JP2002365282A - Composition for separating blood serum or blood plasma - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a composition for separating blood serum or blood plasma not floating an oily component in blood serum or blood plasma after centrifugal separation regardless of a centrifugal separation method. SOLUTION: The composition for separating blood serum or blood plasma consists of 100 pts.wt. of an oligomer A of cyclopentadiene with a softening point of 95-140 deg.C, 5-850 pts.wt. of an oligomer B of cylopentadiene with a softening point of 70-90 deg.C, 50-650 pts.wt. of a plasticizer, 5-85 pts.wt. of fine powdery silica and 0.1-6 pts.wt. of an organic gelling agent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a composition for separating serum or plasma used for centrifugation utilizing the difference in specific gravity of blood components.

[0002]

2. Description of the Related Art Conventionally, there has been known a blood test container in which a thixotropic composition for separating serum or plasma, for example, a mixture of silicone and silica is previously contained in the bottom of a blood collection tube (Japanese Patent Application Laid-Open No. 51-1979). No. 83654). When blood is collected in the blood collection tube, left for a suitable time, and then centrifuged, the centrifugal force causes the gel-like serum or plasma separation composition to have fluidity. Further, since the gel-like composition for separating serum or plasma has a specific gravity of the serum or plasma component and an intermediate specific gravity of that of the clot or blood cell component, it gradually rises in the collected blood from the bottom of the tube to form a serum or plasma layer. And a blood clot or blood cell layer, so that a serum or plasma component and a blood clot or blood cell component can be separated. The serum or plasma component thus separated from the blood clot or blood cell component can be easily removed from the blood collection tube and subjected to various tests, and can be stored without being transferred to another container.

[0003] In addition to the above silicones, α-olefin-maleic diester copolymers (Japanese Patent Application Laid-Open Nos. 56-16956 and 56-16956) are the main components of such thixotropic serum or plasma separating compositions. 2-16815
No. 9), a polyester-based polymer (JP-A-61-2333).
No. 68), acrylic polymers (JP-A-53-42283).
No.), chlorinated polybutene (JP-A-57-9718),
Cyclopentadiene resin (JP-A-1-295163)
No.), hydroxyl group, ester group,
A modified product of cyclopentadiene resin having an ether group, an epoxy group or the like introduced therein (Japanese Patent Laid-Open No. 2-95257) is known. Inorganic fillers that also act as gelling agents, substances having polar groups at both ends of molecules such as propylene glycol and ethylenediamine (JP-A-1-295163), condensates of sorbitol and aromatic aldehydes ( 2-1681
An organic gelling agent such as No. 59) is incorporated as necessary.

[0004] However, silicone is poorly compatible with inorganic fillers and hardly reacts at present due to a curing reaction caused by sterilization by radiation (γ-ray, electron beam, etc.), and α-olefin-maleic acid is not used at present. Diester copolymer, polyester polymer, acrylic polymer,
Those having a polar group, such as a modified product of cyclopentadiene resin, have relatively little effect on the measurement of clinical test items in blood, but have a drug concentration in blood (for example, phenobarbital, which is an antiepileptic drug, Carbamazepine, phenytoin, etc.).

[0005] On the other hand, in the case of using chlorinated polybutene, chlorine gas is generated at the time of incineration disposal after use, so that there is a problem that the incinerator is damaged or the environment is adversely affected.

Japanese Patent Application Laid-Open No. Hei 4-203965 discloses a composition for separating serum or plasma which does not have such disadvantages, and discloses an oligomer of cyclopentadiene and a condensate of sorbitol and an aromatic aldehyde as an organic gelling agent. Have been proposed. This composition for serum or plasma separation solves the above-mentioned problems of the prior art,
Dimethyl sulfoxide (hereinafter abbreviated as DMSO) or N, N-dimethylacetamide (hereinafter abbreviated as DMA) is used as a dispersant for the organic gelling agent. In a composition containing DMSO, radiation is used. (Γ-ray,
When sterilization is performed, there is a problem that dimethyl sulfide is generated due to the decomposition of DMSO and a bad smell is generated. In the case of containing DMA, the DMA may cause hemolysis by contact with blood, There has been a problem that a correct measurement value may not be given in measurement of a specific biochemical test item in blood.

