CN111118111A - Rapid detection method of glucose - Google Patents

Rapid detection method of glucose Download PDF

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CN111118111A
CN111118111A CN201911263333.2A CN201911263333A CN111118111A CN 111118111 A CN111118111 A CN 111118111A CN 201911263333 A CN201911263333 A CN 201911263333A CN 111118111 A CN111118111 A CN 111118111A
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glucose
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李向阳
李宗益
迟静甜
郭曼莉
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Shandong Agricultural University
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Abstract

The invention discloses a rapid detection method of glucose. The invention prepares a nano GOx & AuNCs @ ZIF-8 compound with high catalytic capacity by using a mild and environment-friendly mode; compared with a uniform mixture of two free enzymes, namely AuNCs and GOX, the GOx & AuNCs @ ZIF-8 compound has the advantages that the biological catalytic cascade of the GOx & AuNCs @ ZIF-8 is enhanced by about 19.3 times, the enzyme stability is high, the enzyme consumption is small, the cost is low, efficient and sensitive detection on glucose is realized, the detection condition is mild, the operation is simple and convenient, and the targeting property is strong. The problems of complicated sample pretreatment procedure and expensive instrument in the chromatography are solved; the electrochemical method has poor targeting property and poor sensor stability, and needs special electrodes and electrochemical equipment; the traditional enzymatic colorimetric sensor has the problems of high cost, more enzyme dosage, easy inactivation of enzyme and the like.

Description

Rapid detection method of glucose
Technical Field
The invention relates to the technical field of saccharide analysis and detection, in particular to a rapid detection method of glucose.
Background
The glucose content in serum is an important index for measuring the health state of human bodies, the glucose content in soil is a common index for measuring the characteristics of soil, the glucose index in sewage plays an important role in sewage purification, and the glucose content in foods and medicines is a common index for component analysis, so that the glucose detection technology is widely used in the fields of medicine, biotechnology, soil detection, industrial wastewater treatment, food and medicine processing. The development of a glucose detection technology with simple operation and high sensitivity has become a key focus of attention in the field.
The current methods for detecting glucose mainly comprise a colorimetric method, an electrochemical method, a high performance liquid chromatography and the like, wherein: the sample pretreatment procedure of the high performance liquid chromatography is complicated, the instrument is expensive, and the cost is high; the electrochemical method is the most widely used glucose detection technology in recent years, and has the characteristics of simple operation, wide linear range and high sensitivity, but the electrochemical method has poor selectivity on a detected object, needs special electrodes and electrochemical instruments for auxiliary detection, and most electrochemical sensors have short storage period and relatively high cost.
Compared with an electrochemical method, the colorimetric method has the outstanding advantages of low cost, short color development time, obvious phenomenon, stable condition, convenience, quickness, no need of special electrodes and electrochemical instruments and the like, and occupies an increasingly important position in the field of analysis and detection. The colorimetric chemical sensor is a simple and convenient analysis method for determining the content of a substance to be detected in a sample by using an ultraviolet-visible spectrophotometer or a photoelectric colorimeter and taking Lambert beer's law as a theoretical basis. Colorimetric chemical sensors are broadly classified into two categories: one type is a visual colorimetric chemical sensor, that is, people use eyes to observe and find a standard solution with a color similar to that of a sample solution, so that the content range of a substance to be detected in the sample is conveniently and quickly determined, but because the resolution of human eyes is not high and the influence of different environments is easily caused, the accuracy of the method is relatively low. The other method is a spectrophotometry method, the content of the substance to be detected is determined according to the Lambert beer law by utilizing the absorbance of the substance at a specific wavelength, and the method has higher accuracy compared with a visual colorimetry.
The glucose oxidase method (GOX method) has the advantages of strong specificity, short detection time, environmental protection and less sample consumption, and is a commonly used method for determining glucose by a colorimetric method. However, the conditions for survival of the required glucose oxidase and peroxidase are harsh, the glucose oxidase and peroxidase need to be stored in PBS solution with a certain concentration at a low temperature, the enzyme can be inactivated and decomposed at a high speed after being exposed at room temperature or a high temperature for a long time, the recycling of the enzyme is hindered, the glucose oxidase is expensive and complex to prepare, the dosage of the enzyme for measuring glucose by a GOX method is large, and the detection cost is high.
