Application of amphiphilic colorimetric test paper based on photocatalysis in pyrophosphate radical detection
By forming a colorimetric test strip with a hydrophilic and hydrophobic area on the test strip, combined with the discoloration reaction of zinc reagent, the problems of high detection cost of pyrophosphate and equipment dependence are solved, and a low-cost, fast and sensitive detection effect is achieved.
Patent Information
- Application Number
- CN202510672633.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-05-23
- Publication Date
- 2025-08-26
AI Technical Summary
The existing pyrophosphate detection methods are costly, rely on expensive enzyme preparations and complex equipment, and are difficult to widely use in resource-constrained environments. The traditional paper-based detection platform lacks sensitivity and portability.
A photocatalysis-based colorimetric test strip is used to form a hydrophilic and hydrophobic region on the test strip through template method and semiconductor photocatalytic degradation technology, and a specific color discoloration reaction is carried out in combination with zinc reagent to achieve visualization and quantitative detection.
It realizes low-cost, fast, visual and high-sensitivity pyrophosphate detection, suitable for in-situ analysis and on-site rapid screening, without the need for complex instrument support, the detection range is 20U/L to 800U/L, and the detection limit is 3U/L.
Smart Images
Figure CN120539136A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the field of biological analysis and detection, and particularly relates to the application of an amphiphilic colorimetric test paper based on photocatalysis in detecting pyrophosphate. Background Art
[0002] Inorganic pyrophosphate (PPi) is an important inorganic anion with the chemical formula P2O7 4- , formed by the dehydration condensation of two phosphate ions, is widely present in nature and in living organisms. Since its initial discovery in the early 19th century, research into its physiological functions has continued to deepen. In 1941, Cori et al. first elucidated the formation mechanism of PPi in cellular metabolism, finding that it is primarily derived from the byproduct released during the cleavage of high-energy phosphate bonds of nucleoside triphosphates (such as ATP) by hydrolases.
[0003] Within cells, PPi is not only a metabolite of high-energy molecules such as ATP and GTP but also plays a crucial role in the biosynthesis of macromolecules such as DNA, RNA, and proteins. For example, during DNA replication, deoxynucleoside triphosphates (dNTPs) are incorporated into nucleotide chains via a nucleophilic substitution reaction (S_N2), accompanied by the cleavage of the bond between the α- and β-phosphates, releasing PPi. Subsequently, PPi can be hydrolyzed into two molecules of inorganic phosphate (Pi) under the catalysis of inorganic pyrophosphatase. This process provides the thermodynamic driving force for the synthesis reaction, thereby ensuring the irreversibility of key metabolic reactions such as DNA synthesis. Furthermore, PPi plays a regulatory and auxiliary role in a range of life activities, including cellular energy transduction, protein phosphorylation, signal transduction, enzymatic reactions, material metabolism, and telomere maintenance. Its concentration changes are also closely related to various pathological conditions, such as vascular calcification, articular cartilage damage, renal dysfunction, and certain types of cancer. Therefore, PPi is considered one of the important bioindicators of intracellular metabolic activity and health.
[0004] Traditional methods for detecting inorganic pyrophosphate (PPi) mainly include enzymatic methods and small molecule chemical sensor methods. Enzymatic methods use specific enzymes such as pyrophosphatase to quantify the hydrolysis of PPi, which has high accuracy and specificity. However, its detection process usually relies on expensive enzyme preparations and needs to be operated under strictly controlled temperature and pH conditions. The detection process is relatively cumbersome, and some methods still rely on radioactive or fluorescent markers, which pose biosafety and environmental risks. Small molecule chemical sensor methods, especially fluorescent sensors, have developed rapidly in recent years. Their advantages are high sensitivity, real-time detection, and ease of structural design and functional modification. However, their synthesis process is complex and costly, and they are highly dependent on detection equipment (such as fluorescence spectrometers), which limits their widespread application in resource-constrained environments.
[0005] In contrast, paper-based detection platforms have unique advantages in PPi detection due to their low cost, portability and rapid response. Summary of the Invention
[0006] In response to the problems existing in the prior art, the present invention aims to provide an application of a photocatalytic amphiphilic colorimetric test paper in the detection of pyrophosphate. The test paper with a hydrophobic layer is treated with a template method and semiconductor photocatalytic degradation technology to make the test paper have both hydrophilic and hydrophobic regions. A zinc reagent is added to the hydrophilic region to undergo a specific color change reaction with zinc ions, thereby obtaining an amphiphilic colorimetric test paper that can be used for quantitative detection of PPi, thereby achieving visual, rapid, repeatable and low-cost pyrophosphate detection.
