High-sensitivity glucose detection material and preparation method thereof

Abstract

The invention relates to the technical field of glucose detection, in particular to a high-sensitivity glucose detection material and a preparation method thereof. The preparation raw materials of the high-sensitivity glucose detection material comprise the following components in parts by weight: 1-2 parts of modified glucose oxidase, 6-10 parts of modified porous microspheres, 1-1.5 parts of chloroplatinic acid and 0.6-1 part of palladium acetate. The prepared detection material is high in detection sensitivity and good in repeatability and stability.

Description

Technical Field

[0001] The present invention relates to the technical field of glucose detection, and particularly relates to a highly sensitive glucose detection material and a preparation method thereof. Background Art

[0002] Glucose detection is of great significance in the fields of clinical diagnosis, diabetes monitoring, and biomedical research. Accurate and highly sensitive glucose detection can help doctors promptly grasp the blood glucose levels of patients and formulate reasonable treatment plans, which is crucial for the early diagnosis and condition monitoring of metabolic diseases such as diabetes. Currently, common glucose detection materials include enzyme-based sensor materials, nanomaterials, etc. Although enzyme-based sensor materials have certain specificity, they have problems such as poor stability, being easily affected by environmental factors, and short service life; nanomaterials such as metal nanoparticles and carbon nanomaterials, although improving the detection sensitivity to a certain extent, have complex preparation processes, high costs, and the detection results are easily interfered, making it difficult to meet the requirements of high sensitivity, high stability, and low cost in practical applications. Therefore, how to develop a glucose detection material with high sensitivity and good stability has become an urgent technical problem in this field. Summary of the Invention

[0003] Aiming at the deficiencies of the prior art, the present invention proposes a highly sensitive glucose detection material and a preparation method thereof.

[0004] The present invention is realized through the following technical solutions: A highly sensitive glucose detection material, the preparation raw materials of which include the following components in parts by weight: 1 - 2 parts of modified glucose oxidase, 6 - 10 parts of modified porous microspheres, 1 - 1.5 parts of chloroplatinic acid, and 0.6 - 1 part of palladium acetate.

[0005] Further, the preparation raw materials of the modified glucose oxidase include the following components in parts by weight: 1 - 2 parts of glucose oxidase (GOD) and 2 - 4 parts of mercapto - polyethylene glycol - carboxyl (SH - PEG - COOH).

[0006] Further, the preparation method of the modified glucose oxidase includes the following steps: A1: Add GOD to the PB buffer solution with pH = 6.5 and mix well to obtain a GOD solution. Add SH - PEG - COOH to the PB buffer solution, add EDC and NHS, and stir at 150 - 200 rpm for 2 h to obtain an activation solution; A2: Add the GOD solution obtained in step A1 to the activation solution, stir and react at 200 - 300 rpm for 12 h, centrifuge at 10000 rpm for 5 - 10 min, and wash the precipitate with deionized water to obtain modified GOD.

[0007] Further, in step A1, in the GOD solution, the mass concentration of GOD in the PB buffer solution is 1-2 mg / mL.

[0008] Further, in step A1, the mass concentration of SH-PEG-COOH in the PB buffer solution is 2-3 mg / mL.

[0009] Further, in step A1, the mass ratio of SH-PEG-COOH, EDC and NHS is 10:1.5:1.

[0010] Further, the raw materials for preparing the modified porous microspheres include the following components in parts by weight: 2-3 parts of 1,3-propanedithiol, 1.5-2.1 parts of propargyl glycidyl ether, 0.3-0.5 parts of 1,7-octadiyne, 0.12-0.17 parts of benzoin dimethyl ether (DMAP), 1-1.5 parts of polyethylene glycol (PEG), 4-6 parts of 1,3-diamino-2-propanol (HD), and 1-1.5 parts of 3-maleimidopropionic acid hydroxysuccinimide ester (SMMP).