Disclosure of Invention
Aiming at the prior art, the invention aims to provide a rapid detection method of glucose, which realizes efficient and sensitive detection of glucose; effectively solves the problems of more enzyme dosage, easy enzyme inactivation and the like in the glucose determination by a glucose oxidase method.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a GOx & AuNCs @ ZIF-8 compound, which is prepared by the following steps:
zinc acetate solution, glucose oxidase (GOx) solution, gold nanoclusters (AuNCs) solution and 2-methylimidazole solution are mixed according to the volume ratio of (3-5): (3-5): (3-5): (3-5), stirring for 1-3min, and then standing for 10-16 h; and (4) centrifugally washing to prepare the GOx & AuNCs @ ZIF-8 compound.
Preferably, the concentration of the zinc acetate solution is 0.03-0.05 mol/L; the concentration of the glucose oxidase solution is 0.4-0.6 mg/mL; the concentration of the gold nanocluster solution is 0.6-0.9 mg/mL; the concentration of the 2-methylimidazole solution is 0.13-0.17 mol/L.
Preferably, the gold nanocluster solution is prepared by the following method:
(1) adding HAuCl4Adding the water solution into Bovine Serum Albumin (BSA) solution under vigorous stirring, and continuously stirring for 3-8 min;
(2) adding NaOH solution into the solution obtained in the step (1), and continuously and violently stirring for 20-28 h;
(3) and (3) dialyzing the solution obtained in the step (2) in ultrapure water for 40-50h to prepare a gold nanocluster solution.
More preferably, the HAuCl4The concentration of the aqueous solution is 8-12 mmol/L; the concentration of the BSA solution is 40-60 mg/mL; the concentration of the NaOH solution is 0.5-1.5 mol/L.
More preferably, HAuCl4The volume ratio of the added aqueous solution, BSA solution and NaOH solution is (4-6): (4-6): (0.4-0.6).
In a second aspect of the present invention, there is provided the use of the above GOx & AuNCs @ ZIF-8 complex in 1) or 2) as follows:
1) detecting glucose;
2) preparing reagent for qualitative or quantitative detection of glucose.
In a third aspect of the present invention, a method for rapidly detecting glucose is provided, which comprises the following steps:
(1) dissolving the GOx & AuNCs @ ZIF-8 compound in dual deionized water to obtain a GOx & AuNCs @ ZIF-8 compound solution with the concentration of 30-50 mg/mL; mixing the GOx & AuNCs @ ZIF-8 compound solution with the solution to be detected, and then carrying out water bath;
(2) and adding an acetic acid buffer solution and a TMB solution into the system after water bath, adjusting the pH of the system to 1-3, and measuring the UV-Vis spectrum of the system after 10-15min, thereby realizing the detection of the glucose in the solution to be detected.
Preferably, in the step (1), the temperature of the water bath is 20-40 ℃, and the water bath time is 15-20 min.
Preferably, in the step (2), the pH of the acetic acid buffer solution is 1-3, and the concentration is 0.1 mol/L; the concentration of the TMB solution is 7-9 mmol/L.
Preferably, in the step (2), the UV-Vis spectral range is 400-800 nm.
Preferably, in the step (2), the detection of the glucose in the solution to be detected is qualitative detection or quantitative detection;
when the qualitative detection is carried out, whether the solution to be detected contains glucose is determined as follows:
if the ultraviolet-visible absorption intensity value measured after the solution to be detected is added is higher than the contrast value, the solution to be detected contains or is candidate to contain glucose; if the ultraviolet-visible absorption intensity value measured after the solution to be measured is added is lower than or equal to the comparison value, the solution to be measured does not contain glucose;
the control value is the ultraviolet-visible absorption intensity value measured by carrying out steps (1) to (2) using a solution containing no glucose in place of the solution to be measured.
When the quantitative detection is carried out, determining the content of the glucose in the solution to be detected as follows: introducing the measured ultraviolet-visible absorption intensity into a linear equation to obtain the glucose content of the solution to be measured;
further, the linear equation is obtained as follows: and (3) carrying out the steps (1) to (2) by using a series of glucose standard products with known concentrations to replace the solution to be detected, and measuring the ultraviolet-visible absorption intensity corresponding to the glucose standard products with the concentrations, thereby obtaining a linear equation between the ultraviolet-visible absorption intensity and the glucose concentration.
