CN109115760B - Rapid detection kit and detection method for urine sugar content based on chloroauric acid color reaction - Google Patents
Rapid detection kit and detection method for urine sugar content based on chloroauric acid color reaction Download PDFInfo
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- CN109115760B CN109115760B CN201811023183.3A CN201811023183A CN109115760B CN 109115760 B CN109115760 B CN 109115760B CN 201811023183 A CN201811023183 A CN 201811023183A CN 109115760 B CN109115760 B CN 109115760B
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The invention discloses a rapid detection kit and a detection method for urine glucose content based on chloroauric acid color reaction, wherein the kit comprises a chloroauric acid reagent bottle and a urine glucose concentration colorimetric card, and can also comprise a urine glucose concentration accurate quantitative card.
Description
Technical Field
The invention belongs to the technical field of detection of urine sugar, and particularly relates to a urine sugar rapid detection kit based on color development of gold ions in an oxidation-reduction reaction process, and a method for rapidly and qualitatively detecting the content of urine sugar by using the kit.
Background
Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by a defect in insulin secretion or an impaired biological action, or both. Diabetes mellitus is a long-standing hyperglycemia, causes chronic damage and dysfunction of various tissues, particularly eyes, kidneys, heart, blood vessels and nerves, and is one of the most known complications at present.
At present, the diagnosis and monitoring of diabetes mainly comprises the detection of blood sugar and urine sugar concentration, wherein the blood sugar detection is generally carried out by a hospital for blood drawing test or a household blood glucose meter for self-test, the two methods both need the patients to suffer from the pain of puncturing, and particularly, the patients with confirmed diabetes need to monitor urine sugar every day, which brings extra burden to the bodies and the minds of the patients; the current commonly used method for detecting urine sugar comprises a qualitative detection method of the urine sugar of the speckles, a urine sugar test paper method and the like, and the method is used for carrying out semi-quantitative analysis on the content of glucose in urine by colorimetry. Due to the difference of the color sensitivity of different individuals, the detection method by taking color comparison as a means has strong subjectivity, and particularly has low judgment reliability on the color near the critical value, so that patients with urine glucose values near the critical value need to be rechecked at home or in hospitals, and the time and economic cost of the patients and even medical systems are increased.
Nano-gold particles, also known as colloidal gold. At present, various gold nanoparticles with different sizes (1.0-50nm) are prepared by a chemical agent synthesis method, solutions of the gold nanoparticles sequentially show blue, purple red, peach red and wine red with different degrees according to different sizes of the gold nanoparticles, corresponding peak absorbance can be detected near 520nm by an ultraviolet visible spectrophotometer, and the wavelength corresponding to the peak absorbance is in a linear relation with the particle size of the gold nanoparticles.
Glucose (glucose) is an organic compound of formula C6H12O6The monosaccharide is a monosaccharide which is most widely distributed and important in the nature, is polyhydroxy aldehyde, has reducibility, and can be used as a reducing agent for synthesizing nano gold particles. The glucose solution reacts with gold ions to generate gold nanoparticles with different particle sizes, the glucose concentration and the size of the gold nanoparticles have a linear relation, and the solution shows different colors according to the size of the particles.
Based on the characteristics, most of the existing urine sugar detection methods use a color reaction and a colorimetric card for reading, for example, CN102749327A uses a liquid chemical reagent to react with urine sugar to color to judge the approximate urine sugar concentration, and the sensitivity is 1 mM; CN104535753A was developed by reaction with urine glucose using a test paper and the concentration of urine glucose was judged by combining with a colorimetric plate, and the sensitivity was 0.5 mM. There is no correlation research result for accurately detecting the concentration of urine sugar by using a urine sugar reducing gold ion color reaction and colorimetric qualitative determination of the concentration of urine sugar, or using a curve fitting formula for correlation between the concentration of urine sugar and the size of nano-gold particles.
Disclosure of Invention
The invention aims to provide a rapid urine glucose detection kit based on a chloroauric acid color reaction and a method for qualitatively and quantitatively detecting the urine glucose content by adopting the kit. By adopting the kit and the detection method, the application field of the color reaction for preparing the nano-gold by the chloroauric acid is expanded; meanwhile, the efficiency of qualitative and quantitative detection of the urine glucose is effectively improved.
