A glucose self-calibrating detection method

Abstract

The application discloses a glucose self-calibration detection method, comprising the following steps: (1) mixing a detection solution containing a known concentration of a bisboronic acid molecule and a known concentration of a bisphenol dye with a plurality of known concentration glucose standard solutions within a detectable concentration, respectively performing fluorescence and cyclic voltammetry tests, and establishing a group of calibration curves according to the results; (2) adjusting the concentrations of the bisboronic acid molecule and the bisphenol dye, repeating (1), obtaining a plurality of groups of calibration curves and composing a self-calibration platform; (3) using a detection solution containing an unknown concentration of a bisboronic acid molecule and an unknown concentration of a bisphenol dye to respectively perform fluorescence and cyclic voltammetry tests on a sample to be measured; (4) substituting the obtained results into each group of calibration curves in the self-calibration platform obtained in (2) respectively, calculating corresponding glucose concentrations, selecting a group of calibration curves with the minimum deviation as an optimal group of calibration curves of the sample to be measured, and regarding the average value of the two concentrations as the glucose concentration of the sample to be measured.

Description

Technical Field

[0001] This invention relates to the field of blood glucose detection, and more specifically to a glucose self-calibration detection method. Background Technology

[0002] Diabetes mellitus is a metabolic disease characterized by prolonged hyperglycemia. It is caused by genetic factors, immune dysfunction, endocrine disorders, viral infections, chemical toxins, medications, unhealthy lifestyle habits, and other factors leading to insufficient insulin secretion, insensitivity to insulin, or both. This results in the ineffective utilization and storage of glucose in the blood. Currently, there is no cure for diabetes, only management methods. Existing management plans use real-time blood glucose levels as a reference, employing insulin injections and oral hypoglycemic agents to maintain blood glucose levels within the target range. If diabetes is not well controlled, it can lead to various acute and chronic complications, such as acute diabetic ketoacidosis (DKA), chronic diabetic nephropathy, diabetic retinopathy, diabetic neuropathy, diabetic lower extremity vascular disease, diabetic foot, and cardiovascular disease. Therefore, for diabetic patients, the most effective management method is for them to monitor their glucose levels continuously and in real-time, intervening promptly to control abnormal glucose levels within the target range, reducing the occurrence of complications such as hyperglycemia and hypoglycemia after medication.

[0003] Most currently available continuous glucose monitoring (CGM) meters monitor glucose based on immobilized enzymes. While enzyme detection offers high selectivity and sensitivity, it also suffers from several drawbacks. Enzyme activity decreases over time, enzymes are prone to denaturation during immobilization, and they are susceptible to changes in temperature, humidity, pH, and toxic substances. These inherent factors contribute to the short lifespan and signal drift issues of enzyme-based CGM meters, necessitating daily external finger-prick blood calibration to compensate for numerical deviations over several days or weeks. Furthermore, enzymes have a short shelf life and are relatively expensive. Therefore, developing enzyme-free CGM meters to circumvent these inherent limitations is a new trend in this field. Summary of the Invention

[0004] Boric acid and its derivatives can reversibly bind to 1,2- or 1,3-cis-diols via covalent bonds to form five- or six-membered boronic esters, respectively, causing changes in acidity and conductivity. Introducing fluorescent groups into the structure of boronic acid compounds induces an optical signal response. Furthermore, when boronic acid compounds form complexes with diols, substitution occurs if a stronger diol compound is present. Sugars are polyhydroxy compounds, thus these properties of boronic acid compounds can be utilized for their detection. Moreover, boronic acid compounds offer advantages such as long shelf life, simple synthesis, and low cost. Therefore, boronic acid compounds have significant development value in the field of enzyme-free sugar detection. The currently marketed fully implantable Eversense continuous glucose meter is a fluorescent glucose detection product based on anthracene-cyclobisphenylboronic acid molecules, with a lifespan of up to 12 months. However, daily calibration is still required. In addition to the instability of hardware (such as light sources or photodetectors), signal drift remains a major problem, mainly due to the instability of anthracene bisphenylboronic acid molecules, such as their susceptibility to decomposition by reactive oxygen species and photobleaching of fluorescence.