[0007] The composition for separating serum or plasma is
There has been a problem that an oily component may be separated from the composition during storage (hereinafter, this phenomenon is referred to as phase separation).
This oil component is a low molecular weight component contained in the oligomer of cyclopentadiene in the above composition, and the low compatibility of this component with silica added as an organic gelling agent and / or a specific gravity adjusting agent is low. Cause of separation.
When this phase separation occurs, when the blood collection tube containing the above composition is turned over or turned over, the oily component adheres to the tube wall or plug, and blood collection is performed using this blood collection tube. When serum or plasma is separated from the clot or blood cells by centrifugation or the like, the oily components float in the serum or plasma layer,
As a result, when testing the components contained in serum or plasma,
The component clogs the sampling needle for serum or plasma of the test device and affects the test value. Furthermore, when phase separation occurs, there is a problem that the homogeneity of the composition is impaired and the partition wall stability is reduced.

An example of a composition for separating serum or plasma which can solve such a problem is disclosed in Japanese Patent Application Laid-Open No. 10-010122. According to this means, the phase separation is greatly improved as compared with the conventional composition for serum or plasma separation, but depending on the use conditions such as the centrifugation method, the separated oil component may be contained in the serum or plasma after centrifugation. The problem of floating on the surface could not be solved.

[0009]

DISCLOSURE OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and an object of the present invention is to make it possible for oily components to float in serum or plasma after centrifugation regardless of a centrifugation method. To provide a composition for serum or plasma separation.

[0010]

SUMMARY OF THE INVENTION The present invention provides a softening point of 95 to 95%.
Oligomeric A 100 of cyclopentadiene at 140 ° C.
5 parts by weight, 5 to 850 parts by weight of oligomer B of cyclopentadiene having a softening point of 70 to 90 ° C., 50 to 650 parts by weight of a plasticizer, 5 to 85 parts by weight of fine powder silica, and 0.1 to 6 parts by weight of an organic gelling agent A composition for separating serum or plasma.

The oligomers of cyclopentadiene include oligomers in which cyclopentadiene is multimerized,
And oligomers in which dicyclopentadiene, which is a dimer of cyclopentadiene, is multimerized. The oligomer can be produced by multiplying cyclopentadiene or dicyclopentadiene using, for example, a Diels-Alder reaction or the like. This is sometimes called a dicyclopentadiene resin (DCPD resin). It is preferable that the oligomer is further hydrogenated to saturate the remaining double bond.

The oligomer is JIS K6862.
-1984 “Melt viscosity test method for hot melt adhesives”
Those having a melt viscosity at 180 ° C. of 0.03 to 0.5 Pa · s as measured by the method A in the above are preferred. If the melt viscosity is too low, the viscosity of the present composition will be insufficient, and if it is too high, the viscosity of the present composition will be excessive.

The above oligomer has a specific gravity at 25 ° C. (based on a floating sedimentation test using a copper sulfate solution) of 1.02 to 2.0.
1.10 g / cm ³ , preferably 1.03 to 1.09
g / cm ³ are preferred. Specific gravity 1.02g /
If less than cm ³ or more than 1.10 g / cm ³ ,
It becomes difficult to suitably adjust the specific gravity of the present composition.

The oligomer preferably has a number average molecular weight of 200 to 800, more preferably 500 to 700, and more preferably 500 to 1500 in weight average molecular weight in terms of polystyrene by GPC. Preferably it is 700-1200. The number average molecular weight and the weight average molecular weight are 200 and 50, respectively.
If it is less than 0, phase separation is liable to occur, and each is 80%.
If it exceeds 0, 1500, the viscosity of the composition becomes too high.

The above oligomer preferably has a glass transition point of 15 to 90 ° C. according to the DSC method, more preferably 20 to 80 ° C. If the glass transition point is lower than 15 ° C., phase separation tends to occur, and if it exceeds 90 ° C., the viscosity of the present composition becomes too high.