The invention has the beneficial effects that:
(1) the invention prepares the nano GOx & AuNCs @ ZIF-8 compound with high catalytic capability by using a mild and environment-friendly mode for the first time; compared with a uniform mixture of two free enzymes, namely AuNCs and GOX, the GOx & AuNCs @ ZIF-8 compound has the advantages that the biocatalysis cascade is enhanced by about 19.3 times, the enzyme stability is high, the enzyme dosage is small, the cost is low, the efficient and sensitive detection on glucose is realized, the detection condition is mild, the operation is simple and convenient, and the targeting property is strong. The problems of complicated sample pretreatment procedure and expensive instrument in the chromatography are solved; the electrochemical method has poor targeting property and poor sensor stability, and needs special electrodes and electrochemical equipment; the traditional enzymatic colorimetric sensor has the problems of high cost, more enzyme dosage, easy inactivation of enzyme and the like.
(2) The synthesized GOx & AuNCs @ ZIF-8 compound can be repeatedly used, and the GOx & AuNCs @ ZIF-8 compound can be immediately collected and stored for later use after each detection is finished.
(3) The determination method can rapidly detect whether the sample contains glucose, the detection limit is 0.02 mu mol/L, and the method can be accurately used for detecting the glucose content in serum, soil, food and the like, and is suitable for popularization and application.
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FIG. 1: the invention discloses a schematic diagram for detecting glucose.
FIG. 2: the synthetic effect of the GOx & AuNCs @ ZIF-8 compound is characterized; wherein, A: pure ZIF-8 and the XRD pattern of the GOx & AuNCs @ ZIF-8 complex synthesized by the invention, B: the absorbance of oxTMB at 652nm varied with time over different catalysts.
FIG. 3: the stability of the GOx & AuNCs @ ZIF-8 compound is compared with that of free GOx & AuNCs; in the figure, A: comparing the long-term storage stability of GOx & AuNCs @ ZIF-8 complex cultured in phosphate buffered saline (pH7.4) at room temperature with that of free GOx & AuNCs; b: GOx & AuNCs @ ZIF-8 complexes cultured in pH7.4 phosphate buffered saline containing 0.25 wt% trypsin or 1 wt% EDTA at room temperature were compared to free GOx & AuNCs stability.
FIG. 4: the constructed mimic enzyme detection system is used for linear investigation of glucose detection; wherein, A: according to the ultraviolet-visible spectrophotometric detection results of glucose with different concentrations, the glucose concentrations represented by curves from bottom to top are respectively 0 mu mol/L, 0.2 mu mol/L, 0.5 mu mol/L, 1 mu mol/L, 5 mu mol/L, 10 mu mol/L, 15 mu mol/L, 20 mu mol/L, 25 mu mol/L, 30 mu mol/L, 35 mu mol/L and 40 mu mol/L; b: linear relationship between uv-vis absorption intensity and glucose concentration.
FIG. 5: the mimic enzyme detection system constructed by the invention has the targeting property to glucose.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
As introduced in the background art, compared with the electrochemical method, the colorimetric method has the advantages of low cost, short color development time, obvious phenomenon, stable conditions, convenience, rapidness, no need of special electrodes and electrochemical instruments, and the like, and occupies an increasingly important position in the field of analysis and detection. The glucose oxidase method (GOX method) has the advantages of strong specificity, short detection time, environmental protection and less sample consumption, and is a commonly used method for determining glucose by a colorimetric method. However, the conditions for survival of the required glucose oxidase and peroxidase are harsh, the glucose oxidase and peroxidase need to be stored in PBS solution with a certain concentration at low temperature, and the enzyme can be inactivated and decomposed at a high speed after being exposed at room temperature or at a high temperature for a long time; in addition, the amount of the required glucose oxidase is large, and the detection cost is high.
Based on the above, the invention aims to construct a novel rapid glucose detection method, and effectively solves the problems of high enzyme dosage, easy enzyme inactivation and the like in glucose determination by a glucose oxidase method.
The principle of the present invention for detecting glucose is shown in FIG. 1, where glucose is first oxidized by GOx to produce gluconic acid and H2O2After that, AuNCs catalyzes H2O2Reduction produces OH, resulting in the formation of the coloured product oxTMB. We found that the coloured product oxTMB had a maximum absorption peak at a652nm, thereby enabling glucose-specific detection.
The invention is realized by simply mixing Gox, AuNCs and Zn at room temperature2+And 2-methylimidazole in aqueous solution&AuNCs @ ZIF-8 complex; invention is to GOx&The effect of the synthesis of AuNCs @ ZIF-8 complex was characterized (FIG. 2); determination of the synthesized composite GOx&SEM image of AuNCs @ ZIF-8, finding GOx&AuNCs @ ZIF-8 is a well-dispersed nanoparticle with an average size of 1 μm; compare GOx&The XRD patterns of AuNCs @ ZIF-8 and pure ZIF-8 (FIG. 2A) were found to overlap, demonstrating that the incorporation of GOx and AuNCs had no effect on the crystal structure of ZIF-8; determine GOx&AuNCs @ ZIF-8 was visualized at 488nm as a fluorescence microscope image, and the red fluorescence of AuNCs was observed, confirming that AuNCs were successfully coated with the complex.