[0005] An effective solution to reduce signal drift in borate continuous glucose meters is to use a dual-mode sensor for real-time calibration to continuously correct drift, but this method has not yet been applied to continuous glucose monitoring.

[0006] According to one aspect of this application, a glucose self-calibration detection method is provided, the method comprising:

[0007] (1) Mix the detection solution containing known concentrations of diboric acid molecules and known concentrations of bisphenol dyes with multiple known concentrations of glucose standard solutions within the detectable concentration range, and perform fluorescence testing and cyclic voltammetry testing respectively. Establish a set of calibration curves based on the test results.

[0008] (2) Adjust the concentrations of the diboronic acid molecules and the bisphenol dyes in the detection solution respectively, repeat (1), and obtain multiple sets of calibration curves to form a self-calibration platform;

[0009] (3) The test sample was subjected to fluorescence test and cyclic voltammetry test respectively using a detection solution containing unknown concentrations of diboronic acid molecules and unknown concentrations of bisphenol dyes, and the fluorescence test results and cyclic voltammetry test results of the test sample were obtained.

[0010] (4) Substitute the fluorescence test results and cyclic voltammetry test results of the sample to be tested into the calibration curves of each group in the self-calibration platform obtained in (2) respectively, and calculate the glucose concentration corresponding to each group of calibration curves. Each group of calibration curves contains two concentrations: the glucose concentration calculated from the fluorescence test results of the sample to be tested and the glucose concentration calculated from the cyclic voltammetry test results. Select the calibration curve group with the smallest concentration deviation as the optimal calibration curve group of the sample to be tested, and the average value of the two concentrations is regarded as the glucose concentration of the sample to be tested.

[0011] The multiple sets of calibration curves are at least two sets of calibration curves.

[0012] The diboronic acid molecule is selected from compounds containing two phenylboronic acid structures that can bind to glucose, and the compound has the structure shown in Formula I:

[0013]

[0014] Wherein: R1 and R2 are each independently selected from hydrogen atoms and unsubstituted C1 to C2 atoms. 30 Alkyl groups, substituted C1-C 30 Alkyl groups, unsubstituted C1-C1 30 heteroalkyl, substituted C1-C 30 heteroalkyl groups;

[0015] R3~R 10 Each group is independently selected from hydrogen, halogen, sulfonic acid, azide, cyanate, isocyanate, nitrate, cyano, isocyano, nitrosoxy, nitroso, nitro, aldehyde, acyl halide, carboxylic acid, carboxylic acid, and unsubstituted C1-C. 30 Alkyl groups, substituted C1-C 30 Alkyl groups, unsubstituted C1-C1 30 heteroalkyl, substituted C1-C 30 heteroalkyl groups, unsubstituted C2-C 30 alkenyl, substituted C2-C 30 alkenyl, unsubstituted C2-C 30 heteroene groups, substituted C2-C 30 heteroene group, unsubstituted C2-C 30 alkynyl group, substituted C2-C 30 alkynyl group, unsubstituted C2-C 30 heteroyne group, substituted C2-C 30 heteroyne group, unsubstituted C6-C 50 aryl, substituted C6-C 50 aryl, substituted C6-C 50 Heterocyclic aryl groups, unsubstituted C6-C 50 Heterocyclic aryl groups.