The above-mentioned oligomer A of cyclopentadiene
Has a softening point of 9 as measured by JIS K 6863-1994 "Testing method of softening point of hot melt adhesive".
Limited to 5-140 ° C. When the softening point is less than 95 ° C., the effect of improving the oil component due to the effect of mixing cyclopentadiene with oligomer B cannot be exhibited. On the other hand, if the softening point exceeds 140 ° C., it becomes difficult to melt, and the production becomes difficult.

The above-mentioned oligomer B of cyclopentadiene
Has a softening point of 7 as measured by JIS K 6863-1994 "Testing method for softening point of hot melt adhesive".
Limited to 0-90 ° C. If the softening point is less than 70 ° C., phase separation is likely to occur, and if it exceeds 90 ° C., the effect of improving the oil component due to the effect of mixing cyclopentadiene with oligomer A cannot be exhibited.

The amount of the oligomer B of cyclopentadiene is limited to 5 to 850 parts by weight, preferably 7 to 600 parts by weight, more preferably 10 to 55 parts by weight, based on 100 parts by weight of the cyclopentadiene oligomer A.
0 parts by weight. If the amount is less than 5 parts by weight or exceeds 850 parts by weight, the effect of improving the oil component due to the effect of mixing cyclopentadiene with oligomer A cannot be exhibited.

The plasticizer is preferably a phthalic acid ester from the viewpoint of excellent compatibility with the oligomer of cyclopentadiene. The phthalic acid ester is preferably one in which at least one of the alcohol residues forming two ester groups has 6 or more carbon atoms. Both diesters having 5 or less carbon atoms tend to have reduced compatibility with cyclopentadiene oligomers. Further, the carbon number of each of the alcohol residues forming the two ester groups is preferably 11 or less, since it is difficult to adjust the specific gravity of the present composition to a suitable range if it is too large.

Examples of the plasticizer include butylpentyl phthalate, dipentyl phthalate, butylhexyl phthalate,
Butyl heptyl phthalate, dihexyl phthalate, pentyl heptyl phthalate, butyl nonyl phthalate, pentyl octyl phthalate, xyl heptyl phthalate, diheptyl phthalate, heptyl octyl phthalate, dioctyl phthalate, octyl nonyl phthalate, diisononyl phthalate, Octyldecyl phthalate, diisodecyl phthalate,
Examples include decylundecyl phthalate, diundecyl phthalate, and butylbenzyl phthalate. A particularly preferred plasticizer is a phthalic acid diester having 9 to 11 carbon atoms in each of the alcohol residues forming the two ester groups.

The amount of the plasticizer used is 50 to 650 with respect to 100 parts by weight of the oligomer A of cyclopentadiene.
It is limited to parts by weight, preferably 60 to 600 parts by weight, more preferably 70 to 550 parts by weight. If it is less than 50 parts by weight or exceeds 650 parts by weight, it becomes difficult to suitably adjust the viscosity of the composition.

The above-mentioned finely divided silica is an amorphous silicon dioxide (chemical formula: SiO ₂ .n ((CH ₃ ) ₂ Si) in which most of the hydroxyl groups on the surface of the primary particles of silicon dioxide are substituted with methyl groups.
O)) is preferred. Since the above-mentioned fumed silica has hydrophobicity, it disperses well in oligomers of cyclopentadiene or a plasticizer, but does not dissolve in blood and does not cause hemolysis. And is preferably used.

The specific surface area of the finely divided silica is 10 to 1
000 m ² / g is preferred, more preferably 50 to 50
0 m ² / g. If the specific surface area is less than 10 m ² / g or 10
When it exceeds 00 m ² / g, it becomes difficult to suitably adjust the thixotropic property of the present composition.

The fine particle silica has a primary particle diameter of 1 to 10
It is preferably 0 μm, more preferably 5 to 50 μm. When the primary particle diameter is less than 1 μm or exceeds 100 μm, it becomes difficult to suitably adjust the thixotropic property of the present composition.