Aiming at the purposes, the urine glucose rapid detection kit adopted by the invention comprises a chloroauric acid reagent bottle and a urine glucose concentration colorimetric card; wherein the chloroauric acid reagent bottle is a disposable covered sealed glass bottle filled with 2-5 mL of chloroauric acid color developing solution, and the chloroauric acid color developing solution is prepared by adjusting the pH value of 0.5mM chloroauric acid aqueous solution to 7-9 by 0.2M sodium hydroxide aqueous solution; the urine glucose concentration colorimetric card consists of a color block A, a color block B, a color block C, a color block D, a color block E, a color block F, a color block G, a color block H and a color block I, wherein subscripts of the color block A and the color block B are more than or equal to 12mM, and the corresponding urine glucose concentration is more than or equal to 12 mM; 11-10mM is marked below the color blocks B and C, and the corresponding urine glucose concentration is 10-11 mM; 9-8mM is marked below the color blocks D and E, and the corresponding urine glucose concentration is 8-9 mM; 7-6mM is marked below the color blocks F and G, and the corresponding urine glucose concentration is 6-7 mM; 5-4mM is marked below the color blocks G and H, and the concentration of corresponding urine sugar is 4-5 mM; the color block H and the color block I are marked with less than or equal to 3mM below and the corresponding urine glucose concentration is less than or equal to 3 mM.
The manufacturing method of the urine glucose concentration colorimetric card comprises the following steps:
1. dissolving glucose dry powder with normal human urine to prepare 100mM glucose stock solution for diabetes, then diluting the glucose stock solution with normal human urine to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12mM respectively, and then sequentially diluting the glucose stock solution with deionized water by 20 times to prepare the glucose standard solution for urine.
2. According to the volume ratio of the urine sugar standard solution to the chloroauric acid color developing solution of 1:15, preheating the urine sugar standard solution through a boiling water bath, adding the preheated urine sugar standard solution into a chloroauric acid reagent bottle preheated through the boiling water bath, rapidly stirring for 5-10 seconds, and continuing to keep the boiling water bath for heating reaction for 5-6 minutes to obtain a nano gold solution; and manufacturing the urine glucose concentration colorimetric card according to the colors of the nano-gold solutions corresponding to the urine glucose standard solutions with different concentrations.
The capacity of the disposable capped and sealed glass bottle is 8mL, the height of the glass bottle is 70-80 mm, the diameter of the glass bottle is 10-15 mm, the glass bottle is made of a screw quartz glass material, and a heat-resistant leakage-proof gasket is additionally arranged inside the heat-resistant plastic material of the bottle cap.
In order to further accurately detect the content of the urine sugar, the kit can further comprise a urine sugar concentration accurate quantitative card, and the accurate quantitative card is written with a formula: Y-3E +44X-15.95(in the formula, E is a scientific notation, E + n represents the power of 10 to the power of n, and the formula expression can also be:
) The formula is obtained by the following method: dripping 1.5-2 mL of the nanogold solution reacted in the step 2 into a quartz cuvette, detecting the wavelength corresponding to the peak absorbance of the nanogold solution within the range of 500-600 nm by using an ultraviolet-visible spectrophotometer, and drawing a correlation curve of the wavelength corresponding to the peak absorbance and the glucose concentration to obtain the formula Y-3E +44X-15.95Wherein X represents the wavelength corresponding to the peak absorbance of the nano-gold solution within the range of 500-600 nm, and Y represents the concentration of glucose in the urine to be detected.
The method for rapidly detecting the content of the urine sugar by adopting the kit comprises the following steps: diluting the urine to be detected by 20 times by using deionized water to obtain a diluted solution of the urine to be detected; preheating the urine diluent to be detected by using a boiling water bath, adding the preheated urine diluent to be detected into a chloroauric acid reagent bottle preheated by the boiling water bath according to the volume ratio of the urine diluent to be detected to the chloroauric acid developing solution of 1:15, rapidly stirring for 5-10 seconds, then continuously keeping the boiling water bath for heating and reacting for 5-6 minutes to enable glucose in the urine and the chloroauric acid to generate oxidation-reduction developing reaction, comparing the color of the obtained nanogold solution with a urine glucose concentration colorimetric card after the reaction is finished, and judging the concentration range of the glucose.
After the concentration range of glucose is judged, 1.5-2 mL of nano-gold solution is dripped into a quartz cuvette, an ultraviolet-visible spectrophotometer is adopted to detect the wavelength corresponding to the peak absorbance within the range of 500-600 nm, and the formula Y on the accurate urine glucose concentration quantification card is 3E +44X-15.95To be detectedThe glucose concentration in the urine was quantitatively calculated.
The invention has the following beneficial effects:
1. according to the method, glucose in urine is used for reducing the tetrachloroauric acid, the content of urine glucose can be rapidly and qualitatively determined by comparing a colorimetric card according to different colors developed by the nano-gold solution, the steps are simple, and the reaction speed is high. The chloroauric acid reagent bottle is stored in a shade dark place and can be stored for half a year.
2. When the urine glucose content needs to be accurately detected, an ultraviolet-visible spectrophotometer is adopted, the glucose content in urine is accurately calculated through a correlation fitting formula of the wavelength of the peak absorbance of the nanogold solution and the glucose concentration in the urine, and the sensitivity is 0.01 mM.
3. The invention completes qualitative and quantitative detection of two kinds of urine sugar contents through one-time chemical color reaction, and effectively improves the urine sugar detection efficiency.
Drawings
FIG. 1 is a photograph of a urine glucose concentration colorimetric card in example 1.
FIG. 2 is a graph showing the results of absorbance detection of the nano-gold solution in the range of 450 to 700nm by an ultraviolet-visible spectrophotometer corresponding to the standard solutions of glucose in urine of different concentrations in example 1.
FIG. 3 is a graph of the correlation between the peak absorbance of the nanogold solution and the wavelength of the nanogold solution corresponding to the standard solution of urine glucose at different concentrations in example 1, and the fitting curve.
Detailed Description
The invention will be further described in detail with reference to the following figures and examples, but the scope of the invention is not limited to these examples.
Example 1
The kit for rapidly detecting the urine glucose content of the embodiment consists of a chloroauric acid reagent bottle, a urine glucose concentration colorimetric card and a urine glucose concentration accurate quantification card.
The chloroauric acid reagent bottle is a disposable capped sealed glass bottle filled with 3mL of chloroauric acid color developing solution, the chloroauric acid color developing solution is prepared by adjusting the pH value of 0.5mM aqueous solution of chloroauric acid to 8 with 0.2M aqueous solution of sodium hydroxide, the disposable capped sealed glass bottle has the capacity of 8mL, the height of 75mM and the diameter of 12mM, is made of a screw quartz glass material, and the bottle cap is made of a heat-resistant plastic material and is internally provided with a heat-resistant leakage-proof gasket.
The urine glucose concentration colorimetric card is shown in figure 1 and comprises a color block A, a color block B, a color block C, a color block D, a color block E, a color block F, a color block G, a color block H and a color block I, wherein the subscripts of the color block A and the color block B are more than or equal to 12mM, and the corresponding urine glucose concentration is more than or equal to 12 mM; 11-10mM is marked below the color blocks B and C, and the corresponding urine glucose concentration is 10-11 mM; 9-8mM is marked below the color blocks D and E, and the corresponding urine glucose concentration is 8-9 mM; 7-6mM is marked below the color blocks F and G, and the corresponding urine glucose concentration is 6-7 mM; 5-4mM is marked below the color blocks G and H, and the concentration of corresponding urine sugar is 4-5 mM; the color block H and the color block I are marked with less than or equal to 3mM below and the corresponding urine glucose concentration is less than or equal to 3 mM. The urine glucose concentration colorimetric card is manufactured by the following method:
1. weighing 0.18g of glucose dry powder, dissolving the glucose dry powder by using 10mL of normal human urine to prepare 100mM of grape diabetes stock solution, then respectively diluting the grape diabetes stock solution to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12mM by using the normal human urine, and sequentially diluting the grape diabetes stock solution by using deionized water for 20 times to prepare the urine glucose standard solution.