[0016] The bisphenol dye is selected from compounds that exhibit spectral changes upon binding with phenylboronic acid, and these compounds have the structure shown in Formula II:

[0017]

[0018] Where: R 11 ~R 14 Each group is independently selected from hydrogen, halogen, sulfonic acid, azide, cyanate, isocyanate, nitrate, cyano, isocyano, nitrosoxy, nitroso, nitro, aldehyde, acyl halide, carboxylic acid, carboxylic acid, and unsubstituted C1-C. 30 Alkyl groups, substituted C1-C 30 Alkyl groups, unsubstituted C1-C1 30 heteroalkyl, substituted C1-C 30 heteroalkyl groups, unsubstituted C2-C 30 alkenyl, substituted C2-C 30 alkenyl, unsubstituted C2-C 30 heteroene groups, substituted C2-C 30 heteroene group, unsubstituted C2-C 30 alkynyl group, substituted C2-C 30 alkynyl group, unsubstituted C2-C 30 heteroyne group, substituted C2-C 30 heteroyne group, unsubstituted C6-C 50 aryl, substituted C6-C 50 aryl, substituted C6-C 50 Heterocyclic aryl groups, unsubstituted C6-C 50 Heterocyclic aryl groups.

[0019] The alkyl group includes straight-chain alkyl, branched alkyl, and cyclic alkyl;

[0020] The alkenyl group includes straight-chain alkenyl, branched alkenyl, and cyclic alkenyl;

[0021] The alkynyl group includes straight-chain alkynyl, branched-chain alkynyl, and cyclic alkynyl;

[0022] The aryl group includes branched aryl, monocyclic aryl, and polycyclic aryl;

[0023] The molar ratio of the diboronic acid molecules to the bisphenol dye in the detection solution is 5:1 to 1:5.

[0024] The concentration range of the diboronic acid molecules in the multiple calibration curves of the self-calibration platform is 10 μM to 100 mM;

[0025] The concentration range of bisphenol dyes shown in the multiple calibration curves in the self-calibration platform is 10 μM to 100 mM.

[0026] The detectable concentration of the glucose detection method is 1–30 mM.

[0027] The purpose of this invention is to provide a self-calibrating glucose detection method.

[0028] The objective of this invention is achieved as follows: Diboronic acid molecules can form reversible covalent complexes with bisphenol dyes, causing fluorescence enhancement and a decrease in the redox peak of the bisphenol dye. Upon addition of glucose molecules, glucose competes with the bisphenol dye for binding to diboronic acid molecules. As the glucose concentration increases, the bisphenol dye in the complex is gradually replaced by glucose molecules, leading to a weakening of the fluorescence signal and an increase in the redox peak of the bisphenol dye. A glucose concentration-dependent standard curve is constructed using both fluorescence and cyclic voltammetry modes, serving as a set of calibration curves. Furthermore, by adjusting the concentrations of both diboronic acid molecules and bisphenol dye, the potential molecular decomposition or photocatalytic degradation that may occur in actual detection is simulated. The process involves bleaching, establishing at least two sets of calibration curves, and forming a self-calibration platform. Using detection reagents containing unknown concentrations of bisboronic acid molecules and bisphenol dyes, fluorescence and cyclic voltammetry tests are performed on the sample to be tested, obtaining the fluorescence and cyclic voltammetric signal values. These two signal values ​​are then substituted into all calibration curve sets within the established self-calibration platform. Each calibration curve set yields two glucose concentrations: the glucose concentration corresponding to the fluorescence signal and the glucose concentration corresponding to the cyclic voltammetric signal. The calibration curve set with the smallest concentration deviation is selected as the optimal calibration curve set for the sample, and the average of the two concentrations is considered the glucose concentration of the sample. This dual-mode optical and electrochemical glucose detection method eliminates the need for external calibration and has broad application prospects in the field of long-term stable glucose monitoring.

[0029] The specific steps include:

[0030] (1) Select a pair of diboronic acid molecules and bisphenol dye and mix them as the detection solution;

[0031] (2) Mix the detection solution containing known concentrations of diboric acid molecules and bisphenol dyes with multiple known concentrations of glucose standard solutions within the detectable concentration range, and perform fluorescence testing and cyclic voltammetry testing respectively. Establish a set of calibration curves based on the test results.

