Extraction and preservation method of human hemoglobin

Abstract

The invention belongs to the technical field of human hemoglobin, and particularly relates to a human hemoglobin extraction and preservation method which comprises the following steps: S1, washing and separating red blood cells: standing collected whole blood, removing upper-layer yellow liquid, adding a sodium chloride solution, uniformly mixing, standing, and centrifuging to remove the upper-layer yellow liquid; s2, erythrocyte lysis: adding a hemolytic agent, stirring, and releasing hemoglobin; s3, hemoglobin collection: centrifuging the oscillated solution, taking the supernatant, and centrifuging again to take the supernatant; s4, preparing a hemoglobin preservation solution; s5, preparing a low-concentration hemoglobin solution: diluting high-concentration hemoglobin with normal saline and hemoglobin preserving fluid respectively and then preserving the high-concentration hemoglobin, by using the method provided by the invention, the hemoglobin extraction process is simple, the consumed time is short, the purity of the obtained hemoglobin is high, meanwhile, the stability of the low-concentration hemoglobin can be improved, and the method is suitable for industrial production. The kit can be used for matched detection of the fecal occult blood detection kit.

Description

Technical Field

[0001] The present invention belongs to the technical field of human hemoglobin, and particularly relates to a method for extracting and preserving human hemoglobin. Background Art

[0002] Human hemoglobin, commonly known as blood pigment, is an important protein component in red blood cells. Its main function is to bind and transport oxygen to various tissues in the human body, and at the same time bring carbon dioxide in the tissues back to the lungs for excretion. In a healthy human body, the content and activity of hemoglobin are maintained within a relatively stable range to ensure that all parts of the body can continuously obtain sufficient oxygen supply. Hemoglobin participates in a series of complex physiological and pathological processes, such as immune response, oxidative stress, etc. The normal functioning of its function is crucial for maintaining the stability of the internal environment of the body. Therefore, the research on hemoglobin not only helps to deeply understand the physiological mechanism of the human body, but also provides an important scientific basis for the diagnosis and treatment of diseases. It plays an important role especially in the diagnosis and evaluation of fecal occult blood.

[0003] Fecal occult blood refers to a small amount of bleeding in the digestive tract that causes hemorrhagic lesions. The red blood cells are digested and destroyed, and the appearance of the feces has no abnormal changes, and the bleeding cannot be confirmed by the naked eye and under the microscope. In the early stage of clinical digestive tract malignancies, 20% of patients may show a positive occult blood test. The positive rate of occult blood in advanced patients can reach more than 90%, and it can be continuously positive; for digestive tract bleeding, the fecal occult blood test of patients with digestive tract ulcers is mostly intermittently positive; dysentery, rectal polyps, hemorrhoids bleeding, etc. can also cause more red blood cells in the feces, resulting in a positive occult blood test. Therefore, fecal occult blood examination can be used as an important test for detecting digestive tract bleeding caused by various reasons, and it is a relatively effective method. At present, the main clinical methods for detecting fecal occult blood are colloidal gold immunochromatography and chemical methods.

[0004] Colloidal gold immunochromatography is widely used in the fields of medical health, food safety, environmental monitoring, etc. due to its advantages of (1) high sensitivity, with a detection limit that can reach the ng level, (2) intuitive detection results, and (3) mature technology and low cost. For the detection of fecal occult blood, colloidal gold immunochromatography uses colloidal gold as an indicator label and applies the technical principle of antigen-antibody immune reaction to quickly detect human hemoglobin in feces. Currently, there are already many products on the market. According to the products of colloidal gold detection reagents on the market, the lowest detection limit for hemoglobin is 50 ng / mL to 200 ng / mL. This requires technical personnel to use human hemoglobin solutions with a concentration lower than 50 ng / mL for research during the R & D process. Research shows that the stability of low-concentration hemoglobin solutions is extremely poor, and there are few hemoglobin quality control products supporting fecal occult blood reagents on the market and they are expensive. After enterprises purchase them, they still need to dilute them into low-concentration solutions for preservation. Therefore, the acquisition and preservation of low-hemoglobin solutions have become urgent problems for enterprises to solve.