2. Preheating the standard urine sugar solution in a boiling water bath for 5 minutes, adding 200 mu L of the standard urine sugar solution into a chloroauric acid reagent bottle preheated in the boiling water bath for 5 minutes, rapidly stirring for 5 seconds, continuing to keep the boiling water bath for heating and reacting for 6 minutes until the color of the solution is stable, and ending the reaction, wherein the color of the mixture is changed from yellow to colorless, blue or red, which indicates that the nano gold solution is prepared; according to the color of the nano-gold solution corresponding to the standard urine glucose solution with different concentrations, a urine glucose concentration colorimetric card is manufactured, as shown in fig. 1.
The accurate quantitative card for the concentration of urine glucose is written with the formula: Y-3E +44X-15.95The formula is obtained according to the following method:
taking 1.5mL of each of the nanogold solutions reacted in the step 2, respectively dripping the nanogold solutions into a clean quartz cuvette, sequentially detecting the absorbance of the nanogold solutions within the wavelength range of 450-700 nm by using an ultraviolet-visible spectrophotometer (see fig. 2), and drawing a correlation curve (see fig. 3) of the wavelength corresponding to the peak absorbance within the wavelength range of 500-600 nm and the glucose concentration to obtain the formula Y & lt 3E & gt +44X-15.95，R20.9822, wherein X represents the wavelength corresponding to the peak absorbance of the nano-gold solution in the range of 500-600 nm, and Y represents the glucose concentration in the urine to be detected. As can be seen from fig. 3, the fitting formula has a higher goodness of fit with the correlation curve.
Example 2
The method for rapidly detecting the content of urine sugar by using the kit of the embodiment 1 comprises the following steps:
diluting the urine to be detected by 20 times by using deionized water to obtain a diluted solution of the urine to be detected; preheating the urine diluent to be detected by using a boiling water bath, adding 200 mu L of the urine diluent to a chloroauric acid reagent bottle preheated by the boiling water bath for 5 minutes, rapidly stirring for 5 seconds, continuously keeping the boiling water bath for heating and reacting for 6 minutes, enabling glucose in the urine and chloroauric acid to generate an oxidation-reduction color reaction, comparing the color of the obtained nanogold solution with a urine glucose concentration colorimetric card after the reaction is finished, and judging the glucose concentration range.
After the concentration range of glucose is judged, 1.5mL of the nanogold solution obtained after the reaction is finished is dripped into a quartz cuvette, the absorbance of the nanogold solution in the wavelength range of 450-700 nm is detected by adopting an ultraviolet-visible spectrophotometer, the wavelength corresponding to the peak absorbance in the wavelength range of 500-600 nm is obtained, and the formula Y on the urine glucose concentration accurate quantitative card is 3E +44X-15.95And quantitatively calculating the concentration of glucose in the urine to be detected.
