Emergency biochemical microsphere, preparation method thereof and application of emergency biochemical microsphere in vacuum blood collection tube

Abstract

The invention discloses emergency biochemical microspheres, a preparation method thereof and application of the emergency biochemical microspheres in a vacuum blood collection tube, and belongs to the technical field of medical instruments. The biochemical microspheres for emergency treatment are prepared from heparin, an excipient and a protective agent through the steps of solution preparation, drop bead balling, freeze drying and screening, and the diameter of the microspheres is 2.0-6.0 mm. The microspheres have the advantages of uniform particle size, good stability and stable anticoagulation effect, can simplify the production process, reduce the energy consumption and improve the blood test accuracy when being applied to the vacuum blood collection tube, and are suitable for large-scale production.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medical devices, in particular to an emergency biochemical microsphere, a preparation method thereof and application thereof in a vacuum blood collection tube. BACKGROUND

[0002] A vacuum blood collection tube is an indispensable blood collection tool in clinical testing, and the inner wall thereof is usually coated with an anticoagulant to prevent blood from coagulating. Among them, a vacuum blood collection tube added with heparin (sodium heparin or lithium heparin) is mainly used for emergency biochemical and blood gas testing. Heparin is widely used due to its good anticoagulation effect and small interference to blood components.

[0003] However, in the production process of a conventional heparin vacuum blood collection tube, heparin needs to be prepared into a liquid reagent, and then sprayed or added to the inner wall of the test tube and then dried at a high temperature. This process is complex, time-consuming, energy-consuming and low in production efficiency. More importantly, in the high-temperature drying process, the sulfate ester bond, glycoside bond and the like in the heparin molecule are easily broken, resulting in damage to the molecular structure and reduction in the anticoagulation activity. In addition, after drying, heparin is tightly attached to the tube wall, and if mixing is not sufficient after blood collection, local blood clotting may occur, affecting the accuracy of the test results. SUMMARY

[0004] The present application aims to provide an emergency biochemical microsphere, a preparation method thereof and application thereof in a vacuum blood collection tube. Heparin is prepared into a microsphere, which is then placed in a vacuum blood collection tube. The microsphere has the advantages of uniform particle size, good stability, stable anticoagulation effect, simple production process, low cost and the like, and can be better applied to the production and manufacturing of vacuum blood collection tubes.

[0005] To achieve the above-mentioned purpose, the present application adopts the following technical solutions: The emergency biochemical microsphere provided by the present application is composed of the following components: Heparin: sodium heparin or lithium heparin; Excipient: one or more selected from trehalose, sucrose and mannitol; Protective agent: one or both of glycine and bovine serum albumin; The diameter of the microsphere is 2.0mm-6.0mm.

[0006] Heparin as an anticoagulant component can inhibit the activity of thrombin in blood, thereby preventing blood clotting, and heparin sodium or heparin lithium is commonly used in clinic. The role of the excipient is to form a porous framework for the microspheres, increase the mechanical strength and stability of the microspheres, prevent the microspheres from collapsing, maintain the spherical structure and porosity, and at the same time form a glassy matrix during the freeze-drying process to protect the activity of heparin from damage; the role of the protective agent is to prevent the microspheres from aggregation and adhesion during preparation and storage, prevent denaturation, aggregation or adsorption to the surface of the container during the freeze-drying process, and improve the biological activity of heparin and the stability of the microspheres.

[0007] Preferably, the content of heparin is such that the final microspheres provide an anticoagulant strength of 12-25 IU / mL blood, the content of the excipient is 2.0-10.0%, and the content of the protective agent is 0.2-1.0%, based on the total mass of the microspheres.

[0008] The application provides a preparation method of the emergency biochemical microspheres. Step 1: solution preparation; heparin, an excipient and a protective agent are added to deionized water, stirred and dissolved to form a uniform mixed solution; Step 2: droplet formation; the mixed solution is dropped into liquid nitrogen through a droplet machine, and the droplets are rapidly frozen at ultra-low temperature to form microsphere precursors; the droplet volume is 5-50 μL / drop, and the liquid discharge speed is 1-3 s / drop; Step 3: freeze-drying; the microsphere precursors are subjected to vacuum freeze-drying, including a pre-freezing stage, a sublimation drying stage and a desorption drying stage, to remove water and obtain freeze-dried microspheres; Step 4: screening; microspheres with a diameter of 2.0-6.0 mm are screened out through a screening device.