[0005] At present, the extraction of hemoglobin mainly lyses red blood cells by mechanical or non-mechanical fragmentation methods to release hemoglobin. Among them, the mechanical methods include homogenization, ultrasonic fragmentation, extrusion, pulverization, etc., and the non-mechanical methods are mainly chemical methods and bioenzymatic methods, etc. The simplest method is to use the principle of red blood cells swelling and bursting by absorbing water to prepare a cell lysate to cause the lysis of red blood cells and release hemoglobin. However, in the prior art, the cell lysate has problems of complex operation, insufficient lysis and long time in actual application.

[0006] At present, there are few studies on improving the stability of hemoglobin. In existing studies, adding buffers, protein stabilizers, surfactants, etc. to the hemoglobin solution can improve the stability of the hemoglobin solution, but new influencing factors may be introduced during the fecal occult blood test, affecting the test results. Summary of the Invention

[0007] In order to solve the problems in the prior art, the present invention provides a method for extracting and preserving human hemoglobin, achieving the purpose of simple extraction of hemoglobin and improving the preservation stability.

[0008] The present invention solves its technical problems by adopting the following technical solutions:

[0009] The present invention aims to provide a method for extracting and preserving human hemoglobin, comprising the following steps:

[0010] S1. Wash and separate red blood cells: After standing the collected whole blood, remove the upper yellow liquid, then add sodium chloride solution, mix well and stand, and centrifuge to remove the upper yellow liquid;

[0011] S2. Lyse red blood cells: Add a hemolytic agent and stir to release hemoglobin;

[0012] S3. Collect hemoglobin: Centrifuge the shaken solution, take the supernatant, and centrifuge again to take the supernatant;

[0013] S4. Prepare a hemoglobin preservation solution;

[0014] S5. Prepare a low-concentration hemoglobin solution: Dilute the high-concentration hemoglobin with physiological saline and a hemoglobin preservation solution respectively and then preserve it.

[0015] Furthermore, the volume of the collected whole blood is 2-10 mL.

[0016] Furthermore, in S1, add 0.9% sodium chloride solution with the same volume as the whole blood, mix well and stand for 0.5 h, and the centrifugation conditions are 1000 rpm / min, 5 min, 4 °C, repeat 3-5 times until there is no yellow liquid on the upper layer.

[0017] Furthermore, the components of the hemolytic agent include 0.05 - 2 g / L of buffer, 10 - 20 g / L of anionic surfactant, and 10 - 20 mL / L of protein precipitant. The solvent of the above hemolytic agent is pure water after high-pressure sterilization.

[0018] Furthermore, the buffer adopts phosphate buffer solution of 0.05 - 2 g / L of sodium dihydrogen phosphate and 0.05 - 2 g / L of potassium dihydrogen phosphate. The anionic surfactant adopts sodium dodecyl sulfonate, and the protein precipitant adopts one or several of absolute ethanol, methanol, and acetone.

[0019] Furthermore, in S2, after adding the hemolytic agent, it is stirred with a magnetic stirrer at a rotation speed of 600 rpm for 30 min at 4°C.

[0020] Furthermore, in S3, the centrifugation conditions are all 12,000 rpm / min, 30 min, and 4°C.

[0021] Furthermore, the hemoglobin preservation solution includes sodium chloride solution, trehalose, glycerol, BSA, surfactant, and preservative.

[0022] Furthermore, the concentration of the sodium chloride solution is 0.9%, the concentration of trehalose is 10 - 100 g / L, the concentration of glycerol is 5% - 20%, the concentration of BSA is 0.5 - 2.0 g / L. The surfactant adopts S9 and / or S24, the concentration of S9 is 0.1% - 1%, the concentration of S24 is 0.1% - 1%, and the preservative adopts sodium diacetate with a concentration of 0.05% - 0.2%.

[0023] Furthermore, the diluted hemoglobin solution is stored at 4°C and -20°C.

[0024] Compared with the prior art, the beneficial technical effects of the present invention are as follows:

[0025] 1. For the method for extracting and preserving human hemoglobin of the present invention, the hemoglobin extraction process is simple and time-consuming. It can completely lyse red blood cells to release hemoglobin, and the obtained hemoglobin has high purity and does not contain white blood cell components. At the same time, it can improve the stability of low-concentration hemoglobin and can be used for the supporting detection of fecal occult blood test kits (colloidal gold method).

[0026] 2. The surfactants S9 and S24 in the hemoglobin preservation solution of the present invention belong to non-hemolytic surfactants, which can improve the fluidity and wettability of the solution, maintain a stable state, and the added preservative sodium diacetate can continuously inhibit the activity of microorganisms, reduce the influence of microorganisms on the solution quality, thereby achieving the effect of improving the stability of the hemoglobin dilution solution.

