Glycosylated hemoglobin detection method based on magnetic bead affinity adsorption

Abstract

The invention relates to the technical field of glycosylated hemoglobin detection, in particular to a glycosylated hemoglobin detection method based on magnetic bead affinity adsorption, which comprises the following steps: A, sample pretreatment; b, total hemoglobin detection: detecting the absorbance H1 of the sample after hemolysis; c, carrying out affinity reaction; d, magnetic separation: starting an electromagnet tightly attached to the wall of the cuvette, and adsorbing the functionalized magnetic beads to the wall of the cuvette by adopting the electromagnet; e, non-glycosylated hemoglobin detection: detecting the absorbance H2 of the supernatant under the same optical path; and F, result calculation: calculating according to a formula HbA1c% = (H1-H2) / H1 * 100% to obtain a detection value of glycosylated hemoglobin. The glycosylated hemoglobin detection method disclosed by the invention is simple and convenient to operate, low in cost and very easy to realize automation. Compared with other high performance liquid chromatography methods for detecting glycosylated hemoglobin, the cost of a detection instrument is reduced by 70%, and the cost of a single test reagent is reduced by 30%.

Description

Technical Field

[0001] This invention relates to the field of glycated hemoglobin detection technology, specifically to a glycated hemoglobin detection method based on magnetic bead affinity adsorption. Background Technology

[0002] Glycated hemoglobin (HbA1c) is a product of the binding of hemoglobin in red blood cells with glucose in human blood. HbA1c was first isolated from other types of hemoglobin using chromatography in 1958 and classified as a glycoprotein in 1968. In 1969, it was discovered that HbA1c levels increased in diabetic patients. In 1975, researchers derived the reaction formula for the formation of HbA1c. The formation of HbA1c from the binding of hemoglobin in red blood cells is an irreversible reaction, directly proportional to blood glucose concentration, and remains stable for approximately 120 days, allowing for the observation of blood glucose concentrations from 120 days prior. The abbreviation for HbA1c is HBA1C. HbA1c testing typically reflects a patient's blood glucose control over the past 8–12 weeks. Elevated glycated hemoglobin (HbA1c) levels have multifaceted effects on the human body. It can alter the affinity of red blood cells for oxygen, leading to tissue and cellular hypoxia and accelerating the formation of cardiovascular and cerebrovascular complications. It can also cause glomerular thickening, inducing diabetic nephropathy (DN). Furthermore, it can increase blood lipids and blood viscosity, a significant factor in the development of cardiovascular disease. Therefore, monitoring HbA1c is crucial for disease control, complication prediction, and patient screening in diabetic patients. From a detection perspective, existing HbA1c detection methods, such as ion exchange chromatography and high-performance liquid chromatography (HPLC), are expensive and complex. Immunoassays (fluorescence immunochromatography and chemiluminescence) require antibodies, resulting in high costs, and the antigen-antibody binding titer leads to a large coefficient of variation (CV), failing to meet the national standard requirement of less than 3%. Some methods based on boric acid affinity may also have cumbersome or inefficient separation steps. Summary of the Invention

[0003] In order to overcome the shortcomings and deficiencies of the existing technology, the purpose of this invention is to provide a glycated hemoglobin detection method based on magnetic bead affinity adsorption.

[0004] The objective of this invention is achieved through the following technical solution: a method for detecting glycated hemoglobin based on magnetic bead affinity adsorption, comprising the following steps:

[0005] A. Sample pretreatment: Add hemolysin and blood sample to a dilution cup and mix well to dissolve the hemolysin;

[0006] B. Total hemoglobin detection: The absorbance H1 of the hemolyzed sample was detected using an LED light source and a silicon photodiode detector.

[0007] C. Affinity reaction: Add phenylboronic acid magnetic bead suspension and react gently with stirring in a buffer system with pH 7.5-8.5;

[0008] D. Magnetic separation: Activate the electromagnet that is in close contact with the wall of the cuvette to attract the functionalized magnetic beads to the wall of the cuvette;

[0009] E. Non-glycated hemoglobin detection: Detect the absorbance H2 of the supernatant under the same optical path;

[0010] F. Result Calculation: The glycated hemoglobin value can be obtained by calculating HbA1c% = (H1-H2) / H1 × 100%.

[0011] Preferably, in step A, the volume ratio of hemolysin to blood sample is 30-500:1; in step C, the amount of phenylboronic acid magnetic bead suspension added is 3-20 times the volume of blood sample.

[0012] Preferably, in step B, the wavelength of the LED light source is 415-630nm.

