JP3237106B2 - Blood collection tube with anticoagulant - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a blood collection tube in which a stable anticoagulant is encapsulated.

[0002]

2. Description of the Related Art In a blood test, particularly in the case of an emergency patient, the measurement must be performed immediately after blood collection, so that it is necessary to suppress the activation of blood coagulation factors. Therefore Various strategies for providing anti-blood <br/> coagulant has been conventionally proposed, typical anticoagulants (hereinafter referred to as anticoagulant) as there are such as heparin. Heparin is composed of sulfated D-glucosamine, D-glucuronic acid and L-iduronic acid to form a polymer, and has a molecular weight of 5,000 to 30,000.
Is a mucopolysaccharide. The blood coagulation factor ATIII bound to heparin changes its conformation, and activates thrombin, IX
It works on both factor X, factor XI, and factor XII and shows strong anticoagulant action. Japanese Patent Application Laid-Open No. Hei 4-9143 discloses a blood collection tube using such heparin as an anticoagulant.
In the above, a carrier is coated or impregnated with sodium heparin or lithium heparin, and a certain amount of an anticoagulant is sealed in a blood collection tube. However, unlike proteins such as thrombin, heparin sodium or lithium heparin have no autolytic properties,
Since the stability to heat is not so good, when stored for a long period of one to two years, the heparin activity value may be reduced and the anticoagulant effect may not be sufficiently exhibited.

[0003]

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above circumstances, and provides a blood collection tube containing an anticoagulant containing a stabilizer so that heparin activity is maintained even after storage for a certain period of time. The purpose is to provide.

[0004]

SUMMARY OF THE INVENTION The present invention relates to a blood collection tube comprising a bottomed tube having one end opened and the other end closed, and a plug for sealing the opening of the bottomed tube. Alternatively, the present invention provides a blood collection tube containing an anticoagulant, which contains lithium heparin and a polypeptide fraction derived from collagen as a stabilizer .

In the present invention, a collagen-derived polypeptide fraction having an average molecular weight of 10,000 to 100,000 as a stabilizer is present in a blood collection tube. When the average molecular weight is less than 10,000, the stabilizing effect is weakened, which is not preferable.
If it exceeds 30, the water solubility will decrease, and if it is stored at a low temperature, it will be cloudy or those having a particularly large molecular weight will cause cloudy gelation, which is not preferable.

[0006] As the polypeptide fraction, highly purified collagen from pig-derived skin collagen or bovine tendon-derived collagen is preferably used. This polypeptide fraction is
0.01mg to 25mg per 1ml of blood to be collected
It is preferable to be enclosed in a blood collection tube together with an anticoagulant. If it is less than 0.01 mg, it is difficult to maintain the stability of sodium heparin or lithium heparin, and if it exceeds 25 mg , the liquid viscosity of the polypeptide fraction becomes high, affecting test item data such as proteins. There is a fear.

[0007] In the present invention, the amount of heparin sodium or lithium heparin sealed in the blood collection tube as an anticoagulant is preferably 5 to 30 units per ml of blood to be collected. If the amount is less than 5 units, the ability to anticoagulate blood is reduced, and if it exceeds 30 units , blood test data may be affected. As the heparin sodium or lithium heparin used in the present invention, those derived from cattle are generally used safely, but animals derived from animals such as pigs and dogs or humans can also be used.

[0008] The bottomed tube in the blood collection tube of the present invention includes:
Glass, homopolymers or copolymers such as polyethylene terephthalate , polybutylene terephthalate , polyethylene naphthalate , and polyethylene isophthalate, acrylic resins, polyethylene, polypropylene, polycarbonate, polyacetal, polyamide, and polyurethane are preferred.

The plug for sealing the open end of the bottomed tube in the blood collection tube of the present invention is preferably a plug made of rubber, an elastomer, an aluminum vapor-deposited film, or the like. Examples of the rubber include chlorinated butyl rubber, brominated butyl rubber, isoprene rubber, and natural rubber, and examples of the elastomer include olefin-based, polyester-based, and polyurethane-based. Further , as the plug, there is a form in which a rubber button such as isoprene is provided on an aluminum vapor-deposited film so that a needle can be punctured, and the bottomed tube and the film are bonded.