[0027] The above description is only an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention, it can be implemented according to the content of the specification. And in order to make the above content, objects, features and advantages of the present invention more obvious and understandable, the specific embodiments of the present invention are given below. Specific Embodiments

[0028] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are only illustrative explanations of the present invention and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0029] In addition, unless otherwise specifically stated, various raw materials, reagents, instruments and equipment used in the present invention can be obtained through market purchase or prepared by existing methods.

[0030] Example 1

[0031] A method for extracting and preserving human hemoglobin:

[0032] (1) Obtain a human whole blood sample: Collect 2 mL of fresh and healthy human EDTA-anticoagulated whole blood.

[0033] (2) Wash and separate red blood cells: Add 2 mL of 0.9% sodium chloride solution, mix well, let stand for 0.5 h, centrifuge at 1000 rpm / min for 5 min at 4°C, and repeat 3 times.

[0034] (3) Lyse red blood cells: Add a hemolytic agent, use a magnetic stirrer, stir at 600 rpm at 4°C for 30 min to release hemoglobin.

[0035] Among them, the components of the hemolytic agent include: sodium dihydrogen phosphate: 0.05 g / L; potassium dihydrogen phosphate: 0.05 g / L; sodium dodecyl sulfate: 10 g / L; absolute ethanol: 10 mL / L; purified water: make up to 1 L after dissolution.

[0036] (4) Collect hemoglobin: Centrifuge the shaken solution, take the supernatant. Centrifuge again and take the supernatant. The centrifugation conditions are both 12000 rpm / min for 30 min at 4°C.

[0037] (5) Measure the hemoglobin concentration: Use an automatic blood cell analyzer to measure the hemoglobin concentration and analyze the purity.

[0038] (6) Prepare a hemoglobin preservation solution: The preservation solution includes sodium chloride solution, trehalose, glycerol, BSA, surfactant, and preservative.

[0039] Sodium chloride solution: 9 g / L;

[0040] Trehalose: 10 g / L;

[0041] Glycerol: 5% (V / V);

[0042] BSA: 0.5 g / L;

[0043] Surfactants: S9 0.1% (V / V), S24 0.1% (V / V);

[0044] Preservative sodium diacetate: 0.05% (V / V).

[0045] (7) Preparation of low-concentration hemoglobin solution: Dilute high-concentration hemoglobin to 2.0 μg / mL, 0.2 μg / mL, and 0.02 μg / mL respectively using normal saline and hemoglobin preservation solution, and store at 4°C and -20°C.

[0046] (8) Stability test of hemoglobin solution: Take out the hemoglobin solution stored at 4°C every 2 days and the hemoglobin solution stored at -20°C every month, and use a fecal occult blood detection reagent (colloidal gold method) to detect the performance of the hemoglobin solution.

[0047] Test the purity and stability of the extracted hemoglobin according to the scheme in Example 1: After the hemoglobin extraction is completed, use an automatic blood cell analyzer to detect its purity and test the stability of the hemoglobin. The results are shown in Table 1 and Table 2 below:

[0048] Table 1: Results of detecting the extracted hemoglobin by an automatic blood cell analyzer

[0049]

[0050]

[0051] Note: * indicates not detected due to too low concentration.

[0052] Table 2: Stability test results of low-concentration hemoglobin Under the storage condition of 4°C:

[0053]

[0054] Under the storage condition of -20°C:

[0055]

[0056]

[0057] As can be seen from Table 1, when hemoglobin is extracted according to the method of the present invention, red blood cells can be completely lysed to release hemoglobin, and the obtained hemoglobin has high purity and almost no white blood cell components; as can be seen from Table 2, the hemoglobin preservation solution prepared according to the method of the present invention can improve the stability of low-concentration hemoglobin and can be used for the supporting detection of fecal occult blood test kits (colloidal gold method).

[0058] Example 2

[0059] A method for extracting and preserving human hemoglobin:

[0060] (1) Obtain a human whole blood sample: Collect 5 mL of fresh and healthy human EDTA-anticoagulated whole blood.

[0061] (2) Wash and separate red blood cells: Add 5 mL of 0.9% sodium chloride solution and mix well. The standing time is 0.5 h, the centrifugation conditions are 1000 rpm / min, 5 min, 4°C, and the number of repetitions is 4 times.