[0013] Preferably, in step F, considering the slight change in the overall solution volume caused by the addition of phenylboronic acid magnetic bead suspension, a volume calibration coefficient δ is introduced, and the formula is as follows: HbA1c%=(H1-H2×δ) / H1×100%.

[0014] Preferably, in step F, considering the influence of other factors, a calibration factor K can be introduced, and the formula is as follows: HbA1c%=K×(H1-H2×δ) / H1×100%.

[0015] The objective of this invention can also be achieved through the following technical solution: a method for detecting glycated hemoglobin based on magnetic bead affinity adsorption, comprising the following steps:

[0016] a. Sample pretreatment: Add hemolysin and blood sample to a dilution cup and mix well to dissolve the hemolysin;

[0017] b. Total hemoglobin detection: The absorbance H1 of the hemolyzed sample was detected using an LED light source and a silicon photodiode detector;

[0018] c. Affinity reaction: Add phenylboronic acid magnetic bead suspension and react gently with stirring in a buffer system with pH 7.5-8.5;

[0019] d. Magnetic separation: Activate the electromagnet that is in close contact with the cuvette wall to attract the functionalized magnetic beads to the cuvette wall;

[0020] e. Detection of total hemoglobin and magnetic bead background scattering: Detect the absorbance H2 of the total hemoglobin and magnetic bead mixture under the same optical path;

[0021] f. Calculate the background scattering factor of magnetic beads: Calculate the influence factor of magnetic beads on hemoglobin detection through H1 and H2, ΔH_magnetic_bead=H2-H1, where ΔH_magnetic_bead is the signal increment after the addition of magnetic beads;

[0022] g. Washing of non-glycated hemoglobin: Drain the supernatant through the drain hole at the bottom of the dilution cup, add the washing solution to the dilution cup, turn off the magnetic force, stir, and then use the magnetic force to attract the phenylboronic acid beads to the cup wall, and drain the washing solution; repeat this process 3-5 times to thoroughly wash away the non-glycated hemoglobin.

[0023] h. Calculate the net signal of glycated hemoglobin: Add the same volume of diluent as when detecting total hemoglobin to the dilution cup, disconnect the magnet, mix thoroughly, and then detect the absorbance H3 under the same optical path. H3 contains glycated hemoglobin and magnetic bead background. The magnetic bead background in H3 is the same as that in H2. Then the net signal of glycated hemoglobin is calculated according to the formula: H_glycated_corrected=H3-(H2-H1);

[0024] i. Result calculation: The glycated hemoglobin value can be obtained by calculating HbA1c% = (H_glycated_corrected / H1) × 100%.

[0025] Preferably, in step A, the volume ratio of hemolysin to blood sample is 30-500:1; in step C, the amount of phenylboronic acid magnetic bead suspension added is 3-20 times the volume of blood sample.

[0026] Preferably, in step B, the wavelength of the LED light source is 415-630nm.

[0027] Preferably, in step i, considering the slight change in the overall solution volume caused by the addition of phenylboronic acid magnetic bead suspension, a volume calibration parameter α is introduced, i.e., the formula is: HbA1c%=(H_glycated_corrected / H1)×100%×α; the final calculation formula can be simplified as follows: HbA1c%=(H3-H2+H1) / H1×100%×α.

[0028] Preferably, in step i, if the background of the magnetic bead is found to be unstable, background subtraction can be performed using dual-wavelength detection, that is, introducing a reference wavelength of 700-900nm for simultaneous detection. The specific calculation formula is as follows:

[0029] H1_corrected=H1_main-H1_ref;

[0030] H2_corrected=H2_main-H2_ref;

[0031] H3_corrected=H3_main-H3_ref;

[0032] HbA1c%=(H3_corrected-H2_corrected+H1_corrected) / H1_corrected×100%;

[0033] Wherein: H1_main, H2_main, and H3_main are the total Hb, total Hb+ magnetic beads, and glycosylated Hb+ magnetic beads detected by the main wavelength, respectively; H1_ref, H2_ref, and H3_ref are the total Hb, total Hb+ magnetic beads, and glycosylated Hb+ magnetic beads detected by the reference wavelength, respectively.

