not peel off from the inner wall surface of the blood collection tube.
I couldn't inspect.

(Comparative Example 3-2) In Example 3-1,
A vinylpyrrolidone-vinyl acetate random copolymer is used instead of a blood test container consisting of a polyethylene terephthalate blood collection tube containing a vinylpyrrolidone-vinyl acetate random copolymer and silica. Example 3-1 without containing silica or silica
The same test was conducted and the results are shown in Table 5.

(Comparative Example 3-3) In Example 3-1,
Instead of the purified aqueous solution of vinylpyrrolidone-vinyl acetate random copolymer, a purified aqueous solution of polyether-modified silicone oil-based nonionic surfactant (SH3749 manufactured by Toray Dow Silicone Co., Ltd.) having a concentration of 1.0% by weight is used. A blood test container was obtained in the same manner as in Example 3-1 except that it was used, and was tested in the same manner as in Example 3-1, and the results are shown in Table 5.

[0063]

[Table 5]

The results of Examples 2 and 3 and Comparative Examples 2 and 3 are the results of polypropylene blood collection tubes (Comparative Example 2-2) and polyethylene terephthalate blood collection tubes in which only silica, which is a blood coagulation promoting substance, is sprayed on the inner wall surface. (Comparative Example 3-1)
However, as compared with the case where the inner wall surface is not subjected to any treatment, it causes more remarkable blood clot adhesion and serum is not recovered at all, whereas when silica is used in combination with the blood component adhesion preventing agent of the present invention, It shows that good separability can be obtained without inhibiting the coagulation promoting activity. In addition, the results of Example 3 and Comparative Example 3 show that the anti-adhesion agent for blood components of the present invention has two types of items which are immunoserologic tests, that is, HB.
It has been shown that s-Ab and Free T4 do not induce false positives, whereas conventional nonionic surfactants induce false positives.

Example 4 Vinylpyrrolidone-vinyl acetate random copolymer (BAS) as a blood component adhesion preventive agent
Using a trade name "Rubiscol VA28" manufactured by Company F and a vinyl acetate content of about 84 mol%, a methyl alcohol solution was prepared to a concentration of 0.02% by weight, and a 10 ml volume (inner diameter) was prepared. A 16 mm × 100 mm long polyethylene terephthalate (PET) resin tube was spray-coated on the inner wall surface and air-dried. The amount attached to the inner wall surface of the container per unit area was 5 × 10 ⁻⁷ g / cm ² of vinylpyrrolidone-vinyl acetate random copolymer.

Furthermore, as a plasma separating agent, dicyclopentadiene (DCPD) liquid resin (manufactured by Exxon Co., trade name "E"
CR-327 ") was mixed with fine powder silica (manufactured by Nippon Aerosil Co., Ltd., trade name" Aerosil A-200 ") with stirring to prepare 1.2 g of a product prepared to have a specific gravity of 1.05. Then, 120 U of sodium heparin was further contained as a blood anticoagulant to manufacture a blood test container.

8 m of fresh human blood was placed in the obtained blood test container.
l, inject tightly, mix by inverting 3 times, and mix at 10 ° C for 10
After allowing to stand for a minute, centrifugation was performed at a rotation speed of 3000 rpm for 5 minutes to observe the separated state of plasma, and at the same time, 1/2 volume of the supernatant plasma was immediately pipetted to obtain a sample after centrifugation. Furthermore, the blood test container after centrifugation was stored at 4 ° C., and after 24 hours, the supernatant plasma was pipetted to obtain a sample after storage for 24 hours. The concentrations of lactate dehydrogenase (LDH), creatine kinase (CPK) and potassium (K) were measured for the sample after centrifugation and the sample after storage for 24 hours.
It was shown to. The measured values shown in Table 6 are numerical values obtained by comparing the measured values of the sample after centrifugation for 24 hours with the measured value of the sample after centrifugation as 100.

(Comparative Example 4-1) 10 ml capacity (inner diameter 16
A blood test container was manufactured by containing 120 U of sodium heparin as a blood anticoagulant in a glass tube (mm x 100 mm) (a vinylpyrrolidone-vinyl acetate random copolymer and a plasma separating agent were not used). ). The performance evaluation of this blood test container was carried out in accordance with the performance evaluation of Example 4 in which “the blood test container after centrifugation was stored at 4 ° C., and after 24 hours, the supernatant plasma was collected with a pipette and stored for 24 hours. Instead of being "sampled,"
"We collected all the plasma from the blood test container after centrifugation,
½ of this was used as the sample after centrifugation, the rest was transferred to another glass container and stored at 4 ° C. for 24 hours to obtain a sample after storing for 24 hours. ” The performance evaluation of No. 4 was performed, and the results are shown in Table 6.

(Comparative Example 4-2) Except that the vinylpyrrolidone-vinyl acetate random copolymer was not used,
A blood test container was manufactured in the same manner as in Example 4 (that is, containing only the plasma separating agent and the blood anticoagulant). Using this blood test container, the performance was evaluated in the same manner as in Example 4, and the results are shown in Table 6.

