A selective detection method for non-therapeutic isomers of glucose

Abstract

This invention discloses a selective detection method for isomers of glucose for non-therapeutic purposes. The selective detection method includes: mixing a mass spectrometry probe with methanol and a glucose-containing sample, followed by testing using high-resolution electrospray mass spectrometry (HS-MS). The selective detection of glucose is achieved based on the characteristic mass-to-charge ratio signal peak generated after the mass spectrometry probe binds to glucose. The selective detection method provided by this invention has advantages such as fast analysis speed, high detection sensitivity, accurate detection of compound molecular weight, and simple spectral analysis. It can also achieve highly selective detection of glucose in complex systems. This detection method does not require sample separation and can directly detect glucose in human urine, fruit juice, and honey, exhibiting advantages such as high selectivity, high reproducibility, and ease of operation.

Description

Technical Field

[0001] This invention belongs to the technical field, specifically relating to a selective detection method for isomer glucose for non-diagnostic purposes. Background Technology

[0002] Glucose is one of the most widely distributed and most important monosaccharides in nature. It is the primary energy source for living cells and plays a vital role in biology. The concentration of glucose in body fluids is an important indicator for the clinical diagnosis and monitoring of diabetes. Furthermore, the determination of glucose content in fruits, vegetables, and foods is also an indicator for judging their quality and can serve as a reference for fruit and vegetable cultivation and food quality evaluation. Therefore, glucose detection is of great significance to clinical diagnosis, the food industry, and other related sectors.

[0003] In light of this, various methods have been developed for the highly selective and sensitive detection of glucose, such as electrochemical methods, colorimetric analysis, fluorescent probe methods, and nuclear magnetic resonance probe methods. However, electrochemical methods are mainly based on glucose enzyme analysis and are easily affected by the surrounding chemical environment; colorimetric analysis and fluorescent probe methods require probe molecules to have corresponding chromophores, and the synthesis process is relatively cumbersome; while the detection sensitivity of nuclear magnetic resonance technology needs to be improved.

[0004] High-resolution mass spectrometry (HMS) can rapidly and accurately identify the molecular weight of organic compounds with high sensitivity, and is widely used in the structural identification of organic compounds. Mass spectrometry does not require a luminescent group; it completes qualitative and quantitative analysis of target compounds through accurate determination of the mass-to-charge ratio. Furthermore, mass spectrometry avoids interference from factors such as autofluorescence and light scattering in spectral analysis, reducing false positive results. However, HMS cannot distinguish organic compounds with the same molecular formula but identical molecular weight. For example, sugars with the same molecular weight as glucose include galactose, mannose, and fructose, which coexist with glucose in human fluids or food; therefore, glucose cannot be directly detected by HMS. Therefore, providing a method for selectively detecting isomers of glucose using HMS is an urgent problem to be solved. Summary of the Invention

[0005] The main objective of this invention is to provide a selective detection method for isomer glucose for non-diagnostic purposes, in order to overcome the shortcomings of the prior art.

[0006] To achieve the aforementioned objectives, the technical solution adopted by this invention includes:

[0007] This invention provides a method for selective detection of glucose isomers for non-diagnostic purposes, comprising: mixing a mass spectrometry probe with methanol and a sample containing glucose, and then performing a test using high-resolution electrospray mass spectrometry; and achieving selective detection of glucose based on the characteristic mass-to-charge ratio signal peak generated after the mass spectrometry probe binds to glucose.

[0008] The mass spectrometry probe has a structure as shown in formula (I):

[0009]

[0010] Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7.

[0011] This invention also provides a mass spectrometry probe for detecting isomer glucose, the mass spectrometry probe having a structure as shown in formula (I):

[0012]

[0013] Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7.

[0014] The present invention also provides a method for preparing the aforementioned mass spectrometry probe for detecting isomer glucose, comprising: reacting R-3-aminophenylboronic acid with diacyl chloride to obtain the mass spectrometry probe for detecting isomer glucose.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0016] (1) The selective detection method for isomer glucose provided by the present invention has the advantages of fast analysis speed, high detection sensitivity, accurate detection of molecular weight of compound and simple spectrum analysis. At the same time, it can realize the high selectivity detection of glucose in complex systems. The detection method does not require sample separation or other operations, and can directly detect glucose in human urine, fruit juice and honey. It has the advantages of high selectivity, fast analysis speed, high linear reproducibility and simple operation.

