JP5822385B2 - Blood collection tube - Google Patents

Description

  The present invention relates to a blood collection tube used for measuring a blood glucose level, and more particularly to a blood collection tube containing a drug containing a glycolysis inhibitor.

  In order to diagnose diabetes and the like, blood glucose concentration, that is, blood glucose level is measured. However, when the collected blood is stored at room temperature, there is a problem that the blood sugar level gradually decreases. Therefore, a blood collection tube containing a pharmaceutical composition containing a glycolysis inhibitor is widely used.

  As the glycolysis inhibitor, fluoride salts such as NaF and KF are used. However, it takes 2-3 hours until the glycolysis inhibitory effect by the fluoride salt appears. Therefore, when the blood is left at room temperature as it is after blood collection, a glycolytic reaction proceeds by a group of glycolytic enzymes contained in cells such as erythrocytes in the blood. Therefore, when the above fluoride salt is used as a glycolysis inhibitor, the amount of glucose measured 2 hours after blood collection is said to be about 87 to 93% of the amount of glucose immediately after blood collection (Non-patent Document 1). ). That is, only using a fluoride salt could not sufficiently suppress a decrease in glucose concentration immediately after blood collection.

  Patent Document 1 below discloses a method of directly coating D-mannose on a blood collection tube in which a glycolysis inhibitor in which D-mannose and / or a derivative thereof is held in a carrier is enclosed in a blood collection tube, or on the inner wall of the blood collection tube Is described. However, in these methods, hemolysis was observed in the blood sample.

  Patent Document 2 below discloses a blood collection tube in which a film made of D-mannose alone or a mixture of D-mannose and sodium fluoride is formed at the bottom of the blood collection tube. In this blood collection tube, the dissolution of the glycolysis inhibitor in the blood sample is delayed by forming a film of the glycolysis inhibitor on the bottom. It is said that hemolysis can be prevented thereby. However, even with this method, hemolysis could not be sufficiently suppressed.

Japanese Patent Laid-Open No. 01-127959 Japanese Patent Laid-Open No. 03-118474 Clinical Chemistry, 28, 1982, 190-192.

  An object of the present invention is to provide a blood collection tube that prevents hemolysis of collected blood and that can effectively suppress a decrease in blood glucose level and that can stably measure the blood glucose level.

  As a result of studying various uses of D-mannose, fluoride salts, and the like as glycolysis inhibitors, the present inventors have included a specific amount of D-mannose and a specific amount of fluoride salt, and as a blood anticoagulant, EDTA And / or a water-soluble drug containing heparin is applied to the inner wall of the blood collection tube by spraying and dried, and then the osmotic pressure when the drug is dissolved in the same amount of water as the collected blood is specified It was found that adjusting the amount of the drug so that it falls within the range can effectively suppress both the decrease in blood sugar level and hemolysis, and the present invention has been achieved.

  That is, as described above, in the prior art using D-mannose and / or a derivative thereof as a glycolysis inhibitor, hemolysis could not be ignored, but according to the present invention, D-mannose was used. In spite of the use, hemolysis can be suppressed and a decrease in blood glucose level can be effectively suppressed.

  The present invention is a blood collection tube used to measure a blood glucose level in blood and collects a predetermined amount of blood, and has a bottomed cylindrical blood collection tube with one end opened and the other closed And a particulate drug attached to the inner wall of the blood collection tube main body, wherein the drug is a water-soluble drug including a glycolysis inhibitor and a blood anticoagulant, The blood anticoagulant contains at least one of EDTA and / or heparin, containing 1 to 5 mg of D-mannose and 0.3 to 1.0 mg of fluoride salt in terms of NaF amount per 1 mL of collected blood. The drug is adhered to the inner wall of the blood collection tube main body by spraying and drying, and the blood collection tube main body is filled with a predetermined amount of water collected in the blood collection tube Of aqueous solution when drug attached to the inner wall There is a range of 30~80mOsm.

  In the blood collection tube according to the present invention, since the particulate drug adheres to the inner wall of the blood collection tube main body, unlike the case of a granular or powdered drug, the drug is not unevenly distributed at the bottom of the blood collection tube. Therefore, the drug can be dissolved more rapidly by blood.

  In a specific embodiment of the blood collection tube according to the present invention, the pH of the blood sample when a predetermined amount of blood is collected is in the range of 7.3 to 8.8. In this case, hemolysis can be more reliably suppressed.

  In another specific aspect of the blood collection tube according to the present invention, the maximum particle size of the particulate drug is in the range of 50 μm to 1500 μm. If the maximum particle size of the adhering drug is within this range, the drug can be dissolved more rapidly. Therefore, the number of times of inversion mixing for dissolving the drug in blood can be reduced. In addition, hemolysis can be more effectively suppressed.