[0032] (3) Adjust the concentrations of the diboronic acid molecules and the bisphenol dyes in the detection solution respectively, repeat (2), and obtain multiple sets of calibration curves to form a self-calibration platform;

[0033] (4) The test sample was subjected to fluorescence test and cyclic voltammetry test respectively using a detection solution containing unknown concentrations of diboronic acid molecules and bisphenol dyes to obtain the fluorescence test results and cyclic voltammetry test results of the test sample.

[0034] (5) Substitute the fluorescence test results and cyclic voltammetry test results of the sample to be tested into the calibration curves of each group in the self-calibration platform obtained in (3), and calculate the glucose concentration corresponding to each calibration curve. Each calibration curve contains two concentrations: the glucose concentration calculated from the fluorescence test results and the glucose concentration calculated from the cyclic voltammetry test results. Select the calibration curve group with the smallest concentration deviation as the optimal calibration curve group for the sample to be tested, and the average value of the two concentrations is regarded as the glucose concentration of the sample to be tested.

[0035] The beneficial effects of this invention are:

[0036] The self-calibrating dual-mode continuous glucose monitoring system established in this invention will significantly improve service life and ensure accuracy while eliminating the pain of finger-prick calibration. This will provide a theoretical model and foundation for the construction of an internal calibration platform for glucose monitoring, and offer new ideas and methods for developing next-generation continuous glucose monitoring technology. Attached Figure Description

[0037] Figure 1 A shows the fluorescence spectra of PBS, PBS+ARS, PBS+ARS+DBA2+ and PBS+ARS+DBA2++Glucose; Figure 1 B represents the fluorescence spectra of different concentrations of glucose added to the complex formed by 100 μM ARS and 50 μM DBA2+. Figure 1 C represents the quantitative standard curve of glucose standard sample fluorescence mode plotted with fluorescence intensity at 600 nm as the ordinate;

[0038] Figure 2 A represents the cyclic voltammetry curves of PBS, PBS+ARS, PBS+ARS+DBA2+ and PBS+ARS+DBA2++Glucose; Figure 2 B represents the cyclic voltammetry curves of different concentrations of glucose added to the complex formed by 100 μM ARS and 50 μM DBA2+. Figure 2 C represents the electrochemical mode quantitative standard curve of glucose standard sample plotted with the current value at +0.43V as the ordinate;

[0039] Figure 3 For ARS / DBA2+ glucose self-calibration system; Figure 3 A represents glucose testing under different combinations of ARS and DBA2+: C1 (100 μM ARS and 50 μM DBA2+), C2 (80 μM ARS and 50 μM DBA2+), plotting the fluorescence intensity at 600 nm against the current at 0.43 V, and fitting the polynomial curve. Figure 3B is the fluorescence-electrochemical dual-mode quantitative curve of glucose standard sample under 100μM ARS and 50μM DBA2+ conditions (used for C1 calibration curve plotting); Figure 3 C represents the fluorescence-electrochemical dual-mode quantitative curve of glucose standard samples under 80 μM ARS and 50 μM DBA2+ conditions (used for C2 calibration curve plotting). Detailed Implementation

[0040] The present invention can be further described through the following embodiments; however, the scope of the present invention is not limited to the following embodiments. Those skilled in the art will understand that various changes and modifications can be made to the present invention without departing from its spirit and scope.

[0041] This invention provides a general and / or specific description of the materials and methods used in the experiments. While many of the materials and methods of operation used to achieve the objectives of this invention are well known in the art, they are still described in as much detail as possible herein.

