A high-sensitivity glucose detection material and preparation method thereof

Abstract

The present invention relates to the field of glucose detection technology, and more specifically to a highly sensitive glucose detection material and a method for preparing the same. The highly sensitive glucose detection material comprises the following components by weight: 1-2 parts modified glucose oxidase, 6-10 parts modified porous microspheres, 1-1.5 parts chloroplatinic acid, and 0.6-1 parts palladium acetate. The resulting detection material exhibits high sensitivity, good repeatability, and good stability.

Description

Technical Field

[0001] The present invention relates to the technical field of glucose detection, and in particular to a high-sensitivity glucose detection material and a preparation method thereof. Background Art

[0002] Glucose detection is of great significance in clinical diagnosis, diabetes monitoring, and biomedical research. Accurate and highly sensitive glucose detection can help doctors understand patients' blood sugar levels in a timely manner and formulate reasonable treatment plans, which is crucial for the early diagnosis and disease monitoring of metabolic diseases such as diabetes. At present, 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, susceptibility to environmental factors, and short service life; nanomaterials such as metal nanoparticles and carbon nanomaterials, although they have improved the detection sensitivity to a certain extent, the preparation process is complicated, the cost is high, and the detection results are easily interfered with, 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 a technical problem that needs to be solved urgently in this field. Summary of the Invention

[0003] In view of the shortcomings of the existing technology, the present invention provides a high-sensitivity glucose detection material and a preparation method thereof.

[0004] The present invention is achieved through the following technical solutions:

[0005] A high-sensitivity glucose detection material is prepared by raw materials comprising 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.

[0006] Furthermore, the raw materials for preparing the modified glucose oxidase include the following components in parts by weight: 1-2 parts of glucose oxidase (GOD) and 2-4 parts of thiol-polyethylene glycol-carboxyl (SH-PEG-COOH).

[0007] Furthermore, the preparation method of the modified glucose oxidase comprises the following steps:

[0008] A1: Add GOD to PB buffer (pH 6.5) and mix well to obtain GOD solution. Add SH-PEG-COOH to the PB buffer, add EDC and NHS, and stir at 150-200 rpm for 2 h to obtain activation solution.

[0009] A2: Add the GOD solution obtained in step A1 to the activation solution, stir at 200-300 rpm for 12 h, centrifuge at 10,000 rpm for 5-10 min, and wash the precipitate with deionized water to obtain modified GOD.

[0010] Furthermore, in step A1, in the GOD solution, the mass concentration of GOD in PB buffer is 1-2 mg / mL.

[0011] Furthermore, in step A1, the mass concentration of the SH-PEG-COOH in the PB buffer is 2-3 mg / mL.

[0012] Furthermore, in step A1, the mass ratio of SH-PEG-COOH, EDC and NHS is 10:1.5:1.

[0013] Furthermore, 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-octanediyne, 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-hydroxysuccinimide maleimidopropionate (SMMP).

[0014] Furthermore, the preparation method of the modified porous microspheres comprises the following steps:

[0015] B1: 1,3-Propanedithiol, propargyl glycidyl ether, and 1,7-octanediyne were mixed, DMPA, chloroform, and PEG were added, and the mixture was stirred and added dropwise to a 5 wt% aqueous solution of sodium dodecylbenzenesulfonate (SDBS). The mixture was sealed and aerated with high-purity nitrogen for 40-60 min. The mixture was then placed in an ice-water bath and irradiated with continuous ultraviolet light (λ = 365 nm) for 2 h. The mixture was washed with water, tetrahydrofuran (THF), and methanol in sequence, and dried in a vacuum at 30-40°C to obtain epoxy-based porous microspheres.

[0016] B2: Ultrasonic dispersion of the epoxy-based porous microspheres obtained in step B1 in a methanol / chloroform mixture (v:v = 5:1) was performed, HD and triethylamine were added, and the mixture was stirred at 80°C and 200-300 rpm for 4 h. The mixture was filtered, and the filter cake was washed with methanol and THF and dried in vacuo.

[0017] B3: The product obtained in step B2 and SMMP were added to PB buffer solution with pH = 7.4, stirred at 200-300 rpm for 4 h at room temperature, centrifuged at 8000 rpm for 10-15 min, and the precipitate was washed with DMF and deionized water, and dried to obtain modified porous microspheres.

[0018] Furthermore, in step B1, the mass ratio of chloroform to PEG is 4:1.

[0019] Furthermore, in step B1, the ratio of the SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g.

[0020] Furthermore, in step B2, the mass concentration of HD in the methanol / chloroform mixture is 40-50 mg / mL.

[0021] Furthermore, in step B2, the amount of triethylamine used is 3-5% of the volume of the methanol / chloroform mixture.

[0022] Furthermore, in step B3, the mass concentration of the SMMP in the PB buffer is 1 mg / mL.

[0023] Furthermore, the present invention also provides a method for preparing the highly sensitive glucose detection material, comprising the following steps:

[0024] S1: The modified porous microspheres were uniformly dispersed in a 25 vol% ethanol aqueous solution to obtain a dispersion. Chloroplatinic acid and palladium acetate were dissolved in THF, respectively, added to the dispersion, stirred for 2 h, and then a 0.5 M NaBH4 aqueous solution was added. The mixture was stirred for 4 h and centrifuged at 8000 rpm for 8-12 min. The precipitate was washed with ethanol and dried in vacuo.

[0025] S2: The product obtained in step S1 and the modified glucose oxidase were added to a PBS buffer solution at pH = 7.4. The mixture was stirred at 100-200 r / min for 24 h under nitrogen protection. The mixture was centrifuged at 8000-10000 rpm for 10 min. The precipitate was washed with PBS buffer solution and freeze-dried to obtain a highly sensitive glucose detection material.

[0026] Furthermore, in step S1, the mass concentration of the modified porous microspheres in the ethanol aqueous solution is 2-5 mg / mL.

[0027] Furthermore, in step S1, the mass concentration of chloroplatinic acid in THF is 20 mg / mL.

[0028] Furthermore, in step S1, the mass concentration of palladium acetate in THF is 20 mg / mL.

[0029] Furthermore, in step S1, the ratio of palladium acetate to NaBH4 aqueous solution is 1 g:50 mL.

[0030] Furthermore, in step S2, the mass concentration of the modified glucose oxidase in the buffer solution is 2-3 mg / mL.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] The present invention provides a kind of high-sensitive glucose detection material, compared with the prior art, in terms of detection sensitivity, stability and cost control, there are significant advantages, can effectively meet the actual needs of the fields of clinical diagnosis, diabetes monitoring. The present invention is modified by glucose oxidase, adopts thiol-polyethylene glycol-carboxyl (SH-PEG-COOH) to react with glucose oxidase (GOD), utilizes the flexible chain segment of polyethylene glycol to increase the steric hindrance of GOD, reduces its aggregation, makes its active center better exposed, is conducive to fully contact with glucose; On the other hand, the connection of SH-PEG-COOH can introduce thiol functional group, can react with maleimide group, fix glucose oxidase to modified porous microspheres, improve the catalytic efficiency of glucose. The present invention prepares modified porous microspheres, provides rich specific surface area and porous structure for detection material, provides more active sites, increases the contact opportunity with glucose, and its unique pore structure contributes to the diffusion and enrichment of glucose molecules, further improves the sensitivity of detection. The present invention utilizes the epoxy group of epoxy porous microspheres to 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, and the maleimide group can be successfully grafted, thereby connecting with the modified glucose oxidase. The nanoparticles formed by the reduction of chloroplatinic acid and palladium acetate during the preparation process have excellent catalytic properties and electronic conductivity, so that the detection material of the present invention can also show high sensitivity when detecting low-concentration glucose, and the effective detection limit is significantly lower than the glucose content in serum. Metal particles and glucose oxidase are jointly loaded on the porous microspheres, which can effectively improve the detection sensitivity and improve stability. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only for the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0034] Figure 1 This is the standard curve of the test material in Example 1 of the present invention;

[0035] Figure 2 The sensitivity of the detection materials described in Example 2 and Comparative Examples 1-3 of the present invention;

[0036] Figure 3The stability of the detection materials described in Example 3 and Comparative Examples 1-3 of the present invention. DETAILED DESCRIPTION

[0037] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific examples, but the present invention is not limited to the following examples. It should be noted that, unless otherwise specified, the chemical reagents involved in the present invention were purchased through commercial channels.

[0038] Example 1: A high-sensitivity glucose detection material is prepared by preparing 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.

[0039] 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).

[0040] The preparation method of modified glucose oxidase comprises the following steps:

[0041] A1: GOD was added to PB buffer at pH 6.5 and mixed to obtain a GOD solution. SH-PEG-COOH was added to the PB buffer, and EDC and NHS were added. The mixture was stirred at 200 rpm for 2 h to obtain an activation solution. In the GOD solution, the mass concentration of GOD in PB buffer was 2 mg / mL; the mass concentration of SH-PEG-COOH in PB buffer was 3 mg / mL; the mass ratio of SH-PEG-COOH, EDC, and NHS was 10:1.5:1.

[0042] A2: Add the GOD solution obtained in step A1 to the activation solution, stir at 300 rpm for 12 h, centrifuge at 10,000 rpm for 10 min, and wash the precipitate with deionized water to obtain modified GOD.

[0043] 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).

[0044] The preparation method of modified porous microspheres comprises the following steps:

[0045] B1: 1,3-Propanedithiol, propargyl glycidyl ether, and 1,7-octanediyne were mixed, DMPA, chloroform, and PEG were added, and the mixture was stirred and added dropwise to a 5 wt% sodium dodecylbenzenesulfonate (SDBS) aqueous solution. The mixture was sealed and aerated with high-purity nitrogen for 60 min, then placed in an ice-water bath and irradiated with continuous ultraviolet light (λ = 365 nm) for 2 h. The mixture was washed with water, tetrahydrofuran (THF), and methanol in sequence, and dried in vacuo at 40°C to obtain epoxy porous microspheres. The mass ratio of chloroform to PEG was 4:1; the ratio of SDBS aqueous solution to 1,3-Propanedithiol was 25 mL:1 g.

[0046] B2: Ultrasonic dispersion of the epoxy-based porous microspheres obtained in step B1 in a methanol / chloroform mixture (v:v = 5:1) was performed. HD and triethylamine were added and stirred at 80°C and 300 rpm for 4 h. The mixture was filtered and the filter cake was washed with methanol and THF and dried under vacuum. The concentration of HD in the methanol / chloroform mixture was 50 mg / mL. The amount of triethylamine used was 5% of the volume of the methanol / chloroform mixture.

[0047] B3: The product obtained in step B2 and SMMP were added to PB buffer at pH = 7.4, stirred at 300 rpm for 4 h at room temperature, centrifuged at 8000 rpm for 15 min, and the precipitate was washed with DMF and deionized water and dried to obtain modified porous microspheres; the mass concentration of SMMP in PB buffer was 1 mg / mL.

[0048] This embodiment also provides a method for preparing the highly sensitive glucose detection material, comprising the following steps:

[0049] S1: The modified porous microspheres were uniformly dispersed in a 25 vol% ethanol aqueous solution to obtain a dispersion. Chloroplatinic acid and palladium acetate were separately dissolved in THF, added to the dispersion, and stirred for 2 h. A 0.5 M NaBH4 aqueous solution was added, stirred for 4 h, and centrifuged at 8000 rpm for 12 min. The precipitate was washed with ethanol and dried in vacuo. The mass concentration of the modified porous microspheres in the ethanol aqueous solution was 5 mg / mL; the mass concentration of chloroplatinic acid in THF was 20 mg / mL; the mass concentration of palladium acetate in THF was 20 mg / mL; and the amount ratio of palladium acetate to NaBH4 aqueous solution was 1 g:50 mL.

[0050] S2: The product obtained in step S1 and the modified glucose oxidase were added to a PBS buffer solution at pH = 7.4. The mixture was stirred at 200 r / min under nitrogen protection for 24 h. The mixture was centrifuged at 10,000 rpm for 10 min. The precipitate was washed with PBS buffer and freeze-dried to obtain a highly sensitive glucose detection material. The mass concentration of the modified glucose oxidase in the buffer solution was 3 mg / mL.

[0051] Example 2: A high-sensitivity glucose detection material is prepared by preparing raw materials including 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 parts of palladium acetate.

[0052] The raw materials for preparing the modified glucose oxidase include the following components in parts by weight: 1 part of glucose oxidase (GOD) and 2 parts of thiol-polyethylene glycol-carboxyl (SH-PEG-COOH).

[0053] The preparation method of modified glucose oxidase comprises the following steps:

[0054] A1: GOD was added to PB buffer at pH 6.5 and mixed to obtain a GOD solution. SH-PEG-COOH was added to the PB buffer, and EDC and NHS were added. The mixture was stirred at 150 rpm for 2 h to obtain an activation solution. In the GOD solution, the mass concentration of GOD in PB buffer was 1 mg / mL; the mass concentration of SH-PEG-COOH in PB buffer was 2 mg / mL; the mass ratio of SH-PEG-COOH, EDC, and NHS was 10:1.5:1.

