JPH0145866B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a blood sample separation agent. To perform blood analysis, generally whole blood is separated into serum (or plasma) and clot (or blood cells) by centrifugation, the serum (or plasma) is pipetted out, and this is used for clinical analysis. It has been used for However, in this case, the operation is complicated and time-consuming, and at the same time, when pipetting out the serum, blood cell components may be mixed in and affect the analytical values, so a person skilled in the operation is required. There are many disadvantages such as blood cell components contaminating the serum (or plasma portion) due to impact during storage and transportation. Conventionally, attempts have been made to phase-separate liquids with different specific gravities by centrifugation to facilitate various analyses. When separating serum (or plasma) from blood clots (or blood cell components), serum (or plasma) and blood clots (blood cell components) are separated.
By using a separation composition that has a specific gravity between the It is known that this can be done only by tilting the For example, Japanese Patent Application Laid-Open No. 49-89389 and US Pat. No. 3,780,935 disclose the use of a gel-like material consisting of a mixture of silicone fluid and hydrophobic silica fine powder as a separation composition. This gel-like substance is given thixotropy by finely powdered silica mixed with a silicone fluid, and when centrifugation is performed, it flows and moves to the phase to be separated, and at the end of centrifugation, it is subjected to a tilting operation, A stable barrier is formed that is not easily destroyed by weak impacts. However, this composition causes the silicone fluid to react with the silica surface, promoting a decrease in the composition's viscosity over time, until it no longer has the ability to form a stable barrier, and due to tilting, the barrier is disrupted and blood cells are transferred to the serum portion. JP-A No. 51-83654 describes in detail that there is a possibility that the serum may be contaminated and have an adverse effect on the accuracy of serum analysis. JP-A-53-51189 also discloses the use of a composition using liquid epoxidized or maleated polybutadiene, finely powdered silica, and a small amount of triethylene glycol as a structure-forming agent. However, even in this case, since modified polybutadiene is used, although it is improved over unmodified polybutadiene, the disadvantage is that the viscosity of the composition changes when stored at room temperature in the air. Since a structure-forming agent such as a low-molecular-weight diol is required for reinforcement, it is possible that it may elute into the serum portion and adversely affect the accuracy of serum analysis. Furthermore, JP-A-53-146795 discloses (a) a saturated aliphatic dicarboxylic acid having 4 to 12 carbon atoms;
(b) polymeric fatty acids containing 75% by weight or more of C ₃₆ dibasic acids, and (c) branched chain aliphatic dihydric alcohols having 3 to 8 carbon atoms, such branched a mixture of chain dihydric alcohols or such branched chain aliphatic dihydric alcohols containing at least 50
essentially stoichiometric amounts of the reaction product of a mixture with a linear aliphatic dihydric alcohol having 2 to 8 carbon atoms containing % by weight, such that equivalents of (a) to (b) 210 with an average molecular weight of about 2,000 to 8,000 and an average molecular weight of about 2,000 to 8,000 centistokes.
Also disclosed are copolyester compositions useful as barrier agents in blood separation devices having kinematic viscosities and densities in the range of 1.015 to 1.060 g/cm ³ . However, in this case as well, it is difficult to completely control the molecular weight and viscosity of the target polyester because branched low molecular weight diols such as neopentyl glycol and 1,2-propanediol are used. The thixotropic properties of the composition may be subject to fluctuations. The purpose of the present invention is to analyze serum (or plasma) parts and blood clots (or blood cells) in analyzing blood components.
The aim is to improve the efficiency of serum (or plasma) separation operations and prevent contamination with blood cells during storage and transportation by using polyether ester, which has a specific gravity between the blood parts and is inert to the blood parts. . The present inventors have made extensive studies to achieve these objectives, and as a result, have arrived at the present invention. That is, the present invention is obtained from a polyoxyalkylene glycol having a number average molecular weight of 600 to 8000, a dicarboxylic acid having 6 to 80 carbon atoms or a derivative thereof, and optionally a low molecular weight diol, and a polyoxyalkylene glycol having a number average molecular weight of 1500 to 8000.