[0007] The purpose of the present invention is achieved through the following technical solutions:
[0008] The present invention provides an application of a photocatalytically based amphiphilic colorimetric test paper in detecting pyrophosphate. The invention utilizes the photocatalytic effect of a titanium-based metal organic framework (TiMOFs) material and a template method to simultaneously form hydrophilic and hydrophobic regions on the test paper. The hydrophilic region is color-modified to construct a color-developing region to form the amphiphilic colorimetric test paper. A pyrophosphate sample is dripped into the color-developing region of the amphiphilic colorimetric test paper. The pyrophosphate is quantitatively detected in combination with a visual colorimetric method or a computer RGB channel separation method.
[0009] Furthermore, the application of the photocatalytic amphiphilic colorimetric test paper in the detection of pyrophosphate comprises the following specific steps:
[0010] (1) preparing a titanium-based metal organic framework-test paper composite;
[0011] (2) hydrophobic treatment of the titanium-based metal organic framework-test paper complex to form a titanium-based metal organic framework-test paper complex with a hydrophobic layer of phosphate groups and alkyl chains;
[0012] (3) punching holes in the light-shielding template according to the design of the hydrophilic region, and then covering the light-shielding template on the titanium-based metal organic framework-test paper composite with the phosphate group and alkyl chain hydrophobic layer in step (2), and performing light treatment. The area punched in the light-shielding template photodegrades the hydrophobic groups of the hydrophobic layer. After cleaning and drying, a concave hydrophilic colorimetric hole is formed. The light-shielding template is removed, and the area other than the hydrophilic colorimetric hole is the hydrophobic area;
[0013] (4) Evenly spread the zinc reagent-zinc ion in the colorimetric well. The red zinc reagent undergoes a specific color change reaction with the zinc ion to form a blue color development area, indicating that the zinc ion is successfully bound to the colorimetric well. After drying, the amphiphilic colorimetric test paper is obtained.
[0014] (5) The pyrophosphate sample is dropped into the color-developing area of the amphiphilic colorimetric test paper. The color of the color-developing area will change with the pyrophosphate sample. The pyrophosphate is quantitatively detected by visual colorimetry or computer RGB channel separation.
[0015] Furthermore, step (1) is to obtain Ti-O cluster powder by a mixed reaction of tetraisopropoxytitanium (Ti(OiPr)4), acetic acid and acetonitrile, and add the Ti-O cluster powder, 2,2'-bipyridine-5,5'-dicarboxylic acid (BPYDC), histidine (His) and trifluoroacetic acid to N,N-dimethylformamide (DMF) solution, and transfer it into a closed reactor. At the same time, the test paper is placed in the closed reactor, and amino-functionalized titanium-based metal organic framework material (NH2-Ti-MOFs) is in situ grown on the test paper. After the reaction is completed, it is washed with N,N-dimethylformamide and ethanol, and vacuum dried to obtain a titanium-based metal organic framework material-test paper complex.
[0016] Furthermore, in step (2), the hydrophobic treatment is to mix and dissolve dodecylphosphonic acid (DDPA) and 11-phosphonoundecanoic acid (11-PUA) in toluene, and then fix the titanium-based metal organic framework material-test paper complex on the top of the bottom of the closed container, with the active substance titanium-based metal organic framework material facing downward, and heat and keep it warm so that the toluene evaporates rapidly and fills the closed reactor, and the phosphate groups attach to the titanium-based metal organic framework material-test paper complex to form a titanium-based metal organic framework material-test paper complex with a phosphate group and an alkyl chain hydrophobic layer.
[0017] Furthermore, in step (3), the light-shielding template is made of black cardboard, the light treatment is performed by mercury lamp irradiation in a photochemical reactor, and the cleaning is performed by a buffer solution with pH=8.5.
[0018] Furthermore, in step (4), a polydiallyldimethylammonium chloride (PDDA) aqueous solution is dripped into the colorimetric well, and after drying, it is washed with a pH = 8.5 buffer solution, blown dry, and then a red zinc reagent is dripped in. After drying, the unbound zinc reagent is washed away with a pH = 8.5 buffer solution, blown dry, and a hexahydrate zinc nitrate aqueous solution is dripped into the colorimetric well to form a blue color development area; the role of PDDA is to promote the uniform spreading of the zinc reagent and the hexahydrate zinc nitrate aqueous solution in the colorimetric well.
