JPH085629A - Method for preserving blood or blood cell - Google Patents

Abstract

PURPOSE:To realize stabilized preservation of white blood cell, especially granulocyte, by adding a polyoxyethylene based nonionic surfactant to blood or blood cell. CONSTITUTION:Polyoxyethylene based nonionic surfactant is preferably employed as a preservative for blood or blood cell. A low pressure blood sampling tube 1 encapsulating the preservative 6 comprises a bottomed tube 2 and a hermetic sealing member 3 and the internal pressure is reduced by an amount corresponding to a predetermined blood sampling quantity. The hermetic sealing member 3 comprises a gas barrier member 4 which exhibits high airtightness before being stuck with a hollow needle and seals the opening of the bottomed tube hermetically. A sealing member 5 is fitted to the area of the member 4 being stuck with a hollow needle in order to choke the needle hole upon removal of the hollow needle. The effective quantity of preservative 6 in the blood sampling tube 1 is preferably set to correspond with the sampling quantity of blood and an anticoagylant is preferably encapsulated. When such blood sampling tube 1 encapsulating the preservative 6 is employed, white blood cell, especially the granulocyte, in the sampled blood can be preserved stably without requiring any subsequently operation.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for preserving blood or blood cells collected in clinical tests.

[0002]

2. Description of the Related Art When collecting blood from a patient for a clinical examination, a blood collecting instrument including a blood collecting tube having a decompressed blood collecting tube and a puncture needle is often used.

[0003] It is preferable to carry out the test immediately after blood collection, but when the test is requested to an external testing organization, the test is not always possible on that day. In that case, the reliability of the measured value over time becomes a problem.

As an example, CPD (Citrate Phosph)
It has been reported that the number of white blood cells in stored blood and the osmotic resistance of red blood cells decrease daily. On the other hand, it has been reported that vitamin E has an inhibitory effect (Reference: Vitamins 39 p86-90 1969, Table 5-50).
0319)). However, Vitamin E lacks stability,
Further, since it is fat-soluble, it is difficult to dissolve it in blood.

On the other hand, a method and a reagent using a polyoxyethylene type nonionic surfactant as a reagent for automatic blood cell measurement are known (Japanese Patent Laid-Open No. 252556/1993, Japanese Patent Publication No. Sho 6).
3-809504). These inventions are reagents for measuring the number of blood cells that are added at the time of measurement, which lyse red blood cells and act on white blood cells to classify and measure the white blood cells. Therefore, it has not been clarified at all about the preservative action / effect of the polyoxyethylene nonionic surfactant on blood or blood cells, and that it can be a preservative added at the time of blood collection, and it is still unsolved.

[0006]

[Means for Solving the Problems] As a result of intensive studies to solve the above problems, the above objects can be achieved by the following methods for preserving blood or blood cells, preservatives, and blood collection tubes containing preservatives. This has led to the completion of the present invention. (1) A method for preserving blood or blood cells, which comprises adding a polyoxyethylene-based nonionic surfactant to blood or blood cells.

(2) The method for preserving blood or blood cells according to (1), wherein the polyoxyethylene nonionic surfactant is polyoxyethylene castor oil or polyoxyethylene hydrogenated castor oil.

(3) The method for preserving blood or blood cells according to (1), wherein the polyoxyethylene nonionic surfactant is a compound represented by the general formula or:

[0009]

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[0010]

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(4) A preservative for blood or blood cells, which comprises a polyoxyethylene nonionic surfactant.

(5) The preservative for blood or blood cells according to claim 4, wherein the polyoxyethylene-based nonionic surfactant is polyoxyethylene castor oil or polyoxyethylene hydrogenated castor oil.

(6) The preservative for blood or blood cells according to (4), wherein the polyoxyethylene nonionic surfactant is a compound represented by the general formula or. (7) A blood collection tube comprising a bottomed tube having one end opened and the other end closed, and a sealing member for sealing the opening of the bottomed tube, wherein at least the polyoxyethylene nonionic ion is contained in the reduced pressure blood collection tube. A blood collection tube containing a surfactant.

(8) The blood collection tube according to (7), wherein the polyoxyethylene nonionic surfactant is polyoxyethylene castor oil or polyoxyethylene hydrogenated castor oil.

(9) The blood collection tube according to (7), wherein the polyoxyethylene nonionic surfactant is a compound represented by the general formula or:

(10) The blood collection tube according to any one of (7) to (9), wherein an anticoagulant is further enclosed in the reduced pressure blood collection tube.

[0017]

In the blood to which the preservative of the present invention is added, the number of white blood cells is less likely to decrease as compared with the blood stored under the same conditions without addition. The effect shows the same white blood cell count as that at the time of blood collection up to 24 hours, and there is a clear difference as compared with the case of no addition. Even at 48 hours, the rate of decrease is lower than in the case of no addition. Especially, a remarkable effect is recognized in the number of granulocytes. Further, it is also effective in the function of cells.