[0010] As a method for encapsulating the anticoagulant heparin sodium or heparin lithium and stabilizers which polypeptide fraction of the blood collection tube is, blood sodium heparin or lithium heparin and polypeptides fraction solution
After injecting into the tube, freeze drying, there is a method in which Ru is dried such as by vacuum drying or spray after hot-cold air.

Further, by enclosing sodium heparin or lithium heparin and a polypeptide fraction in a blood collection tube, sealing the open end of the bottomed tube with a stopper, and sealing it with an aluminum vapor-deposited film or a plastic packaging material,
Further, it is protected from heat and moisture for a long period of time, and can increase the stability of heparin.

[0012] Before the presence of heparin sodium or heparin lithium and polypeptides fractionation blood collection tube of the present invention, having a bottom inner surface on the blood of the abnormal coagulation by applying a silicone coating, and the this to prevent a deposited it can.

[0013] Furthermore, the blood collection tube of the present invention, the presence of heparin sodium or heparin lithium and-polypeptide fractions, by filling the serum separating agent, it is with this to simplify the inspection.

Hereinafter, the present invention will be described in more detail by way of examples.

EXAMPLES [Example 1] As a stabilizer, an average molecular weight of 2
000 collagen-derived polypeptide fraction (PEN
A 10% aqueous solution of TAPHARM (PRIONEX) was prepared. Beef heparin sodium (SPL)
396 mg of lyophilized powder was weighed, and 9.6 g of a 10% aqueous solution of the above-mentioned polypeptide fraction was added thereto to make the total amount about 10 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin sodium in the blood collection tube had an activity of 24.0 units per ml of blood. In addition, the above-mentioned stabilizer was added to 0.24
mg.

Example 2 As a stabilizer, an average molecular weight of 2
000 collagen-derived polypeptide fraction (PEN
A 10% aqueous solution of TAPHARM (PRIONEX) was prepared. 402 mg of a lyophilized powder of lithium bovine heparin (SPL) was weighed, and 9.6 g of a 10% aqueous solution of the above polypeptide fraction was added thereto.
0 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin lithium in the blood collection tube had an activity of 22.2 units per ml of blood. The stabilizer was contained in an amount of 0.24 mg per 1 ml of blood.

Comparative Example 1 A collagen-derived polypeptide fraction having an average molecular weight of 20,000 (PENTAPARM)
A 10% aqueous solution of PRIONEX) was prepared. 396 mg of lyophilized powder of bovine heparin sodium (SPL) was weighed, and 0.32 g of a 10% aqueous solution of a polypeptide fraction and distilled water were added thereto, and the total amount was 10 g.
And In the blood collection tube whose inner surface is coated with silicone,
After spraying the solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin sodium in the blood collection tube had an activity of 23.2 units per ml of blood. The stabilizer is 0.008 mg per 1 ml of blood.
Was included.

Comparative Example 2 A collagen-derived polypeptide fraction having an average molecular weight of 20,000 (PENTAPHARM)
A 10% aqueous solution of PRIONEX) was prepared. 396 mg of lyophilized powder of bovine heparin lithium (SPL) was weighed, and 1 part of the polypeptide fraction was added thereto.
0.32 g of 0% aqueous solution and distilled water were added to make the total amount 10 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin lithium in the blood collection tube had an activity of 22.1 units per ml of blood. The stabilizer was contained in an amount of 0.008 mg per 1 ml of blood.

Comparative Example 3 The effect of stabilization was measured using gelatin (Nitta Gelatin) having an average molecular weight of 6,000, which is a purified collagen product. 396 mg of a lyophilized powder of bovine heparin sodium (SPL) was weighed, and a 10% aqueous solution of gelatin was added thereto.
0 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin sodium in the blood collection tube had an activity of 22.2 units per ml of blood.

Comparative Example 4 The effect of stabilization was measured using gelatin having a mean molecular weight of 6,000 (Nitta Gelatin), which is a purified collagen product. Bovine heparin lithium (S.
P. L. 396 mg of freeze-dried powder of
A 10% aqueous solution of gelatin was added thereto to make the total amount 10 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin lithium in the blood collection tube had an activity of 24.0 units per ml of blood.