[0034] The beneficial effects of this invention are as follows: The glycated hemoglobin detection method of this invention involves labeling Wulff-type phenylboronic acid onto carboxyl-containing Fe3O4@MPS nanospheres, then adding these functionalized magnetic beads to a diluted sample after hemolysin infusion. The functionalized magnetic beads then specifically bind to glycated hemoglobin. After the glycated hemoglobin has fully reacted with the functional magnetic beads, the beads are adsorbed onto the wall of a cuvette using an open-ring symmetric electromagnet. At this point, the supernatant contains non-glycated hemoglobin. The amount of glycated hemoglobin is calculated by detecting both total hemoglobin and the non-glycated hemoglobin supernatant. Alternatively, the amount of glycated hemoglobin can be calculated by washing the supernatant first, adding a diluent, and then detecting the absorbance.

[0035] The core of the glycated hemoglobin detection method of this invention lies in the specific binding of glycated hemoglobin to Wulff-type phenylboronic acid functionalized magnetic beads. The HbA1c percentage is calculated through magnetic separation and the difference in absorbance detection. Wulff-type phenylboronic acid functionalized magnetic beads bind to glycated hemoglobin via covalent bonds, making them more stable than physical adsorption or simple embedding, and resistant to repeated washing. Furthermore, Wulff-type phenylboronic acid operates at a pH closer to physiological levels and exhibits stronger binding force. In addition, the reaction site is supported by a spacer arm, reducing spatial phase and ensuring free reaction between the reaction site and the specific cis-diol groups of glycated hemoglobin, resulting in a significantly higher reaction efficiency than conventional phenylboronic acid.

[0036] The glycated hemoglobin detection method of this invention is simple to operate, low in cost, and very easy to automate. Compared with other methods such as high-performance liquid chromatography (HPLC) for glycated hemoglobin detection, the cost of the detection instrument is reduced by 70%, and the cost of a single test reagent is reduced by 30%.

[0037] The glycated hemoglobin detection method of the present invention has the following advantages:

[0038] 1. It avoids the use of expensive antibodies or complex chromatography equipment.

[0039] 2. The magnetic beads are separated efficiently and thoroughly, reducing interference.

[0040] 3. The difference calculation method effectively reduces systematic errors and improves accuracy.

[0041] 4. The entire process is easy to integrate into fully automated analytical instruments. Attached Figure Description

[0042] Figure 1 This is a fitting curve graph of samples with different concentrations detected in Example 1; where AE represents the test samples with HGB solubility of 70g / L, 150g / L, 165g / L, 256g / L and 301g / L respectively, and F represents all samples.

[0043] Figure 2 This is a fitting curve graph of samples with different contents detected in Example 1.

[0044] Figure 3 This is a fitting curve graph of samples with different concentrations detected in Example 2; where AE represents the test samples with HGB solubility of 70g / L, 150g / L, 165g / L, 256g / L and 301g / L respectively, and F represents all samples.

[0045] Figure 4 This is a fitting curve graph of samples with different contents detected in Example 2. Detailed Implementation

[0046] To facilitate understanding by those skilled in the art, the following description is provided in conjunction with embodiments and appendices. Figure 1-4 The invention will be further described below, and the content mentioned in the embodiments is not intended to limit the invention.

[0047] To facilitate understanding by those skilled in the art, the present invention will be further described below with reference to embodiments. The content mentioned in the embodiments is not intended to limit the present invention.

[0048] The Wulff-type phenylboronic acid functionalized magnetic beads used in this embodiment can be prepared according to the invention patent with publication number CN108569772A. The phenylboronic acid magnetic bead suspension can be prepared by dispersing the Wulff-type phenylboronic acid functionalized magnetic beads in an appropriate amount of buffer solution or deionized water.

[0049] Example 1

[0050] A method for detecting glycated hemoglobin based on magnetic bead affinity adsorption includes the following steps:

[0051] A. Sample pretreatment: Add hemolysin and blood sample to a dilution cup and mix well to dissolve the hemolysin;

[0052] B. Total hemoglobin detection: The absorbance H1 of the hemolyzed sample was detected using an LED light source and a silicon photodiode detector.

[0053] C. Affinity reaction: Add phenylboronic acid magnetic bead suspension and react gently with stirring in a buffer system with pH 7.5-8.5;

[0054] D. Magnetic separation: Activate the electromagnet that is in close contact with the wall of the cuvette to attract the functionalized magnetic beads to the wall of the cuvette;

[0055] E. Non-glycated hemoglobin detection: Detect the absorbance H2 of the supernatant under the same optical path;

[0056] F. Result Calculation: The glycated hemoglobin value can be obtained by calculating HbA1c% = (H1-H2) / H1 × 100%.