The amount of the fine silica powder is 5 to 85 parts by weight based on 100 parts by weight of the oligomer A of cyclopentadiene.
Parts by weight, preferably 8 to 40 parts by weight,
More preferably, it is 10 to 35 parts by weight. When the use amount of the fine powder silica is less than 5 parts by weight or exceeds 85 parts by weight,
It becomes difficult to suitably adjust the specific gravity or thixotropic property of the present composition.

The organic gelling agent is preferably a condensate of sorbitol and an aromatic aldehyde, and examples thereof include dibenzylidene sorbitol, tribenzylidene sorbitol, and methyl-substituted dibenzylidene sorbitol. Since they do not have water absorption or water solubility, even when they come into contact with blood for a long time, the present composition does not become cloudy due to water absorption and has no side effects such as blood concentration. Among these, dibenzylidene sorbitol is particularly preferred from the viewpoint of exhibiting the thixotropic property of the present composition well.

The amount of the organic gelling agent used is 0.1 to 100 parts by weight of cyclopentadiene oligomer A.
It is limited to 6 parts by weight, preferably 0.1 to 4 parts by weight. When the amount of the organic gelling agent is less than 0.1 part by weight or exceeds 6 parts by weight, it becomes difficult to suitably adjust the thixotropic property of the present composition.

If necessary, a dispersant of an organic gelling agent may be added to the composition of the present invention.

The dispersant for the above-mentioned organic gelling agent is preferably 1.0 to 9.0, more preferably 4.0 to 6.0.
Is selected from the group consisting of polyoxyethylene-polyoxypropylene block copolymers having an HLB value of 1-methyl-2-pyrrolidone and mixtures thereof.

When the HLB value of the above block copolymer is too low, the dispersing effect of the organic gelling agent becomes insufficient, and the thixotropic property of the present composition decreases. If it is too high, the hydrophobicity is insufficient, and the composition may be dissolved in blood at the time of use of the composition to lyse the blood, and components in red blood cells may be mixed into serum or plasma to give an incorrect test result.

The amount of the block copolymer to be used is 0.1 to 100 parts by weight of cyclopentadiene oligomer A.
It is preferably 1 to 15 parts by weight, more preferably 0.1 to 5 parts by weight.
Parts by weight. If the amount of the block copolymer is too small, the effect of dispersing the organic gelling agent becomes insufficient, and the thixotropic property of the present composition decreases. If the amount is too large, the compatibility of the cyclopentadiene with the oligomer decreases, and the thixotropic property of the present composition decreases.

The above-mentioned 1-methyl-2-pyrrolidone dissolves the organic gelling agent satisfactorily, does not react with blood to cause hemolysis, and decomposes the composition by irradiation for the purpose of sterilization. It is preferably used because it does not have any.

The specific gravity of the composition of the present invention composed of the above-mentioned components is preferably intermediate between the specific gravity of serum or plasma and that of blood clots or blood cells. preferably 1.08 g / cm ^3, more preferably 1.04~1.06g / cm ^3.

The viscosity of the present composition can be determined by a conventional centrifugation operation, such that the present composition is located in the middle of a serum or plasma layer and a clot or blood cell layer, or to a blood test container such as a vacuum blood collection tube. In view of the ease of filling work, the pressure is preferably 50 to 1000 Pa · s at 25 ° C., more preferably 60 to 500 Pa · s.

The method for producing the composition of the present invention is not particularly limited, and is carried out by a usual high-viscosity type mixing apparatus. The mixing order is not particularly limited, but it is preferable to add finely divided silica at the end. At this time, if the viscosity is too high, uniform mixing becomes difficult, so that the heating is usually performed at 50 to 150 ° C, preferably at 70 to 120 ° C.
Further, it is preferable that the organic gelling agent and the dispersant of the organic gelling agent are mixed in advance. Further, the mixing may be performed under reduced pressure using a vacuum pump or the like. By performing the treatment under reduced pressure, air bubbles can be prevented from being mixed into the composition, and air dissolved in the components can be removed. The degree of pressure reduction is preferably 0 to 13.3 kPa as the pressure in the mixing step in order to effectively remove air.

