JPS6116023B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a separating agent used when separating a blood sample into serum or plasma and blood cells by centrifugation. In recent years, testing of blood components in clinical laboratory departments has become extremely important, and the number of tests conducted has continued to increase. Among these, many serological tests use only serum or plasma as a sample, and it is necessary to separate serum or plasma from blood cells as a pre-test procedure. For this reason,
Conventionally, serum or blood cells have been collected by precipitating the blood cell portion only by centrifugation and then sucking it up with a pipette. However, this method provides insufficient separation and is very time-consuming, so recently various methods have been devised to allow serum or plasma collection to be carried out easily and with high yield. Ta. One method is to add a substance having a specific gravity intermediate between that of serum or plasma and blood cells to a blood sample, and use centrifugation to position the substance between the two to form a partition wall. According to this method, it is possible to separate serum or plasma only by decantation, and time and labor can be reduced. The form of this material for forming partition walls is solid form,
It is classified into three types: liquid form and mixtures thereof. Solid forms include, for example, polystyrene powder or pellet materials, but when these are used, the collection rate of serum or plasma may be unavoidably reduced, or the function as a septum may fail. Since this is sufficient, there are various drawbacks such as contamination of blood cells into serum or plasma and the tendency for the septum itself to be destroyed. In addition, in liquid form,
Acrylic polymers are known. However, in general, when trying to obtain a highly viscous liquid polymer from monomers such as acrylic polymers, there are difficult manufacturing problems such as the removal of impurities depending on the reaction control and reaction method. many. In addition, as a mixture of solid and liquid, for example, there is a mixture of silicone and silica powder added as a viscosity and specific gravity adjusting agent, but silicone is expensive and silica is difficult to measure in serum or plasma. There is also a risk of adsorbing components. In view of the current state of serum or plasma separation agents as described above, the present invention allows serum or plasma separation to be easily and accurately performed without affecting serum test values, and at the same time, is easy to manufacture. The purpose of this work was to provide a separating agent for obtaining polyptene, which has a specific gravity of less than 1.03 at 25°C, and a specific gravity of 1.03 to 1.08 at 25°C and a viscosity of 5000 to 5000. A serum or plasma separating agent characterized in that it consists of a liquid chlorinated product with a concentration of 500,000 centipoise. The liquid polyptene in the present invention has a specific gravity of less than 1.03 at 25°C, does not react directly with serum or plasma, and is given fluidity by centrifugation. However, since its specific gravity at 25° C. is smaller than 1.03, when it is centrifuged and flowed, it cannot be moved to an intermediate position between serum and blood cells to form a partition wall. If centrifugal separation is to be used to move the blood cells to a position intermediate between serum and blood cells, the specific gravity should be adjusted to 1.03 to 1.08. For this purpose, it is conceivable, for example, to add a suitable filler to the liquid polyptene in order to adjust the specific gravity. However, adding a filler may cause separation of the liquid polypten and the filler during centrifugation. As a result of various studies taking this point into consideration, the present inventors have found that by chlorinating the liquid polyptene, its specific gravity increases and at the same time its viscosity also increases, yet it does not cause any reaction with serum or plasma. It was found that there was no effect on serum test values. That is, in the present invention
Among the chlorinated products obtained by chlorinating liquid polyptene with a specific gravity of less than 1.03 at 25℃, the specific gravity at 25℃ is 1.03 ~
A liquid chloride with a specific gravity of 1.08 is selectively used because a liquid chloride with a specific gravity in this range creates a barrier between the blood and the serum or plasma from which the blood has been separated by centrifugation. This is because it can be formed. The viscosity of the liquid chloride used as a serum or plasma separating agent in the present invention must be 5,000 to 500,000 centipoise at 25°C, and within this range the Spitz tube used when centrifuging sample blood must be The optimum viscosity range can be determined as appropriate according to the material, pipe diameter, usage conditions of the separating agent, etc. In addition, to carry out the chlorination, any conventionally known method for chlorinating polymers can be adopted, but catalysts, solvents, etc. It is preferable to employ a method that allows the above-mentioned chlorination to be carried out under conditions that do not introduce impurities. For example, a method of chlorinating liquid polyptene by blowing chlorine gas into it under irradiation with ultraviolet rays or radiation can be suitably employed. In the above method, the only impurity mixed in the chlorinated product is unreacted chlorine, but since this is a gas, it can be easily removed by operations such as depressurization. In the present invention, in order to use the liquid chlorinated product obtained by chlorinating liquid polyptene as a serum or plasma separating agent, the chlorinated product is added alone to the sample blood when the sample blood is separated by centrifugation. You may. Alternatively, if necessary, other liquid substances having a specific gravity and viscosity similar to those of the chlorinated product may be mixed and used, or granular materials having a specific gravity similar to those of the chlorinated product, for example, a specific gravity of 1.00 at 25°C. It is also possible to mix and use particulate materials such as polystyrene, acrylonitrile-styrene copolymer, polyphenylene oxide resin, etc. in the range of 1.10 to 1.10, or hollow glass microspheres. According to the serum or plasma separating agent of the present invention, when it is added to sample blood and separated into serum or plasma and blood cells by centrifugation, the interface is not destroyed by decantation by the separating agent of the present invention. A partition wall is formed, and serum or plasma can be easily and precisely extracted. Furthermore, the separating agent of the present invention does not affect serum test values in any way. Furthermore, the separating agent of the present invention can be produced with high purity by simply chlorinating liquid polyptene without requiring any particular purification. Examples of the present invention will be described below. Note that the specific gravity and viscosity are both values at 25°C. Example 1 100g of liquid polyptene with a specific gravity of 0.860 and a viscosity of 400 centipoise was charged into a flask, and the liquid was
Chlorine gas was blown in at a flow rate of 60 ml, the temperature was maintained at 70 to 75°C while stirring with a magnetic stirrer, and ultraviolet rays were irradiated. After reacting in this state for 8 hours, dechlorination was carried out under reduced pressure at about 80° C. for 4 hours to obtain a chlorinated liquid polyptene as a reaction product having a specific gravity of 1.057 and a viscosity of 85,000 centipoise. This about 2
Put g into a polystyrene Spitz tube and add sample blood 8
ml and centrifuged at 2500 rpm for 5 minutes, the septum formed between the serum and blood clot was not easily destroyed by decantation, making it easy to separate the serum. It was hot. In addition, blood tests were conducted on 26 items such as proteins, serum enzymes, and inorganic ions using blood collected from the subjects, with and without using the reaction products, and these serum test values were calculated. When the results were compared, almost identical results were obtained, and it was found that the use of the reaction product had no effect on serum test values. Among the test values obtained, the values for the main items are shown in the table.