[0011] Further, the method for preparing the modified porous microspheres includes the following steps: B1: Mix 1,3-propanedithiol, propargyl glycidyl ether, and 1,7-octadiyne, add DMPA, chloroform, and PEG, stir and mix evenly, dropwise add to a 5wt% sodium dodecylbenzenesulfonate (SDBS) aqueous solution, seal, introduce high-purity nitrogen for 40-60 min, then place in an ice-water bath and irradiate continuously with ultraviolet light (λ = 365 nm) for 2 h, wash successively with water, tetrahydrofuran (THF), and methanol, and dry in vacuum at 30-40 °C to obtain epoxy group porous microspheres; B2: Ultrasonically disperse the epoxy group porous microspheres obtained in step B1 in a methanol / chloroform mixed solution (v:v = 5:1), add HD and triethylamine, stir and react at 80 °C and 200-300 rpm for 4 h, filter by suction, wash the filter cake with methanol and THF, and dry in vacuum; B3: Add the product obtained in step B2 and SMMP to a PB buffer solution with pH = 7.4, stir and react at room temperature at 200-300 rpm for 4 h, centrifuge at 8000 rpm for 10-15 min, wash the precipitate with DMF and deionized water, and dry to obtain the modified porous microspheres.

[0012] Further, in step B1, the mass ratio of chloroform to PEG is 4:1.

[0013] Further, in step B1, the dosage ratio of the SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g.

[0014] Further, in step B2, the mass concentration of HD in the methanol / chloroform mixture is 40 - 50 mg / mL.

[0015] Further, in step B2, the dosage of triethylamine is 3 - 5% of the volume of the methanol / chloroform mixture.

[0016] Further, in step B3, the mass concentration of SMMP in the PB buffer solution is 1 mg / mL.

[0017] Further, the present invention also provides a preparation method of the high-sensitivity glucose detection material, comprising the following steps: S1: Uniformly disperse the modified porous microspheres in a 25 vol% aqueous ethanol solution to obtain a dispersion liquid. Dissolve chloroplatinic acid and palladium acetate in THF respectively, add them to the dispersion liquid, stir for 2 h, add a 0.5 M aqueous solution of NaBH4, stir for 4 h, centrifuge at 8000 rpm for 8 - 12 min, wash the precipitate with ethanol, and dry it under vacuum; S2: Add the product obtained in step S1 and the modified glucose oxidase to a PBS buffer solution with pH = 7.4, stir and react at 100 - 200 r / min under nitrogen protection for 24 h, centrifuge at 8000 - 10000 rpm for 10 min, wash the precipitate with the PBS buffer solution, and freeze-dry to obtain the high-sensitivity glucose detection material.

[0018] Further, in step S1, the mass concentration of the modified porous microspheres in the aqueous ethanol solution is 2 - 5 mg / mL.

[0019] Further, in step S1, the mass concentration of chloroplatinic acid in THF is 20 mg / mL.

[0020] Further, in step S1, the mass concentration of palladium acetate in THF is 20 mg / mL.

[0021] Further, in step S1, the dosage ratio of palladium acetate to the aqueous NaBH4 solution is 1 g:50 mL.

[0022] Further, in step S2, the mass concentration of the modified glucose oxidase in the buffer solution is 2 - 3 mg / mL.

[0023] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a highly sensitive glucose detection material, which has significant advantages in detection sensitivity, stability, and cost control compared with the prior art, and can effectively meet the actual needs in the fields of clinical diagnosis, diabetes monitoring, etc. In the present invention, glucose oxidase is modified, and thiol-polyethylene glycol-carboxyl (SH-PEG-COOH) reacts with glucose oxidase (GOD). The flexible chain segment of polyethylene glycol is used to increase the steric hindrance of GOD, reduce its aggregation, and better expose its active center, which is beneficial to full contact with glucose. On the other hand, the connection of SH-PEG-COOH can introduce thiol functional groups, which can react with maleimide groups to immobilize glucose oxidase on the modified porous microspheres, improving the catalytic efficiency for glucose. The present invention prepares modified porous microspheres, which provide a rich specific surface area and porous structure for the detection material, provide more active sites, increase the contact opportunity with glucose, and its unique pore structure helps the diffusion and enrichment of glucose molecules, further improving the detection sensitivity. In the present invention, the epoxy groups of epoxy-based porous microspheres react with 1,3-diamino-2-propanol to successfully introduce amino and hydroxyl groups. On the one hand, the hydroxyl and amino groups can coordinate with metal ions, which is beneficial to the subsequent loading of metal ions. On the other hand, the amino group can be modified by SMMP to successfully graft maleimide groups, which are then connected to the modified glucose oxidase. The nanoparticles formed by the reduction of chloroplatinic acid and palladium acetate during the preparation process have excellent catalytic performance and electron conduction ability, enabling the detection material of the present invention to exhibit high sensitivity even when detecting low-concentration glucose, and the effective detection limit is significantly lower than the glucose content in serum. The metal particles and glucose oxidase are co-loaded on the porous microspheres, which can effectively improve the detection sensitivity and stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following will briefly introduce the drawings required for the description of the embodiments or the prior art. Obviously, the drawings in the following description are only those of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0025] Figure 1 It is the standard curve of the detection material in Example 1 of the present invention; Figure 2 It is the sensitivity of the detection materials described in Example 2 and Comparative Examples 1-3 of the present invention; Figure 3 It is the stability of the detection materials described in Example 3 and Comparative Examples 1-3 of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0026] In order to make the purpose, technical scheme and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific embodiments, but the present invention is not limited to the following embodiments. It should be noted that, unless otherwise specified, the chemical reagents involved in the present invention are purchased through commercial channels.