(Function) As the cyclopentadiene oligomer used in the present composition, the compatibility between the cyclopentadiene oligomer and the plasticizer can be improved by using at least two oligomers having different softening points. By this effect, the phase separation of the plasticizer, which has been generated in a very small amount from the present composition by the centrifugal action at the time of centrifugation, can be suppressed. Therefore, regardless of the centrifugation method, the oil component does not float in the serum or plasma after the centrifugation.

[0037]

BEST MODE FOR CARRYING OUT THE INVENTION In the following Examples and Comparative Examples, the materials used as components of the compositions are as follows. Cyclopentadiene oligomer A • Cyclopentadiene oligomer A1: softening point 101
C., Tonex Corporation, ECR251.Cyclopentadiene oligomer A2: softening point 125
C., Tonex Corporation, ECR251 Cyclopentadiene oligomer B • Cyclopentadiene oligomer B1: softening point 86
5 ° C., manufactured by Tonex Corporation, 5380. Cyclopentadiene oligomer B2: softening point 73
5 ° C, 5380 plasticizer manufactured by Tonex, Inc. ・ Phthalic acid diester in which the alcohol residue is an alkyl group having 9 to 11 carbon atoms, PL-200 fine powdered silica manufactured by Mitsubishi Gas Chemical Company, Inc. ・ Specific surface area 250 m ² / g, Tokuyama Corporation Made, Leolo Seal DM30S

Example 1 (Mixture of organic gelling agent and dispersant of organic gelling agent) In a 2 liter glass beaker, 100 parts by weight of 1-methyl-2-pyrrolidone (manufactured by BASF), dibenzylidene sorbitol (Shin Nippon Rika Co., Ltd., Gel All D) 25 parts by weight, and using a magnetic stirrer at room temperature 2
After stirring for an hour, the dibenzylidene sorbitol was dissolved. (Mixing of this composition) Cyclopentadiene oligomer A1 100 parts by weight Cyclopentadiene oligomer B1 22 parts by weight Plasticizer 88 parts by weight Fine powder silica 9.4 parts by weight Mixture of the above gelling agent and dispersant of gelling agent 0 0.8 parts by weight 10 mmHg while maintaining the above at 105 ± 5 ° C.
Under reduced pressure to prepare a composition for serum or plasma separation. The specific gravity at 25 ° C. of the obtained composition is 1.05 g.
/ Cm ³ , the viscosity at 25 ° C. was 160 Pa · s.

Example 2 (mixture of the present composition) 100 parts by weight of oligomer A1 of cyclopentadiene 54 parts by weight of oligomer B1 of cyclopentadiene 107 parts by weight of plasticizer 11.7 parts by weight of fine powdered silica 11.7 parts by weight The same gel as in Example 1 0.9 parts by weight of a mixture of a dispersant of a gelling agent and a gelling agent A composition for separating serum or plasma was prepared in the same manner as in Example 1, except that the composition of the composition was changed as described above. . The specific gravity of the obtained composition at 25 ° C. is 1.05 g /
The viscosity at cm ³ and 25 ° C. was 150 Pa · s.

Example 3 (Blending of the present composition) 100 parts by weight of oligomer A1 of cyclopentadiene 107 parts by weight of oligomer B1 of cyclopentadiene 140 parts by weight of plasticizer 15.4 parts by weight of fine powdered silica Gel similar to that in Example 1 1.2 parts by weight of a mixture of a dispersant of a gelling agent and a gelling agent A composition for separating serum or plasma was prepared in the same manner as in Example 1 except that the composition of the composition was changed as described above. . The specific gravity of the obtained composition at 25 ° C. is 1.05 g /
The viscosity at cm ³ and 25 ° C. was 160 Pa · s.