Claims (3)
1. A rapid detection kit for urine sugar content based on chloroauric acid color reaction is characterized in that: the kit comprises a chloroauric acid reagent bottle and a urine glucose concentration colorimetric card; wherein the chloroauric acid reagent bottle is a disposable covered sealed glass bottle filled with 2-5 mL of chloroauric acid color developing solution, and the chloroauric acid color developing solution is prepared by adjusting the pH value of 0.5mM chloroauric acid aqueous solution to 7-9 by 0.2M sodium hydroxide aqueous solution; the urine glucose concentration colorimetric card consists of a color block A, a color block B, a color block C, a color block D, a color block E, a color block F, a color block G, a color block H and a color block I, wherein subscripts of the color block A and the color block B are more than or equal to 12mM, and the corresponding urine glucose concentration is more than or equal to 12 mM; 11-10mM is marked below the color blocks B and C, and the corresponding urine glucose concentration is 10-11 mM; 9-8mM is marked below the color blocks D and E, and the corresponding urine glucose concentration is 8-9 mM; 7-6mM is marked below the color blocks F and G, and the corresponding urine glucose concentration is 6-7 mM; 5-4mM is marked below the color blocks G and H, and the concentration of corresponding urine sugar is 4-5 mM; the lower parts of the color blocks H and I are marked with less than or equal to 3mM, and the corresponding urine glucose concentration is less than or equal to 3 mM;
the manufacturing method of the urine glucose concentration colorimetric card comprises the following steps:
(1) dissolving glucose dry powder with normal human urine to prepare 100mM glucose stock solution for diabetes, then diluting the glucose stock solution with normal human urine to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12mM respectively, and then sequentially diluting the glucose stock solution with deionized water by 20 times to prepare a glucose standard solution;
(2) according to the volume ratio of the urine sugar standard solution to the chloroauric acid color developing solution of 1:15, preheating the urine sugar standard solution through a boiling water bath, adding the preheated urine sugar standard solution into a chloroauric acid reagent bottle preheated through the boiling water bath, rapidly stirring for 5-10 seconds, and continuing to keep the boiling water bath for heating reaction for 5-6 minutes to obtain a nano gold solution; preparing a urine glucose concentration colorimetric card according to the colors of the nano-gold solutions corresponding to the urine glucose standard solutions with different concentrations;
the kit also comprises a urine glucose concentration accurate quantitative card, and a formula is written on the accurate quantitative card: y =3 × 1044X-15.95(ii) a The formula on the urine glucose concentration accurate quantitative card is obtained by the following method: dripping 1.5-2 mL of the nanogold solution reacted in the step (2) into a quartz cuvette, detecting the wavelength corresponding to the peak absorbance of the nanogold solution within the range of 500-600 nm by using an ultraviolet-visible spectrophotometer, and drawing a correlation curve of the wavelength corresponding to the peak absorbance and the glucose concentration to obtain the formula Y =3 × 1044X-15.95Wherein X represents the wavelength corresponding to the peak absorbance of the nano-gold solution within the range of 500-600 nm, and Y represents the concentration of glucose in the urine to be detected.
2. The kit for rapidly detecting the content of urine sugar according to claim 1, wherein: the disposable capped and sealed glass bottle has the capacity of 8mL, the height of 70-80 mm and the diameter of 10-15 mm, is made of a screw quartz glass material, and the bottle cap is made of a heat-resistant plastic material, and is internally provided with a heat-resistant leakage-proof gasket.
3. A method for the detection of urine glucose content for non-disease diagnostic purposes using the kit of claim 1 or 2, characterized in that: diluting the urine to be detected by 20 times by using deionized water to obtain a diluted solution of the urine to be detected; preheating a urine diluent to be detected by using a boiling water bath, adding the preheated urine diluent to be detected into a chloroauric acid reagent bottle preheated by the boiling water bath according to the volume ratio of the urine diluent to be detected to the chloroauric acid developing solution of 1:15, rapidly stirring for 5-10 seconds, then continuously keeping the boiling water bath for heating and reacting for 5-6 minutes to enable glucose in the urine and the chloroauric acid to generate oxidation-reduction developing reaction, comparing the color of the obtained nanogold solution with a urine glucose concentration colorimetric card after the reaction is finished, and judging the concentration range of the glucose; after the concentration range of glucose is judged, 1.5-2 mL of nano-gold solution is dripped into a quartz cuvette, an ultraviolet-visible spectrophotometer is adopted to detect the wavelength corresponding to the peak absorbance within the range of 500-600 nm, and the formula Y =3 × 10 on a urine glucose concentration accurate quantitative card is adopted44X-15.95And quantitatively calculating the concentration of glucose in the urine to be detected.
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| CN113945708A (en) * | 2021-10-15 | 2022-01-18 | 南方医科大学 | Urine detection method and system |
| CN114414507B (en) * | 2021-12-23 | 2024-01-12 | 中国石油大学(华东) | Glucose colorimetric detection method based on artificial intelligence |
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| CN104535541A (en) * | 2015-01-20 | 2015-04-22 | 西南大学 | Method for detecting cell glycosylation level |
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| Matteo Scampicchio，et al..Sugar determination via the homogeneous reduction of Au salts:A novel optical measurement.《Talanta》.2009,第79卷 * |
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