JP2000249700A - Anticoagulant composition and its manufacture - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent hemolyzation from occurring when mixed with blood by combining an anticoagulant with a binder for making solid. SOLUTION: The composition can be obtained by drying a solution with a pH of for example 5-10 containing an anticoagulant and a binder for preventing blood from being coagulated. The anticoagulant is selected from a group consisting of a calciumchelate agent, a thrombin inhibitor, and a protein synthesis inhibitor and is used in concentration for preventing the blood from being coagulated. The binder includes gelatin, carboxymethylcellulose, or the like, and polyvinylalcohol is preferable among them. Also, a drying method includes vacuum drying, hot-air drying, or the like. Among them, freezing drying and atomizing drying are preferable, where the atomizing drying is most preferable since it can efficiently prevent hemolyzation.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an anticoagulant composition used for anticoagulation of blood in a clinical test.

[0002]

2. Description of the Related Art Various anticoagulants are used in blood tests. For example, EDTA salt is mainly used for a blood cell count test, and citrate is mainly used for a blood coagulation test. In addition, an anticoagulant suitable for each of various test items such as heparin and CTAD is used. Vacuum blood collection tubes containing these anticoagulants are commercially available. These anticoagulants are solid or liquid.

[0003]

By the way, these solid anticoagulants are apt to cause hemolysis at the moment when they are mixed with blood and dissolved, the osmotic pressure locally increases. However, hemolysis is not preferable because it adversely affects the measurement of various test items. In order to prevent this, various measures have been taken such as granulating the anticoagulant or coating the blood collection tube wall, but this is not sufficient.

An object of the present invention is to provide a solidified anticoagulant composition which does not cause hemolysis when mixed with blood.

[0005]

The anticoagulant composition of the present invention is characterized in that it is solidified containing an anticoagulant capable of preventing blood from coagulating and a binder.

[0006]

BEST MODE FOR CARRYING OUT THE INVENTION Anticoagulants used for medical purposes include calcium chelating agents,
It is selected from the group consisting of thrombin inhibitors and protein synthesis inhibitors. Examples of calcium chelators include E
DTA salts, citrates, CTADs and the like. Examples of thrombin inhibitors include heparin and hirudin, and examples of the protein synthesis inhibitors include DFP (diisopropylfluorophosphate) and PMSF (phenylmethylsulfonylfluro). Ride).

[0007] The amount of the anticoagulant is used at a concentration that can prevent blood from coagulating. The required amount per ml of blood varies depending on the type of anticoagulant to be used. For example, about 0.5 to 2 mg for EDTA salt, about 2 to 5 mg for citrate, and about 0.01 to 5 mg for heparin.
0.1 mg and 1.6 mg to 6.4 mg in CTAD are suitably used.

[0008] As the binder, those used in the granulation are suitable, for example, gelatin, dextrin, potato starch, sodium alginate, gum arabic, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose. , Glucose, sorbit, guar gum, polyvinyl alcohol and polyvinyl pyrrolidone, with polyvinyl alcohol being preferred.

The amount of the binder used varies depending on the type of the binder. In the case of methylcellulose, about 0.2 w / v% of the aqueous solution of the anticoagulant composition is used. In the case of polyvinyl alcohol, the amount of the aqueous solution of the anticoagulant composition is 0%. 0.2-5.0
It is preferably used in w / v%.

Although the mechanism of action of the binder in the present invention is not clear, the solidification of the binder with the anticoagulant slows the dissolution rate of the anticoagulant into the blood when it comes into contact with blood. As a result, it is considered that the osmotic pressure can be prevented from locally increasing.

The anticoagulant composition of the present invention has a pH of 5 to 1 containing an anticoagulant capable of preventing blood from coagulating and a binder.
It can be obtained by drying an aqueous solution having a pH of 0, preferably pH 7 to 9.

The pH can be adjusted with an acid or an alkali, but it is also possible to use a combination of the same kind of acid and an alkaline salt, such as a combination of citric acid and sodium citrate.

Depending on the anticoagulant, there is a case where the component cannot be dissolved even if it is directly dissolved in the above pH range. In such a case, the insoluble constituents are once dissolved in a water-soluble organic solvent, for example, ethanol of 20 to 40 (v / v)% of the prepared amount of an aqueous solution of the anticoagulant composition, and then the solution is dissolved. May be mixed with an aqueous solution containing other components.

Drying methods include freeze drying, vacuum drying,
Hot air drying, spray drying and the like can be mentioned. Among them, freeze drying and spray drying are preferable, but spray drying is most preferable because hemolysis can be efficiently prevented. Spray drying is a method of converting a solution into a solid (powder or granule) with a spray drier.

For drying, after the aqueous solution containing the anticoagulant composition of the present invention is dried, a necessary amount may be weighed in a test tube, or the anticoagulant composition of the present invention may be previously placed in a test tube. After dispensing a predetermined amount of the aqueous solution, the aqueous solution in the test tube may be dried. Alternatively, drying may be performed while spraying a predetermined amount into a test tube.