Example 4 Oligomer A of cyclopentadiene in Example 2
1 Same as Example 2 except that 100 parts by weight of cyclopentadiene oligomer A2 was used in place of 100 parts by weight, and 54 parts by weight of cyclopentadiene oligomer B1 was used in place of 54 parts by weight of oligomer B2 of cyclopentadiene. To prepare a composition for separating serum or plasma. The specific gravity of the obtained composition at 25 ° C is 1.05 g / cm ³ , and the viscosity at 25 ° C is 170 Pa
-It was s.

Comparative Example 1 (Mixing of Compositions) Cyclopentadiene Oligomer A1 100 parts by weight Plasticizer 74 parts by weight Fine powdered silica 6.9 parts by weight Same gelling agent and gelling agent dispersant as in Example 1. 0.6 part by weight of a mixture of the above, except that the composition of the composition was changed as described above, to prepare a serum or plasma separation composition in the same manner as in Example 1. The specific gravity of the obtained composition at 25 ° C. is 1.05 g /
The viscosity at cm ³ and 25 ° C. was 160 Pa · s.

Comparative Example 2 (Mixing of Compositions) Cyclopentadiene Oligomer B1 100 parts by weight Plasticizer 61 parts by weight Fine powdered silica 7.2 parts by weight Gelling agent and dispersant for gelling agent as in Example 1 0.6 part by weight of a mixture of the above, except that the composition of the composition was changed as described above, to prepare a serum or plasma separation composition in the same manner as in Example 1. The specific gravity of the obtained composition at 25 ° C. is 1.05 g /
The viscosity at cm ³ and 25 ° C. was 150 Pa · s.

Comparative Example 3 (Mixing of Compositions) Cyclopentadiene Oligomer A1 100 parts by weight Cyclopentadiene Oligomer B1 880 parts by weight Plasticizer 605 parts by weight Fine powdered silica 72 parts by weight The same gelling agent as in Example 1 5.7 parts by weight of a mixture of a gelling agent and a dispersant A composition for separating serum or plasma was prepared in the same manner as in Example 1 except that the composition was changed as described above. The specific gravity of the obtained composition at 25 ° C. is 1.05 g /
The viscosity at cm ³ and 25 ° C. was 160 Pa · s.

[Performance Test] The following performance tests were performed on the serum or plasma separation compositions obtained in the above Examples and Comparative Examples. (Blood separation test) 1.35 g of the present composition was weighed 16 mm in outer diameter
A glass test tube having an inner diameter of 13 mm and a length of 100 mm and having a round bottom was filled so that air bubbles did not enter the bottom, and 2 ml of fresh human blood was added. The opening of the glass tube was sealed with a butyl rubber stopper, gently mixed by inverting 5 times, and then left at room temperature for 3 hours. Thereafter, the mixture was centrifuged at 1800 G for 10 minutes. After centrifugation, the serum layer was observed to confirm the presence or absence of oil-phase-separated substances.

As a result, no suspension of oily substance was observed in any of Examples 1 to 4, whereas in Comparative Example, 1
浮遊 2 oil droplet-like floating substances were observed to float.

[0047]

The composition of the present invention comprises a specific amount of a cyclopentadiene oligomer A having a specific softening point, a cyclopentadiene oligomer B, a plasticizer, finely divided silica and an organic gelling agent. It can be suitably used when separating serum or plasma by centrifugation.

 ────────────────────────────────────────────────── ─── Continuing on the front page (72) Inventor Hironobu Ishikawa 4560 Kaiseicho, Shinnanyo-shi, Yamaguchi Sekisui Chemical Co., Ltd. F-term (reference) 2G045 AA01 CA25 HB12 4J002 BK00W BK00X CE00W CE00X DJ017 EC058 EH146 FD026 FD208 GB00

Claims (1)

[Claims] 1. A cyclopentadiene oligomer A having a softening point of 95 to 140 ° C., 100 parts by weight, a softening point of 70 to 90 ° C.
5 to 850 parts by weight of a cyclopentadiene oligomer B, 50 to 650 parts by weight of a plasticizer,
A composition for separating serum or plasma, which comprises parts by weight and 0.1 to 6 parts by weight of an organic gelling agent.