The blood or blood cell preservative of the present invention is preferably a polyoxyethylene-based surfactant, more preferably polyoxyethylene castor oil or polyoxyethylene hydrogenated castor oil, and particularly preferably the above-mentioned general formula or Is a compound represented by. The polyoxyethylene castor oil and polyoxyethylene hydrogenated castor oil are castor oil and hydrogenated castor oil to which ethylene oxide is added. The polyoxyethylene castor oil or the polyoxyethylene hydrogenated castor oil as a preservative for blood or blood cells is preferably 10 to 100 mol adduct of ethylene oxide, more preferably 30 to 100 mol adduct, particularly preferably 40 mol. To 60 mol adduct. D of the compound represented by the general formula or
e, f, x, y, z are positive integers, and their sum is 1
0 to 100 is preferable, and 30 to 10 is more preferable.
It is 0, particularly preferably 40 to 60.

The amount of the preservative of the present invention added to blood or blood cells is not uniform because it depends on the effective dose of the compound. For example, the amount of polyoxyethylene hydrogenated castor oil added with 60 mol of ethylene oxide added to blood is 10-
100 μ / ml is preferred, more preferably 15-50
μ / ml. The amount of polyoxyethylene hydrogenated castor oil added with 40 mol of ethylene oxide added to the blood is 10-
300 μ / ml is preferable, and more preferably 10 to 10
0 μ / ml, particularly preferably 15 to 50 μ / ml
Is. Compounds that are solid or pasty at room temperature are physiological saline or PBS (phosphate buffered saline)
It is preferable to add it to blood or blood cells after dissolving it in a buffer solution such as.

The preservative of the present invention may be a polyoxyethylene-based surfactant alone. Furthermore, anticoagulants, nutrients, inorganic salts, antibiotics, etc. may be added. As the anticoagulant, citrate phosphate dextrose, citrate phosphate dextrose adenine, heparin sodium, EDTA-2Na, sodium citrate and the like are preferable. As nutrients, adenine, dextrose,
Mannitol, glucose, gluconic acid and the like are preferable. As the inorganic salts, phosphoric acid, sodium acetate and the like are preferable.

FIG. 1 is a sectional view of a reduced pressure blood collection tube which is an embodiment of the blood collection tube of the present invention. The reduced pressure blood collection tube 1 of the present invention is
Bottomed tube 2 and sealing member 3 for sealing the opening of the bottomed tube 2
And the inside is depressurized by a predetermined blood sampling amount.

The material of the bottomed tube 2 is plastic or glass. As the plastic, one having a high gas barrier property is preferable. The sealing member 3 is composed of a gas barrier member 4 and a sealing member 5. The gas barrier member 4 is made of an aluminum foil film or the like, has high airtightness before the hollow needle is punctured, and seals the opening of the bottomed tube 2. The seal member 5 is made of polyisopropylene rubber or the like, and is attached to at least the hollow needle puncture region of the gas barrier member 4 so that the needle hole after the punctured hollow needle is removed can be closed. The sealing member 3 may be a plug body made of polyisopropylene rubber or the like in which a gas barrier member and a sealing member are integrated. Inside the vacuum blood collection tube 1,
The preservative 6 of the present invention is enclosed. The amount of the internal preservative is not uniform because it depends on the effective dose of the compound,
It is preferable that the effective amount corresponds to a predetermined blood collection amount.
Further, in order to prevent blood from coagulating, it is preferable to encapsulate the anticoagulant inside. By using such a vacuum blood collection tube containing a preservative of the present invention, leukocytes can be stabilized and stored in the collected blood without post-treatment.

[0023]

[Example] Polyoxyethylene hydrogenated castor oil was added to human blood, and the time-dependent change in the number of granulocytes was examined.

<Method> Polyoxyethylene hydrogenated castor oil (HCO-60 (for pharmaceuticals) manufactured by Nikko Chemicals Co., Ltd.) containing 60 mol of ethylene oxide was dissolved in physiological saline to prepare a 10% solution. 300 μl of this 10% solution was added to 2 ml of human blood and left at room temperature. As a control, physiological saline was similarly added and compared. The number of granules of the sample was measured with an automatic blood cell counter [TOA Medical Electronics Co., Ltd.] at 0 (immediately after blood collection) and 24 and 48 hours later. The results were expressed by the rate of change in the number of cells with 100% as the number of granulocytes immediately after blood collection.

<Results> As shown in FIG. 2, the control group was 2
The number of granulocytes decreased at 4 and 48 hours, but in the polyoxyethylene hydrogenated castor oil-added group, it did not decrease and was similar to that immediately after blood collection, and a significant effect was observed (Student's t-test; p <0.01).

[0026]

According to the method for preserving blood or blood cells of the present invention, leukocytes, particularly granulocytes, can be preserved without being stabilized or reduced. Further, by using the blood collection tube containing the preservative of the present invention, leukocytes, particularly granulocytes, can be stabilized and stored without post-treatment of the collected blood.

[Brief description of drawings]

FIG. 1 is a cross-sectional view of a reduced pressure blood collection tube which is an embodiment of the blood collection tube of the present invention.

FIG. 2 is a graph showing the effect of polyoxyethylene hydrogenated castor oil on the change with time of the number of granulocytes in human blood.

[Brief description of reference numerals]

 1 Decompression blood collection tube 2 Bottomed tube 3 Sealing member 4 Barrier member 5 Sealing member 6 Preservative

Claims (1)

[Claims] 1. A method for preserving blood or blood cells, which comprises adding a polyoxyethylene-based nonionic surfactant to blood or blood cells.