[0019] Furthermore, in step (4), the zinc reagent-zinc ions in the colorimetric wells carry positive charges, and self-assemble with the negatively charged phosphate groups in the colorimetric wells to form a molecular layer, which is conducive to more zinc reagent-zinc ion binding, thereby improving the sensitivity of color change.
[0020] Furthermore, in step (4), the concentration of the zinc solvent is 0.05 g / L, and the concentration of zinc nitrate hexahydrate is 500 μM.
[0021] Furthermore, in the visual colorimetric method in step (5), artificial serum is first used to prepare standard solutions with pyrophosphate concentrations of 20U / L, 40U / L, 80U / L, 200U / L, 400U / L, 600U / L and 800U / L, and 20 μL of each is pipetted and dropped onto different colorimetric wells in the color development area of the parent colorimetric test paper obtained in step (4), and wait for 2 minutes. The colors displayed by each colorimetric well in the color development area form a standard colorimetric card; then 20 μL of the serum to be tested is dropped into the colorimetric wells in the color development area of another new parent colorimetric test paper, and the displayed color is compared with the standard colorimetric card to determine the concentration of pyrophosphate.
[0022] Furthermore, in the computer RGB channel separation method in step (5), first, artificial serum is used to prepare standard solutions with pyrophosphate concentrations of 20U / L, 40U / L, 80U / L, 200U / L, 400U / L, 600U / L and 800U / L, and 20 μL of each solution is pipetted and dropped onto different colorimetric wells in the color development area of the parent colorimetric test paper obtained in step (4), and waited for 2 minutes. The colors displayed by the colorimetric wells in the color development area form a standard colorimetric card, and then 20 μL of the serum to be tested is dropped onto the colorimetric wells in the color development area of another new parent colorimetric test paper, and waited for 2 minutes;
[0023] At the same time, the color-developing areas of the standard colorimetric card and the serum sample to be tested are photographed. The red and blue channel grayscale values of the standard colorimetric card are separated by computer RGB. Two standard curves are formed by computer processing. The grayscale value of the red channel is directly proportional to the pyrophosphate concentration, and the grayscale value of the blue channel is inversely proportional to the pyrophosphate concentration. The red and blue channel grayscale values of the color-developing area of the serum sample to be tested are then separated by computer RGB. The concentrations corresponding to the grayscale values are found on the two standard curves respectively, and the average value is taken, which is the concentration of pyrophosphate in the analyte.
[0024] Advantages and effects of the present invention:
[0025] The paper-based material of the present invention has a good hydrophilic and hydrophobic surface, and can realize the transmission and reaction of liquid samples without the need for external power. In addition, the paper-based material of the present invention is combined with the colorimetric method, and the concentration of PPi in the sample can be quantitatively evaluated by comparing the colored area with the standard colorimetric card. It is more suitable for in situ analysis and on-site rapid screening, does not require complex instrument support, has good portability and user-friendliness, and is conducive to application in the sinking market. The colored area can also be separated into RGB channels by a computer, and the grayscale value after separation can accurately and quantitatively evaluate PPi expression. The detection range is 20U / L to 800U / L, and the detection limit is 3U / L. When the concentration is not within the detection range, it can be adjusted by concentration or dilution. Due to the stability of the amphiphilic colorimetric pore structure and the high selectivity of the zinc reagent, the amphiphilic colorimetric test paper of the present invention has the potential for long-term use and can be applied to the fields of clinical and family health management self-examination. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] Figure 1 The figure shows the preparation process and detection principle of the amphiphilic colorimetric test paper in Examples 1 and 2. DETAILED DESCRIPTION
[0027] The present invention is described in detail below with reference to the embodiments.
[0028] Example 1
[0029] The present invention provides an application of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate. Figure 1 The preparation process shown in the figure and the visual colorimetric method are used for detection. The specific steps are as follows:
[0030] (1) Preparation of titanium-based metal organic framework-test paper composite:
[0031] 0.4 mL of Ti(OiPr)4, 2 mL of acetic acid, and 12 mL of acetonitrile were mixed evenly and transferred to a 50 mL sealed container. The mixture was reacted at 100 °C for 24 h, and then centrifuged and washed, and dried at 70 °C for 12 h to obtain Ti-O cluster powder.
[0032] 100 mg of Ti-O cluster powder, 80 mg of BPYDC, 50 mg of His and 1 mL of trifluoroacetic acid were added to 15 mL of DMF, ultrasonically mixed and then transferred to a 50 mL sealed container. The test paper was placed in the sealed container and reacted at 150°C for 24 hours to in situ grow NH2-Ti-MOFs on the test paper. After the reaction was completed, the obtained sample was washed with DMF and ethanol, and finally placed in a vacuum drying oven at 80°C for 12 hours to obtain the TiMOFs-test paper composite.