[0027] Example 1: A high-sensitivity glucose detection material, prepared by raw materials including the following components in parts by weight: 2 parts of modified glucose oxidase, 10 parts of modified porous microspheres, 1.5 parts of chloroplatinic acid, and 1 part of palladium acetate.

[0028] The raw materials for preparing the modified glucose oxidase include the following components in parts by weight: 2 parts of glucose oxidase (GOD) and 4 parts of thiol-polyethylene glycol-carboxyl (SH-PEG-COOH).

[0029] The preparation method of modified glucose oxidase comprises the following steps: A1: Add GOD to PB buffer at pH=6.5 and mix well to obtain GOD solution, add SH-PEG-COOH to PB buffer, add EDC and NHS, stir at 200 rpm for 2 h to obtain activation solution; in the GOD solution, the mass concentration of GOD in PB buffer is 2 mg / mL; the mass concentration of SH-PEG-COOH in PB buffer is 3 mg / mL; the mass ratio of SH-PEG-COOH, EDC and NHS is 10:1.5:1; A2: Add the GOD solution obtained in step A1 to the activation solution, stir the reaction at 300 rpm for 12 h, centrifuge at 10,000 rpm for 10 min, wash the precipitate with deionized water to obtain modified GOD.

[0030] The raw materials for preparing the modified porous microspheres include the following components in parts by weight: 3 parts of 1,3-propanedithiol, 2.1 parts of propargyl glycidyl ether, 0.5 parts of 1,7-octanediyne, 0.17 parts of benzoin dimethyl ether (DMAP), 1.5 parts of polyethylene glycol (PEG), 6 parts of 1,3-diamino-2-propanol (HD), and 1.5 parts of 3-hydroxysuccinimide maleimidopropionate (SMMP).

[0031] The preparation method of modified porous microspheres comprises the following steps: B1: Mix 1,3-propanedithiol, propargyl glycidyl ether, and 1,7-octadiyne, add DMPA, chloroform, and PEG, stir and mix well, dropwise add to an aqueous solution of 5 wt% sodium dodecylbenzenesulfonate (SDBS), seal, introduce high-purity nitrogen for 60 min, then place in an ice-water bath and irradiate continuously with ultraviolet light (λ = 365 nm) for 2 h. Wash successively with water, tetrahydrofuran (THF), and methanol, and dry in vacuum at 40 °C to obtain epoxy-group porous microspheres; the mass ratio of chloroform to PEG is 4:1; the dosage ratio of the SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g; B2: Ultrasonically disperse the epoxy-group porous microspheres obtained in step B1 in a methanol / chloroform mixture (v:v = 5:1), add HD and triethylamine, stir and react at 80 °C and 300 rpm for 4 h, filter by suction, wash the filter cake with methanol and THF, and dry in vacuum; the mass concentration of HD in the methanol / chloroform mixture is 50 mg / mL; the dosage of triethylamine is 5% of the volume of the methanol / chloroform mixture; B3: Add the product obtained in step B2 and SMMP to a PB buffer solution with pH = 7.4, stir and react at 300 rpm at room temperature for 4 h, centrifuge at 8000 rpm for 15 min, wash the precipitate with DMF and deionized water, and dry to obtain modified porous microspheres; the mass concentration of SMMP in the PB buffer solution is 1 mg / mL.