[Comparative Example 5] The measurement was carried out for the case where no stabilizer was added. Bovine heparin sodium (SP.
L. 396 mg of the freeze-dried powder of the above was weighed, and distilled water was added thereto to make the total amount 10 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin sodium in the blood collection tube was 2 per ml of blood.
8.2 units of activity.

[Comparative Example 6] The measurement was carried out for the case where no stabilizer was added. Bovine heparin lithium (SP
L. Was weighed, and distilled water was added thereto to make the total amount 10 g. A blood collection tube an inner surface and silicone coating, after spraying the aqueous solution 10 mg, dried by predetermined drying method, then blood 4ml was depressurized to allow blood sampling. At this time, bovine heparin lithium in the blood collection tube was 27.27 ml / ml of blood.
2 units of activity.

[Comparative Example 7] A commercially available product in which the inner surface of a blood collection tube was subjected to anticoagulation treatment with sodium heparin was used.

[Comparative Example 8] A commercially available product obtained by subjecting the inner surface of a blood collection tube to anticoagulation treatment with lithium heparin was used.

The above blood collection tubes were stored at 60 ° C. and 75% for 2 weeks, and the activity values of various heparin salts were measured using human blood.
(Unit) was measured. Table 1 shows the results.

[0025]

[Table 1]

[0026] Table 1 Examples 1 and 2 but the activity of the heparin salt, even after 14 days was maintained from Comparative Examples 1 and 2 is reduced to the active value of the heparin half after 14 days. This indicates that stabilizing effects are not obtained when the amount of polypeptide is less than 0.01 per ml of blood. Similarly, from Examples 1 and 2 and Comparative Examples 3 and 4, the heparin activity value is reduced to half after 14 days. Therefore, if the molecular weight of the polypeptide is less than 10,000, there is no stabilizing effect. You can see that. In Comparative Examples 5 and 6 in which a stabilizer was not added to the heparin salt , the activity was almost impaired after a storage period of 2 weeks. Even in Comparative Examples 7 and 8 using a commercially available blood collection tube containing heparin, the activity was 14 days later. Has been impaired.

From the above results, in order to preserve the blood collection tube to which heparin salt has been added for a long period of time, it is necessary to add a stabilizer. As the stabilizer, a polypeptide having a molecular weight of 10,000 or more is used in an amount of 0.01 or more. It can be said that the addition is more effective.

Furthermore, Table 2 shows the results of visual observation of the anticoagulant performance after the blood collection tube was stored at 60 ° C. and 75% for 2 weeks. The anticoagulant performance was judged based on the occurrence of fibrin. A sample in which plasma was successfully collected without generation of fibrin was indicated by a circle, and a sample in which fibrin was generated was indicated by a cross.

[0029]

[Table 2]

From the results shown in Table 2, it was found that Examples 1 and 2 of the present invention
When the blood collection tube 2 is used, plasma can be collected well even after 14 days. On the other hand, in Comparative Examples 1 to 8, the occurrence of fibrin was observed in all blood collection tubes, indicating that heparin activity was reduced during the storage period.

By using the blood collection tube of the present invention, the activation of coagulation factors in blood can be suppressed, so that blood can be provided in a good condition in a blood test in an emergency or the like. Further, the unstable heparin salt can be stabilized in the blood collection tube, and the quality can be maintained for a long period of time.

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int.Cl. ⁷ , DB name) A61B 5/15 G01N 33/48-33/49

Claims (2)

(57) [Claims] A bottomed pipe having one end open and the other end closed;
In a blood collection tube comprising a stopper for sealing the opening of the bottomed tube, sodium or heparin lithium and an average molecular weight of 10,000 to 100,000 as a stabilizer.
An anticoagulant-containing blood collection tube , wherein a collagen-derived polypeptide fraction of 0 is encapsulated. 2. The blood collection tube containing an anticoagulant according to claim 1, wherein the collagen-derived polypeptide fraction is enclosed in an amount of 0.01 to 25 mg per ml of blood to be collected.