[0033] (2) The TiMOFs-test paper composite was hydrophobically treated, and DDPA and 11-PUA were mixed at a molar ratio of 1:4 (X COOH :X CH3 ) were dissolved in toluene to a total concentration of 0.1 mM. The TiMOFs-test paper complex was fixed to the top of the bottom of a sealed container with the active substance TiMOFs facing downward. The reactor was kept at 60°C for 2 hours to allow the toluene solution to evaporate rapidly and fill the reactor. The phosphate groups attached to the TiMOFs-test paper complex, and the alkyl chains acted as hydrophobic groups, forming a TiMOFs-test paper complex with a hydrophobic layer of phosphate groups and alkyl chains.
[0034] (3) The hydrophilic area is designed as an array hole. The black cardboard is punched with an array hole. The black cardboard is then covered on the TiMOFs-test paper complex with a hydrophobic layer of phosphate groups and alkyl chains in step (2). The black cardboard is then placed in a photochemical reactor and illuminated under a 500W mercury lamp for 7 minutes. The alkyl chains of the hydrophobic layer in the array hole area on the black cardboard are photodegraded. The hydrophilic area is washed with a pH = 8.5 buffer solution and blown dry to form a concave colorimetric well without a hydrophobic layer. The black cardboard is removed, and the area other than the hydrophilic colorimetric well is the hydrophobic area.
[0035] (4) 7 μL of 0.2% PDDA aqueous solution was dripped into the colorimetric well, dried at 80°C for 1 hour, washed three times with a pH = 8.5 buffer solution, and blown dry; then 7 μL of 0.05 g / L red zinc reagent solution was dripped into the well, dried at 80°C for 1 hour, and unbound zinc reagent was washed away with a pH = 8.5 buffer solution, blown dry, and finally 20 μL of 500 μM hexahydrate zinc nitrate aqueous solution was removed with a pipette and dripped into the well to form a blue color development area. After drying at 80°C for 1 hour, the amphiphilic colorimetric test paper was obtained for use.
[0036] (5) Detection was performed using a visual colorimetric method. Standard solutions of artificial serum pyrophosphate concentrations of 20 U / L, 40 U / L, 80 U / L, 200 U / L, 400 U / L, 600 U / L, and 800 U / L were prepared, and 20 μL of each solution was dropped onto different colorimetric wells in the color development area of the double-parent colorimetric test paper obtained in step (4). After waiting for 2 minutes, the colors displayed by the colorimetric wells in the color development area formed a standard colorimetric card. Then, 20 μL of the serum to be tested was dropped onto the colorimetric wells in the color development area of another new double-parent colorimetric test paper, and the displayed colors were compared with the standard colorimetric card to determine the concentration of pyrophosphate.
[0037] Example 2
[0038] The present invention provides an application of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate. Figure 1The preparation process shown and the computer RGB channel separation method are used for detection. The specific steps are as follows:
[0039] (1) Preparation of titanium-based metal organic framework-test paper composite:
[0040] 0.4 mL of Ti(OiPr)4, 2 mL of acetic acid, and 12 mL of acetonitrile were mixed evenly and transferred to a 50 mL sealed container. The mixture was reacted at 100 °C for 24 h, and then centrifuged and washed, and dried at 70 °C for 12 h to obtain Ti-O cluster powder.
[0041] 100 mg of Ti-O cluster powder, 80 mg of BPYDC, 50 mg of His and 1 mL of trifluoroacetic acid were added to 15 mL of DMF, ultrasonically mixed and then transferred to a 50 mL sealed container. The test paper was placed in the sealed container and reacted at 150°C for 24 hours to in situ grow NH2-Ti-MOFs on the test paper. After the reaction was completed, the obtained sample was washed with DMF and ethanol, and finally placed in a vacuum drying oven at 80°C for 12 hours to obtain the TiMOFs-test paper composite.
[0042] (2) The TiMOFs-test paper composite was hydrophobically treated, and DDPA and 11-PUA were mixed at a molar ratio of 1:4 (X COOH :X CH3 ) were dissolved in toluene to a total concentration of 0.1 mM. The TiMOFs-test paper complex was fixed to the top of the bottom of a sealed container with the active substance TiMOFs facing downward. The reactor was kept at 60°C for 2 hours to allow the toluene solution to evaporate rapidly and fill the reactor. The phosphate groups attached to the TiMOFs-test paper complex, and the alkyl chains acted as hydrophobic groups, forming a TiMOFs-test paper complex with a hydrophobic layer of phosphate groups and alkyl chains.