50000 and a viscosity of 10 to 3000 poise (20°C) containing an inert filler and optionally a surfactant and a structure forming agent, and a density of 1.010 to 1.070 g/cm ³ This is a blood sample separation agent characterized by the following. Put an appropriate amount of the blood sample separation agent of the present invention (hereinafter abbreviated as separation agent) into a suitable blood collection tube (for example, put 2.0 g of separation agent into a 10 ml blood collection tube for serum separation), collect blood, and obtain serum. If you want to leave the blood until it coagulates and then perform a centrifugation operation, the separating agent, which has a specific gravity between those of serum and blood clots, will move to the middle of the serum and blood clots during the centrifugation operation, causing a drop in the phase between the two. Forms a stable partition wall. Therefore, there is no need to pipette out to collect serum, and serum can be obtained simply by tilting the blood collection tube, and fluctuations in analytical values due to contamination of blood cell components due to phase disturbance during pipetting out can be avoided. Furthermore, the blood sample separation agent of the present invention has excellent viscosity stability. The blood sample separation agent of the present invention is a serum and plasma separation agent that improves the conventional drawbacks and can be manufactured at low cost. The polyether ester used in the present invention has a molecular weight of
1500-50000 (preferably 3000-20000),
The viscosity is 10 to 3000 poise (preferably 15 to 1000 poise). Also, 0.850 to 1.050 (preferably
It is desirable to have a specific gravity of 0.900 to 1.045 g/cm ³ ), and it is desirable to be a homogeneous polyetherester that is inert to blood components. The polyether ester used in the present invention is produced, for example, by reacting 1 mol of polyoxyalkylene glycol with a number average molecular weight of 600 to 8,000 and 0.50 to 0.98 mol of a dicarboxylic acid having 8 to 80 carbon atoms. Polyoxyalkylene glycols include polypropylene glycol and propylene oxide.
Ethylene oxide copolymer, polytetramethylene glycol, polypentamethylene glycol,
Examples include polyhexamethylene glycol. These polyalkylene glycols may be used alone or in combination of two or more types. Particularly preferred are polypropylene glycol and polytetramethylene glycol, each alone, or a mixture of the two. The number average molecular weight of polyoxyalkylene glycol is 600 to 8000, especially 600 to 8000.
Preferably it is 3000. The dicarboxylic acid having 6 to 80 carbon atoms is an aliphatic, alicyclic or aromatic dicarboxylic acid, such as adipic acid, terephthalic acid, isophthalic acid,
O-phthalic acid, naphthalene dicarboxylic acid, succinic acid, azelaic acid, sebacic acid, undecanedioic acid, dodecanedioic acid and oleic acid, linoleic acid, linoleic acid, eleostearic acid and similar olefinic acids having 8 to 40 carbon atoms Carbon number 16 obtained by polymerization of unsaturated monocarboxylic acid
~80 dimer acids, etc. Particularly preferred are oleic acid, linoleic acid, and dicarboxylic acids obtained by dimerizing linoleic acid. In some cases, 20% by weight of polyether ester can be used to obtain low molecular weight diols having 2 to 16 carbon atoms such as ethylene glycol, propylene glycol, butanediol, hexanediol, etc.
The following may be copolymerized. The molar ratio of the total diol component (adding the polyoxyalkylene glycol or polyoxyalkylene glycol and a small amount of low-molecular-weight diol) to the molar ratio of dicarboxylic acid is 1.0:0.5 to 1.0:0.5.