[0070]

[Table 6]

(Example 5-1) Vinylpyrrolidone-vinyl acetate random copolymer (B
ASF Co., Rubiscol VA64, vinyl acetate content approx. 46 mol%), using heparin lithium as blood anticoagulant, 1.0 wt%, 4000 IU / ml, respectively
A purified water mixed solution of was prepared. About 25 μl of this solution was sprayed onto a 7 ml-volume blood collection tube made of polyethylene terephthalate and dried by a blow dryer at 60 ° C. Subsequently, about 1 g of a paste-like serum / plasma separating agent (manufactured by Sekisui Chemical Co., Ltd., trade name "ESCOLECT") was injected into the bottom of the dried blood collection tube to obtain a blood test container.

6 ml of fresh rabbit blood in this blood test container
Blood is collected, mixed thoroughly by inversion, and kept at 25 ° C for 1300
Centrifugation was performed under the conditions of G and 5 minutes, and the separation state was visually observed. Immediately thereafter, about half of the plasma was sampled and transferred to a clean hard glass test tube (sample for initial value),
The rest was left as it was in a polyethylene terephthalate blood collection tube, and both were refrigerated together at 4 ° C. for 24 hours. After 24 hours, the remaining plasma stored in the polyethylene terephthalate blood collection tube as it was was collected and transferred to a clean hard glass test tube (24-hour storage value sample). The effects on the blood chemistry test items (LDH as the enzyme and K as the electrolyte) were examined for the two types of plasma samples transferred to the hard glass test tube, and the results are shown in Table 7.

(Example 5-2) In Example 5-1:
Instead of the vinylpyrrolidone-vinyl acetate random copolymer, a vinyl alcohol-vinyl acetate random copolymer (Unitika Povar UMR-30L, vinyl acetate content of about 60 mol%) was used. A blood test container was manufactured in the same manner as in Example 5-1, and Example 5 was used.
The test was conducted in the same manner as in -1, and the results are shown in Table 7.

Comparative Example 5-1 A blood test container (ie, a blood test container) was prepared in the same manner as in Example 5-1 except that a vinylpyrrolidone-vinyl acetate random copolymer was not used as an agent for preventing blood component adhesion. Lithium heparin and serum / plasma separating agent are used). Using this blood test container, the same test as in Example 5-1 was conducted, and the results are shown in Table 7.
It was shown to.

(Comparative Example 5-2) About 25 μl of 4000 IU / ml purified aqueous solution of lithium heparin as a blood anticoagulant was sprayed on a clean hard glass blood collection tube having a volume of 7 ml, and dried with a blast dryer at 60 ° C. A blood test container was obtained (note that the serum / plasma separating agent was not injected). 6 ml of fresh rabbit blood was collected in this blood test container, mixed thoroughly by inversion, and kept at 25 ° C, 1300G, 5
Centrifugation was performed under the condition of minute, and the separation state after centrifugation was observed in the same manner as in Example 5-1. Then, the whole amount of plasma was immediately transferred to another clean hard glass test tube (sample for initial value) and refrigerated at 4 ° C. for 24 hours. 24
After the lapse of time, the same test as in Example 5-1 was conducted, and the results are shown in Table 7.

[0076]

[Table 7]

The results of Example 5 and Comparative Example 5 show that in a blood collection tube made of polyethylene terephthalate in which only the anticoagulant heparin salt was sprayed on the inner wall surface, adhesion of platelets occurred and therefore the sample was stored at 4 ° C. It shows that these values in plasma are elevated due to LDH and K, etc. leaking from platelets. However, even in such a case, it can be seen that the blood component adhesion preventive agent of the present invention effectively suppresses the adhesion of platelets and keeps the test value stable.
Further, Example 4 and Comparative Example 4 also show similar results.

Example 6 Vinylpyrrolidone-vinyl acetate random copolymer (BAS) as a blood component adhesion preventive agent
Manufactured by Company F, rubiscol VA64, content of vinyl acetate is about 46 mol%), and Example 2-1 as a blood coagulation promoting substance
Using the same fine powder silica as above, a methanol suspension was prepared so that the respective concentrations were 1.0% by weight and 2.0% by weight. The suspension was coated on polystyrene pellets having a diameter of about 3 mm while mixing and stirring in an explosion-proof blower dryer at 60 ° C., and dried to prepare a blood component adhesion-preventing carrier. The amount of each component adhering to the surface of the pellet was about 2 × when the vinylpyrrolidone-vinyl acetate random copolymer was used.
10 ⁻⁵ g / cm ² , fine silica powder is about 7 × 10 ⁻⁵ g / c
It was m ² .