[0032] This example also provides a preparation method of the high-sensitivity glucose detection material, including the following steps: S1: Uniformly disperse the modified porous microspheres in a 25 vol% ethanol aqueous solution to obtain a dispersion. Dissolve chloroplatinic acid and palladium acetate in THF respectively, add to the dispersion, stir for 2 h, add a 0.5 M aqueous solution of NaBH4, stir for 4 h, centrifuge at 8000 rpm for 12 min, wash the precipitate with ethanol, and dry in vacuum; the mass concentration of the modified porous microspheres in the ethanol aqueous solution is 5 mg / mL; the mass concentration of chloroplatinic acid in THF is 20 mg / mL; the mass concentration of palladium acetate in THF is 20 mg / mL; the dosage ratio of palladium acetate to the NaBH4 aqueous solution is 1 g:50 mL; S2: Add the product obtained in step S1 and the modified glucose oxidase to a PBS buffer solution with pH = 7.4, stir and react at 200 r / min under nitrogen protection for 24 h, centrifuge at 10000 rpm for 10 min, wash the precipitate with the PBS buffer solution, and freeze-dry to obtain the high-sensitivity glucose detection material; the mass concentration of the modified glucose oxidase in the buffer solution is 3 mg / mL.

[0033] Example 2: A highly sensitive glucose detection material, the preparation raw materials of which include the following components in parts by weight: 1 part of modified glucose oxidase, 6 parts of modified porous microspheres, 1 part of chloroplatinic acid, and 0.6 part of palladium acetate.

[0034] The preparation raw materials of modified glucose oxidase include the following components in parts by weight: 1 part of glucose oxidase (GOD), and 2 parts of mercapto-polyethylene glycol-carboxyl (SH-PEG-COOH).

[0035] The preparation method of modified glucose oxidase includes the following steps: A1: Add GOD into PB buffer solution with pH = 6.5 and mix evenly to obtain GOD solution. Add SH-PEG-COOH into PB buffer solution, add EDC and NHS, and stir at 150 rpm for 2 h to obtain an activation solution; in the GOD solution, the mass concentration of GOD in PB buffer solution is 1 mg / mL; the mass concentration of SH-PEG-COOH in PB buffer solution is 2 mg / mL; the mass ratio of SH-PEG-COOH, EDC and NHS is 10:1.5:1; A2: Add the GOD solution obtained in step A1 into the activation solution, stir and react at 200 rpm for 12 h, centrifuge at 10000 rpm for 5 min, and wash the precipitate with deionized water to obtain modified GOD.

[0036] The preparation raw materials of modified porous microspheres include the following components in parts by weight: 2 parts of 1,3-propanedithiol, 1.5 parts of propargyl glycidyl ether, 0.3 part of 1,7-octadiyne, 0.12 part of benzoin dimethyl ether (DMAP), 1 part of polyethylene glycol (PEG), 4 parts of 1,3-diamino-2-propanol (HD), and 1 part of 3-maleimidopropionic acid hydroxysuccinimide ester (SMMP).

[0037] The preparation method of modified porous microspheres includes the following steps: B1: Mix 1,3-propanedithiol, propargyl glycidyl ether, and 1,7-octadiyne, add DMPA, chloroform and PEG and stir evenly, dropwise add it into 5wt% sodium dodecylbenzenesulfonate (SDBS) aqueous solution, seal, introduce high-purity nitrogen for 40 min, then place it in an ice-water bath and irradiate with continuous ultraviolet light (λ = 365 nm) for 2 h, wash successively with water, tetrahydrofuran (THF) and methanol, and vacuum dry at 30 - 40 °C to obtain epoxy group porous microspheres; the mass ratio of chloroform to PEG is 4:1; the dosage ratio of SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g; B2: Ultrasonically disperse the epoxy group porous microspheres obtained in step B1 in a methanol / chloroform mixture (v:v = 5:1), add HD and triethylamine, stir and react at 80 °C and 200 rpm for 4 h, filter by suction, wash the filter cake with methanol and THF, and dry in vacuum; the mass concentration of HD in the methanol / chloroform mixture is 40 mg / mL; the dosage of triethylamine is 3% of the volume of the methanol / chloroform mixture; B3: Add the product obtained in step B2 and SMMP to a PB buffer solution with pH = 7.4, stir and react at 200 rpm at room temperature for 4 h, centrifuge at 8000 rpm for 10 min, wash the precipitate with DMF and deionized water, and dry to obtain the modified porous microspheres; the mass concentration of SMMP in the PB buffer solution is 1 mg / mL.