Normal esterification and/or at a ratio of 1.0:0.98
Or using conventional esterification methods and equipment in the presence of a polycondensation catalyst. Stir at 150-250℃ under a nitrogen stream and heat while removing water generated by the reaction, then reduce the pressure to 0.1-50mmHg at 200-250℃ to promote esterification and reduce the acid value to an appropriate value. let Examples of the esterification catalyst include phosphoric acid, sulfuric acid, p-toluenesulfonic acid, methanesulfonic acid, stannous oxalate, alkyl lead oxide, tetrabutyl titanate, zinc acetate, and sodium carbonate. The amount and type of catalyst are not limited and can be varied, but usually the amount of catalyst is 0.001 to 0.8
Approximately % by weight is used. The polyether ester used in the present invention has a number average molecular weight of 1,500 to 50,000, preferably 5,000 to 30,000. If the average molecular weight is less than 1500, the amount of oligomers will increase, which will adversely affect the analytical results and at the same time weaken the barrier strength formed by the resulting composition, thus defeating the purpose. If it exceeds 50,000, the floatability becomes small and the particles do not move smoothly between serum and blood cells during centrifugation. The polyetherester used in the present invention has a viscosity of 10 to 3000 poise at 20°C, preferably 15 to 3000 poise.
It is 1000 poise. Also, the density is 0.850~1.050g/
cm ³ , preferably 0.900 to 1.045 g/cm ³ . The blood sample separating agent of the present invention contains 1 to 40 parts by weight (preferably 5 to 30 parts by weight) of an inert filler per 100 parts by weight of the polyester for the purpose of adjusting the specific gravity and imparting thixotropy to the polyether ester. part) include. The density of the resulting composition is 1.010-1.070 g/cm ³
(preferably 1.025 to 1.065 g/cm ³ ). Viscosity (20℃) is 1000~30000 poise (preferably 2000~
10000 poise) is desirable. In the present invention, the viscosity is measured using a B-type rotational viscometer by placing 200 g of the composition in a 200 ml beaker at a temperature of 20°C. Inert fillers used in the present invention include various amorphous silicas such as precipitated silica, vaporized silica, and hydrophobic silica treated with silanes or polysiloxanes, alumina, talc, other silicates, bentonites, titanium oxides, and minerals. Examples include clay. Particularly preferred are amorphous silicas such as precipitated silica, vaporized silica and hydrophobic silica. If the density of the composition of the present invention is less than 1.010 g/cm ³ , it will be lighter than the serum part, and if it is more than 1.070 g/cm ³ , it will be heavier than the blood clot, and even if centrifugation is performed, the serum part and blood will be lighter. The ingredients do not migrate to the middle of the mochi portion, so it does not work as a separating agent. Furthermore, although the viscosity of the composition of the present invention is not particularly limited, if it is less than 1000 poise, the strength of the barrier formed between the serum and the blood clot by the composition will be weakened, and the barrier will be broken by a small impact.
If the concentration exceeds 30,000 poise, the composition may not float to the intermediate layer in practical centrifugation operations, but may remain at the bottom of the test tube and have no effect as a separating agent. The blood sample separation agent of the present invention, which is composed of a polyether polyester composition separated from an inert filler, is easily located at the interface between the serum (or plasma) portion and the blood clot (or blood cell) portion, forming a barrier. so that it flows during the centrifugation process. After centrifugation, the composition easily develops thixotropy and forms a continuous and stable barrier with high viscosity. The composition of the present invention may contain 10% by weight or less of a surfactant, such as polyoxyethylene hydrogenated castor oil monolaurate, polyoxyethylene hydrogenated castor oil tristearate, etc., in order to improve floating properties. As the surfactant, polyoxyethylene hydrogenated castor oil tristearate is particularly preferred. The composition of the present invention contains two structure-forming agents such as polydimethylsiloxane-polyethylene glycol block copolymer to improve the barrier strength of the composition.
It may be contained in an amount of % by weight or less. Since the blood sample separating agent of the present invention contains the above-mentioned polyether ester as a main component, it is possible to separate serum and blood clots, and there is little change in viscosity, so that it can be left for a long period of time. Furthermore, it is inert to blood and does not cause errors in blood component analysis. Hereinafter, the present invention will be specifically explained with reference to Examples. In the examples, parts simply mean parts by weight. Example 1 1,192 parts of 1,2-polypropylene glycol with a number average molecular weight of 1000, 608 parts of a dicarboxylic acid having 34 carbon atoms obtained by dimerization of oleic acid (hereinafter referred to as dimer acid, molecular weight 561.7), and 0.103 parts of tetrabutyl titanate. and 0.085 parts of trimethyl thiphosphate were placed in an autoclave of 3 and heated to 160 to 230°C while stirring at normal pressure under a nitrogen stream.
