JPS58137757A - Separating agent of blood sample - Google Patents

Abstract

PURPOSE:To make the sepn. operation of blood sera efficient and to prevent intrusion of blood cells, etc. during preservation and transfer, by using polyether ester which has the specific gravity between the specific gravities in a serum part and a clot part and is inert to blood components. CONSTITUTION:Polyether ester having about 1,500-50,000mol.wt., and 10-3,000 poises viscosity is produced by causing 1mol polyoxy alkylene glycol of 200- 8,000mol.wt. and 0.50-0.98mol dicarboxylic acid of 8-80 carbon atoms to react with each other. An inert filler, a surfactant and a structure forming agent are contained in such polyether ester, whereby a separating agent for blood samples is prepd. The separating agent is put into a blood letting tube and blood is drawn therein. If the blood is subjected to centrifugal sepn. operation after allowing the blood to stand until the same clots, the separating agent which is the intermediate of the serum and the clot in specific gravity forms a stable barrier between the phases of the serum and the clot; therefore, the serum is obtained simply by tilting the blood letting tube, and the fluctuation in the analytical value by the intrusion of blood cell components on account of disturbance between the phases in the stage of pipetting out is prevented.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a blood sample separation agent. In blood analysis, generally, whole blood is centrifuged to separate serum (or plasma) and blood clots (or blood cells). The serum (or plasma) portion was pipetted out and used for clinical analysis. but,
In this case, the operation was complicated and time-consuming, and at the same time, when pipetting out the serum, blood cell components could be mixed in and affect the analytical values, so nine people were required to be skilled in the operation.9 However, 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 blood plasma) from blood clots (or blood cell components), etc., it has a specific gravity between that of serum (or plasma) and blood clots (blood cell components), and creates a barrier between the two components. It is known to prevent contamination of blood cell components into serum (or plasma) by using a separation composition that is formed, and at the same time to take out the serum (or plasma) portion only by tilting it.

For example, Tokushu 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 separating composition. This gel-like substance is given thixotropic properties by finely powdered silica mixed with silicone fluid, and is transferred directly to the phase to be cut and separated by centrifugation operation, and upon completion of centrifugation.

A stable barrier is formed that is not easily destroyed by tilting operations or mild impacts. However, in this composition, the silicone fluid reacts with the silica surface, accelerating the viscosity reduction of the composition over an hour, and the K's ability to form a stable barrier is compromised, and the slope disturbs the barrier and prevents serum Blood cells are mixed into the part.

JP-A No. 51-83654 describes in detail that the accuracy of serum analysis may be adversely affected.

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. But even in this case.

Although it is an improvement over polybutadiene, which does not have f properties, because it uses a modified polybutadiene.

When stored at room temperature in air, the viscosity of the composition changes, and IIa-forming agents such as low molecular weight diols are required to enhance thixotropy. This may have a negative impact on the analysis accuracy.

Furthermore, JP-A-53-146795 K (a)
(b) a polymeric fatty acid containing 75% by weight or more of Cs@dibasic acid; and (c) a saturated aliphatic dicarboxylic acid having 4 to 12 carbon atoms; a branched aliphatic dihydric alcohol, a mixture of such branched dihydric alcohols, or a branched aliphatic dihydric alcohol having at least 50% of such branched aliphatic dihydric alcohols;
consisting essentially of the reaction product of a chemical chemistries, of a mixture with a straight chain aliphatic dihydric alcohol having 2 to 8 carbon atoms.

Therefore, the equivalent ratio of (a) to (b) is 0.80:0.2
In the range of 0 to 0.97:0.03, the polyester has an average molecular weight of about 2.000 to s, o o o, a viscosity of 210°F of 8,000 centistokes, and a viscosity of 1. 015 to 1
．． Copolyester compositions useful as barrier agents in blood separation assembly devices are also disclosed, but again with neopentyl glycol, 1,2-propane In rounding using branched low molecular weight diols such as diols, it is difficult to completely control the molecular weight and viscosity of the target polyester, which may lead to fluctuations in the chicken-tropic properties of the composition.

The purpose of the present invention is to analyze blood components by using a polyether ester that has a specific gravity between that of serum (or plasma) and blood clot (or blood cells) and is inert to blood components. The goal is to improve the efficiency of separation operations (or plasma), and to prevent contamination with blood cells during storage and transportation.

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 provides polyoxyalkylene glycol having a molecular weight of 200 to 8,000, a dicarboxylic acid having 6 to 80 carbon atoms or a derivative thereof, and optionally a low molecular weight diol, and having an average molecular weight of about 1,500 to 50,000. Yes, viscosity is 10
~3,000 poise (20°C) polyether ester containing an inert filler and optionally a surfactant and a structure forming agent, and a density of 1.010 to 1.0
70f/-.