  In still another specific aspect of the blood collection tube according to the present invention, the drug adheres to the inner wall of the blood collection tube main body at a position below the blood surface when the predetermined amount of blood is collected. In this case, even if no inversion mixing is performed after blood collection, if the bottom of the blood collection tube is turned down and it is kept upright, the collected blood will come into contact with most of the medicine in the blood collection tube and the drug will be promptly Can be dissolved in blood. Therefore, the number of times of overturning can be further reduced.

  According to the blood collection tube according to the present invention, despite the use of a glycolysis inhibitor containing D-mannose, the drug can be quickly dissolved in the collected blood, and the generation of hemolysis is effectively prevented. In addition to being able to suppress, the number of inversion mixing can be reduced. In addition, a decrease in blood glucose level over time can be effectively suppressed.

  That is, according to the blood collection tube of the present invention, it is possible to effectively suppress both hemolysis suppression and blood glucose level reduction. Therefore, the blood sugar level can be accurately measured.

It is a typical front view showing a blood collection tube concerning one embodiment of the present invention. It is a schematic block diagram for demonstrating the spray coating apparatus used in order to obtain the blood-collecting tube of one embodiment of this invention. It is a partial notch expansion front sectional view which shows the principal part of the spray nozzle part of the spray coating apparatus shown in FIG. It is sectional drawing which follows the ZZ line | wire of FIG. It is the photograph which expands and shows the chemical | medical agent adhering to the inner wall of the blood collection tube main body of the blood collection tube in one embodiment of this invention.

  Hereinafter, the present invention will be clarified by describing the blood collection tube of the present invention and specific embodiments thereof.

(Drug)
The drug used in the blood collection tube according to the present invention contains 1 to 5 mg of D-mannose and 0.3 to 1.0 mg of fluoride salt in terms of NaF amount per 1 mL of blood as a glycolysis inhibitor. A water-soluble drug containing EDTA and / or heparin as a blood anticoagulant. Hereinafter, the amount of the fluoride salt described in the present specification is converted to the amount of NaF unless otherwise specified.

  In the blood collection tube, the amount of blood collected is determined according to the size of the blood collection tube. For example, a 5 mL blood collection tube is a blood collection tube that collects 5 mL of blood. That is, a predetermined amount of blood is collected in the blood collection tube. Therefore, 1 to 5 mg of D-mannose and 0.3 to 1.0 mg of fluoride salt with respect to 1 mL of the blood, for example, in the blood collection tube for collecting 5 mL of blood, the drug is 5 to 25 mg. It is meant to contain D-mannose and 1.5-5.0 mg fluoride salt.

  More generally expressed, in the case of a blood collection tube for collecting blood of nmL (n is an integer), the drug is 1n-5n (mg) of D-mannose, and 0.3n to 1.0n (mg ) Fluoride salt.

(Glycolysis inhibitor)
The glycolysis inhibitor consists of 1 to 5 mg of D-mannose and 0.3 to 1.0 mg of fluoride salt per 1 mL of blood.

  When the amount of D-mannose is less than 1 mg per mL of blood, blood glucose stability immediately after blood collection is deteriorated, and when the amount of D-mannose exceeds 5 mg, hemolysis is likely to occur.

  The amount of D-mannose is more preferably 1.5 to 4 mg per mL of blood. In this case, the stability of the blood sugar level can be further increased, and hemolysis can be more effectively suppressed.

  When the amount of the fluoride salt is less than 0.3 mg per mL of blood, blood glucose stability is deteriorated, and when the amount of the fluoride salt is more than 1 mg, hemolysis is likely to occur. Further, the amount of the fluoride salt is more preferably 0.5 to 0.8 mg per mL of blood. In this case, the stability of the blood sugar level can be further increased, and hemolysis can be more effectively suppressed.

  Examples of the fluoride salt that can be used in the present invention include NaF and KF.

(Blood anticoagulant)
The blood anticoagulant used in the present invention is EDTA and / or heparin.

  As EDTA, EDTA salt is preferably used, and it is preferable to use an amount corresponding to 1 mg to 2 mg in terms of EDTA amount per 1 mL of collected blood. Thereby, blood coagulation can be more effectively suppressed.

  As the EDTA salt, for example, EDTA-2Na or EDTA-2K can be used. In the case of EDTA-2K, it is desirable to use 1.25 mg to 2.50 mg per 1 mL of collected blood.

  Heparin may be used in an amount of 10 to 20 U per 1 mL of collected blood, and heparin sodium, heparin lithium, or the like can be used. In addition, when EDTA and heparin are used in combination as blood anticoagulants, they have blood anticoagulant ability, and when they are mixed by inversion immediately after blood collection and centrifuged at 1500 g × 10 minutes, each amount is an amount that does not cause hemolysis. It is desirable to do.