[0057] In step A, the volume ratio of hemolysin to blood sample is 100:1; in step C, the amount of phenylboronic acid magnetic bead suspension added is 5 times the volume of blood sample.

[0058] In step B, the wavelength of the LED light source is 540nm.

[0059] In step F, considering the slight change in the overall solution volume caused by the addition of phenylboronic acid magnetic bead suspension, a volume calibration coefficient δ is introduced, and the formula is as follows: HbA1c%=(H1-H2×δ) / H1×100%.

[0060] In step F, considering the influence of other factors, a calibration factor K can be introduced, and the formula is as follows: HbA1c%=K×(H1-H2×δ) / H1×100%.

[0061] Example 2

[0062] A method for detecting glycated hemoglobin based on magnetic bead affinity adsorption includes the following steps:

[0063] a. Sample pretreatment: Add hemolysin and blood sample to a dilution cup and mix well to dissolve the hemolysin;

[0064] b. Total hemoglobin detection: The absorbance H1 of the hemolyzed sample was detected using an LED light source and a silicon photodiode detector;

[0065] c. Affinity reaction: Add phenylboronic acid magnetic bead suspension and react gently with stirring in a buffer system with pH 7.5-8.5;

[0066] d. Magnetic separation: Activate the electromagnet that is in close contact with the cuvette wall to attract the functionalized magnetic beads to the cuvette wall;

[0067] e. Detection of total hemoglobin and magnetic bead background scattering: Detect the absorbance H2 of the total hemoglobin and magnetic bead mixture under the same optical path;

[0068] f. Calculate the background scattering factor of magnetic beads: Calculate the influence factor of magnetic beads on hemoglobin detection through H1 and H2, ΔH_magnetic_bead=H2-H1, where ΔH_magnetic_bead is the signal increment after the addition of magnetic beads;

[0069] g. Washing of non-glycated hemoglobin: Drain the supernatant through the drain hole at the bottom of the dilution cup, add the washing solution to the dilution cup, turn off the magnetic force, stir, and then use the magnetic force to attract the phenylboronic acid beads to the cup wall, and drain the washing solution; repeat this process 3-5 times to thoroughly wash away the non-glycated hemoglobin.

[0070] h. Calculate the net signal of glycated hemoglobin: Add the same volume of diluent as when detecting total hemoglobin to the dilution cup, disconnect the magnet, mix thoroughly, and then detect the absorbance H3 under the same optical path. H3 contains glycated hemoglobin and magnetic bead background. The magnetic bead background in H3 is the same as that in H2. Then the net signal of glycated hemoglobin is calculated according to the formula: H_glycated_corrected=H3-(H2-H1);

[0071] i. Result calculation: The glycated hemoglobin value can be obtained by calculating HbA1c% = (H_glycated_corrected / H1) × 100%.

[0072] In step A, the volume ratio of hemolysin to blood sample is 100:1; in step C, the amount of phenylboronic acid magnetic bead suspension added is 5 times the volume of blood sample.

[0073] In step B, the wavelength of the LED light source is 540nm.

[0074] In step i, considering the slight change in the overall solution volume caused by the addition of phenylboronic acid magnetic bead suspension, a volume calibration parameter α is introduced, i.e., the formula is: HbA1c%=(H_glycated_corrected / H1)×100%×α; the final calculation formula can be simplified as follows: HbA1c%=(H3-H2+H1) / H1×100%×α.

[0075] In step i, if the background of the magnetic bead is found to be unstable, background subtraction can be performed using dual-wavelength detection. That is, a reference wavelength of 700-900nm is introduced for simultaneous detection. The specific calculation formula is as follows:

[0076] H1_corrected=H1_main-H1_ref;

[0077] H2_corrected=H2_main-H2_ref;

[0078] H3_corrected=H3_main-H3_ref;

[0079] HbA1c%=(H3_corrected-H2_corrected+H1_corrected) / H1_corrected×100%;

[0080] Wherein: H1_main, H2_main, and H3_main are the total Hb, total Hb+ magnetic beads, and glycosylated Hb+ magnetic beads detected by the main wavelength, respectively; H1_ref, H2_ref, and H3_ref are the total Hb, total Hb+ magnetic beads, and glycosylated Hb+ magnetic beads detected by the reference wavelength, respectively.

[0081] The detection results of Examples 1-2 are as follows:

[0082] I. Optical System Parameters and Sample Parameters

[0083]

[0084] II. Detection Results of Example 1

[0085] 1. Feasibility verification

[0086] Following the detection method in Example 1, samples with HBA1c% of 3.0%, 6.0%, 10.0%, 15.0%, and 20.0%, respectively, including low hemoglobin samples, normal hemoglobin samples, high hemoglobin samples, and very high hemoglobin samples, were tested and calculated. The results are shown in the table below, and the fitted curves are shown in the figure. Figure 1 As shown.