[0016]

EXAMPLE Preparation of CTAD Anticoagulant Composition 6.4184 g of sodium citrate and 37.000 citric acid in 20% (v / v) water of the aqueous solution of the anticoagulant composition.
Dissolve 0 mg. Separately, polyvinyl alcohol (Wako Pure Chemical Industries, Ltd., degree of polymerization 500) was used as a binder in 20% (v / v)% water of the prepared aqueous solution of the anticoagulant composition.
After adding 2-5.0 g, theophylline and adenine, further adding 40 (v / v)% ethanol and dipyridamole of the prepared aqueous solution of the anticoagulant composition, dissolving the mixture, mixing with the citrate solution, Preparation amount of aqueous solution of coagulant composition 2
0 (v / v)% water is added to obtain 100 ml of a CTAD anticoagulant composition at pH 7.3.

Drying of CTAD Anticoagulant Composition The anticoagulant composition obtained above was spray-dried using a granulator GB-21 (manufactured by Yamato Scientific Co., Ltd.). The resulting dried product contains a small amount of 2-3 μm containing a small particle size of 5-10 μm.
It was a powder having a particle size of m.

Preparation of blood collection tube The dried product of the anticoagulant composition obtained above was mixed with 1 ml of blood.
The CTAD amount was set to 3.2 mg per test tube for each 5 ml to prepare a blood collection tube.

Confirmation of Degree of Hemolysis Blood collected with a syringe was collected into the blood collection tube obtained above and mixed well. After that, the sample was
After centrifugation for 0 minutes, the absorbance of the supernatant at the absorption wavelength (576.6 nm) of oxyhemoglobin was measured with a spectrophotometer. As controls, EDTA-2K and 3.8% sodium citrate were used.

Blood treated with CTAD that does not contain polyvinyl alcohol (PVA) causes hemolysis and the absorbance of the supernatant increases under the influence of the hemolysis, but the anticoagulant composition of the present invention (CTAD containing PVA) does not. It was confirmed that the treated blood had a low absorbance of the supernatant and could prevent hemolysis.
(Fig. 1)

Influence on blood cell count test data and changes over time In the above examination, a blood collection tube containing an anticoagulant composition (CTAD PVA 1.0
%), The blood collected by syringe was collected, and measurement was performed with a multi-item automatic blood cell analyzer SE-9000 (Sysmex Corporation) to examine the influence on test data and changes over time. The same blood treated with EDTA-2K was used as a control.

[0022]

[Table 1]

As shown in Table 1, the anticoagulant composition of the present invention showed the same test data and change over time as the control,
It was confirmed that the measurement was not adversely affected.

Time-dependent changes in the degree of hemolysis The same operations as those performed during the preparation of the blood collection tube were performed, and the time-dependent changes in the degree of hemolysis were examined. Anticoagulant composition of the present invention (CTAD
(PVA 1.0%) was confirmed to be less likely to cause hemolysis than the control. (Fig. 2)

Effect on Coagulation Test Data Blood collection tubes containing the anticoagulant composition of the present invention (CTAD PV
A1.0%), and collected by syringe.
After centrifugation at 0 rpm for 10 minutes, measurement was performed with a fully automatic blood coagulation analyzer CA-1000 (Sysmex Corporation). The same blood treated with 3.8% sodium citrate was used as a control.

The anticoagulant composition of the present invention comprises PT, APT
For both T and Fbg, data equivalent to the control was obtained.
(FIGS. 3 to 5)

[0027]

According to the anticoagulant composition of the present invention, when mixed with blood, hemolysis can be avoided and the measured data is not adversely affected. In addition, hemolysis can be suppressed even when blood is stored.

[Brief description of the drawings]

FIG. 1 is a diagram confirming the degree of hemolysis of an anticoagulant composition of the present invention.

FIG. 2 is a graph showing the change over time in the degree of hemolysis of the anticoagulant composition of the present invention.

FIG. 3 is a correlation diagram of the anticoagulant composition of the present invention and a control in PT measurement.

FIG. 4 is a correlation diagram of the anticoagulant composition of the present invention and a control in APTT measurement.

FIG. 5 is a correlation diagram of the anticoagulant composition of the present invention and a control in Fbg measurement.

Claims (5)

[Claims] 1. A solidified anticoagulant composition comprising an anticoagulant capable of preventing blood from coagulating and a binder. 2. The method according to claim 1, wherein the binder is gelatin, dextrin,
The anticoagulant composition according to claim 1, wherein the composition is selected from the group consisting of potato starch, sodium alginate, gum arabic, carboxymethylcellulose, hydroxypropylcellulose, methylcellulose, glucose, sorbitol, guar gum, polyvinyl alcohol and polyvinylpyrrolidone. . 3. The anticoagulant composition according to claim 1, wherein the anticoagulant is selected from the group consisting of a calcium chelating agent, a thrombin inhibitor and a protein synthesis inhibitor. 4. The anticoagulant composition according to claim 1, wherein the solidified state is powdery or granular. 5. A method for producing an anticoagulant composition, comprising spray-drying or freeze-drying an aqueous solution containing an anticoagulant capable of preventing blood from coagulating and a binder.