[0062] (3) Lyse red blood cells: Add a lysing agent and use a magnetic stirrer to stir at a speed of 600 rpm at 4°C for 30 min to release hemoglobin.

[0063] Among them, the components of the lysing agent include: sodium dihydrogen phosphate: 1.0 g / L; potassium dihydrogen phosphate: 1.0 g / L; sodium dodecyl sulfate: 15 g / L; absolute ethanol: 15 mL / L; purified water: make up to 1 L after dissolution.

[0064] (4) Collect hemoglobin: Centrifuge the shaken solution and take the supernatant. Centrifuge again and take the supernatant. The centrifugation conditions are both 12000 rpm / min, 30 min, 4°C.

[0065] (5) Measure the hemoglobin concentration: Use an automatic blood cell analyzer to measure the hemoglobin concentration and analyze the purity.

[0066] (6) Prepare a hemoglobin preservation solution: The preservation solution includes sodium chloride solution, trehalose, glycerol, BSA, surfactant, and preservative.

[0067] Sodium chloride solution: 9 g / L;

[0068] Trehalose: 50 g / L;

[0069] Glycerol: 10% (V / V);

[0070] BSA: 1.0 g / L;

[0071] Surfactant: S9 0.5% (V / V), S24 0.5% (V / V);

[0072] Sodium diacetate as preservative: 0.1% (V / V).

[0073] (7) Preparation of low-concentration hemoglobin solutions: Dilute high-concentration hemoglobin to 2.0 μg / mL, 0.2 μg / mL, and 0.02 μg / mL respectively using normal saline and hemoglobin preservation solution (the concentration is set according to the detection limit of 0.2 μg / mL for fecal occult blood detection reagent), and store at 4 °C and -20 °C.

[0074] (8) Stability test of hemoglobin solutions: Take out the hemoglobin solutions stored at 4 °C every 2 days and those stored at -20 °C monthly, and use fecal occult blood detection reagent (colloidal gold method) to detect the performance of the hemoglobin solutions.

[0075] Test the purity and stability of the extracted hemoglobin according to the scheme in Example 2: After the extraction of hemoglobin, use an automatic blood cell analyzer to detect its purity and test the stability of hemoglobin. The results are shown in Table 3 and Table 4 below:

[0076] Table 3: Results of detecting the extracted hemoglobin by an automatic blood cell analyzer

[0077]

[0078] Note: * indicates not detected due to too low concentration.

[0079] Table 4: Stability test results of low-concentration hemoglobin under the condition of storage at 4 °C:

[0080]

[0081] Under the condition of storage at -20 °C:

[0082]

[0083] As can be seen from Table 3, by extracting hemoglobin according to the method of the present invention, red blood cells can be completely lysed to release hemoglobin, and the obtained hemoglobin has high purity and does not contain white blood cell components; as can be seen from Table 4, the hemoglobin preservation solution prepared according to the method of the present invention can improve the stability of low-concentration hemoglobin and can be used for the supporting detection of fecal occult blood detection kits (colloidal gold method).

[0084] Example 3

[0085] A method for extracting and preserving human hemoglobin:

[0086] (1) Obtain human whole blood samples: Collect 10 mL of fresh and healthy human EDTA-anticoagulated whole blood.

[0087] (2) Wash and separate red blood cells: Add 2 mL of 0.9% sodium chloride solution and mix well. Let it stand for 0.5 h. Centrifuge at 1000 rpm / min for 5 min at 4°C. Repeat 5 times.

[0088] (3) Lyse red blood cells: Add lysing agent and use a magnetic stirrer to stir at 600 rpm at 4°C for 30 min to release hemoglobin.

[0089] Among them, the components of the lysing agent include: sodium dihydrogen phosphate: 2.0 g / L; potassium dihydrogen phosphate: 1.0 g / L; sodium dodecyl sulfate: 20 g / L; absolute ethanol: 20 mL / L; purified water: make up to 1 L after dissolution.

[0090] (4) Collect hemoglobin: Centrifuge the shaken solution and take the supernatant. Centrifuge again and take the supernatant. The centrifugation conditions are both 12000 rpm / min for 30 min at 4°C.

[0091] (5) Determine hemoglobin concentration: Use a fully automatic blood cell analyzer to determine the hemoglobin concentration and analyze the purity.