0.6 g of the obtained blood component adhesion-preventing carrier
Was collected in a clean hard glass blood collection tube having a capacity of 10 ml to obtain a blood test tube for blood test. 4 ml of fresh rabbit blood was collected in this blood test tube and left at room temperature of 23 to 25 ° C. After measuring the coagulation time of blood,
Centrifugation was performed under the conditions of 0 G for 5 minutes to visually observe the separation state of serum and to measure the yield of serum. As a result, the pellet was buried in the clot so that it was scattered around the head of the clot, but clear serum without hemolysis was obtained.
The coagulation time and serum yield were as shown in Table 8.

Comparative Example 6-1 A polystyrene pellet coated only with fine powder silica was obtained in the same manner as in Example 6 except that the vinylpyrrolidone-vinyl acetate random copolymer was not used. The adhered amount of finely divided silica was about 5 × 10 ⁻⁵ g / cm ² . The polystyrene pellets (0.6 g) thus obtained were housed in a clean hard glass blood collection tube having a volume of 10 ml to obtain a blood test blood collection tube. Collect 4 ml of rabbit fresh blood into this blood test tube,
In the same manner as in Example 6, blood coagulation time, serum separation status, and serum yield were measured. As a result, the pellet was buried in the vicinity of the head of the blood clot as in Example 6, and although the serum yield itself was comparable to that in Example 6, remarkably strong hemolysis was observed. The coagulation time and serum yield were as shown in Table 8.

(Comparative Example 6-2) 0.6 g of polystyrene pellets not coated with anything was housed in a clean hard glass blood collection tube having a capacity of 10 ml to obtain a blood collection tube for blood test. 4 ml of rabbit fresh blood was collected in the blood collecting tube for blood test, and blood coagulation time, serum separation status, and serum yield were measured in the same manner as in Example 6. As a result, the pellet was buried in the vicinity of the head of the blood clot as in Example 6, and the serum yield itself was comparable to Example 6, but
Intense hemolysis was seen. The coagulation time and serum yield were as shown in Table 8.

[0082]

[Table 8]

[0083]

The anti-adhesion agent for blood components of the present invention can effectively prevent the blood components from adhering to the inner wall surface of the test container without causing false positives in immunoserologic tests. By itself, or minerals such as glass, kaolin, bentonite, silica, and celite, or substances having an organic blood coagulation-promoting property such as ellagic acid, and further anti-oxidants such as ethylenediaminetetraacetic acid salt, citrate salt, and heparin salt. When used in combination with a coagulant, a fluorinated salt, a glycolytic inhibitor such as mannose, etc., it is extremely useful in improving the accuracy of tests such as blood chemistry, immune serology, hematology tests.

The blood test container of the present invention has the anti-adhesion agent for blood components of the present invention on its inner wall surface, and does not cause false positive immunoserologic tests, and is a blood component test container. Since it can effectively prevent the adhesion to the inner wall surface of itself, glass, kaolin, bentonite, silica, mineral substances such as celite, or a substance having an organic blood coagulation promoting property such as ellagic acid, Furthermore, by using in combination with anticoagulants such as ethylenediaminetetraacetate, citrate, heparin salts, and anti-glycolytic agents such as fluoride, mannose, etc. It is extremely useful in improving.

The anti-adhesion carrier for blood components of the present invention has the anti-adhesion agent for blood components of the present invention on the surface thereof, and the carrier surface for blood components does not cause false positives in immunoserologic tests. Since it can effectively prevent adhesion to
By itself, or minerals such as glass, kaolin, bentonite, silica, and celite, or substances having an organic blood coagulation-promoting property such as ellagic acid, and further anti-oxidants such as ethylenediaminetetraacetic acid salt, citrate salt, and heparin salt. When used in combination with a coagulant, a fluorinated salt, a glycolytic inhibitor such as mannose, etc., it is extremely useful in improving the accuracy of tests such as blood chemistry, immune serology, hematology tests.

Claims (3)

(57) [Claims] 1. A monomer component homopolymer exhibits water soluble and (a), a random copolymer homopolymer is further a monomer component showing a water-insoluble (b), monomer Formation
Minute (a) is vinyl alcohol, vinylpyrrolidone, ethi
Renoxide, acrylate, styrene sulfonic acid
Salt, vinylphosphonate, allylamine salt, hydroxy
Methyl (meth) acrylate, glycosylethyl (meth)
) Acrylate, sugars such as glucose, and glutamine
At least one selected from the group consisting of amino acids such as acids
And the monomer component (b) is ethylene, propylene,
Ropylene oxide, vinyl acetate, vinyl chloride, alk
(Meth) acrylate, styrene, acrylonitrile
And at least one selected from the group consisting of acrolein
And the content of the monomer component (b) is 10
A blood component adhesion preventive agent comprising the copolymer in an amount of ˜90 mol%. 2. A blood test container, wherein the blood component adhesion preventing agent according to claim 1 is present on the inner wall surface. 3. A blood component adhesion-preventing carrier, wherein the blood component adhesion preventing agent according to claim 1 is present on the surface.