The present invention compares the time-dependent absorbance change at 652nm of oxTMB using different catalyst detection systems (fig. 2B) and found that when AuNCs @ ZIF-8 and free GOx were introduced into the system, the rates were approximately equal to the rates of the free GOx and AuNCs mixtures, and the ZIF-8 coating had no significant effect on the catalytic activity of GOx and AuNCs. When GOx @ ZIF-8 and free AuNCs were introduced into the system at a rate 4.0 times that of free GOx and AuNCs, while when GOx & AuNCs @ ZIF-8 (i.e., 5 in FIG. 2B) was used as the catalyst, the biocatalytic cascade was enhanced by about 19.3 times compared to the free GOx and AuNCs mixture. The results show that the ZIF-8 encapsulation significantly improves the catalytic activity of GOx @ AuNCs.
The invention also characterizes the stability of GOx and AuNCs @ ZIF-8: the stability of enzymes in biological systems is crucial for glucose detection, and the invention compares the long-term stability of a homogeneous mixture of two free enzymes, AuNCs and GOX, incubated in a pH7.4 phosphate buffered saline solution at room temperature with the GOx & AuNCs @ ZIF-8 complex as shown in FIG. 3A. after 2 days of storage at room temperature, GOx & AuNCs @ ZIF-8 retains 90% of the initial total activity, whereas a homogeneous mixture of two free enzymes, AuNCs and GOX, retains only 10% of the initial total activity. The stability of the free enzyme and GOx & AuNCs @ ZIF-8 complex incubated in a pH7.4 phosphate buffered saline solution containing 0.25 wt% trypsin or 1 wt% EDTA, as shown in FIG. 3B, the excellent storage stability of GOx & AuNCs @ ZIF-8 was also found, due to the cross-linking of the enzyme to the ZIF-8 support, after 30 minutes of tryptic digestion at 37 ℃, GOx & AuNCs @ ZIF-8 and the homogeneous mixture of two free enzymes AuNCs and GOx maintained about 90% and about 60% of the initial total activity, respectively, while the GOx & AuNCs @ ZIF-8 complex in the presence of 1 wt% of EDTA retained 80% of its original activity, and the homogeneous mixture of two free enzymes, AuNCs and GOX, lost 100% of its original activity, demonstrating that the ZIF-8 scaffold has excellent protection effect on the enzyme, and the storage stability of GOx & AuNCs @ ZIF-8 is significantly higher than that of the homogeneous mixture of two free enzymes, AuNCs and GOX.
The invention also verifies the selectivity of GOx & AuNCs @ ZIF-8 to glucose, compares the catalytic effects of GOx & AuNCs @ ZIF-8 to fructose, mannose, maltose, lactose and glucose solutions with the same concentration, and finds that GOx & AuNCs @ ZIF-8 has obvious targeting property to the selection of glucose (figure 5). Therefore, the ZIF-8 is preferably used as a coating material of GOx @ AuNCs to obtain a novel nano enzyme system GOx & AuNCs @ ZIF-8.
Therefore, the 2-methylimidazole zinc salt MAF-4(ZIF-8) with high porosity is used for the first time to wrap the bovine serum albumin functionalized gold nanoclusters (AuNCs) and glucose oxidase (GOx) to form a GOx & AuNCs @ ZIF-8 nano-scale glucose detection enzyme linked system, and the system can be used for quickly and sensitively detecting glucose in a sample.
The invention also inspects the pH and temperature conditions of the glucose detection system, respectively compares the effect of the pH value (1, 2, 3, 4, 5, 6, 7, 8, 9) and the solution temperature (10, 20, 30, 40, 50, 60 ℃) of the detection system solution on the catalytic activity of GOx & AuNCs @ ZIF-8, and determines the pH value and the temperature range when the catalytic activity of GOx & AuNCs @ ZIF-8 is better. GOx & AuNCs @ ZIF-8 is suitably at a pH in the range of (1-3) and a temperature in the range of (20-40 ℃).
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention, which were not specifically described, were all those conventional in the art and commercially available.