[0017] (2) The preparation method of the mass spectrometry probe used in this invention is simple and efficient. The reaction can be carried out at room temperature and has the advantages of easy separation and purification and readily available raw materials. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is the HRMS spectrum of the mass spectrometry probe 1 prepared in Example 1 of this invention;

[0020] Figures 2a-2c This is the HRMS spectrum of glucose in methanol solution, mass spectrometry probe 1 mixed with various sugars in Example 5 of the present invention;

[0021] Figure 3 This is a standard curve of the peak intensity of glucose concentration versus characteristic mass-to-charge ratio (m / z 515.1776±0.0025) in Example 6 of the present invention. Detailed Implementation

[0022] In view of the deficiencies of the prior art, the inventors of this invention, through long-term research and extensive practice, have proposed the technical solution of this invention. The main point is that the target analyte is simply added to a mass spectrometry probe solution that can selectively detect glucose for high-resolution mass spectrometry testing. The presence of glucose can be accurately and quickly identified by the generated characteristic mass-to-charge ratio (M+Na+108, where M is the molecular weight of the probe). The concentration of glucose can be calculated by the characteristic peak intensity-concentration relationship standard curve.

[0023] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0024] Specifically, as one aspect of the technical solution of this invention, a method for selective detection of isomer glucose for non-diagnostic purposes includes:

[0025] The mass spectrometry probe is mixed with methanol and a glucose-containing sample, and then tested using high-resolution electrospray mass spectrometry. Based on the characteristic mass-to-charge ratio signal peak generated after the mass spectrometry probe binds to glucose, selective detection of glucose is achieved.

[0026] The mass spectrometry probe has a structure as shown in formula (I):

[0027]

[0028] Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7.

[0029] In some preferred embodiments, the selective detection method specifically includes:

[0030] Using the standard curve method, a series of glucose solutions of different concentrations were added to a methanol solution containing a mass spectrometry probe, and then tested by high-resolution electrospray mass spectrometry (HRMS-ESI) to obtain the characteristic mass-to-charge ratio signal peak intensity (m / z M+Na+108, where M is the molecular weight of the probe) generated by the binding of the mass spectrometry probe and glucose. By utilizing the relationship between glucose concentration and characteristic mass-to-charge ratio signal peak intensity, a standard curve of glucose concentration-characteristic mass-to-charge ratio signal peak intensity was obtained.

[0031] The sample containing glucose was mixed with a methanol solution containing a mass spectrometry probe, and then detected using a high-resolution electrospray mass spectrometer under the same conditions. The intensity of the characteristic mass-to-charge ratio signal peak of the sample was obtained and compared with the standard curve to obtain the concentration of glucose in the sample.

[0032] In some preferred embodiments, the concentration of the mass spectrometry probe in the methanol solution containing the mass spectrometry probe is 1–10 mM;

[0033] In some preferred embodiments, when using high-resolution electrospray mass spectrometry for testing, the injection volume and injection rate of the mass spectrometer injection needle are the same each time.

[0034] Furthermore, the injection volume is 20–300 μL.

[0035] Furthermore, the injection rate is 20–400 μL / h.

[0036] In some preferred embodiments, the sample to be tested includes, but is not limited to, fruit juice, honey, or human bodily fluids.

[0037] Furthermore, the sample to be tested is honey.

[0038] The mass spectrometry probe prepared in this invention links two phenylboronic acid molecules together via amide bonds. Without altering the probe's selectivity, this increases both its solubility and stability after binding to glucose. This is because atoms in the amide bond units of the linker in the resulting cyclic phenylboronic ester (the source of the characteristic peak) can form intramolecular hydrogen bonds with glucose units, which is beneficial to the stability of the cyclic phenylboronic ester. Furthermore, the R groups in the probe molecule are all electron-donating substituents. The introduction of these substituents increases the electron cloud density in the system, alleviating the electron-deficient state of boron atoms in the corresponding cyclic phenylboronic ester, thus further promoting its stability. Through the combined stabilizing effect of the linker and the corresponding substituents on the corresponding cyclic phenylboronic ester, a stable glucose characteristic signal peak (M+Na+108, where M is the probe molecular weight) is generated in the mass spectrometer, enabling highly selective recognition of glucose.

[0039] Another aspect of the present invention provides a mass spectrometry probe for detecting isomer glucose, the mass spectrometry probe having a structure as shown in formula (I):

[0040]

[0041] Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7.

[0042] In this invention, the synthesis route of the mass spectrometry probe with the structure shown in formula (I) is as follows:

[0043]

[0044] Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7.