[0042] Example 1

[0043] 1) Synthesis of diboronic acid molecule DBA2+

[0044] The synthesis of DBA2+ can be performed according to the method described in the literature (Angew. Chem. Int. Ed., 2019, 58, 10612-10615). A mixture of 60 mL of 2M dimethylamine (dissolved in tetrahydrofuran) in dry ice and acetone was placed in a bath for approximately 10 min. After cooling to -78°C, 20 mL of 1,4-dibromotoluene (5.3 g, 20 mmol, dissolved in tetrahydrofuran) was slowly added. The mixture was then brought back to room temperature, and after reacting for 1 h, it was poured into a mixture of 400 mL of ethyl acetate and 100 mL of 1M K2CO3 aqueous solution. The mixture was stirred vigorously, and after the solution separated into layers, the organic phase was collected and dried over Na2SO4. The organic solvent was removed by rotary evaporation to obtain the crude product 1,4-bis(dimethylaminomethyl)benzene, with a yield of 95%. 1,4-bis(dimethylaminomethyl)benzene and 2-bromomethylphenylboronic acid (12.9 g, 60 mmol) were dissolved in 40 mL of anhydrous DMF, bubbled with argon for 20 min, and heated at 60 °C for 24 h. The mixture was precipitated with 200 mL of ethyl acetate, and the precipitate was washed twice with 20 mL of ethyl acetate and dried under vacuum. The dried product was purified by C18 reversed-phase chromatography with water:methanol = 9:1 as the eluent. After removing the methanol, the product was freeze-dried to give DBA2+Br as a white solid (3.8 g, 30%).

[0045] 2) Construct a testing system

[0046] Mix 50 μM DBA2+ with 100 μM ARS (or other concentration combinations) in 1x PBS (pH 7.4) and shake at room temperature for 3 min to obtain the detection system containing the DBA2+ and ARS complex.

[0047] 3) Competitive displacement reaction of glucose

[0048] Add glucose of different concentrations to the above detection system and shake at room temperature for 3 minutes to complete the replacement.

[0049] 4) Fluorescence test

[0050] Using 460 nm as the excitation wavelength, the fluorescence spectra of the detection system and its reaction solutions after the addition of different concentrations of glucose were tested in the wavelength range of 500–700 nm. The results showed that ARS itself had almost no fluorescence, but the fluorescence was significantly enhanced after forming a complex with DBA2+. The fluorescence of the solution weakened after the addition of glucose, indicating that ARS was replaced. Figure 1 A), and as the concentration of added glucose gradually increases, the fluorescence of the solution gradually decreases. Figure 1 B), therefore, the relationship between fluorescence intensity and glucose concentration can be found ( Figure 1 C).

[0051] 5) Electrochemical testing

[0052] Measurements were performed on a CHI660E electrochemical workstation. Cyclic voltammetry (CV) curves of the DBA²⁺-ARS complex and its reaction solutions with different concentrations of glucose were scanned in the range of 0–0.8 V, using Ag / AgCl electrode, platinum wire electrode, and gold disk electrode as reference, counter, and working electrodes, respectively. The results showed that ARS exhibited an oxidation peak at +0.43 V. When the complex was formed with DBA²⁺, the amount of free ARS decreased, and the oxidation peak current decreased. The addition of glucose enhanced the peak current, indicating that ARS was replaced by glucose and released into the solution. Figure 2 A), and as glucose concentration increases, more ARS is released, and the +0.43V oxidation peak current continuously increases. Figure 2 ).

[0053] 6) Establish a self-calibration curve

[0054] The detection system uses only one detection solution, called the original solution. The initial concentrations of diboronic acid molecules and bisphenol dyes in the original solution are fixed, ranging from 10 μM to 100 mM. However, as time progresses, both may degrade or photobleach, causing their concentrations to change. If a fixed quantitative curve is used for glucose quantification, the calculated glucose value will be off-target and inaccurate. By constructing multiple calibration curves, each representing a possible change in the concentration of diboronic acid molecules or bisphenol dyes, the drift in glucose values ​​can be corrected, resulting in a more accurate glucose concentration value. In actual measurements, the more calibration curves constructed, the more accurate the calculation results. The following example of constructing two calibration curves illustrates the method of this invention. A 2× solution containing 200 μM MARS and 100 μM DBA2+ was used as the original solution (2×C1), and a 2× solution containing 160 μM MARS and 100 μM DBA2+ (2×C2) was used as a simulated solution for 20% ARS degradation. The fluorescence intensity at 600 nm and the current at 0.43 V for different concentrations of glucose standard solutions were obtained through the above fluorescence and electrochemical tests (3B&C). The current value was plotted on the x-axis and the fluorescence intensity value on the y-axis. Two curves were obtained for the two solutions, and a polynomial function was used for fitting, which served as calibration curve C1 and calibration curve C2, respectively. Figure 3 A).