The temperature was raised over time to remove water produced by the reaction. Then 0.5mm from 760mmHg at 230℃
The pressure was gradually reduced to Hg over 1 hour. moreover
The temperature was raised to 240°C, and the reaction was further carried out for 4 hours under reduced pressure of 0.5 mmHg while removing water produced by the reaction. The resulting polyetherester has a density of 0.995
g/cm ³ , viscosity 50 poise/20°C, and hydroxyl value
11.2, and had an acid value of 3.8. 100 parts of this polyetherester and hydrophobic Aerosil R972 (trade name,
A thixotropic substance having a density of 1.048 g/cm ³ and a viscosity of 11,000 poise/20° C. was obtained by mixing with 12 parts of hydrophobic silica fine powder (manufactured by Nippon Aerosil Co., Ltd.). Table 1 shows the change in viscosity when the obtained composition was stored at room temperature in air and in vacuum.

[Table] Next, this composition was heated to 50 to 70°C, and 1.5 g was dispensed into a 10 c.c. serum separation blood collection tube using a dispenser with an appropriate nozzle, followed by a vacuum process. I made a breakout. The blood collection tube was then sealed with a rubber stopper in a vacuum set to a blood collection volume of 10 c.c. in a vacuum process. After blood collection using these blood collection tubes, centrifugation (2500 rpm x 10 minutes, arm length 12
cm), a barrier layer was formed between the serum and blood cells in all cases, and the barrier was not destroyed at all even if these blood collection tubes were left upside down for several days. Also, remove the serum by decantation,
When a clinical test was performed, each test item was consistent with the analytical values obtained when serum was pipetted out without using a serum separating agent. From this, it is clear that the polyetherester composition of the present invention is inactive against blood. Example 2 700 parts of 1,2-polypropylene glycol having a number average molecular weight of 700, 505 parts of dimer acid, 0.075 parts of tetrabutyl titanate and 0.064 parts of trimethyl phosphate were placed in 3 autoclaves, and polyether ester was prepared under the same conditions as in Example 1. was manufactured. The resulting polyetherester has a density of 0.990
g/cm ³ , viscosity 60 poise/20℃, and hydroxyl value
12.3, and had an acid value of 4.05. 100 parts of this polyether ester, 8 parts of hydrophobic Aerosil R972, and 4 parts of Aerosil 200 (trade name, manufactured by Nippon Aerosil Co., Ltd.)
A thixotropic substance having a density of 1.045 g/cm ³ and a viscosity of 15,300 poise/20°C was obtained. Table 2 shows the change in viscosity when the resulting composition was stored at room temperature in air and in vacuum.

[Table] Next, this composition was heated to 50 to 70°C, and the following operations were performed in the same manner as in Example 1. Serum was taken out and clinical tests were performed. The analysis values when taken out matched with each test item. From this, it is clear that the polyetherester composition of the present invention is inactive against blood. Example 3 1000 parts of polytetramethylene glycol having a number average molecular weight of 1000, 505 parts of dimer acid, 0.075 parts of tetrabutyl titanate, and 0.064 parts of trimethyl phosphate were placed in 3 autoclaves to produce polyether ester under the same conditions as in Example 1. did. Density of the obtained polyetherester
It had a viscosity of 0.985 g/cm ³ , a viscosity of 63 poise/20°C, a hydroxyl value of 10.3, and an acid value of 1.7. 100 parts of this polyether ester and 14 parts of hydrophobic Aerosil were mixed to give a density of 1.045 g/cm ³ and a viscosity of 9300 poise/20°C.
A thixotropic substance was obtained. Table 3 shows the viscosity change when the obtained composition was stored at room temperature in air and in vacuum.