[0045] In some preferred embodiments, the mass spectrometry probe has any of the following structures:

[0046]

[0047] Another aspect of the present invention provides a method for preparing the aforementioned mass spectrometry probe for detecting isomer glucose, comprising: reacting R-3-aminophenylboronic acid with diacyl chloride to obtain the mass spectrometry probe for detecting isomer glucose.

[0048] In some preferred embodiments, the preparation method of the mass spectrometry probe includes: mixing R-3-aminophenylboronic acid with a first solvent and cooling to -10 to 10°C to form an R-3-aminophenylboronic acid solution; simultaneously mixing diacyl chloride with a second solvent to form a diacyl chloride solution; then adding the diacyl chloride solution dropwise to the R-3-aminophenylboronic acid solution and stirring at 0 to 50°C for 12 to 24 hours; followed by post-processing to obtain the mass spectrometry probe for detecting the isomer glucose. In some preferred embodiments, the R-3-aminophenylboronic acid has the structure shown in formula (II):

[0049]

[0050] R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl.

[0051] In some preferred embodiments, the diacyl chloride has the structure shown in formula (III):

[0052]

[0053] Where n is selected from positive integers from 1 to 7.

[0054] In some preferred embodiments, the first solvent includes any one or a combination of two or more of the following: aqueous sodium hydroxide solution, aqueous potassium hydroxide solution, aqueous sodium carbonate solution, and aqueous potassium carbonate solution, and is not limited thereto.

[0055] In some preferred embodiments, the second solvent includes any one or a combination of two or more of methanol, ethanol, acetonitrile, diethyl ether, toluene, and tetrahydrofuran, and is not limited thereto.

[0056] In some preferred embodiments, the molar ratio of the diacyl chloride to R-3-aminophenylboronic acid is 1:2 to 1:3.

[0057] In some preferred embodiments, the post-processing includes: after the reaction is completed, adjusting the pH of the obtained mixture to 1-3, and then washing and drying it to obtain the mass spectrometry probe for detecting isomer glucose.

[0058] Furthermore, the pH of the obtained mixture is adjusted to 1-3 using an acidic substance, wherein the acidic substance includes hydrochloric acid.

[0059] Furthermore, the solvent used for the washing includes ultrapure water.

[0060] Furthermore, the drying process is carried out at a temperature of 25–200°C for a duration of 12–48 hours.

[0061] The technical solution of the present invention will be further described in detail below with reference to several preferred embodiments and accompanying drawings. This embodiment is implemented on the premise of the technical solution of the invention, and provides detailed implementation methods and specific operation processes. However, the protection scope of the present invention is not limited to the following embodiments.

[0062] Unless otherwise specified, the experimental materials used in the examples below can be purchased from conventional biochemical reagent companies.

[0063] Example 1

[0064] 3-Amino-5-methylphenylboronic acid (181.16 mg, 1.2 mmol) was dissolved in 1 M NaOH aqueous solution and cooled to 0 °C. Succinyl chloride (33 μL, 0.3 mmol) was dissolved in THF solution and then added dropwise to the NaOH solution. The mixture was then stirred at room temperature for 12 h. The pH of the solution was adjusted to 2 with 1 M HCl, resulting in a light brown precipitate. This precipitate was filtered, washed three times with water, and then dried under vacuum at 80 °C for 12 h to obtain mass spectrometry probe 1 with a yield of 86%. Mass spectrometry probe 1 has the structure shown in the following formula:

[0065]

[0066] Performance characterization:

[0067] 1 H NMR (400MHz, DMSO): δ10.03(s,2H),8.05(s,4H),7.54-7.05(m,6H)2.65(s,4H),2.36(s,6H).

[0068] 13 C NMR (100MHz, DMSO): δ177.4,138.6,138.4,131.6,130.7,125.4,117.6,31.6,21.7.

[0069] 11 B NMR (128MHz, DMSO): δ 19.8.

[0070] HRMS(ESI)(m / z):C 22 H 30 B₂N₂O₆Na[M+4CH₃OH-4H₂O+Na] + Calculated value: 463.2188, Actual value: 463.2176.

[0071] The HRMS spectrum of mass spectrometry probe 1 prepared in this embodiment is shown in the figure below. Figure 1 As shown.