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 of the separation agent into a 10 m blood collection tube for serum separation).

If you collect blood and want to obtain serum, leave it until the blood coagulates and then perform centrifugation. During centrifugation, the separating agent, whose specific gravity is between that of serum and blood clots, moves to the middle of serum and blood clots. A stable partition wall is formed in the correlation between the two. Therefore, there is no need to pipette out to collect serum, and serum can be obtained by simply tilting the blood collection tube, and f# in the analysis value due to contamination of blood cell components due to turbulence when pipetting out can be avoided. .

Furthermore, the blood sample separating agent of the present invention has excellent viscosity stability.

The blood sample separation agent of the present invention improves the conventional drawbacks.

Moreover, it is a serum and plasma separation agent that can be manufactured at low cost.

The polyether ester used in the present invention has a molecular weight of approximately 1.5.
00-50,000 (preferably 3,000-20.
000) and a viscosity of 10 to 3,000 poise (preferably 15 to 1,000 poise). Also, 0.8
50 to 1.050 (preferably 0.900 to 1.045
It is desirable to have a specific gravity of f /d ), and it is desirable to be a homogeneous polyetherester that is inert to blood components.

An example of the polyether ester used in the present invention is 1 mole of polyoxyalkylene glycol having a molecular weight of 1,200 to 8,000 and a dicarboxylic acid having a carbon number of 8 to 80.
>98 mol is reacted and manufactured by one company.

Examples of polyoxyalkylene glycol include polypropylene glycol, propylene oxide-ethylene oxide copolymer, polytetramethylene glycol, and polybentamethylene 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/or polytetramethylene glycol. The molecular weight of polyoxyalkylene glycol is 2
00~s, ooo.

In particular, it is preferably 300 to a, ooo.

The dicarboxylic acid having 6 to 80 carbon atoms is an aliphatic, cyclic or aromatic dicarboxylic acid, examples of which are adipic acid, terephthalic acid and isophthalic acid.

1-phthalic acid, naphthalene dicarboxylic acid, colic acid, azefinic acid, sebacic acid, undecanedioic acid, dodecanedioic acid and oleic acid, liluic acid, linoleic acid, eleostearic acid and similar carbon atoms of 8 to 40
Examples include di-positional acids having 16 to 80 carbon atoms obtained by polymerization of 0-olefinic unsaturated monocarboxylic acids.

Particularly preferred are dicarboxylic acids prepared from oleic acid, lyluic acid, and linole M.

In some cases, polyether esters capable of obtaining low molecular weight diols having 2 to 16 carbon atoms, such as ethylene glycol, propylene glycol, butanediol, hexanediol, etc., may be copolymerized with 20 or less polymers.

The molar ratio of the total diol component containing the polyoxyalkylene glycol or polyoxyalkylene glycol and a small amount of low-molecular-weight diol added and the molar ratio of dicarboxylic acid is 1.0:0.5 to 1.0:0.
Using conventional esterification methods and equipment in the presence of conventional esterification and/or polycondensation catalysts in a proportion of 98%. Stir at 150-250°C under a nitrogen stream and heat while removing water produced by the reaction, then heat to 200-250°C
The pressure is reduced to 0.1-50 HP at 0°C to promote esterification and reduce the acid value to an appropriate value.

Examples of the esterification catalyst include phosphoric acid, sulfuric acid, p-toluenesulfonic acid, methanesulfonic acid, stannous oxalate, alkyl carrier oxide, tetrabutyl titanate, zinc acetate, and sodium carbonate.

The amount and type of catalyst are not limited and can be varied; however, the amount of catalyst used is usually about 0.001 to 0.8 times.

The polyetherester used in the present invention has an average molecular weight of about 1,500 to 50,000, preferably 5.000 to 50,000.
30,000. If the average molecular weight is less than 1,500, the oligomer density will increase, which will adversely affect the results of analysis 1. At the same time, the barrier strength formed by the resulting composition will be weakened, and the purpose will not be achieved.

If it exceeds so, o o o, the floating property becomes small and the particles tend to move smoothly between serum and blood cells during centrifugation operation.

The polyetherester used in the present invention has a viscosity of about 10 to 3,000 poise at 20°C, preferably 15 to 1
．． It is 000 poise. Also, the density is 0.850 to 1.0
50 f/ad, preferably 0.900-1.045
It is desirable that it be F/-.

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 of the polyetherester and imparting thixotropic properties. part) include.

The density of the resulting composition is 1.010 to 1.070 f/-
(preferably 1.025 to 1.065 t/Cd)
It is.

The viscosity (at 20° C.) is preferably 1,000 to 30,000 poise (preferably 2,000 to 10,000 poise).

In the present invention, viscosity is measured using a B-type rotational viscosity needle.
00g/200t of one composition was put into a beaker, and the test was carried out at a humidity of 20°C.

---2 Inert fillers used in the present invention include various amorphous silicas such as precipitated silica, vaporized silica, and hydrophobic silica treated with silane or polysiloxane, alumina,
Talc and other silicates.

Examples include bentonite, titanium oxide, and mineral 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 1.011/- or less, it will be lighter than the serum part, and if it is greater than 1,070 y/d, it will be heavier than the blood clot part, and C, which performs the centrifugation operation, will also be the serum part. The components do not migrate to the middle of the blood clot, so it does not work as a separating agent.

In addition, the viscosity of the composition of the present invention is not particularly limited, but 1
When it is less than 30,000 poise, the strength of the barrier formed between the serum and blood clot by one composition becomes weaker, and the barrier becomes damaged by a small impact, and when it is more than 30,000 poise, SJ! In conventional centrifugation operations, the composition may not float to the surface and remain at the bottom of the test tube, failing to act 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 uses a surfactant 1 to improve floating properties.
For example, polyoxyethylene hydrogenated castor oil monolaurate, polyoxyethylene hydrogenated castor oil tristearate, etc. may be contained in an amount of not more than 10% by weight. As the surfactant, polyoxyethylene hydrogenated castor oil tristearate is particularly preferred.

The composition of the present invention may contain up to 2 mm5 of a structure forming agent such as a polydimethylsiloxane-polyethylene glycol block copolymer to improve the barrier value of the composition.

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 the analysis of blood components.

Hereinafter, the present invention will be specifically explained with reference to Examples.

In the examples, parts simply mean parts by weight.

Example 1 An excellent dicarboxylic acid having 34 carbon atoms (hereinafter referred to as dimer acid with a molecular weight of 561.7) was obtained by dimerizing 1,192 parts of 1,2-polypropylene glycol with a number average molecular weight i, o o o and oleic acid (hereinafter referred to as dimer acid, 1 molecular weight 561.7). 1 part, 0.103 part of tetrabutyl titanate, and 0.085 part of trimethylnaphosphate were placed in a 3J autoclave, and heated to 160 to 230°C over 3 hours while stirring at normal pressure under a nitrogen stream, resulting in one reaction. The reaction was further carried out for 4 hours while removing the water produced by 1 incubation of F5 under reduced pressure with fresh Ht.

The obtained polyether ester t[Jto, 995f/
-1 viscosity 50 poise/20℃, hydroxyl value 11.2
．． It has an acid value of 3.8. 100 parts of this polyether ester and 12 parts of hydrophobic Aerosil R972 (product name 1, hydrophobic silica fine powder manufactured by Nippon Aerosil Co., Ltd.) were mixed, and the mixture had a density of 1.048 f/d and a viscosity of 11,000 poise/2.
A chicken tropic substance at 0'C was obtained. Table 1 shows the change in viscosity when the obtained composition was stored at room temperature in air and in vacuum.

Chapter 1 Table This composition was then heated to 50-70°C.

Using a dispenser with an appropriate nozzle, 1.5 was dispensed into a 10 (!1) serum separation blood collection tube, and then defoamed using a vacuum process.

The blood collection tube was then sealed with a rubber stopper in a vacuum set at 1.1 1 Occ for blood collection. After blood collection using these blood collection tubes, centrifugation (2500 rpm x I 3 minutes, arm length 1
21), a barrier layer was formed between the serum and blood cells in all cases, and the barrier was not broken 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, the analytical values obtained when serum was extracted by pipetting out without using a serum separating agent and each test item were consistent. 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 0.0 parts of trimethyl phosphate.
06411 was placed in a 3J autoclave, and polyether ester was produced under the same conditions as in Example 1.

Obtained polyether Nistelha density 0.990 f/e
ta, a density of 60 poise/20°C, and a hydroxyl value of 12.
3. It had an acid value of 4.05. 100 parts of this polyether ester, 8 parts of hydrophobic Axyl R972, and Ae Rosil 200 (trade name, manufactured by Nippon Aerosil Co., Ltd.) 4
s, density 1.045tlaA, viscosity 15.3
A chicken tropic substance of 00 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 2 This composition was then heated to 50-70°C for 7JIM.

Thereafter, the procedure was performed in the same manner as in Example 1, and the serum was expelled from the throat.

When a clinical test was performed, the analytical values obtained when serum was extracted by pipetting out without using a serum separating agent and each test item were consistent. 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 1.000, 505s of dimer acid and 0.075 parts of tetrabutyl titanate and 0 parts of trimethyl phosphate.
．． Take 064 parts into 31 autoclave.

A polyetherester was produced under the same conditions as in Example 1. Density of the obtained polyetherester: 0.985
f/ad, viscosity 63 poise/20°C.

It had a hydroxyl value of 10.3% and an acid value of 1.7. ・100 parts of this polyetherester and 14 parts of hydrophobic Aerosil were mixed to have a density of 1.045 f/A and a viscosity of 9,300 voids/thixotropy at 20°C. Obtained a sexual substance.

Table 3 shows the viscosity change when the obtained composition was stored at room temperature in air and in vacuum.

Table 3 This composition was then heated to 50-70°C for 7JIML.

The procedure was then carried out in the same manner as in Example 1, and the serum was taken out.

When a clinical test was performed, each test item was consistent with the analytical values obtained when the serum was taken out by pipetting out without using a serum separating agent. From this fact, 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 humidified by 60°C, and using a dispenser with a single nozzle, the compositions were moistened to 110CC (inner diameter 13cm).
After dispensing 2 f of each sample into serum separation blood collection tubes with a height of 100 cm), defoaming was performed in a vacuum process. The blood collection tube was then sealed with a rubber seal in a vacuum process with the blood collection amount set at 10 cc. Using these blood collection tubes, 1 sample each from the same person.
After collecting 0 cc of blood, centrifugation (250 rpm, 10 minutes,
As a result of carrying out arm length 121), a serum separation layer with a thickness of 8 to 12 ai was formed between the serum and the blood clot, and a single agent barrier layer was formed between the serum and the blood clot, 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 of one analysis in which serum was separated by pipetting out without using a serum separating agent.

Table 4 As shown in Table 4, the polymer of the present invention did not affect the test results at all in clinical tests and was proven to be inactive against blood components.

(Note) BIJN (blood urea nitrogen)
est) Diacetyl monooxime method UA (uric acid test): Reduction method Crea (creatinine test): Yatsufue reaction TP (total protein) r B
iuret method Alb (alubumin test)
: HABCA method IgG (immunoglobuli
n G test): 5RID method IgM (#
M//):# GOT (glutamate oxaloacetate)
e transaminase test) s UV method GPT (glutamate pyruvate tr
ansaminase test): alpha method LDH (lactate dehydrogenase test)
test) r UV method LAP (leucine a
minopeptidase test) :Bass
y-Lowry method ALP (alkaline phosphotase
test) : Goldberg decision r-GTP(r-
glutamyl transpeptidase
test): UV method β-1ipo (β-1ipo-protein te
st): immunocrite method PL (phos
pholipid test): Enzymatic method T-Ch (t
otal cholesterol test): Enzyme method TG (triglyceride test):
savdesai-manning method FAA (fr
ee fatty acid test): Cu
- Pack Proin Method Na (sodium test)
:Flame absorption spectroscopy K (potassium test
): pCl (chlorine test)
: Titration method Fe (iron test) : Matsubara method Comparative example 1 Polypropylene glycol 220 with a number average molecular weight of 200
A polyether ester was produced under the same conditions as in Example 1 using 145 parts of adipic acid and 0.150 parts of tetrabutyl titanate. The density of the obtained polyetherester was 1.098t/CIA, and the hydroxyl value was 31.
0, acid value 15.3. Viscosity 130 poise (20
℃).

This polyether ester t-50 to 70'CK was added and operated in the same manner as in Example 1.

Even after centrifugation, the polyether ester did not float between the serum and the blood clot, but remained at the bottom of the test tube. 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 polyetherester obtained in Example 1, and the density was 1.095 t/crA.
, a polyetherester composition having a viscosity of 38,000 voices (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 middle between the serum and the blood clot, and the composition remained at the bottom of the test tube. Friend remaining in. From this, it is clear that this composition does not function as a serum separating agent.

Patent applicant: Toyobo Co., Ltd.

Claims (1)

[Claims] It is obtained from a polyoquine alkylene glycol having a molecular weight of 200 to 8,000, a dicarboxylic acid having 6 to 80 carbon atoms or a derivative thereof, and optionally a low molecular weight diol, and has an average molecular weight of about 1,500 to 5 o, o o o. ,
A polyether ester having a viscosity of 10 to 3,000 poise (20°C) contains an inert filler and optionally a surfactant and a structure forming agent, and has a density of 1.0.
10 to 1.07 Of/CIA. A blood sample separating agent.