[0092] (6) Prepare hemoglobin preservation solution: The preservation solution includes sodium chloride solution, trehalose, glycerol, BSA, surfactant, and preservative.

[0093] Sodium chloride solution: 9 g / L;

[0094] Trehalose: 100 g / L;

[0095] Glycerol: 20% (V / V);

[0096] BSA: 2.0 g / L;

[0097] Surfactant: S9 1% (V / V), S24 0.5% (V / V);

[0098] Preservative sodium diacetate: 0.2% (V / V).

[0099] (7) Prepare low-concentration hemoglobin solutions: Dilute high-concentration hemoglobin to 2.0 μg / mL, 0.2 μg / mL, and 0.02 μg / mL with normal saline and hemoglobin preservation solution respectively, and store at 4°C and -20°C.

[0100] (8) Test the stability of hemoglobin solutions: Take out the hemoglobin solutions stored at 4°C every 2 days and those stored at -20°C every month, and use a fecal occult blood detection reagent (colloidal gold method) to detect the performance of the hemoglobin solutions.

[0101] Comparative Example 1

[0102] Prepare hemoglobin according to Example 1 in CN107266563 A, and dilute the hemoglobin after preparing the hemoglobin diluent according to Example 1 in CN115267209A. After the extraction of hemoglobin is completed, use an automatic blood cell analyzer to detect its purity and test the stability of the hemoglobin.

[0103] (1) Ingredients: Take 1 - 5 ml of fresh whole blood from a healthy person, and add an anticoagulant. The anticoagulant used is EDTAK2. The fresh whole blood from a healthy person is whole blood that is negative for HIV, HCV, HBsAg, and TP.

[0104] (2) Washing and centrifugation: Add physiological saline with a volume 8 - 10 times that of the ingredients to the ingredients. The physiological saline is a 0.9% NaCl solution. Centrifuge at 1000 revolutions per minute for 10 minutes and remove the supernatant. The washing and centrifugation process is repeated 8 - 10 times.

[0105] (3) Obtain erythrocyte suspension: After the washing and centrifugation are completed, add physiological saline to restore to the volume of the fresh blood from a healthy person in step (1) to obtain an erythrocyte suspension.

[0106] (4) Extract hemoglobin: Add polyethylene glycol octylphenyl ether with a volume ratio of 0.5 - 2% to the erythrocyte suspension and mix well on a vortex mixer for 10 - 15 minutes to rupture the erythrocyte membrane and release hemoglobin. Then centrifuge at 3000 revolutions per minute for 20 - 30 minutes, and use a pipette to aspirate 70 - 80% of the upper liquid to obtain a hemoglobin solution, and discard the remaining part.

[0107] (5) Dialysis: Add 3 - 5 times the buffer solution to the hemoglobin solution, then place the hemoglobin solution in a dialysis bag and dialyze it in the dialysis solution. Place the dialysis bag in 3 - 5 L of dialysis solution at 4°C and dialyze for 16 - 24 hours. The dialysis solution includes:

[0108] Buffer solution: 50 - 200 mM, pH = 7.0 - 7.5;

[0109] Sodium chloride: 9 g / L;

[0110] Antioxidant: 50 - 200 mM;

[0111] Mass percentage of preservative: 0.1 - 5%;

[0112] Wherein the above solvent is purified water.

[0113] (6) Sub - packaging: Determine the value of the dialyzed hemoglobin on an automatic blood cell analyzer.

[0114] (7) Prepare hemoglobin diluent:

[0115] 1) Add 100 g / L sucrose to 1X PBS solution and stir until dissolved;

[0116] 2) Add 100 mg / L horseradish peroxidase and stir until dissolved;

[0117] 3) Add 1 g / L BSA and stir until dissolved;

[0118] 4) Then add 10% (v / v) glycerol and stir. The mixing time in step S4 should be at least 20 minutes. After preparation, transfer it to a bottle and store at 2 - 8°C.

[0119] Test the purity and stability of the extracted hemoglobin according to the schemes in Example 3 and Comparative Example 1: After the extraction of hemoglobin, use an automatic blood cell analyzer to detect its purity and test the stability of hemoglobin. The results are shown in Table 5 and Table 6 below:

[0120] Table 5 Results of detecting the extracted hemoglobin by an automatic blood cell analyzer

[0121]

[0122] Note: * indicates that the concentration is too low to be detected.