[0072] Example 2

[0073] 3-Amino-5-hydroxyphenylboronic acid (244.71 mg, 1.6 mmol) was dissolved in 1 M NaOH aqueous solution and cooled to 0 °C. Succinyl chloride (35 μL, 0.4 mmol) was dissolved in THF solution and then added dropwise to the NaOH solution. The mixture was then stirred at room temperature for 24 h. The pH of the solution was adjusted to 2 with 1 M HCl, resulting in a light yellow precipitate. This precipitate was filtered, washed three times with water, and then dried under vacuum at 120 °C for 12 h to obtain mass spectrometry probe 2 with a yield of 96%. Mass spectrometry probe 2 has the structure shown in the following formula:

[0074]

[0075] Performance characterization:

[0076] 1 H NMR (400MHz, DMSO): δ10.01(s,2H),9.45(s,2H),8.02(s,4H),7.14-6.64(m,6H)2.65(s,4H).

[0077] 13 C NMR (100MHz, DMSO): δ176.8,158.7,139.9,133.1,113.2,111.5,105.131.6.

[0078] 11 B NMR (128MHz, DMSO): δ 19.6.

[0079] HRMS(ESI)(m / z):C 20 H 26 B₂N₂O₈Na[M+4CH₃OH-4H₂O+Na] + Calculated value: 463.2188, Actual value: 467.1773.

[0080] Example 3

[0081] 3-Amino-5-methoxyphenylboronic acid (250.46 mg, 1.5 mmol) was dissolved in 1 M KOH aqueous solution and cooled to 0 °C. Succinyl chloride (42 μL, 0.38 mmol) was dissolved in THF solution and then added dropwise to the KOH solution. The mixture was then stirred at room temperature for 24 h. The pH of the solution was adjusted to 2 with 1 M HCl, resulting in a pale yellow precipitate. This precipitate was filtered, washed three times with water, and then dried under vacuum at 100 °C for 12 h to obtain mass spectrometry probe 3 with a yield of 88%. Mass spectrometry probe 3 has the structure shown in the following formula:

[0082] Performance characterization:

[0083] 1 H NMR (400MHz, DMSO): δ10.03(s,2H),8.04(s,4H),7.33-6.79(m,6H),3.81(s,6H),2.65(s,4H).

[0084] 13 C NMR (100MHz, DMSO): δ177.3,160.8,139.5,132.7,112.9,110.2,109.9,55.8,31.6.

[0085] 11 B NMR (128MHz, DMSO): δ 18.6.

[0086] HRMS(ESI)(m / z):C 22 H 30 B₂N₂O₈Na[M+4CH₃OH-4H₂O+Na] + Calculated value: 495.2086, Actual value: 495.2106.

[0087]

[0088] Example 4

[0089] 3-Amino-5-methoxyphenylboronic acid (200.36 mg, 1.2 mmol) was dissolved in 1 M KOH aqueous solution and cooled to 0 °C. Octanoyl chloride (54 μL, 0.3 mmol) was dissolved in THF solution and then added dropwise to the KOH solution. The mixture was then stirred at room temperature for 24 h. The pH of the solution was adjusted to 2 with 1 M HCl, resulting in a pale yellow precipitate. This precipitate was filtered, washed three times with water, and then dried under vacuum at 120 °C for 12 h to obtain mass spectrometry probe 4 with a yield of 75%. Mass spectrometry probe 4 has the structure shown in the following formula:

[0090]

[0091] Performance characterization:

[0092] 1 H NMR (400MHz, DMSO): δ10.03(s,2H),8.04(s,4H),7.36-6.89(m,6H),3.82(s,6H),2.35(t,4H),1.63-1.35(m,8H).

[0093] 13C NMR (100MHz, DMSO): δ179.8,160.6,139.4,132.1,112.3,110.8,109.9,55.5,38.3,27.9,25.6.

[0094] 11 B NMR (128MHz, DMSO): δ 18.8.

[0095] HRMS(ESI)(m / z):C 26 H 38 B₂N₂O₈Na[M+4CH₃OH-4H₂O+Na] + Calculated value: 551.2712, Actual value: 551.2738.

[0096] Example 5

[0097] 100 μL of 6 mM glucose and a mixed solution of mannose, fructose and galactose were added to 500 μL of 6 mM methanol solution of mass spectrometry probe 1 prepared in Example 1. After incubation at room temperature for the same time, high-resolution mass spectrometry experiments were performed (Figure 2). When glucose was added to the probe, a characteristic mass-to-charge ratio m / z of 515.1776 ± 0.0025 (M + Na + 108) was generated after glucose and probe molecule 1 were bound, indicating that the mass spectrometry probe can selectively recognize glucose. Figures 2a-2c The images show HRMS spectra of glucose in methanol solution, mass spectrometry probe 1 + glucose, and mass spectrometry probe 1 + a mixed solution of fructose, galactose, and mannose, respectively.