[Table] Next, this composition was heated to 50 to 70°C, and the following operations were performed in the same manner as in Example 1. Serum was taken out and clinical tests were performed. The analysis values when taken out matched with each test item. From this, it is clear that the polyether ester of the present invention is inert to blood. Example 4 The compositions of Examples 1 to 3 were each heated to 60°C, and using a dispenser with an appropriate nozzle, 10 c.c. (inner diameter
After dispensing 2 g of each into serum separation blood collection tubes (13 mm, height 100 mm), defoaming was performed in a vacuum process.
The blood collection tube was then sealed with a rubber stopper in a vacuum setting at a blood collection volume of 10 c.c. Using these blood collection tubes, we collected 10 c.c. of blood from the same person and centrifuged it (2500 rpm, 10 minutes, arm length 12 cm).
In either case, there is a thickness of 8 to 12 mm between the serum and the clot.
A barrier layer of serum separating agent was formed, and the serum was completely separated by decantation. Table 4 shows the results of clinical analysis of these sera.
The results are shown in comparison with the results obtained by separating serum by pipetting out without using a serum separating agent.

[Table] As shown in Table 4, the polymer of the present invention had no effect on the test results in clinical tests and was proven to be inactive against blood components. (Note) BUN (blood urea nitogen test): Diacetyl monooxime method UA (uric acid test): Reduction method Crea (creatinine test): Yatsufue reaction TP (total protein): Biuret method Alb (alubumin test): HABCA method IgG ( immunoglobulin G test): SRID method IgM (immunoglobulin M test): SRID method GOT (glutamate oxaloacetate transaminase)
test): UV method GPT (glutamate pyruvate transaminase)
test): UV method LDH (lactate dehydrogenase test): UV method LAP (leucine aminopeptidase test): Bassy
−lowry method ALP (alkaline phosphatase test): Goldberg
γ-GTP (γ-glutamyl transpeptidase)
test): UV method β-Lipo (β-lipo-Protein test):
immunocrite method PL (Phospholipid test): Enzyme method T-Ch (total cholesterol test): Enzyme method TG (triglycerid test): Savdesai-manning method FAA (free fatty acid test): Cu-Bassokubroin method Na (sodium test): Flame Absorption spectroscopy K (potassium test): Flame absorption spectroscopy Cl (chlorine test): Titration method Fe (iron test): Matsubara method Comparative example 1 Polypropylene glycol with number average molecular weight 200
A polyether ester was produced under the same conditions as in Example 1 using 220 parts of adipic acid, 145 parts of adipic acid, and 0.150 parts of titanium tetrabutyl. The density of the obtained polyether ester was 1.098 g/ ^cm3 , and the hydroxyl value was
31.0, acid value 15.3, and viscosity 130 poise (20°C). When this polyether ester was heated to 50 to 70°C and operated in the same manner as in Example 1, the polyether ester did not float between the serum and the blood clot even after centrifugation. It remained at the bottom. It was found that this polyether ester had no function as a serum separating agent. Comparative Example 2 25 parts of Aerosil 200 were mixed with 100 parts of the polyether ester obtained in Example 1, and the density was 1.095 g/cm ³ .
A polyetherester composition having a viscosity of 38,000 poise (20°C) was obtained. When this composition was heated to 50 to 70°C and operated in the same manner as in Example 1, it was found that even after centrifugation, the composition did not float to the bottom of the test tube between the serum and the blood clot. It remained in From this, it is clear that this composition does not function as a serum separating agent.

Claims (1)

[Claims] 1 Mainly polyoxyalkylene glycol [A] with a number average molecular weight of 600 to 8000 and a carbon number of 6 to 8000
80 dicarboxylic acid or its derivative [B], has a number average molecular weight of 1,500 to 50,000, and has a viscosity of 10 to 3,000 poise (20°C), and an inert filler [C]. D] and has a density of 1.010 to 1.070 g/cm ³ , and [A]
The molar ratio of and [B] is 1.0:0.5 to 1.0:0.98, and the weight ratio of [C] to [D] is 100:1 to 100:
40. A blood sample separation agent characterized in that: