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CN-122016748-A - Glucose detection method and application

CN122016748ACN 122016748 ACN122016748 ACN 122016748ACN-122016748-A

Abstract

The application relates to the technical field of glucose detection, in particular to a glucose detection method and application. The glucose detection method comprises the following detection steps of S1, preparing a sodium borate solution and a guanidine solution, S2, weighing glucose, preparing glucose working solutions with different concentrations, adding the sodium borate solution and the guanidine solution in the step S1 into the glucose working solution, heating in a water bath, cooling to a constant volume of 10mL, performing fluorescence detection, taking the logarithmic value of concentration and fluorescence intensity as a linear regression equation according to the detection result to obtain a working curve, S3, adding the sodium borate solution and the guanidine solution in the step S1 into the solution to be detected, heating in a water bath, measuring the fluorescence intensity, and substituting the detection result into the working curve in the step S2 to calculate the glucose concentration in the solution to be detected. The glucose detection method has the characteristics of high sensitivity and high accuracy, and is favorable for wide application.

Inventors

  • HUANG ZIXUAN
  • ZHANG HAIRONG
  • ZHANG MINGXIA
  • Gu Wanlu

Assignees

  • 广州华商学院

Dates

Publication Date
20260512
Application Date
20260227

Claims (10)

  1. 1. A method of glucose detection, the method comprising the steps of: s1, preparing a sodium borate solution and a guanidine solution; s2, weighing glucose to prepare glucose working solutions with different concentrations, adding the sodium borate solution and the guanidine solution in the step S1 into the glucose working solution, heating in a water bath, cooling to a constant volume of 10mL, performing fluorescence detection, and taking the logarithmic value of the concentration and the fluorescence intensity as a linear regression equation according to the detection result to obtain a working curve; and S3, adding the sodium borate solution and the guanidine solution in the step S1 into the solution to be detected, heating in a water bath, measuring the fluorescence intensity, substituting the detection result into the working curve in the step S2, and calculating to obtain the glucose concentration in the solution to be detected.
  2. 2. The method according to claim 1, wherein in step S1, the concentration of the sodium borate solution is 0.04 to 0.2mol/L.
  3. 3. The method according to claim 1, wherein in step S1, the concentration of the guanidine solution is 0.002-0.04mol/L.
  4. 4. The method according to claim 1, wherein in step S2 and step S3, the water bath is heated for 30-180min.
  5. 5. The method according to claim 4, wherein in step S2 and step S3, the water bath is heated for 50-150min.
  6. 6. The method according to claim 1, wherein in step S2, the operation curve is InI f = -0.4877InC+9.3259; wherein InI f is fluorescence intensity, and InC is glucose concentration.
  7. 7. The method according to claim 1, wherein in the step S3, the volume ratio of the solution to be measured to the sodium borate solution and the guanidine solution is (1-2): 1:1.
  8. 8. The method according to claim 7, wherein in the step S3, the volume ratio of the solution to be tested to the sodium borate solution and the guanidine solution is 1:1:1.
  9. 9. Use of the glucose assay according to any one of claims 1-8 in food testing.
  10. 10. The method for detecting the glucose content in the food is characterized by comprising the following detection steps of: processing food to be detected to obtain a solution to be detected, detecting by the detection method according to any one of claims 1-8 to obtain the glucose concentration in the solution to be detected, and calculating to obtain the glucose content of the food to be detected.

Description

Glucose detection method and application Technical Field The application relates to the technical field of glucose detection, in particular to a glucose detection method and application. Background Glucose is a substance widely existing in nature, plays a very important role in plant and animal bodies, and has stable content, which is an important factor for whether human body is healthy or not, and if the content is too low, the glucose is low, dizziness and hypodynamia are caused, and if the content is too high, the glucose is the diabetes of people, so that the research of the property of the glucose is of great significance. However, the detection of saccharides is difficult because the saccharides have neither strong UV absorption nor fluorescent groups, and are typically electrically neutral. Detection of saccharides typically requires first derivatization to provide UV absorption, fluorescence properties, or some other measurable physicochemical property. One of the important derivatization is the reaction of glucose and amine substances, and the reaction with amine compounds is one of the first derivatization methods used for determining saccharides, and the principle is that-NH 2 reacts with a functional group of the saccharides, namely carbonyl (aldehyde group or ketone group), to generate-C=N. This type of method can be classified into the following ones according to the derivatizing agents. ⑴ The reaction with alkylamine and aromatic amine, honda, etc. shows for the first time that reducing sugar and ethylenediamine are co-heated in a slightly alkaline phosphate buffer solution to generate strong fluorescence, and this finding can be used for measuring reducing sugar. The detection limit of the derivatization method for most sugars is less than 1nmol. Later, kato and Kinoshita derivatized the sugar with ethanolamine and 2-aminopropionitrile, and then assayed by fluorescence. (2 Reductive amination derivatization reaction: the principle of reductive amination derivatization reaction is that the reduced terminal carbonyl group (aldehyde, ketone) of sugar reacts with aromatic compound containing primary amino group, after Shift base is formed, it is reduced to secondary amine by NaBH 3 CN, and then detected by ultraviolet absorption or fluorescence (3) Reacting with amide compound, amide compound is a derivatization reagent of sugar which is used earlier and has wider application. Of these, 2-cyanoacetamide is typical, which co-produces a strong fluorescent substance with aldose in a weakly alkaline solution, and this reaction is suitable for post-column derivatization, and the product can also be detected by a photometric method, with a limit of detection of aldose of up to 0.1nmol, with which the isomer can be resolved at low temperatures. (4) Derivatization of acid sugar Mechref and EI Rassi propose a method for derivatization of acid sugar based on the principle that amino group of p-aminobenzenesulfonic acid (SA) or 7-aminonaphthalene-1, 3-disulfonic acid (ANDSA) and carbonyl group of acid sugar are condensed into one peptide chain in the presence of water-soluble carbodiimide (HN: C: NH) and detected by UV. (5) Reaction with guanidine and its derivatives guanidine has two-NH 2 and reacts with sugar in the presence of borate to form imidazole ring (as shown in formula 1), and sugar can be detected by fluorescence of imidazole ring. The detection limit was 5pmol of pentose and 6pmol of hexose. The reaction is easier than the reaction of vinylamino derivative and sugar, and has high sensitivity and good reproducibility. (Formula 1). Two compounds with similar structures to guanidine, arginine and benzamidine, are good fluorescent probes for saccharides. Arginine derivatization conditions are mild and similar to the reaction of 2-cyanoacetamide and fatty amine in boric acid medium. The fluorescence intensities of glucose and fructose were similar, the glucose detection limit was 25pmol, sucrose and raffinose gave only 4% fluorescence intensity corresponding to glucose, whereas 2-deoxyglucose did not. Benzamidine and its para-methoxy derivatives are highly sensitive when used as derivatizing agents, but the reaction is carried out in a strong alkaline medium. At present, a glucose detection method with mild reaction conditions and high sensitivity is needed. Disclosure of Invention The application aims to provide a glucose detection method and application thereof, which are used for solving the technical problem of how to efficiently and accurately detect glucose. The detection method provided by the application has the advantages that the fluorescence emission is very good after the derivatization reaction of glucose and guanidine, the concentration of glucose is detected through the fluorescence intensity, the relative standard deviation meets the requirements in the same test, the sensitivity is high, the good linear relationship is shown in a certain concentration range, and the accurate and e