[0038] This example also provides a preparation method of the high-sensitivity glucose detection material, including the following steps: S1: Uniformly disperse the modified porous microspheres in a 25 vol% aqueous ethanol solution to obtain a dispersion. Dissolve chloroplatinic acid and palladium acetate in THF respectively, add them to the dispersion, stir for 2 h, add a 0.5 M aqueous NaBH4 solution, stir for 4 h, centrifuge at 8000 rpm for 8 min, wash the precipitate with ethanol, and dry in vacuum; the mass concentration of the modified porous microspheres in the aqueous ethanol solution is 2 mg / mL; the mass concentration of chloroplatinic acid in THF is 20 mg / mL; the mass concentration of palladium acetate in THF is 20 mg / mL; the dosage ratio of palladium acetate to the aqueous NaBH4 solution is 1 g:50 mL; S2: Add the product obtained in step S1 and the modified glucose oxidase to a PBS buffer solution with pH = 7.4, stir and react at 100 r / min under nitrogen protection for 24 h, centrifuge at 8000 rpm for 10 min, wash the precipitate with the PBS buffer solution, and freeze-dry to obtain the high-sensitivity glucose detection material; the mass concentration of the modified glucose oxidase in the buffer solution is 2 mg / mL.

[0039] Example 3: A high-sensitivity glucose detection material, the preparation raw materials include the following components in parts by weight: 1.5 parts of modified glucose oxidase, 8 parts of modified porous microspheres, 1.2 parts of chloroplatinic acid, and 0.8 part of palladium acetate.

[0040] The preparation raw materials of the modified glucose oxidase include the following components in parts by weight: 1.5 parts of glucose oxidase (GOD), 3 parts of mercapto-polyethylene glycol-carboxyl (SH-PEG-COOH).

[0041] The preparation method of the modified glucose oxidase includes the following steps: A1: Add GOD to the PB buffer solution with a pH of 6.5 and mix well to obtain a GOD solution. Add SH-PEG-COOH to the PB buffer solution, then add EDC and NHS, and stir at 180 rpm for 2 h to obtain an activation solution. In the GOD solution, the mass concentration of GOD in the PB buffer solution is 1.5 mg / mL; the mass concentration of SH-PEG-COOH in the PB buffer solution is 2.5 mg / mL; the mass ratio of SH-PEG-COOH, EDC, and NHS is 10:1.5:1. A2: Add the GOD solution obtained in step A1 to the activation solution, stir and react at 250 rpm for 12 h, centrifuge at 10000 rpm for 8 min, and wash the precipitate with deionized water to obtain modified GOD.

[0042] The raw materials for preparing the modified porous microspheres include the following components in parts by weight: 2.5 parts of 1,3-propanedithiol, 1.8 parts of propargyl glycidyl ether, 0.4 part of 1,7-octadiyne, 0.15 part of benzoin dimethyl ether (DMAP), 1.2 parts of polyethylene glycol (PEG), 5 parts of 1,3-diamino-2-propanol (HD), and 1.2 parts of 3-maleimidopropionic acid N-hydroxysuccinimide ester (SMMP).