【table】

[Table] Comparative example 1 Liquid polyptene with a specific gravity of 0.867 (manufactured by NOF Corporation,
Product number 015SH) 100 parts by weight, the specific gravity after blending is approximately
1.050, hydrophilic silica A (specific gravity 2.65, manufactured by Illinois Mineral Company, trade name Imusil), hydrophilic silica B (specific gravity 2.2, manufactured by Degussa Corporation, trade name Aerosil), 200) or hydrophobic silica (specific gravity 2.2, manufactured by Degussa, trade name Aerosil R972) were mixed and dispersed to prepare four types of separation agents (1-1 to 1-4). The specific gravity was as shown in the same table.

[Table] From each of the four separation agents prepared above,
Two 1 g portions were taken and each 1 g portion was dispensed at the bottom of a test tube. Approximately 5 ml of human blood was added to this, left for 40 minutes to coagulate, and then centrifuged at 3000 rpm for 10 minutes using a centrifuge with an arm length of approximately 10 cm. . When the state of serum separation thus obtained was observed, in all four tests, it was observed that minute oil droplets were present in the serum and an oil film was present on the surface of the serum. Furthermore, in all cases except Test No. 2-3, a small amount of separating agent remained at the bottom of the test tube. Example 2 Liquid polyptene (same as used in Comparative Example 1)
A liquid chlorinated polyptene having a specific gravity of 1.050 was prepared in the same manner as in Example 1. 1 g each of this chlorinated polyptene was taken into two test tubes, and serum separation was performed in the same manner as in Comparative Example 1. When the state of separation was observed, the separating agent formed a clean partition wall, and the oil in the serum was separated. No droplets or oil film on the serum surface were observed. Next, the following test was conducted for the purpose of examining the storage stability of the separation agents prepared in this Example and the Comparative Example. Prepare 1 g each of the separating agents of Example 2 and Comparative Examples 1-1 to 1-4 and dispense them into 4 test tubes for each separating agent, and add 2 tubes for each separating agent. They were divided into two groups. One group at room temperature and the other at 40
Each was stored at ℃ for 2 weeks. After storage, add approximately 5 ml of human blood to each test tube,
Thereafter, serum separation was performed in the same manner as in Comparative Example 1. The state of separation was observed as follows. Example 2 The separation state was good both when stored at room temperature and at 40°C. Comparative Examples No. 1-1 and 1-2 After centrifugation, many minute oil droplets were floating in the serum and on the surface of the serum, and the separation agent remained at the bottom of the test tube. The amount of separation agent interposed between the serum and the blood clot was smaller than that in the Examples. The amount of oil droplets and tube bottom residue was more significant in the samples stored at 40°C. Comparative Example No. 1-3 Most of the separation agent remained at the bottom of the tube, and there was no interposition between the serum and blood clot, and many minute oil droplets were present in the serum. Comparative Example No. 1-4 Most of the separation agent was present between the serum and the blood clot, but minute oil droplets were floating in the serum. The amount of oil droplets was more prominent in the samples stored at 40°C.

Claims (1)

[Claims] 1 Liquid polyptene, whose specific gravity at 25℃ is less than 1.03, has a specific gravity at 25℃ due to chlorination.
A serum or plasma separating agent comprising a liquid chloride having a viscosity of 1.03 to 1.08 and a viscosity of 5,000 to 500,000 centipoise.