(Drug spraying and drying)
The blood collection tube of the present invention has a bottomed cylindrical shape in which an aqueous solution obtained by dissolving a drug containing the above-mentioned specific amounts of glycolysis inhibitor and blood anticoagulant in water is opened at one end and closed at the other end. It is made by spraying and drying on the inner wall surface of the blood collection tube body. Further, the osmotic pressure when the drug is dissolved with a predetermined amount of water collected in the blood collection tube is 30 to 80 mOsm.

  In the present invention, an aqueous solution in which a drug containing the glycolysis inhibitor and the blood anticoagulant is dissolved in water is prepared by spray-coating on the bottomed cylindrical inner wall surface of the blood collection tube body and drying it. In a state where NaF which is difficult to dissolve in water and D-mannose which is easily dissolved in water are mixed, they adhere to the inner wall of the blood collection tube as fine droplets and are dried. Accordingly, the minute droplets are dried, and the drug adheres to the inner wall of the blood collection tube main body in a particulate form.

  For this reason, the drug surface area of the inner wall of the blood collection tube of the present invention is larger than the surface area of the drug coated on the inner wall of the blood collection tube. Therefore, according to the present invention, contrary to the above prior art, the drug can be dissolved quickly in the blood. Thereby, it is considered that both the avoidance of hemolysis and the prevention of a decrease in blood glucose level can be achieved.

  In addition, about the method of spray-applying a pharmaceutical composition to the inner wall of a blood-collecting blood vessel main body, various spray application apparatuses can be used. Examples of such a spray coating device include a device using a spray nozzle that sprays a pharmaceutical composition from the tip as an aerosol, a spray coating device using a piezo-type jet dispenser, and the like. Preferably, a spray coating apparatus described with reference to FIGS.

(Osmotic pressure of aqueous drug solution)
The osmotic pressure of the aqueous drug solution is 30-80 mOsm when the drug is dissolved by adding a predetermined amount of water to the blood collection tube of the present invention to an amount equal to that of the collected blood.

  When the drug osmotic pressure is less than 30 mOsm, the blood anticoagulant ability and the stability of blood glucose level are insufficient. Further, when the drug osmotic pressure is higher than 80 mOsm, hemolysis is likely to occur.

  This osmotic pressure can be controlled by adjusting the amounts of the glycolysis inhibitor and blood anticoagulant. It should be noted that the addition of other compounds is not excluded as long as there is no adverse effect.

(PH of collected blood sample)
In the blood collection tube according to the present invention, the pH of the blood sample when the predetermined amount of blood is collected is preferably in the range of 7.3 to 8.8. Usually, the pH of the collected blood is known to be higher than the blood pH in the body because the carbon dioxide partial pressure of the blood is lowered. However, if the pH is within the above range, hemolysis is more likely to occur. It can suppress more effectively.

  In addition, control of pH can be achieved by adjusting the composition of the drug.

(Particle size of sprayed and dried drug)
The maximum particle size of the drug when the water-soluble drug according to the present invention is spray-coated and dried is preferably 50 μm to 1500 μm. Furthermore, it is preferable that the maximum particle size be 50 μm to 1000 μm. If the maximum particle size is smaller than 50 μm, the area required to apply the drug increases, the size of the blood collection tube increases compared to the amount of blood collected, and the number of inversions increases to dissolve all the drug in the blood. There is.

  In addition, when the maximum particle size of the drug obtained by spray-coating and drying the water-soluble drug of the present invention is larger than 1500 μm, the drug solubility may be lowered and hemolysis may easily occur.

(Drug adherence to the inner wall of the blood collection tube)
In the present invention, it is preferable that the drug is fixed to the inner wall of the blood collection tube main body as fine particles. In that case, it is difficult for the drug to be unevenly distributed locally in the blood collection tube. In addition, the concentration of the drug in the collected blood is difficult to increase locally. When stored without being fixed, the fine powder may be localized at the bottom of the blood collection tube, etc., and when blood is collected in that state, the concentration of the drug in some blood in contact with the fine powder is locally Since it becomes high, there is a risk of hemolysis.

(Drug application position)
In the present invention, the water-soluble drug is attached to the inner wall of the blood collection tube main body, and the position where the drug is attached is preferably lower than the blood surface. The blood surface means the blood surface when the aforementioned predetermined amount of blood is collected in a blood collection tube. When the drug attachment position is below the blood surface, the collected blood and the drug come into contact more rapidly. Therefore, all drugs are easily dissolved. Therefore, even if the number of times of overturning is reduced, it is possible to more effectively suppress a decrease in blood glucose level.