[0123] Table 6 Stability test results of low - concentration hemoglobin under the storage condition of 4°C:

[0124]

[0125] Under the storage condition of - 20°C:

[0126]

[0127] As can be seen from Table 5, the purity of the hemoglobin solution prepared by Comparative Example 1 and the hemoglobin prepared by Example 3 of the present invention method is relatively high. However, the dialysis operation in the preparation process of Comparative Example 1 takes 16 - 24 hours, while dialysis is not required in the present invention, which greatly shortens the preparation time and saves time cost. As can be seen from Table 6, both the hemoglobin diluent prepared by Comparative Example 1 and the hemoglobin diluent prepared by Example 3 of the present invention method can improve the stability of hemoglobin with a concentration of 0.02 - 2 μg, but the stability of the present invention is higher. The surfactants S9 and S24 involved in the present invention belong to non - hemolytic surfactants, which can improve the fluidity and wettability of the solution, maintain a stable state, and the added preservative sodium diacetate can continuously inhibit the activity of microorganisms, reduce the influence of microorganisms on the solution quality, thereby achieving the effect of improving the stability of the hemoglobin diluent.

[0128] The serial numbers of the above - mentioned embodiments of the present invention are only for description and do not represent the advantages or disadvantages of the embodiments.

[0129] The embodiments of the present invention have been described above. However, the present invention is not limited to the above specific embodiments. The above specific embodiments are merely illustrative rather than restrictive. Under the inspiration of the present invention, those of ordinary skill in the art can also make many forms without departing from the spirit of the present invention and the scope protected by the claims. All of these fall within the protection scope of the present invention.

Claims

1. A method for extracting and preserving human hemoglobin, characterized in that: The following steps are involved: S1. Washing and separation of red blood cells: After the collected whole blood is allowed to stand, the upper yellow liquid is removed, and then sodium chloride solution is added, mixed and allowed to stand, and the upper yellow liquid is removed by centrifugation; S2, red blood cell lysis: add hemolytic agent, stir and release hemoglobin; S3. Hemoglobin collection: centrifuge the shaken solution, take the supernatant, and centrifuge again to take the supernatant; S4. Prepare hemoglobin preservation solution; S5. Preparation of low-concentration hemoglobin solution: dilute high-concentration hemoglobin with normal saline and hemoglobin preservation solution respectively and then store it.

2. A method for extracting and preserving human hemoglobin as claimed in claim 1, characterized in that: Add 0.9% sodium chloride solution in the same volume as whole blood to S1, mix well and let stand for 0.5 h. Centrifuge at 1000 rpm / min, 5 min, 4°C, and repeat 3 to 5 times.

3. A method for extracting and preserving human hemoglobin as claimed in claim 1, characterized in that: The components of the hemolytic agent include 0.05-2 g / L of a buffer, 10-20 g / L of an anionic surfactant, and 10-20 mL / L of a protein precipitant.

4. A method for extracting and preserving human hemoglobin as claimed in claim 3, characterized in that: The buffer is a phosphate buffer of sodium dihydrogen phosphate and potassium dihydrogen phosphate, the anionic surfactant is sodium dodecyl sulfate, and the protein precipitant is one or more of anhydrous ethanol, methanol and acetone.

5. A method for extracting and preserving human hemoglobin as claimed in claim 4, characterized in that: In S2, after adding the hemolytic agent, a magnetic stirrer was used to stir at a speed of 600 rpm at 4°C for 30 min.

6. A method for extracting and preserving human hemoglobin as claimed in claim 1, characterized in that: In S3, the centrifugation conditions were 12000 rpm / min, 30 min, and 4°C.

7. A method for extracting and preserving human hemoglobin as claimed in claim 1, characterized in that: The hemoglobin preservation solution includes sodium chloride solution, trehalose, glycerol, BSA, surfactant and preservative.

8. A method for extracting and preserving human hemoglobin as claimed in claim 7, characterized in that: The concentration of sodium chloride solution is 0.9%, the concentration of trehalose is 10-100 g / L, the concentration of glycerol is 5%-20%, the concentration of BSA is 0.5-2.0 g / L, the surfactant is S9 and / or S24, the concentration of S9 is 0.1%-1%, the concentration of S24 is 0.1%-1%, and the preservative is sodium diacetate with a concentration of 0.05%-0.2%.

9. A method for extracting and preserving human hemoglobin as claimed in claim 1, characterized in that: The diluted hemoglobin solution was stored at 4°C and -20°C.

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