[0055] 7) Determine glucose concentration

[0056] The original and simulated solutions were anonymized and shuffled to serve as detection solutions 1 and 2 (only one was used in actual measurements) to illustrate the method for determining glucose concentration. As shown in Table 1, the two detection solutions were used to test the sample. The fluorescence and current values ​​measured for detection solution 1 were 22235 and 157.7, respectively. These two values ​​were substituted into the two calibration curve groups C1 and C2 of the self-calibration platform (if there are multiple calibration curve groups, multiple values ​​are substituted). Each calibration curve group will yield two glucose concentrations: one calculated from the fluorescence value and one calculated from the current value. Ideally, these two glucose concentrations will match one of the calibration curve groups, and the concentration values ​​will be... Similarly, based on this, the calibration curve with the smallest standard deviation of glucose concentration is selected as the glucose concentration calculation curve in the actual measurement, and the average of the two calculated glucose concentrations is taken as the glucose concentration in the sample to be tested. Therefore, for test solution 1, the calculated standard deviation of the C2 calibration curve group (0.09%) is much smaller than the calculated standard deviation of the C1 calibration curve group (44%). Therefore, the average glucose concentration of 6.28 mM calculated by the C2 calibration curve group is selected as the glucose concentration of the sample to be tested. Similarly, the glucose concentration measured by test solution 2 is 6.45 mM, and the glucose concentrations detected by both test solutions are close to the actual concentration of the glucose sample of 6 mM.

[0057] Table 1

[0058]

[0059] Among them, [G F [G] is the glucose concentration calculated by substituting the fluorescence value of the sample into the calibration curve set; C The glucose concentration is calculated by substituting the current value of the sample to be tested into the calibration curve set;

[0060] The standard deviation % is calculated by taking the [G] from each calibration curve. F ] numerical value and [G C The standard deviation of the value divided by the average of the two values ​​multiplied by 100%

[0061] The above description is merely a few embodiments of this application and is not intended to limit this application in any way. Although this application discloses preferred embodiments as described above, it is not intended to limit this application. Any changes or modifications made by those skilled in the art without departing from the scope of the technical solution of this application using the disclosed technical content are equivalent to equivalent implementation cases and fall within the scope of the technical solution.