[0055] A2: Add the GOD solution obtained in step A1 to the activation solution, stir at 200 rpm for 12 h, centrifuge at 10,000 rpm for 5 min, and wash the precipitate with deionized water to obtain modified GOD.

[0056] The raw materials for preparing the 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 parts of 1,7-octanediyne, 0.12 parts 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-hydroxysuccinimide maleimidopropionate (SMMP).

[0057] The preparation method of modified porous microspheres comprises the following steps:

[0058] B1: 1,3-Propanedithiol, propargyl glycidyl ether, and 1,7-octanediyne were mixed, DMPA, chloroform, and PEG were added, and the mixture was stirred and added dropwise to a 5 wt% sodium dodecylbenzenesulfonate (SDBS) aqueous solution. The mixture was sealed and aerated with high-purity nitrogen for 40 min, and then placed in an ice-water bath and irradiated with continuous ultraviolet light (λ = 365 nm) for 2 h. The mixture was washed with water, tetrahydrofuran (THF), and methanol in sequence, and dried under vacuum at 30-40°C to obtain epoxy porous microspheres. The mass ratio of chloroform to PEG was 4:1; the ratio of SDBS aqueous solution to 1,3-Propanedithiol was 25 mL:1 g.

[0059] B2: Ultrasonic dispersion of the epoxy-based porous microspheres obtained in step B1 in a methanol / chloroform mixture (v:v = 5:1) was performed. HD and triethylamine were added and stirred at 80°C and 200 rpm for 4 h. The mixture was filtered and the filter cake was washed with methanol and THF and dried under vacuum. The concentration of HD in the methanol / chloroform mixture was 40 mg / mL. The amount of triethylamine used was 3% of the volume of the methanol / chloroform mixture.

[0060] B3: The product obtained in step B2 and SMMP were added to PB buffer at pH = 7.4, stirred at 200 rpm for 4 h at room temperature, centrifuged at 8000 rpm for 10 min, and the precipitate was washed with DMF and deionized water and dried to obtain modified porous microspheres; the mass concentration of SMMP in PB buffer was 1 mg / mL.

[0061] This embodiment also provides a method for preparing the highly sensitive glucose detection material, comprising the following steps:

[0062] S1: The modified porous microspheres were uniformly dispersed in a 25 vol% ethanol aqueous solution to obtain a dispersion. Chloroplatinic acid and palladium acetate were separately dissolved in THF, added to the dispersion, and stirred for 2 h. A 0.5 M NaBH4 aqueous solution was added, stirred for 4 h, and centrifuged at 8000 rpm for 8 min. The precipitate was washed with ethanol and dried in vacuo. The mass concentration of the modified porous microspheres in the ethanol aqueous solution was 2 mg / mL; the mass concentration of chloroplatinic acid in THF was 20 mg / mL; the mass concentration of palladium acetate in THF was 20 mg / mL; and the amount ratio of palladium acetate to NaBH4 aqueous solution was 1 g:50 mL.

[0063] S2: The product obtained in step S1 and the modified glucose oxidase were added to a PBS buffer solution at pH 7.4. The mixture was stirred at 100 r / min under nitrogen for 24 h, centrifuged at 8000 rpm for 10 min, and the precipitate was washed with PBS buffer and freeze-dried to obtain a highly sensitive glucose detection material. The mass concentration of the modified glucose oxidase in the buffer solution was 2 mg / mL.

[0064] Example 3: A high-sensitivity glucose detection material is prepared using raw materials including 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 parts of palladium acetate.

[0065] The raw materials for preparing the modified glucose oxidase include the following components in parts by weight: 1.5 parts of glucose oxidase (GOD) and 3 parts of thiol-polyethylene glycol-carboxyl (SH-PEG-COOH).

[0066] The preparation method of modified glucose oxidase comprises the following steps:

[0067] A1: GOD was added to PB buffer at pH 6.5 and mixed to obtain a GOD solution. SH-PEG-COOH was added to the PB buffer, followed by EDC and NHS. The mixture was stirred at 180 rpm for 2 h to obtain an activation solution. In the GOD solution, the mass concentration of GOD in PB buffer was 1.5 mg / mL; the mass concentration of SH-PEG-COOH in PB buffer was 2.5 mg / mL; the mass ratio of SH-PEG-COOH, EDC, and NHS was 10:1.5:1.