[0009] Preferably, the freeze-drying conditions are as follows: The pre-freezing stage is carried out at a temperature of-80℃ to-40℃ for 2-6 hours; The sublimation drying stage is carried out at a temperature of-40℃ to 0℃ for 12-48 hours; The desorption drying stage is carried out at a temperature of 0℃ to 60℃ for 2-12 hours.

[0010] The application further provides application of the above-mentioned emergency biochemical microspheres in a vacuum blood collection tube, specifically at least one microsphere is added to the vacuum blood collection tube for anticoagulant treatment of a blood sample.

[0011] Compared with the prior art, the application has the following beneficial effects: (1) By optimizing the component combination and the preparation process, the obtained microspheres have the characteristics of uniform particle size, loose and porous structure, fast dissolution speed and stable anticoagulant effect; (2) The use of drop forming and freeze-drying process avoids the complex process of traditional spraying and high-temperature drying, and improves the production efficiency; (3) The freeze-drying process significantly reduces energy consumption compared to high-temperature drying, and better ensures the activity of heparin; (4) The microspheres dissolve quickly after contacting with blood, and the heparin is evenly distributed, avoiding the coagulation problem caused by insufficient mixing; (5) The microsphere shape is convenient for visual inspection, reducing the quality problems of missing or misadding; (6) Clinical application shows that the blood samples collected by the vacuum blood collection tube containing the microspheres of the application have no significant difference from the traditional blood collection tube in the detection of multiple biochemical indicators. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 The preparation process flow chart of the emergency biochemical microspheres of the application.

[0013] Figure 2 The structure schematic diagram of the emergency biochemical microspheres of the application.

[0014] Figure 3 The structure schematic diagram of the vacuum blood collection tube using the microspheres of the application. DETAILED DESCRIPTION

[0015] The technical solutions in the embodiments of the application will be described clearly and completely below with reference to the drawings in the embodiments of the application. Obviously, the described embodiments are only part of the embodiments of the application, not all the embodiments.

[0016] The information of the equipment and reagents used in the embodiments is as follows: Drop bead machine: SCIENTZ-DQIR, produced by Ningbo Xinzhi Freeze-drying Equipment Co., Ltd.

[0017] Freeze dryer: Scientz-50F / A, produced by Ningbo Xinzhi Freeze-drying Equipment Co., Ltd.

[0018] Sodium heparin: batch number 241215, produced by Wujiang Hehe Biochemical Products Co., Ltd.

[0019] Lithium heparin: batch number 20250307, produced by Wuhan Xizheceng Technology Co., Ltd.

[0020] Trehalose: batch number P3119481, produced by Adamas-beta.

[0021] Sucrose: batch number 180308, produced by Xilong Chemical Co., Ltd.

[0022] Mannitol: batch number 20150206, produced by Shanghai Ruili Spectrum Biotechnology Co., Ltd.

[0023] Glycine: batch number 241211A2, produced by Xi Long Chemical Co., Ltd.

[0024] Bovine serum albumin: batch number 210107, produced by Shanghai Ruili Map Biological Technology Co., Ltd.

[0025] Example 1 Solution preparation: weigh 1.41 g of heparin sodium (160 IU / mg), 5.0 g of sucrose, and 0.5 g of glycine, and place them in a triangular flask. Add about 80 mL of deionized water and stir to dissolve. Dilute to 100 mL.

[0026] Droplet formation: use a droplet machine to drop the solution into liquid nitrogen at 20 μL / drop, and the liquid outlet speed is 1.5 s / drop to form microsphere precursors.

[0027] Freeze-drying: place the microsphere precursors in a freeze-dryer, pre-freeze (-50°C, 2 h) → sublimation drying (-30°C, 15 h; -15°C, 1.5 h; 0°C, 1.5 h) → desorption drying (10°C, 1 h; 20°C, 1 h).

[0028] Screening and sub-packaging: screen microspheres with a diameter of 2.5-3.5 mm, and sub-packaging into 12×75 mm test tubes, 1 particle per tube, vacuum to 3 mL, and seal with a rubber plug.

[0029] According to the calculation, the heparin content of the blood collection tube prepared in this example is 15 IU / mL of blood.

[0030] Example 2 Solution preparation: weigh 2.50 g of lithium heparin (150 IU / mg), 5.0 g of mannitol, and 0.5 g of glycine, and prepare as above; Droplet and freeze-drying conditions are the same as in Example 1; Sub-packaging: add 1 particle of microspheres to each test tube (12×100 mm), and vacuum 5 mL to make a 5 mL blood collection tube.

[0031] According to the calculation, the heparin content of the blood collection tube prepared in this example is 15 IU / mL of blood.