Claims

1. A self-calibrated glucose detection method, characterized in that, The method includes: (1) Mix the detection solution containing known concentrations of diboric acid molecules and known concentrations of bisphenol dyes with multiple known concentrations of glucose standard solutions within the detectable concentration range, and perform fluorescence testing and cyclic voltammetry testing respectively. Establish a set of calibration curves based on the test results. (2) Adjust the concentrations of the diboronic acid molecules and the bisphenol dyes in the detection solution respectively, repeat (1), and obtain multiple sets of calibration curves to form a self-calibration platform; (3) The test sample was subjected to fluorescence test and cyclic voltammetry test respectively using a detection solution containing unknown concentrations of diboronic acid molecules and unknown concentrations of bisphenol dyes, and the fluorescence test results and cyclic voltammetry test results of the test sample were obtained. (4) Substitute the fluorescence test results and cyclic voltammetry test results of the sample to be tested into the calibration curves of each group in the self-calibration platform obtained in (2) respectively, and calculate the glucose concentration corresponding to each group of calibration curves. Each group of calibration curves contains two concentrations: the glucose concentration calculated from the fluorescence test results of the sample to be tested and the glucose concentration calculated from the cyclic voltammetry test results. Select the calibration curve group with the smallest concentration deviation as the optimal calibration curve group of the sample to be tested, and the average value of the two concentrations is regarded as the glucose concentration of the sample to be tested. The diboronic acid molecule is selected from compounds containing two phenylboronic acid structures that can bind to glucose, and the compound has the structure shown in Formula I: Formula I Wherein: R1 and R2 are each independently selected from hydrogen atoms and unsubstituted C1~C atoms. 30 Alkyl groups, substituted C1~C 30 Alkyl groups, unsubstituted C1~C 30 heteroalkyl, substituted C1~C 30 heteroalkyl groups; R3~R 10 Each group is independently selected from hydrogen, halogen, sulfonic acid, azide, cyanate, isocyanate, nitrate, cyano, isocyano, nitrosoxy, nitroso, nitro, aldehyde, acyl halide, carboxylic acid, carboxylic acid, or unsubstituted C1~C. 30 Alkyl groups, substituted C1~C 30 Alkyl groups, unsubstituted C1~C 30 heteroalkyl, substituted C1~C 30 heteroalkyl groups, unsubstituted C2~C 30 alkenyl, substituted C2~C 30 alkenyl, unsubstituted C2~C 30 heteroene groups, substituted C2~C 30 heteroene group, unsubstituted C2~C 30 alkynyl group, substituted C2~C 30 alkynyl group, unsubstituted C2~C 30 heteroyne group, substituted C2~C 30 heteroyne group, unsubstituted C6~C 50 aryl, substituted C6~C 50 aryl, substituted C6~C 50 Heterocyclic aryl groups, unsubstituted C6~C 50 Heterocyclic aryl groups; The bisphenol dye is selected from compounds that exhibit spectral changes upon binding with phenylboronic acid, and these compounds have the structure shown in Formula II: Formula II Where: R 11 ~R 14 Each group is independently selected from hydrogen, halogen, sulfonic acid, azide, cyanate, isocyanate, nitrate, cyano, isocyano, nitrosoxy, nitroso, nitro, aldehyde, acyl halide, carboxylic acid, carboxylic acid, or unsubstituted C1~C. 30 Alkyl groups, substituted C1~C 30 Alkyl groups, unsubstituted C1~C 30 heteroalkyl, substituted C1~C 30 heteroalkyl groups, unsubstituted C2~C 30 alkenyl, substituted C2~C 30 alkenyl, unsubstituted C2~C 30 heteroene groups, substituted C2~C 30 heteroene group, unsubstituted C2~C 30 alkynyl group, substituted C2~C 30 alkynyl group, unsubstituted C2~C 30 heteroyne group, substituted C2~C 30 heteroyne group, unsubstituted C6~C 50 aryl, substituted C6~C 50 aryl, substituted C6~C 50 Heterocyclic aryl groups, unsubstituted C6~C 50 Heterocyclic aryl groups.

2. The glucose self-calibration detection method according to claim 1, characterized in that, The multiple sets of calibration curves are at least two sets of calibration curves.

3. The glucose self-calibration detection method according to claim 1, characterized in that, The alkyl group includes straight-chain alkyl, branched alkyl, and cyclic alkyl; The alkenyl group includes straight-chain alkenyl, branched alkenyl, and cyclic alkenyl; The alkynyl group includes straight-chain alkynyl, branched-chain alkynyl, and cyclic alkynyl; The aryl group includes branched aryl, monocyclic aryl, and polycyclic aryl.

4. The glucose self-calibration detection method according to claim 1, characterized in that, The molar ratio of the diboronic acid molecules and the bisphenol dye in the detection solution is 5:1 to 1:

5.

5. The glucose self-calibration detection method according to claim 1, characterized in that, The concentration range of the diboronic acid molecules in the multiple calibration curves of the self-calibration platform is 10 μM to 100 mM; The concentration range of bisphenol dyes shown in the multiple calibration curves in the self-calibration platform is 10 μM to 100 mM.

6. The glucose self-calibration detection method according to claim 1, characterized in that, The detectable concentration of the glucose self-calibration detection method is 1~30 mM.

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