[0068] A2: Add the GOD solution obtained in step A1 to the activation solution, stir at 250 rpm for 12 h, centrifuge at 10,000 rpm for 8 min, and wash the precipitate with deionized water to obtain modified GOD.

[0069] 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 parts of 1,7-octanediyne, 0.15 parts 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-hydroxysuccinimide maleimidopropionate (SMMP).

[0070] The preparation method of modified porous microspheres comprises the following steps:

[0071] B1: 1,3-Propanedithiol, propargyl glycidyl ether, and 1,7-octanediyne were mixed, DMPA, chloroform, and PEG were added, and the mixture was stirred and added dropwise to a 5 wt% sodium dodecylbenzenesulfonate (SDBS) aqueous solution. The mixture was sealed and aerated with high-purity nitrogen for 50 min, then placed in an ice-water bath and irradiated with continuous ultraviolet light (λ = 365 nm) for 2 h. The mixture was washed with water, tetrahydrofuran (THF), and methanol in sequence, and dried in a vacuum at 35°C to obtain epoxy porous microspheres. The mass ratio of chloroform to PEG was 4:1; the ratio of SDBS aqueous solution to 1,3-Propanedithiol was 25 mL:1 g.

[0072] B2: Ultrasonic dispersion of the epoxy-based porous microspheres obtained in step B1 in a methanol / chloroform mixture (v:v = 5:1) was performed. HD and triethylamine were added, and the mixture was stirred at 80°C and 250 rpm for 4 h. The mixture was filtered, and the filter cake was washed with methanol and THF, and dried under vacuum. The concentration of HD in the methanol / chloroform mixture was 45 mg / mL. The amount of triethylamine used was 4% of the volume of the methanol / chloroform mixture.

[0073] B3: The product obtained in step B2 and SMMP were added to PB buffer at pH = 7.4. The reaction was stirred at 250 rpm at room temperature for 4 h, and centrifuged at 8000 rpm for 12 min. The precipitate was washed with DMF and deionized water, and dried to obtain modified porous microspheres; the mass concentration of SMMP in PB buffer was 1 mg / mL.

[0074] This embodiment also provides a method for preparing the highly sensitive glucose detection material, comprising the following steps:

[0075] S1: The modified porous microspheres were uniformly dispersed in a 25 vol% ethanol aqueous solution to obtain a dispersion. Chloroplatinic acid and palladium acetate were separately dissolved in THF, added to the dispersion, and stirred for 2 h. A 0.5 M NaBH4 aqueous solution was added, stirred for 4 h, and centrifuged at 8000 rpm for 10 min. The precipitate was washed with ethanol and dried in vacuo. The mass concentration of the modified porous microspheres in the ethanol aqueous solution was 3 mg / mL; the mass concentration of chloroplatinic acid in THF was 20 mg / mL; the mass concentration of palladium acetate in THF was 20 mg / mL; and the dosage ratio of palladium acetate to NaBH4 aqueous solution was 1 g:50 mL.

[0076] S2: The product obtained in step S1 and the modified glucose oxidase were added to a PBS buffer solution at pH 7.4. The mixture was stirred at 150 rpm for 24 h under nitrogen protection. The mixture was centrifuged at 9000 rpm for 10 min. The precipitate was washed with PBS buffer and freeze-dried to obtain a highly sensitive glucose detection material. The mass concentration of the modified glucose oxidase in the buffer solution was 2.5 mg / mL.

[0077] The only difference between Comparative Example 1 and Example 1 is that glucose oxidase is used instead of the modified glucose oxidase.

[0078] The only difference between Comparative Example 2 and Example 1 is that palladium acetate is not added.

[0079] The only difference between Comparative Example 3 and Example 1 is that step B3 is not performed, that is, the product obtained in step B2 is used instead of the modified porous microspheres.

[0080] Experimental Example 1: 100 μL of glucose solutions of different concentrations and 100 μL of the high-sensitivity glucose detection material (1 mg / mL) of Example 1 were added to 700 μL of acetic acid-sodium acetate buffer (pH 4). After incubation at 37°C for 20 min, 100 μL of TMB (10 mmol·L -1 ), react for another 30 min, detect the absorbance at 652 nm in the supernatant, and record the relationship between glucose concentration and absorbance. The results are as follows Figure 1 shown.