[0043] (3) The hydrophilic area is designed as an array hole. The black cardboard is punched with an array hole. The black cardboard is then covered on the TiMOFs-test paper complex with a hydrophobic layer of phosphate groups and alkyl chains in step (2). The black cardboard is then placed in a photochemical reactor and illuminated under a 500W mercury lamp for 7 minutes. The alkyl chains of the hydrophobic layer in the array hole area on the black cardboard are photodegraded. The hydrophilic area is washed with a pH = 8.5 buffer solution and blown dry to form a concave colorimetric well without a hydrophobic layer. The black cardboard is removed, and the area other than the hydrophilic colorimetric well is the hydrophobic area.
[0044] (4) 7 μL of 0.2% PDDA aqueous solution was dripped into the colorimetric well, dried at 80°C for 1 hour, washed three times with a pH = 8.5 buffer solution, and blown dry; then 7 μL of 0.05 g / L red zinc reagent solution was dripped into the well, dried at 80°C for 1 hour, and unbound zinc reagent was washed away with a pH = 8.5 buffer solution, blown dry, and finally 20 μL of 500 μM hexahydrate zinc nitrate aqueous solution was removed with a pipette and dripped into the well to form a blue color development area. After drying at 80°C for 1 hour, the amphiphilic colorimetric test paper was obtained for use.
[0045] (5) Using the computer RGB channel separation method, first prepare standard solutions with artificial serum pyrophosphate concentrations of 20 U / L, 40 U / L, 80 U / L, 200 U / L, 400 U / L, 600 U / L, and 800 U / L, and pipette 20 μL of each solution and drop it onto different colorimetric wells in the color development area of the parent colorimetric test paper obtained in step (4). Wait for 2 minutes. The colors displayed by the colorimetric wells in the color development area form a standard colorimetric card. Then, drop 20 μL of the serum to be tested onto the colorimetric wells in the color development area of another new parent colorimetric test paper and wait for 2 minutes.
[0046] At the same time, the color-developing areas of the standard colorimetric card and the serum sample to be tested are photographed. The red and blue channel grayscale values of the standard colorimetric card are separated by computer RGB. Two standard curves are formed by computer processing. The grayscale value of the red channel is directly proportional to the pyrophosphate concentration, and the grayscale value of the blue channel is inversely proportional to the pyrophosphate concentration. The red and blue channel grayscale values of the color-developing area of the serum sample to be tested are then separated by computer RGB. The concentrations corresponding to the grayscale values are found on the two standard curves respectively, and the average value is taken, which is the concentration of pyrophosphate in the analyte.
Claims
1. An application of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate, characterized in that: The photocatalytic effect of titanium-based metal organic framework materials and the template method are used to simultaneously form hydrophilic and hydrophobic regions on the test paper. The hydrophilic region is color-modified to construct a color-developing region to form an amphiphilic colorimetric test paper. The pyrophosphate sample to be tested is dropped into the color-developing region of the amphiphilic colorimetric test paper, and the pyrophosphate is quantitatively detected in combination with visual colorimetry or computer RGB channel separation method.
2. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 1, wherein: The specific steps are as follows: (1) preparing a titanium-based metal organic framework-test paper composite; (2) hydrophobic treatment of the titanium-based metal organic framework-test paper complex to form a titanium-based metal organic framework-test paper complex with a hydrophobic layer of phosphate groups and alkyl chains; (3) punching holes in the light-shielding template according to the design of the hydrophilic region, and then covering the light-shielding template on the titanium-based metal organic framework-test paper composite with the phosphate group and alkyl chain hydrophobic layer in step (2), and performing light treatment. The area punched in the light-shielding template photodegrades the hydrophobic groups of the hydrophobic layer. After cleaning and drying, a concave hydrophilic colorimetric hole is formed. The light-shielding template is removed, and the area other than the hydrophilic colorimetric hole is the hydrophobic area; (4) evenly spreading the zinc reagent-zinc ions in the colorimetric well, the red zinc reagent reacts specifically with the zinc ions to form a blue color-developing area, and after drying, a biparental colorimetric test paper is obtained; (5) The pyrophosphate sample is dropped into the color-developing area of the amphiphilic colorimetric test paper. The color of the color-developing area will change with the pyrophosphate sample. The pyrophosphate is quantitatively detected by visual colorimetry or computer RGB channel separation.
3. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 2, wherein: Step (1) is to obtain Ti-O cluster powder by a mixed reaction of tetraisopropoxytitanium, acetic acid and acetonitrile, add the Ti-O cluster powder, 2,2'-bipyridine-5,5'-dicarboxylic acid, histidine and trifluoroacetic acid into an N,N-dimethylformamide solution, transfer the solution into a closed reactor, and place a test paper in the closed reactor to in situ grow an amino-functionalized titanium-based metal-organic framework material on the test paper. After the reaction is completed, the solution is washed with N,N-dimethylformamide and ethanol, and vacuum dried to obtain a titanium-based metal-organic framework material-test paper composite.
4. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 2, wherein: In step (2), the hydrophobic treatment is to mix and dissolve dodecylphosphonic acid and 11-phosphonoundecanoic acid in toluene, and then fix the titanium-based metal organic framework material-test paper complex on the top of the bottom of the closed container with the active substance titanium-based metal organic framework material facing downward. Heat and keep it warm so that the toluene evaporates rapidly and fills the closed reactor, and the phosphate groups attach to the titanium-based metal organic framework material-test paper complex to form a titanium-based metal organic framework material-test paper complex with a phosphate group and an alkyl chain hydrophobic layer.
5. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 2, wherein: In step (3), the light-shielding template is made of black cardboard, the light treatment is performed by mercury lamp irradiation in a photochemical reactor, and the cleaning is performed by a buffer solution with pH=8.
5.
6. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 2, wherein: Step (4) dripping a polydiallyldimethylammonium chloride aqueous solution into the colorimetric well, drying it, washing it with a pH = 8.5 buffer solution, blowing it dry, then dripping a red zinc reagent, drying it, washing away the unbound zinc reagent with a pH = 8.5 buffer solution, blowing it dry, and dripping a hexahydrate zinc nitrate aqueous solution into the colorimetric well to form a blue color development area.
7. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 2, wherein: In step (4), the zinc reagent-zinc ions in the colorimetric wells carry positive charges, and self-assemble with the negatively charged phosphate groups in the colorimetric wells to form a molecular layer, which is conducive to more zinc reagent-zinc ion binding, thereby improving the sensitivity of color change.
8. The use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 2, wherein: In step (4), the concentration of the zinc solvent is 0.05 g / L, and the concentration of zinc nitrate hexahydrate is 500 μM.
9. Use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 8, characterized in that: In the visual colorimetric method in step (5), first use artificial serum to prepare standard solutions with pyrophosphate concentrations of 20U / L, 40U / L, 80U / L, 200U / L, 400U / L, 600U / L and 800U / L, and pipette 20μL of each solution and drop it on different colorimetric wells in the color development area of the parent colorimetric test paper obtained in step (4). Wait for 2 minutes, and the colors displayed by each colorimetric well in the color development area form a standard colorimetric card; then drop 20μL of the serum to be tested into the colorimetric wells in the color development area of another new parent colorimetric test paper, and compare the displayed color with the standard colorimetric card to determine the concentration of pyrophosphate.
10. Use of a photocatalytic amphiphilic colorimetric test paper in detecting pyrophosphate according to claim 8, characterized in that: In step (5), the computer RGB channel separation method is first used to prepare standard solutions with pyrophosphate concentrations of 20U / L, 40U / L, 80U / L, 200U / L, 400U / L, 600U / L and 800U / L using artificial serum, and 20μL of each solution is dropped onto different colorimetric wells in the color development area of the parent colorimetric test paper obtained in step (4), and wait for 2 minutes. The colors displayed by each colorimetric well in the color development area form a standard colorimetric card, and then 20μL of the serum to be tested is dropped onto the colorimetric wells in the color development area of another new parent colorimetric test paper, and wait for 2 minutes; At the same time, the color-developing areas of the standard colorimetric card and the serum sample to be tested are photographed. The red and blue channel grayscale values of the standard colorimetric card are separated by computer RGB. Two standard curves are formed by computer processing. The grayscale value of the red channel is directly proportional to the pyrophosphate concentration, and the grayscale value of the blue channel is inversely proportional to the pyrophosphate concentration. The red and blue channel grayscale values of the color-developing area of the serum sample to be tested are then separated by computer RGB. The concentrations corresponding to the grayscale values are found on the two standard curves respectively, and the average value is taken, which is the concentration of pyrophosphate in the analyte.