  In addition, since the drug in the blood collection tube of the present invention quickly dissolves in the blood even if the number of inversions after blood collection is small at the drug application position of the present invention, the preferable drug application range is from the bottom of the tube to the blood. It is preferable to be below the surface.

(Preferred example of spray coating apparatus)
A specific example of the spray coating apparatus used for the spray coating will be described with reference to FIGS.

  As shown in FIG. 2, the spray coating apparatus 1 has a chemical tank 2. A chemical solution for spraying is stored in the chemical solution tank 2.

  The chemical tank 2 is connected to the spray nozzle 4 via a pipe 3. A pump 5 is connected in the middle of the pipe 3 in order to send a chemical solution.

  As shown in a front sectional view in FIG. 3, the spray nozzle 4 includes an inner tube 6 and an outer tube 7 into which the inner tube 6 is inserted. The inner tube 6 and the outer tube 7 are integrated on the upper end side of the inner tube 6.

  At the upper end of the spray nozzle 4, an opening 6a of the inner tube is located. The pipe 3 is connected to the opening 6 a, and the spray chemical solution is supplied into the inner pipe 6. At the tip of the inner tube 6, a discharge port 6 b through which a chemical solution is discharged is opened. That is, the inner tube 6 is a member having a chemical liquid flow path that connects the opening 6a and the discharge port 6b.

  The outer tube 7 is provided to supply compressed gas for generating aerosol in the space between the outer wall of the inner tube 6 and the inner surface of the outer tube 7.

  In the present embodiment, the outer tube 7 has a cylindrical shape. Therefore, the space 8 between the outer wall of the inner tube 6 and the inner wall of the outer tube 7 has a substantially circular tubular cross section. The upper end of the space 8 communicates with an opening 9 provided on the side of the outer tube 7.

  In the present embodiment, the tip of the inner tube 6 protrudes to the outside from the tip of the outer tube 7.

  A pipe 10 shown in FIG. 2 is connected to the opening 9. As shown in FIG. 2, the pipe 10 is connected to a compressor C for supplying compressed gas. A valve V for adjusting the flow path of the compressed gas is connected in the middle of the pipe 10.

  Compressed gas is supplied from the compressor C through the pipe 10 to the space 8, and at the tip of the spray nozzle 4, the spray chemical solution discharged from the inner tube 6 is sheared by the compressed gas to form a chemical aerosol.

  As shown in the sectional view taken along the line ZZ in FIG. 3, in the inner tube 6 and the outer tube 7, when the outer diameter of the inner tube 6 is d1 and the inner diameter of the outer tube 7 is D1, the space 8 The distance d2 between the inner wall of the outer tube 7 and the outer wall of the inner tube 6 is preferably in the range of 25 to 75% of the difference ΔD between D1 and d1. Within this range, a uniform chemical aerosol can be more reliably formed.

(Blood collection tube)
The blood collection tube according to the present invention is obtained as described above. FIG. 1 shows a schematic front view of the blood collection tube of one embodiment of the present invention. The blood collection tube 11 has a bottomed cylindrical blood collection body 12. The drug 13 is attached to the inner wall of the blood collection tube main body 12 as schematically shown by cross hatching. In FIG. 1, the drug 13 schematically shows an adhesion region by cross hatching, but actually, the drug 13 is adhered to the inner wall of the blood collection tube main body 12 in a particulate form as described above.

  The material of the blood collection tube main body is not particularly limited, and those used for known blood collection tubes can be used. Preferably, polyethylene terephthalate is used.

  In the blood collection tube according to the present invention, preferably, a closing member is attached so as to close the opening of the blood collection tube main body. As such a blocking member, an appropriate blocking member such as a rubber plug, an overcap type rubber plug, or a film plug in which rubber is laminated on an aluminum foil can be used.

  Preferably, in the present invention, the inside of the blood collection tube body to which the blocking member is attached is preferably decompressed. By reducing the pressure in the blood collection tube, blood can be collected quickly. Note that the degree of decompression may be appropriately set according to the amount of blood collected, that is, according to the predetermined blood volume described above.

[Examples and Comparative Examples]
Next, the effects of the present invention will be clarified by giving specific examples and comparative examples.

  In the following examples and comparative examples, the spray coating apparatus 1 shown in FIGS. 2 to 4 was used. Here, the length of the outer tube, which is a tube for supplying air, was 81.4 mm, the outer diameter was 3.5 mm, and the inner diameter was 1.0 mm. The length of the inner tube for discharging the drug solution was 145 mm, the outer diameter was 0.88 mm, and the inner diameter was 0.55 mm. Also, a spray nozzle was used in which the inner tube was inserted into the outer tube and the tip of the inner tube protruded 0.2 mm from the tip of the outer tube.