[0081] Figure 1 The results showed that there was a good linear relationship between the absorbance value and the glucose concentration (within 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 quantitative analysis of glucose and effectively detect glucose concentration.

[0082] Experimental Example 2: The detection limits of glucose obtained from the detection materials of Example 2 and Comparative Examples 1-3 were calculated based on S / N=3. The results are as follows: Figure 2 shown.

[0083] Figure 2 The results showed that Example 2 and Examples 1-3 all had lower detection limits, among which Example 2 had the best effect and was significantly lower than the glucose content in serum. The detection material of the present invention can effectively detect glucose content with high detection sensitivity.

[0084] Experimental Example 3: The test materials of Example 3 and Comparative Examples 1-3 were placed at 4°C for 14 days, and 1 mM glucose was used as the test object. The test was performed 10 times, and the relative standard deviation was calculated. The results are as follows: Figure 3 shown.

[0085] Figure 3 The results showed that the relative standard deviations of Example 3 and Comparative Examples 1-3 were relatively small, among which the standard deviation data of Example 1 was the best, indicating that the high-sensitivity glucose detection material of the present invention has high detection accuracy and stability.

[0086] Those skilled in the art should understand that the discussion of any of the above embodiments is merely illustrative and is not intended to imply that the scope of the present invention is limited to these examples. Within the scope of the present invention, the technical features in the above embodiments or different embodiments may be combined, the steps may be implemented in any order, and there are many other variations of the different aspects of the present invention as described above, which are not provided in detail for the sake of simplicity.

Claims

1. A high-sensitivity 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 raw materials for preparing 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 comprises the following steps: A1: Add GOD to PB buffer and mix well to obtain GOD solution. Add SH-PEG-COOH to PB buffer, add EDC and NHS, and stir to obtain activation solution. A2: Add the GOD solution obtained in step A1 to the activation solution, stir, centrifuge, and wash to obtain modified GOD; 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-octanediyne, 0.12-0.17 parts of benzoin dimethyl ether, 1-1.5 parts of PEG, 4-6 parts of 1,3-diamino-2-propanol, and 1-1.5 parts of 3-maleimidopropionic acid hydroxysuccinimide ester; The preparation method of the modified porous microspheres comprises the following steps: B1: 1,3-Propanedithiol, propargyl glycidyl ether, and 1,7-octanediyne were mixed, benzoin dimethyl ether, chloroform, and PEG were added, and the mixture was stirred evenly. The mixture was then added dropwise to the SDBS aqueous solution. The mixture was sealed and aerated with high-purity nitrogen for 40-60 minutes. The mixture was then placed in an ice-water bath and continuously irradiated with ultraviolet light, washed, and dried to obtain epoxy-based porous microspheres. B2: ultrasonically disperse the epoxy porous microspheres obtained in step B1 in a methanol / chloroform mixture, add 1,3-diamino-2-propanol and triethylamine, react, wash, and dry; B3: adding the product obtained in step B2 and 3-maleimidopropionic acid hydroxysuccinimide ester to PB buffer, stirring for reaction, washing, and drying to obtain modified porous microspheres; The preparation method of the highly sensitive glucose detection material comprises the following steps: S1: The modified porous microspheres are uniformly dispersed in a 25 vol% ethanol aqueous solution to obtain a dispersion, chloroplatinic acid and palladium acetate are dissolved in THF, respectively, added to the dispersion, stirred, and a NaBH4 aqueous solution is added, stirred, washed, and dried; S2: Add the product obtained in step S1 and the modified glucose oxidase into PBS buffer, stir under nitrogen protection, wash, and dry to obtain a high-sensitivity glucose detection material.

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

1.

3. The high-sensitivity glucose detection material according to claim 2, wherein In step B1, the mass ratio of chloroform to PEG is 4:1; the amount ratio of SDBS aqueous solution to 1,3-propanedithiol is 25 mL:1 g.

4. The high-sensitivity glucose detection material according to claim 3, wherein In step B2, the mass concentration of the 1,3-diamino-2-propanol in the methanol / chloroform mixture is 40-50 mg / mL; the amount of triethylamine used is 3-5% of the volume of the methanol / chloroform mixture.

5. The high-sensitivity glucose detection material according to claim 4, wherein In step B3, the mass concentration of the 3-maleimidopropionic acid hydroxysuccinimide ester in the PB buffer is 1 mg / mL.

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