[0032] Example 3 Solution preparation: weigh 1.41 g of heparin sodium, 5.0 g of trehalose, and 0.5 g of bovine serum albumin, and prepare as above; Droplet and freeze-drying conditions are the same as in Example 1; Sub-packaging: add 1 particle of microspheres to each test tube (12×75 mm), and vacuum 3 mL to make a 3 mL blood collection tube.

[0033] According to the calculation, the heparin content of the blood collection tube prepared in this example is 15 IU / mL of blood.

[0034] Example 4 Solution preparation: 2.50 g of lithium heparin, 5.0 g of mannitol, 0.5 g of bovine serum albumin were weighed and prepared as above; Droplet and freeze-drying conditions were the same as in Example 1. Sub-packing: the same as in Example 2.

[0035] It was calculated that the heparin content of the blood collection tube prepared in this example was 15 IU / mL blood.

[0036] Example 5 Solution preparation: 1.41 g of sodium heparin, 5.0 g of trehalose, 0.3 g of glycine, 0.3 g of bovine serum albumin were weighed and prepared as above; Droplet and freeze-drying conditions were the same as in Example 1. Sub-packing: the same as in Example 1.

[0037] It was calculated that the heparin content of the blood collection tube prepared in this example was 15 IU / mL blood.

[0038] To verify the effectiveness of the product of this example, 6 healthy volunteers were recruited to use the vacuum blood collection tubes prepared in Examples 1-5 above and the vacuum blood collection tubes produced by the conventional process on the market (batch number: 24122719, as a control group) for blood collection. Each person collected 6 tubes, a total of 36 samples. After mixing and centrifuging all samples under the same conditions, the upper serum was sent to a third-party testing laboratory for detection of more than 40 biochemical indicators including alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood glucose (GLU), total cholesterol (TCHO), etc.

[0039] The test results show that in all test items, the deviation of the test results of each example group of the application and the control group is less than 10%, and there is no statistically significant difference. This proves that the emergency biochemical microsphere vacuum blood collection tube provided has the same performance as the traditional product under the premise of ensuring the accuracy of the test results, and has more advantages in production process, energy consumption and quality control.

[0040] The above describes only the preferred specific embodiments of the present application, but the protection scope of the present application is not limited thereto, and any person skilled in the art can make equivalent substitutions or changes to the technical solutions and inventive concepts of the present application within the technical scope disclosed by the present application, which should be covered within the protection scope of the present application.

Claims

1. An emergency biochemical microsphere, characterized in that, The microspheres are heparin freeze-dried microspheres with a diameter of 2.0–6.0 mm; the microspheres are prepared by freeze-drying of raw materials comprising the following components: heparin, excipients, and protectants.

2. The emergency biochemical microsphere according to claim 1, characterized in that, The heparin is either sodium heparin or lithium heparin.

3. The emergency biochemical microspheres according to claim 1, characterized in that, The excipient is selected from one or more of trehalose, sucrose, and mannitol, and its content accounts for 2% to 10% of the total mass of the microspheres.

4. The emergency biochemical microsphere according to claim 1, characterized in that, The protective agent is selected from one or two of glycine and bovine serum albumin, and its content accounts for 0.2% to 1.0% of the total mass of the microspheres.

5. A method for preparing the emergency biochemical microspheres according to any one of claims 1 to 4, characterized in that, Includes the following steps: (a) Dissolve heparin, excipients and protective agents in a solvent to form a mixed solution; (b) The mixed solution is added dropwise to a freezing medium to form a microsphere precursor; (c) The microsphere precursor is subjected to vacuum freeze-drying to obtain the emergency biochemical microspheres.

6. The method according to claim 5, characterized in that, In step (b), the volume of a single drop added is 5 μL to 50 μL; the freezing medium is liquid nitrogen.

7. The method according to claim 5, characterized in that, In step (c), the vacuum freeze-drying includes: Pre-freezing stage: temperature -80℃ to -40℃, time 2 to 6 hours; Sublimation drying stage: temperature -40℃~0℃, time 12~48 hours; Analysis and drying stage: temperature is 0℃~60℃, time is 2~12 hours.

8. The application of the emergency biochemical microspheres according to claim 4 in the preparation of vacuum blood collection tubes.

9. The application according to claim 8, characterized in that, At least one of the aforementioned emergency biochemical microspheres is placed in a test tube of a vacuum blood collection tube and sealed with a rubber stopper; the emergency biochemical microspheres provide an anticoagulant strength of 12–25 IU / mL blood.

Images