(Example 1: 1.00 mg of D-mannose and 0.3 mg of NaF per mL of blood)
As a glycolysis inhibitor, 0.889 g of D-mannose (manufactured by Wako Pure Chemical Industries, Wako Special Grade), 0.267 g of NaF (special grade, manufactured by Wako Pure Chemical Industries, Ltd.), EDTA-2K 1 as a blood anticoagulant .676 g (manufactured by Dojindo Laboratories, for test research) was prepared, and water was added to make the total amount 20 g. These components were sufficiently dissolved to prepare a spray drug solution.

  Next, a chemical solution tank is used to apply the spray drug solution to the inner wall of a polyethylene terephthalate blood collection tube main body (length 75 mm, outer diameter 12.5 mm, inner diameter 10.5 mm) using the spray application device. The drug solution for spraying was contained in Next, an air pressure of 0.20 MPa was applied to aerosolize the drug solution for spraying, and 45 mg was spray coated in the blood collection tube body. Thereafter, air was supplied into the blood collection tube main body, and the spray drug solution was dried to obtain a blood collection tube.

  In this blood collection tube, the amount of blood to be collected is 2 mL, and the amount of drug adhered to the inner wall of the blood collection tube main body is 1.00 mg of D-mannose and 0.3 mg of NaF per 1 mL of collected blood It has become.

  The spray drug solution was uniformly applied and dried within a height of 70 mm from the bottom in the blood collection tube.

(Example 2 to Example 8)
In the following Examples 2 to 8, Example 1 and Example 8, except that the amounts of D-mannose and NaF in the spray drug solution were changed so that the amounts of D-mannose and NaF were as follows. Similarly, a spray drug solution was prepared, and each blood collection tube was prepared.

Example 2: 1.00 mg of D-mannose and 1.0 mg of NaF per mL of blood
Example 3: 1.25 mg of D-mannose and 0.5 mg of NaF per mL of blood
Example 4: D-mannose 2.0 mg / NaF 0.5 mg per mL of blood
Example 5: 4.0 mg of D-mannose and 0.5 mg of NaF per mL of blood
Example 6: 5.0 mg of D-mannose and 0.5 mg of NaF per mL of blood
Example 7: 1.25 mg of D-mannose and 0.8 mg of NaF per mL of blood
Example 8: 2.50 mg of D-mannose per mL of blood, 0.6 mg of NaF

(Example 9: D-mannose 2.50 mg, NaF 0.6 mg per mL of blood)
As a glycolysis inhibitor, 2.222 g of D-mannose, 0.533 g of NaF, 0.844 g of EDTA-2K as a blood anticoagulant and 2,000,000 U of heparin (manufactured by Wako Pure Chemical Industries, for biochemistry) ) Was dissolved in water to prepare a heparin aqueous solution 3000 U / g, and water was added to 1.48 g of the heparin aqueous solution to make a total amount of 20 g. These components were sufficiently dissolved to prepare a spray drug solution.

(Comparative Examples 1-6)
For the following Comparative Examples 1 to 6, except that the amount of D-mannose and NaF in the spray drug solution was changed so that the amounts of D-mannose and NaF per 1 mL of blood were the following amounts, respectively. A spray drug solution was prepared in the same manner as in Example 1 to prepare blood collection tubes for each comparative example.

Comparative Example 1: 0.75 mg of D-mannose and 0.5 mg of NaF per 1 mL of blood
Comparative Example 2: 1.25 mg of D-mannose and 0.1 mg of NaF per mL of blood
Comparative Example 3: D-mannose 10.0 mg / NaF 1.0 mg per mL of blood
Comparative Example 4: 1.25 mg of D-mannose and 1.5 mg of NaF per 1 mL of blood
Comparative Example 5: 0.75 mg of D-mannose and 0.1 mg of NaF per mL of blood
Comparative Example 6: D-Mannose 10.0 mg, NaF 1.5 mg per mL of blood

(Comparative Example 7)
In Comparative Example 7, 45 mg of the drug solution for spraying having the same composition as Example 8 in which D-mannose was 2.50 mg and NaF 0.6 mg per mL of blood was dispensed into a tube, and after adding an equivalent amount of ethanol and mixing, the inner wall of the tube As in Example 8, except that the tube was rotated so as to be uniformly wet, air was blown into the tube, the drug solution was gradually dried, and the inner wall of the blood collection tube was coated with the drug solution to prepare the blood collection tube It was.

(Comparative Example 8: D-mannose 2.50 mg, NaF 0.6 mg per mL of blood)
45 mg of the spray drug solution having the same composition as in Example 8 was placed in the bottom of the blood collection tube and dried to form a film of the drug composition to prepare a blood collection tube.