In order to avoid the influence of impurities such as metal ions in water on the detection effect of the system, all the solvents except TMB use ethanol as a solvent, and all the other solvents use double deionized water which is obtained by double distilling the deionized water.
Example 1: synthesis of GOx & AuNCs @ ZIF-8
(1) Synthesis of AuNCs:
5mL of HAuCl solution with the temperature of 37 ℃ and the concentration of 10mmol/L4The aqueous solution was added to 5mL of a BSA solution having a concentration of 50mg/mL and being vigorously stirred at 37 ℃. After 5 minutes, 0.5mL of a 1.0mol/L NaOH solution was added and the mixture solution was stabilized at 37 ℃ for 24 hours with vigorous stirring. Stirring was stopped when the color of the solution changed from light yellow to dark brown, and the solution was dialyzed in ultrapure water for 48 hours to remove unreacted impurities.
(2) Synthesis of GOx & AuNCs @ ZIF-8:
5mL of zinc acetate solution with the concentration of 0.04mmol/L, 5mL of GOx solution with the concentration of 0.5mg/mL, 4mL of AuNCs solution with the concentration of 0.8mg/mL and 5mL of 2-methylimidazole solution with the concentration of 0.16mmol/L are mixed at room temperature, the mixed solution is stirred for 1 minute and then stands for 12 hours, centrifugal washing is carried out at 3000r/min for 15 minutes, the centrifugal washing step is repeated for 5 times, and the GOx & AuNCs @ ZIF-8 is prepared.
Example 2: establishment of linear equation for glucose detection
(1) GOx & AuNCs @ ZIF-8 (prepared in example 1) solutions were prepared at 40mg/mL concentrations, respectively, of glucose solutions at 0. mu. mol/L, 0.2. mu. mol/L, 0.5. mu. mol/L, 1. mu. mol/L, 5. mu. mol/L, 10. mu. mol/L, 15. mu. mol/L, 20. mu. mol/L, 25. mu. mol/L, 30. mu. mol/L, 35. mu. mol/L, 40. mu. mol/L.
(2) And (2) respectively and uniformly mixing 500 mu L of GOx & AuNCs @ ZIF-8 solution obtained in the step (1) with different gradients of 1mL of glucose solution prepared in the step (1), and carrying out water bath at 37 ℃ for 30 minutes.
(3) 300mL of an acetic acid buffer solution (pH 3.0) having a concentration of 0.1mmol/L and 100. mu.L of a TMB solution having a concentration of 8 mmol/L were sequentially added to the solution obtained in step (2), and after 10 minutes, the UV-Vis spectrum of the solution was measured and the UV-visible absorption intensity of glucose at 652nm was obtained at various concentrations.
(4) Plotting a linear relationship of the ultraviolet-visible absorption intensity (y) to the glucose concentration (x) from the data obtained in step (3) (fig. 4B), and obtaining a linear equation:
y＝0.015x+0.167；(R20.999); the detection limit was 0.02. mu. mol/L.
Example 3: detection of glucose content in a sample
(1) A GOx & AuNCs @ ZIF-8 (prepared as described in example 1) solution was prepared at a concentration of 40 mg/mL.
(2) And (2) uniformly mixing 500 mu L of the GOx & AuNCs @ ZIF-8 solution obtained in the step (1) with 1.0mL of a sample solution to be detected, and carrying out water bath at 37 ℃ for 30 minutes.
(3) 300mL of an acetic acid buffer solution (pH 3.0) having a concentration of 0.1mmol/L and 100. mu.L of a TMB solution having a concentration of 8 mmol/L were sequentially added to the solution obtained in step (2), and after 10 minutes, the UV-Vis spectrum of the solution was measured to obtain A652nm ultraviolet-visible absorption intensity.
(4) And (3) substituting the ultraviolet-visible absorption intensity obtained in the step (3) into a linear equation y of 0.015x +0.167 (obtained in example 2), and calculating the glucose content of the sample to be detected.