[0098] Example 6

[0099] Standard solutions were prepared by adding 100 μL of a series of glucose concentration solutions (6, 12, 18, 24, 30, 36, 48, 60 mM) to 500 μL of a 6 mM methanol solution of the mass spectrometry probe 1 prepared in Example 1, resulting in a series of 5 mM mass spectrometry probe 1 solutions containing different glucose concentrations (1, 2, 3, 4, 5, 6, 8, 10 mM). These solutions were then tested on a high-resolution time-of-flight mass spectrometer. A linear curve relationship was established between the peak intensity of the characteristic mass-to-charge ratio m / z 515.1776 ± 0.0025 (M + Na + 108) after glucose binding to the probe molecule and the glucose concentration, thus obtaining the standard curve. Figure 3 ): y = 3657x – 123.2, correlation coefficient R 2=0.9962, where y is the mass spectrum peak intensity at m / z 515.1776±0.0025; x is the actual concentration of glucose solution after mixing with mass spectrometry probe 1. According to the standard curve, 100 μL of 6, 12, and 24 mM glucose solutions were added to 500 μL of a 6 mM methanol solution of mass spectrometry probe 1 molecules, respectively, to obtain actual glucose concentrations of 1, 2, and 4 mM, under the same test conditions as the standard curve. High-resolution mass spectra were obtained, and the tests were performed in parallel three times. The intensity value of the characteristic mass-to-charge ratio (m / z 515.1776±0.0025) was read from the spectrum, and the concentration of glucose in the aqueous solution was calculated according to the standard curve, as shown in Table 1.

[0100] Table 1. Concentration of glucose in aqueous solution

[0101]

[0102] Example 7

[0103] Prepare 100 mL of a mixture solution containing 100 mM mannose, fructose, and galactose. Then, use this solution to dilute a 600 mM glucose solution to obtain mixed solutions containing 6, 30, and 60 mM glucose. Take 100 μL of each of the diluted mixed solutions and add them to 500 μL of a 6 mM methanol solution of mass spectrometry probe 1. The actual glucose concentrations in the mixed solutions are 1, 5, and 10 mM. Obtain high-resolution mass spectra under the same test conditions as the test standard curve. Perform the test three times in parallel and read the intensity values ​​of the characteristic mass-to-charge ratio from the spectra. Calculate the glucose concentration in the mixed solutions based on the standard curve, as shown in Table 2.

[0104] Table 2. Concentration of glucose in mixed sugar solution

[0105] Theoretical glucose value (mM) Glucose test value (%) RSD / % 1.0 1.01 3.8 5.0 5.13 2.7 10.0 9.98 1.6

[0106] Example 8

[0107] Accurately weigh 1g of honey sample and dilute it with ultrapure water to a 100mL volumetric flask. Add 100μL of this solution to 500μL of a 6mM methanol solution of mass spectrometry probe 1. Obtain high-resolution mass spectra under the same test conditions as the test standard curve. Perform the test in parallel three times, read the intensity value of the characteristic mass-to-charge ratio from the spectrum, calculate the glucose concentration according to the standard curve, and calculate the glucose content in the honey according to the sample weight. Compare the results with the glucose content in the honey sample determined by liquid chromatography (LC). The test results of the two methods are consistent (see Table 3), indicating that this method has good precision and reliability in detecting glucose in honey.

[0108] Table 3. Glucose concentration in urine

[0109]

[0110] Examples 6-8 demonstrate that the detection method provided by the present invention has the advantages of high selectivity and high reproducibility.

[0111] In addition, the inventors of this case also conducted experiments with other raw materials, process operations, and process conditions described in this specification, referring to the aforementioned embodiments, and obtained relatively ideal results in all cases.

[0112] It should be understood that the technical solutions of the present invention are not limited to the specific embodiments described above. Any technical modifications made to the technical solutions of the present invention without departing from the spirit and scope of the claims are within the scope of protection of the present invention.

Claims

1. A method for the selective detection of isomer glucose for non-diagnostic purposes, characterized in that, include: The mass spectrometry probe is mixed with methanol and a glucose-containing sample, and then tested using high-resolution electrospray mass spectrometry. Based on the characteristic mass-to-charge ratio signal peak generated after the mass spectrometry probe binds to glucose, selective detection of glucose is achieved. The isomers of glucose include any one or more combinations of mannose, fructose, and galactose. The mass spectrometry probe has a structure as shown in formula (I): ； Formula (I); Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7.

2. The selective detection method according to claim 1, characterized in that, Specifically, it includes: Using the standard curve method, a series of glucose solutions of different concentrations were added to a methanol solution containing a mass spectrometry probe, and then tested by high-resolution electrospray mass spectrometry to obtain the characteristic mass-to-charge ratio signal peak intensity generated by the binding of the mass spectrometry probe and glucose. By utilizing the relationship between glucose concentration and characteristic mass-to-charge ratio signal peak intensity, a standard curve of glucose concentration-characteristic mass-to-charge ratio signal peak intensity was obtained. The sample containing glucose was mixed with a methanol solution containing a mass spectrometry probe, and then detected using a high-resolution electrospray mass spectrometer under the same conditions. The intensity of the characteristic mass-to-charge ratio signal peak of the sample was obtained and compared with the standard curve to obtain the concentration of glucose in the sample.

3. The selective detection method according to claim 2, characterized in that: The concentration of the mass spectrometry probe in the methanol solution containing the mass spectrometry probe is 1 ~ 10 mM; And / or, when using high-resolution electrospray mass spectrometry for testing, the injection volume and injection rate of the mass spectrometer injection needle are the same for each test; wherein, the injection volume is 20~300 μL; and the injection rate is 20-400 μL / h.

4. The selective detection method according to claim 1, characterized in that: The samples to be tested include fruit juice, honey, or human bodily fluids.

5. A mass spectrometry probe for detecting isomers of glucose, characterized in that, The mass spectrometry probe has the structure shown in formula (I): ； Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxyl, and n is selected from positive integers from 1 to 7; wherein, isomers of glucose include any one or more combinations of mannose, fructose, and galactose.

6. The mass spectrometry probe for detecting isomer glucose according to claim 5, characterized in that, The mass spectrometry probe has any of the compounds with the structure shown in the following formula: 。 7. The method for preparing the mass spectrometry probe for detecting isomer glucose as described in claim 5 or 6, characterized in that, include: The mass spectrometry probe for detecting the isomer glucose was prepared by reacting R-3-aminophenylboronic acid with diacyl chloride.

8. The preparation method according to claim 7, characterized in that, Specifically, it includes: R-3-aminophenylboronic acid was mixed with a first solvent and cooled to -10 to 10°C to form an R-3-aminophenylboronic acid solution. Simultaneously, diacyl chloride was mixed with a second solvent to form a diacyl chloride solution. The diacyl chloride solution was then added dropwise to the R-3-aminophenylboronic acid solution and stirred at 0 to 50°C for 12 to 24 hours. After post-processing, the mass spectrometry probe for detecting isomer glucose was obtained.

9. The preparation method according to claim 8, characterized in that, The R-3-aminophenylboronic acid has the structure shown in formula (II): ； Wherein, R is selected from any one of methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, and hydroxy; And / or, the diacyl chloride has the structure shown in formula (III): ； Where n is selected from positive integers from 1 to 7; And / or, the first solvent includes any one or a combination of two or more of the following: aqueous sodium hydroxide solution, aqueous potassium hydroxide solution, aqueous sodium carbonate solution, and aqueous potassium carbonate solution; And / or, the second solvent includes any one or a combination of two or more of methanol, ethanol, acetonitrile, diethyl ether, toluene, and tetrahydrofuran.

10. The preparation method according to claim 7, characterized in that: The molar ratio of the diacyl chloride to R-3-aminophenylboronic acid is 1:2 to 1:

3.

11. The preparation method according to claim 8, characterized in that, The post-processing includes: after the reaction is completed, adjusting the pH of the obtained mixture to 1-3, and then washing and drying it to obtain the mass spectrometry probe for detecting isomer glucose.

12. The preparation method according to claim 11, characterized in that: The pH of the obtained mixture is adjusted to 1-3 using an acidic substance, wherein the acidic substance includes hydrochloric acid.

13. The preparation method according to claim 11, characterized in that: The solvent used for the washing includes ultrapure water.

14. The preparation method according to claim 11, characterized in that: The drying process is carried out at a temperature of 25~200℃ for a time of 12~48h.

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