(Comparative Example 9: D-mannose 2.50 mg, NaF 0.6 mg per mL of blood)
After 45 mg of the spray drug solution having the same composition as in Example 8 was placed in the bottom of the blood collection tube and dried to form a film of the drug composition, the film was pulverized with a spoon to obtain a fine powder.

(Comparative Example 10: Blood sample pH 7.1 with a drug containing 2.50 mg of D-mannose, 0.6 mg of NaF, and 0.65 mg of citric acid per mL of blood)
The same as Example 1 except for the preparation of the following drug solution for spraying.

  As a glycolysis inhibitor, D-mannose 2.222 g, NaF 0.533 g, blood anticoagulant EDTA-2K 1.676 g, citric acid 0.578 g with water added to 20 g, the drug was sufficiently dissolved, A spray drug solution was prepared.

(Comparative Example 11: Blood sample pH 9.0 with a drug containing 2.50 mg of D-mannose and 0.6 mg of NaF per mL of blood)
The same as Example 1 except for the preparation of the following drug solution for spraying.

  As a glycolysis inhibitor, D-mannose 2.222 g, NaF 0.533 g, blood anticoagulant EDTA-2K 1.676 g, trisodium phosphate dodecahydrate 1.90 g (manufactured by Wako Pure Chemical Industries, Ltd.) added water The drug was sufficiently dissolved to prepare a spray drug solution.

(Comparative Example 12)
A blood glucose tube (product number: Benogect II, code number: VP-FC052K), which is a blood collection tube for blood glucose level measurement, manufactured by Terumo Corporation was prepared.

(Evaluation of blood collection tubes prepared in each Example and Comparative Example)
1) Evaluation of drug particle size The maximum particle size of the drugs in the blood collection tubes of Examples 1 to 9 and Comparative Examples 1 to 6 and 9 to 12 was measured using a microscope (manufactured by Nikon, MEASUREING MICROSCOPE MM-60).

2) Evaluation of osmotic pressure of blood collection tubes 2 mL of water for injection (manufactured by Otsuka Pharmaceutical Co., Ltd.) is added to the blood collection tubes of Examples 1 to 9 and Comparative Examples 1 to 12, and mixed by inversion, and all the drugs in each blood collection tube are mixed. After dissolution, the osmotic pressure of the drug solution was measured with an osmometer (manufactured by Arkray, OSMOSTAT OM-6040).

3) Evaluation of blood collection tubes using human blood (evaluation of blood glucose stability and hemolysis)
(1) Evaluation of blood glucose level To each of the three blood collection tubes obtained in Examples 1 to 9 and Comparative Examples 1 to 12, 2 mL of healthy human blood was added and mixed by inversion 10 times to mix the blood and the drug. did. Thereafter, all blood collection tubes were immediately centrifuged at 1500 g × 10 minutes (room temperature). Among them, the blood glucose level (blood glucose level immediately after blood collection) in the plasma of each of the blood collection tubes of Examples 1 to 9 and Comparative Examples 1 to 12 was measured.

  One of the remaining two blood collection tubes was stored at room temperature for 2 hours, and the other one was stored at room temperature for 1 day, and then the blood glucose level in the plasma was measured.

  The method for measuring the blood glucose level is as follows.

  A 7170S type Hitachi automatic analyzer (manufactured by Hitachi, Ltd.) was used as the measuring apparatus, and L type Wako GLU2 (manufactured by Wako Pure Chemical Industries, Ltd.) was used as the blood glucose level measuring reagent. The blood glucose level of plasma obtained from each blood collection tube was measured immediately after blood collection, 2 hours after room temperature storage, and 1 day after room temperature storage. The blood glucose level of the blood sample immediately after blood collection was taken as 100%, and the blood glucose level after storage was shown as a relative value. The results are shown in Table 1.

(2) Hemolysis evaluation For blood samples collected in the blood collection tubes of Examples 1 to 9 and Comparative Examples 1 to 12, after measuring the blood glucose level after 1 day of storage at room temperature, the absorbance at 415 nm of the plasma of each blood sample was measured And the presence or absence of hemolysis was evaluated. The results are shown in Table 1 below.

  The absorbance at 415 nm of a blood glucose tube (Benject II, code number: VP-FC052K) manufactured by Terumo Corporation, which is Comparative Example 12, is defined as 100%, and hemolysis evaluations of Examples 1-9 and Comparative Examples 1-11 are expressed as relative values. It is shown in 1.

(3) Measurement of blood sample pH The blood collection tubes of Examples 1 to 9 and Comparative Examples 1 to 12 were centrifuged at 1500 g × 10 minutes immediately after blood collection, and after measuring the blood glucose level immediately after blood collection, a small pH meter (Horiba A blood sample pH (plasma pH) of each blood collection tube was measured using a twin pH meter B-212) manufactured by Seisakusho, and the results are shown in Table 1.