Example 4: targeted investigation of glucose detection methods
In order to evaluate the targeting of the glucose detection method of the invention, the invention researches the influence of fructose, mannose, maltose and lactose on the glucose detection. Specifically, fructose, mannose, maltose and lactose solutions with the same concentration are prepared, the ultraviolet visible absorption intensity of A652nm is measured by the method of example 3, and the result shows that the detection system has obvious selectivity on glucose (figure 5). We also investigate the detection results of a detection system in which fructose, mannose, maltose and lactose are respectively added to the glucose solution as interferents, and find that the addition of fructose, mannose, maltose and lactose does not significantly affect the ultraviolet-visible absorption intensity, and is completely consistent with the results of a single quantitative measurement of the glucose solution. That is, when glucose coexists with other saccharides, the detection system of the invention can still realize accurate detection of glucose in a sample.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims (10)

1. The GOx & AuNCs @ ZIF-8 compound is characterized by being prepared by the following method:
zinc acetate solution, glucose oxidase solution, gold nanocluster solution and 2-methylimidazole solution are mixed according to the volume ratio of (3-5): (3-5): (3-5): (3-5), stirring for 1-3min, and then standing for 10-16 h; and (4) centrifugally washing to prepare the GOx & AuNCs @ ZIF-8 compound.
2. The GOx & AuNCs @ ZIF-8 complex according to claim 1, wherein the concentration of the zinc acetate solution is 0.03-0.05 mol/L; the concentration of the glucose oxidase solution is 0.4-0.6 mg/mL; the concentration of the gold nanocluster solution is 0.6-0.9 mg/mL; the concentration of the 2-methylimidazole solution is 0.13-0.17 mol/L.
3. The GOx & AuNCs @ ZIF-8 complex of claim 1, wherein the gold nanocluster solution is prepared by a method comprising:
(1) adding HAuCl4Adding the aqueous solution into the BSA solution under the condition of vigorous stirring, and continuously stirring for 3-8 min;
(2) adding NaOH solution into the solution obtained in the step (1), and continuously and violently stirring for 20-28 h;
(3) and (3) dialyzing the solution obtained in the step (2) in ultrapure water for 40-50h to prepare a gold nanocluster solution.
4. GOx according to claim 3&AuNCs @ ZIF-8 complex, characterized in that said HAuCl4The concentration of the solution is 8-12 mmol/L; the concentration of the BSA solution is 40-60 mg/mL; the concentration of the NaOH solution is 0.5-1.5 mol/L.
5. GOx according to claim 4&AuNCs @ ZIF-8 complex characterized by HAuCl4The volume ratio of the solution, the BSA solution and the NaOH solution is (4-6): (4-6): (0.4-0.6).
6. Use of a GOx & AuNCs @ ZIF-8 complex as defined in any one of claims 1-5 in 1) or 2) as follows:
1) detecting glucose;
2) preparing reagent for qualitative or quantitative detection of glucose.
7. A rapid detection method of glucose is characterized by comprising the following steps:
(1) dissolving the GOx & AuNCs @ ZIF-8 complex of any one of claims 1-5 in dual deionized water to obtain a solution of GOx & AuNCs @ ZIF-8 complex at a concentration of 30-50 mg/mL; mixing the GOx & AuNCs @ ZIF-8 compound solution with the solution to be detected, and then carrying out water bath;
(2) and adding an acetic acid buffer solution and a TMB solution into the system after water bath, and measuring the UV-Vis spectrum of the system after 10-15min, thereby realizing the detection of the glucose in the solution to be detected.
8. The detection method according to claim 7, wherein in the step (1), the temperature of the water bath is 20-40 ℃, and the time of the water bath is 15-20 min.
9. The detection method according to claim 7, wherein in the step (2), the pH of the acetic acid buffer solution is 1 to 3, and the concentration is 0.1 mol/L; the concentration of the TMB solution is 7-9 mmol/L.
10. The detection method according to claim 7, wherein in the step (2), the detection of the glucose in the solution to be detected is a qualitative detection or a quantitative detection;
when the qualitative detection is carried out, whether the solution to be detected contains glucose is determined as follows:
if the ultraviolet-visible absorption intensity value measured after the solution to be detected is added is higher than the contrast value, the solution to be detected contains or is candidate to contain glucose; if the ultraviolet-visible absorption intensity value measured after the solution to be measured is added is lower than or equal to the comparison value, the solution to be measured does not contain glucose;
the control value is the ultraviolet-visible absorption intensity value measured by carrying out steps (1) to (2) using a solution containing no glucose in place of the solution to be measured.
When the quantitative detection is carried out, determining the content of the glucose in the solution to be detected as follows: introducing the measured ultraviolet-visible absorption intensity into a linear equation to obtain the glucose content of the solution to be measured;
preferably, the linear equation is obtained as follows: and (3) carrying out the steps (1) to (2) by using a series of glucose standard products with known concentrations to replace the solution to be detected, and measuring the ultraviolet-visible absorption intensity corresponding to the glucose standard products with the concentrations, thereby obtaining a linear equation between the ultraviolet-visible absorption intensity and the glucose concentration.
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