[0043] The preparation method of the modified porous microspheres includes the following steps: B1: Mix 1,3-propanedithiol, propargyl glycidyl ether, and 1,7-octadiyne, add DMPA, chloroform, and PEG, stir and mix well, dropwise add to a 5wt% sodium dodecylbenzenesulfonate (SDBS) aqueous solution, seal, introduce high-purity nitrogen for 50 min, then place in an ice-water bath and irradiate with continuous ultraviolet light (λ = 365 nm) for 2 h. Wash successively with water, tetrahydrofuran (THF), and methanol, and dry in vacuo at 35 °C to obtain epoxy-group porous microspheres; the mass ratio of chloroform to PEG is 4:1; the dosage ratio of the SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g. B2: Ultrasonically disperse the epoxy-group porous microspheres obtained in step B1 in a methanol / chloroform mixed solution (v:v = 5:1), add HD and triethylamine, stir and react at 80 °C and 250 rpm for 4 h, filter by suction, wash the filter cake with methanol and THF, and dry in vacuo; the mass concentration of HD in the methanol / chloroform mixed solution is 45 mg / mL; the dosage of triethylamine is 4% of the volume of the methanol / chloroform mixed solution. B3: Add the product obtained in step B2 and SMMP to the PB buffer solution with a pH of 7.4, stir and react at 250 rpm at room temperature for 4 h, centrifuge at 8000 rpm for 12 min, wash the precipitate with DMF and deionized water, and dry to obtain the modified porous microspheres; the mass concentration of SMMP in the PB buffer solution is 1 mg / mL.

[0044] This embodiment also provides a method for preparing the high-sensitivity glucose detection material, which includes the following steps: S1: Uniformly disperse the modified porous microspheres in an ethanol aqueous solution of 25 vol% to obtain a dispersion. Dissolve chloroplatinic acid and palladium acetate in THF respectively, add them to the dispersion, stir for 2 h, add a 0.5 M aqueous solution of NaBH4, stir for 4 h, centrifuge at 8000 rpm for 10 min, wash the precipitate with ethanol, and dry it under vacuum; the mass concentration of the modified porous microspheres in the ethanol aqueous solution is 3 mg / mL; the mass concentration of chloroplatinic acid in THF is 20 mg / mL; the mass concentration of palladium acetate in THF is 20 mg / mL; the dosage ratio of palladium acetate to the aqueous solution of NaBH4 is 1 g:50 mL; S2: Add the product obtained in step S1 and the modified glucose oxidase to a PBS buffer solution with pH = 7.4, stir and react at 150 r / min for 24 h under nitrogen protection, centrifuge at 9000 rpm for 10 min, wash the precipitate with the PBS buffer solution, and freeze-dry to obtain the high-sensitivity glucose detection material; the mass concentration of the modified glucose oxidase in the buffer solution is 2.5 mg / mL.

[0045] The difference between Comparative Example 1 and Example 1 is only that glucose oxidase is used to replace the modified glucose oxidase.

[0046] The difference between Comparative Example 2 and Example 1 is only that palladium acetate is not added.

[0047] The difference between Comparative Example 3 and Example 1 is only that step B3 is not carried out, that is, the product obtained in step B2 is used to replace the modified porous microspheres.

[0048] Experimental Example 1: Add 100 μL of glucose solutions with different concentrations and 100 μL of the high-sensitivity glucose detection material of Example 1 at 1 mg / mL to 700 μL of acetic acid-sodium acetate buffer solution (pH 4). After incubating at 37 °C for 20 min, add 100 μL of TMB (10 mmol·L -1 ), react for another 30 min, detect the absorbance at 652 nm in the supernatant, record the relationship between the glucose concentration and the absorbance, and the results are as Figure 1 shown.

[0049] Figure 1 The results show that there is a good linear relationship between the absorbance value and the glucose concentration (in the range of 0.02 - 2 mmol / L), R 2 = 0.9943, indicating that the drawn standard curve has good accuracy. The detection material of the present invention can be used for the quantitative analysis of glucose and effectively detect the glucose concentration.

[0050] Experimental Example 2: Calculate the detection limit of glucose for the detection materials obtained in Example 2 and Comparative Examples 1-3 according to S / N = 3. The results are as Figure 2 shown.

[0051] Figure 2 The results show that Example 2 and Examples 1-3 all have relatively low detection limits. Among them, Example 2 has the best effect and is significantly lower than the glucose content in serum. The detection material of the present invention can effectively detect the glucose content with high detection sensitivity.

[0052] Experimental Example 3: Place the detection materials of Example 3 and Comparative Examples 1-3 at 4 °C for 14 days. Using 1 mM glucose as the detection object, detect 10 times and calculate the relative standard deviation. The results are as Figure 3 shown.