[0087]

[0088] 2. Accuracy Verification

[0089] According to the detection method in Example 1, six groups of samples with HBA1c% of 3.0%, 5.0%, 7.0%, 10.0%, 14.0%, and 20.0% were tested and calculated respectively. Each group of samples was tested five times, and the average value was taken. The results are shown in the table below.

[0090]

[0091] 3. Intra-batch precision verification

[0092] Following the detection method in Example 1, three testing personnel performed twenty tests and calculations on three groups of samples with HBA1c% of 4.0%, 6.9%, and 18.3%, respectively. The results are shown in the table below.

[0093]

[0094]

[0095]

[0096] 4. Linearity range verification

[0097] Following the detection method of Example 1, thirty-seven groups of samples with HBA1c% of 3.0%, 3.5%, 4.0%, 5.5%...20.0%, 20.5%, and 21.0% were tested and calculated. Each group of samples was tested three times, and the average value was taken. The results are shown in the table below, and the fitted curve is shown in the figure below. Figure 2 As shown.

[0098]

[0099]

[0100]

[0101] 5. Sensitivity Verification

[0102] Following the detection method of Example 1, twenty groups of samples were tested, and the detection sensitivity was calculated. The results are shown in the table below.

[0103] III. Detection Results of Example 2

[0104] 1. Feasibility verification

[0105] Following the detection method in Example 2, samples with HBA1c% of 3.0%, 6.0%, 10.0%, 15.0%, and 20.0%, respectively, including low hemoglobin samples, normal hemoglobin samples, high hemoglobin samples, and very high hemoglobin samples, were tested and calculated. The results are shown in the table below, and the fitted curves are shown in the figure. Figure 3 As shown.

[0106]

[0107] 2. Accuracy Verification

[0108] According to the detection method in Example 2, six groups of samples with HBA1c% of 3.0%, 5.0%, 7.0%, 10.0%, 14.0%, and 20.0% were tested and calculated respectively. Each group of samples was tested five times, and the average value was taken. The results are shown in the table below.

[0109]

[0110] 3. Intra-batch precision verification

[0111] According to the detection method in Example 2, three testing personnel performed twenty tests and calculations on three groups of samples with HBA1c% of 4.0%, 6.9%, and 18.3%, respectively. The results are shown in the table below.

[0112]

[0113]

[0114]

[0115]

[0116] 4. Linearity range verification

[0117] Following the detection method of Example 2, thirty-seven groups of samples with HBA1c% of 3.0%, 3.5%, 4.0%, 5.5%...20.0%, 20.5%, and 21.0% were tested and calculated. Each group of samples was tested three times, and the average value was taken. The results are shown in the table below, and the fitted curve is shown in the figure below. Figure 4 As shown.

[0118]

[0119]

[0120]

[0121] 5. Sensitivity Verification

[0122] Following the detection method of Example 2, twenty groups of samples were tested, and the detection sensitivity was calculated. The results are shown in the table below.

[0123]

[0124] The glycated hemoglobin detection method of this invention is simple to operate, low in cost, and very easy to automate. Compared with other methods such as high-performance liquid chromatography (HPLC) for glycated hemoglobin detection, the cost of the detection instrument is reduced by 70%, and the cost of a single test reagent is reduced by 30%.

[0125] The above embodiments are preferred implementations of the present invention. In addition, the present invention can be implemented in other ways. Any obvious substitutions without departing from the concept of the present invention are within the protection scope of the present invention.

Claims

1. A method for detecting glycated hemoglobin based on magnetic bead affinity adsorption, characterized in that: Includes the following steps: A. Sample pretreatment: Add hemolysin and blood sample to a dilution cup and mix well to dissolve the hemolysin; B. Total hemoglobin detection: The absorbance H1 of the hemolyzed sample was detected using an LED light source and a silicon photodiode detector. C. Affinity reaction: Add phenylboronic acid magnetic bead suspension and react gently with stirring in a buffer system with pH 7.5-8.5; D. Magnetic separation: Activate the electromagnet that is in close contact with the wall of the cuvette to attract the functionalized magnetic beads to the wall of the cuvette; E. Non-glycated hemoglobin detection: Detect the absorbance H2 of the supernatant under the same optical path; F. Result Calculation: The glycated hemoglobin value can be obtained by calculating HbA1c% = (H1-H2) / H1 × 100%.

2. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 1, characterized in that: In step A, the volume ratio of hemolysin to blood sample is 30-500:1; in step C, the amount of phenylboronic acid magnetic bead suspension added is 3-20 times the volume of blood sample.

3. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 1, characterized in that: In step B, the wavelength of the LED light source is 415-630nm.

4. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 1, characterized in that: In step F, considering the slight change in the overall solution volume caused by the addition of phenylboronic acid magnetic bead suspension, a volume calibration coefficient δ is introduced, and the formula is as follows: HbA1c%=(H1-H2×δ) / H1×100%.

5. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 4, characterized in that: In step F, considering the influence of other factors, a calibration factor K can be introduced, and the formula is as follows: HbA1c%=K×(H1-H2×δ) / H1×100%.

6. A method for detecting glycated hemoglobin based on magnetic bead affinity adsorption, characterized in that: Includes the following steps: a. Sample pretreatment: Add hemolysin and blood sample to a dilution cup and mix well to dissolve the hemolysin; b. Total hemoglobin detection: The absorbance H1 of the hemolyzed sample was detected using an LED light source and a silicon photodiode detector; c. Affinity reaction: Add phenylboronic acid magnetic bead suspension and react gently with stirring in a buffer system with pH 7.5-8.5; d. Magnetic separation: Activate the electromagnet that is in close contact with the cuvette wall to attract the functionalized magnetic beads to the cuvette wall; e. Detection of total hemoglobin and magnetic bead background scattering: Detect the absorbance H2 of the total hemoglobin and magnetic bead mixture under the same optical path; f. Calculate the background scattering factor of magnetic beads: Calculate the influence factor of magnetic beads on hemoglobin detection through H1 and H2, ΔH_magnetic_bead=H2-H1, where ΔH_magnetic_bead is the signal increment after the addition of magnetic beads; g. Washing of non-glycated hemoglobin: Drain the supernatant through the drain hole at the bottom of the dilution cup, add the washing solution to the dilution cup, disconnect the magnetic force, stir, and then use the magnetic force to adsorb the phenylboronic acid beads to the cup wall, and drain the washing solution. This process is repeated 3-5 times to thoroughly wash away the non-glycated hemoglobin; h. Calculate the net signal of glycated hemoglobin: Add the same volume of diluent as when detecting total hemoglobin to the dilution cup, disconnect the magnet, mix thoroughly, and then detect the absorbance H3 under the same optical path. H3 contains glycated hemoglobin and magnetic bead background. The magnetic bead background in H3 is the same as that in H2. Then the net signal of glycated hemoglobin is calculated according to the formula: H_glycated_corrected=H3-(H2-H1); i. Result calculation: The glycated hemoglobin value can be obtained by calculating HbA1c% = (H_glycated_corrected / H1) × 100%.

7. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 6, characterized in that: In step A, the volume ratio of hemolysin to blood sample is 30-500:1; in step C, the amount of phenylboronic acid magnetic bead suspension added is 3-20 times the volume of blood sample.

8. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 6, characterized in that: In step B, the wavelength of the LED light source is 415-630nm.

9. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 6, characterized in that: In step i, considering the slight change in the overall solution volume caused by the addition of phenylboronic acid magnetic bead suspension, a volume calibration parameter α is introduced, i.e., the formula is: HbA1c%=(H_glycated_corrected / H1)×100%×α; the final calculation formula can be simplified as follows: HbA1c%=(H3-H2+H1) / H1×100%×α.

10. The method for detecting glycated hemoglobin based on magnetic bead affinity adsorption according to claim 6, characterized in that: In step i, if the background of the magnetic bead is found to be unstable, background subtraction can be performed using dual-wavelength detection. That is, a reference wavelength of 700-900nm is introduced for simultaneous detection. The specific calculation formula is as follows: H1_corrected=H1_main-H1_ref; H2_corrected=H2_main-H2_ref; H3_corrected=H3_main-H3_ref; HbA1c%=(H3_corrected-H2_corrected+H1_corrected) / H1_corrected×100%; Wherein: H1_main, H2_main, and H3_main are the total Hb, total Hb+ magnetic beads, and glycosylated Hb+ magnetic beads detected by the main wavelength, respectively; H1_ref, H2_ref, and H3_ref are the total Hb, total Hb+ magnetic beads, and glycosylated Hb+ magnetic beads detected by the reference wavelength, respectively.

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