(Stability of hemolysis and blood sugar level)
In Examples 1 to 9, even if the blood was collected and centrifuged at room temperature for 1 day, there was no hemolysis and the stability of the blood glucose level was very good at 97.0 to 99.5%.

  On the other hand, in Comparative Examples 1, 2 and 5 where the amount of the glycolysis inhibitor is small, there was no hemolysis after 1 day of room temperature storage, but the stability of the blood glucose level was 94.0 and 94.0, respectively. And 91.0%, which was worse than the examples.

  In other Comparative Examples 3, 4, and 6 to 11, the stability of the blood glucose level at 1 day after storage at room temperature was as good as 96.0 to 99.5%. Was recognized.

  In Comparative Example 12, the stability of the blood glucose level at room temperature for 1 day after blood collection was as good as 99.5%, but hemolysis was observed.

(Maximum particle size of drug and hemolysis)
In Examples 1 to 9, the maximum particle size of the drug was 330 to 890 μm.

  On the other hand, in Comparative Examples 3 and 6 having a large amount of D-mannose, the maximum particle size of the drug was 1600 and 1800 μm, respectively, which were larger than those in Examples 1-9. The main cause is that the solid content of the drug is large. Therefore, since it becomes difficult for the medicine to dissolve in blood, hemolysis was slightly observed as compared with Examples 1-9.

  In Comparative Example 4 having a large amount of NaF, hemolysis was also observed. In Comparative Example 12, immediately after blood collection, inversion mixing was performed 10 times as described in the product box, but hemolysis was severely observed. This hemolysis is considered to be caused by the erythrocyte membrane being damaged by the acidic drug.

(Drug storage method and hemolysis)
In Examples 1 to 9, hemolysis was not observed because the drug was sprayed and dried in a range of 70 mm in height from the bottom of the blood collection tube main body.

  On the other hand, in Comparative Example 7 (the drug was coated and dried on the entire inner wall of the blood collection tube main body) and Comparative Example 8 (the drug was dried in the form of a film on the bottom of the blood collection tube), the surface area of the drug was smaller than that in Example, so Difficult to dissolve in blood. For this reason, it is considered that the drug concentration in the blood in contact with the drug locally increased and hemolysis occurred.

  Moreover, in Comparative Example 9 (fine powder), since there is fine powder locally at the bottom of the blood collection tube, the concentration of the drug in the blood in contact with the drug is locally increased, so it is considered that hemolysis occurred.

(PH of blood sample and hemolysis)
The blood samples of Examples 1 to 9 had a pH of 7.8 to 8.0 and no hemolysis was observed.

  On the other hand, the blood samples of Comparative Examples 10 and 11 had a pH of 7.1 and 9.0, respectively, and hemolysis was observed. In Comparative Example 10, the cause of hemolysis was that the pH of the blood sample was slightly acidified by adding citric acid, and in Comparative Example 11, the pH of the blood sample was slightly shifted to alkali by adding trisodium phosphate. Thus, it is considered that the red blood cell membrane was damaged and hemolyzed.

(Examples 10-12 and Comparative Examples 13-17)
(Example 10)
As Example 10, the blood collection tube of Example 8 described above was prepared.

(Example 11)
45 mg of the spray drug solution obtained in Example 8 was applied uniformly between the bottom of the blood collection tube main body used in Example 1 at a height of 28 mm and dried, as in Example 8. It was. Therefore, the height of the blood surface when a predetermined blood volume is collected is 29 mm, whereas the upper end of the drug application position is located below the blood surface.

(Example 12)
Example 11 was the same as Example 11 except that 45 mg of the spray drug solution used in Example 9 was used.

(Comparative Examples 13, 15 and 16)
The same blood collection tubes as those of Comparative Examples 7, 8, and 9 were designated as Comparative Examples 13, 15, and 16, respectively.

(Comparative Example 14)
45 mg of the same spray drug solution used in Example 8 was coated at a position 28 mm in height from the bottom in the blood collection tube main body in the same manner as in Comparative Example 7 to obtain a blood collection tube.

(Comparative Example 17)
The above-described Comparative Example 12, that is, Terumo's blood sugar tube (product number: Benogect II, code number: VP-FC052K) was prepared.

(Evaluation method)
2 mL of blood was collected in the blood collection tubes of Examples 10 to 12 and Comparative Examples 13 to 17, and the influence of the number of inversions on hemolysis and blood glucose stability was evaluated.