[0053] Figure 3 The results show that the relative standard deviations of Example 3 and the groups of Comparative Examples 1-3 are all relatively small. Among them, the standard deviation data of Example 1 is the best, indicating that the high-sensitivity glucose detection material of the present invention has high detection accuracy and stability.

[0054] Those of ordinary skill in the art should understand that the discussion of any of the above embodiments is only exemplary and is not intended to imply that the scope of the present invention is limited to these examples; under the concept of the present invention, the technical features in the above embodiments or different embodiments can also be combined, and the steps can be implemented in any order, and there are many other variations in different aspects of the present invention as described above, which are not provided in detail for the sake of brevity.

Claims

1. A highly sensitive glucose detection material, characterized in that, The preparation raw materials include the following components in parts by weight: 1-2 parts of modified glucose oxidase, 6-10 parts of modified porous microspheres, 1-1.5 parts of chloroplatinic acid, and 0.6-1 part of palladium acetate; The preparation raw materials of the modified glucose oxidase include the following components in parts by weight: 1-2 parts of GOD, and 2-4 parts of SH-PEG-COOH; The preparation method of the modified glucose oxidase includes the following steps: A1: Add GOD to PB buffer solution and mix evenly to obtain GOD solution. Add SH-PEG-COOH to PB buffer solution, add EDC and NHS, and stir to obtain an activation solution; A2: Add the GOD solution obtained in step A1 to the activation solution, stir, centrifuge, and wash to obtain modified GOD; The preparation raw materials of the modified porous microspheres include the following components in parts by weight: 2-3 parts of 1,3-propanedithiol, 1.5-2.1 parts of propargyl glycidyl ether, 0.3-0.5 parts of 1,7-octadiyne, 0.12-0.17 parts of DMAP, 1-1.5 parts of PEG, 4-6 parts of HD, and 1-1.5 parts of SMMP; The preparation method of the modified porous microspheres includes the following steps: B1: Mix 1,3-propanedithiol, propargyl glycidyl ether, and 1,7-octadiyne, add DMPA, chloroform, and PEG, stir and mix evenly, dropwise add to the SDBS aqueous solution, seal, introduce high-purity nitrogen for 40-60 min, place in an ice-water bath, continuously irradiate with ultraviolet light, wash, and dry to obtain epoxy group porous microspheres; B2: Ultrasonically disperse the epoxy group porous microspheres obtained in step B1 in a methanol / chloroform mixed solution, add HD and triethylamine, react, wash, and dry; B3: Add the product obtained in step B2 and SMMP to PB buffer solution, stir and react, wash, and dry to obtain modified porous microspheres.

2. The highly sensitive glucose detection material according to claim 1, characterized in that, In step A1, in the GOD solution, the mass concentration of GOD in PB buffer solution is 1-2 mg / mL; the mass concentration of SH-PEG-COOH in PB buffer solution is 2-3 mg / mL; the mass ratio of SH-PEG-COOH, EDC, and NHS is 10:1.5:

1.

3. The highly sensitive glucose detection material according to claim 2, characterized in that, In step B1, the mass ratio of chloroform to PEG is 4:1; the dosage ratio of SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g.

4. The highly sensitive glucose detection material according to claim 3, characterized in that, In step B2, the mass concentration of HD in the methanol / chloroform mixed solution is 40-50 mg / mL; the dosage of triethylamine is 3-5% of the volume of the methanol / chloroform mixed solution.

5. The highly sensitive glucose detection material according to claim 4, characterized in that, In step B3, the mass concentration of SMMP in PB buffer solution is 1 mg / mL.

6. A preparation method of the highly sensitive glucose detection material according to any one of claims 1-5, characterized in that, It includes the following steps: S1: Uniformly disperse the modified porous microspheres in a 25 vol% ethanol aqueous solution to obtain a dispersion liquid. Dissolve chloroplatinic acid and palladium acetate in THF respectively, add them to the dispersion liquid, stir, add an aqueous solution of NaBH4, stir, wash, and dry; S2: Add the product obtained in step S1 and the modified glucose oxidase to PBS buffer solution, stir under nitrogen protection, wash, and dry to obtain a highly sensitive glucose detection material.

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