(Stability of hemolysis and blood sugar level)
In Example 10, no hemolysis occurred at room temperature for 1 day after blood collection and centrifugation, regardless of the number of times of inversion mixing. In addition, regarding the stability of the blood glucose level at room temperature for 1 day, the relative% to the blood glucose level immediately after blood collection was 93.0% even without inversion mixing. When the inversion mixing number increased to 0, 2 and 5 times, the relative% was improved to 93.0, 99.0 and 99.5%, respectively.

  In Examples 11 and 12, since the drug is located below the blood surface, the stability of the blood glucose level is not affected by the number of inversions compared to Example 10, and the room temperature storage day The blood glucose stability was as good as 99.0 to 99.5%.

  On the other hand, in Comparative Examples 13-15, since the solubility of a chemical | medical agent was bad, the blood glucose stability of the room temperature preservation | save day without inversion mixing was 81-88% compared with Examples 10-12.

  In Comparative Example 16, the blood glucose stability for 1 day at room temperature without inversion was 91%.

  However, hemolysis was observed in any of Comparative Examples 13-16.

  Further, in Comparative Example 17, that is, Terumo's blood glucose tube (Benoject II, code number: VP-FC052K), the blood glucose level at room temperature for 1 day when the number of inversions immediately after blood collection was changed to 0, 2, and 5 times. The stability of the samples varied greatly as 68.0%, 97.0% and 98.0%, respectively. The cause of this is that in the product box of Terumo's blood glucose tube, in the section “Precautions related to the method of use”, “inverted blend with glycemic inhibitor varieties FC tube should be mixed 10 times or more. If it is not sufficiently mixed due to the reagent characteristics, hemolysis may occur]], because the solubility of the granular drug (maximum particle size 700 μm) is very poor, This is probably because the concentration of the drug dissolved in the blood greatly varied depending on the number of times of overturning.

(Maximum particle size of drug)
In Examples 10-12, the maximum particle size of the drug was 510-630 μm.

  On the other hand, in Comparative Example 16, the maximum particle size was 490 μm.

(Drug storage method and hemolysis)
In Example 10 and Examples 11 and 12, since the drug was spray-coated and dried in the range of 70 mm and 28 mm in height from the bottom in the blood collection tube, the drug had good solubility and no hemolysis was observed.

  On the other hand, in Comparative Examples 13 and 14 (the drug was coated and dried on the entire inner wall of the blood collection tube) and Comparative Example 15 (the drug was dried in the form of a film on the bottom of the blood collection tube), the surface area of the drug was small compared to Examples 10-12. Therefore, the drug is difficult to dissolve in the blood. Therefore, it is considered that the drug concentration in the blood in contact with the drug locally increased and hemolysis occurred.

  Further, in Comparative Example 16 (fine powder), since there is fine powder locally at the bottom of the blood collection tube main body, the concentration of the drug in the blood in contact with the drug is locally increased, so it is considered that hemolysis occurred.

(Blood sample pH)
The blood sample pHs of Examples 10 to 12 and Comparative Examples 13 to 16 were all 7.9.

DESCRIPTION OF SYMBOLS 1 ... Spray coating device 2 ... Chemical solution tank 3 ... Pipe 4 ... Spray nozzle 5 ... Pump 6 ... Inner pipe 6a ... Opening part 6b ... Discharge port 7 ... Outer pipe 8 ... Space 9 ... Opening part 10 ... Pipe 11 ... Blood collection tube 12 ... blood collection tube body 13 ... drug C ... compressor d1 ... outer diameter d2 ... distance D1 ... inner diameter

Claims (4)

A blood collection tube that is used to measure blood glucose levels in blood and collects a predetermined amount of blood,
A bottomed tubular blood collection tube body with one end open and the other end closed;
A particulate drug attached to the inner wall of the blood collection tube main body,
The drug is a water-soluble drug containing a glycolysis inhibitor and a blood anticoagulant, and the glycolysis inhibitor is 1 to 5 mg of D-mannose and NaF in terms of 1 mL of collected blood. 3 to 1.0 mg of fluoride salt, and the blood anticoagulant is at least one of EDTA and / or heparin,
The drug is attached in the form of particles by spraying and drying on the inner wall of the blood collection tube body,
A blood collection tube having an osmotic pressure of an aqueous solution in a range of 30 to 80 mOsm when a drug attached to the inner wall of the blood collection tube main body is dissolved with a predetermined amount of water collected in the blood collection tube.   The blood collection tube according to claim 1, wherein the pH of the blood sample when the predetermined amount of blood is collected is in the range of 7.3 to 8.8.   The blood collection tube according to claim 1 or 2, wherein the maximum particle size of the particulate drug is in the range of 50 µm to 1500 µm.   The blood collection tube according to any one of claims 1 to 3, wherein the drug is attached to an inner wall of the blood collection tube main body at a position below a blood surface when the predetermined amount of blood is collected. .

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