Preparation and application of cellulose hydrogen peroxide colorimetric test paper

Abstract

The application discloses a kind of preparation and application of cellulose hydrogen peroxide colorimetric test paper, and the colorimetric test paper is obtained by the self-assembly of gold nanoclusters on cellulose membrane.The colorimetric test paper prepared by the application shows peroxidase simulation behavior, can catalyze hydrogen peroxide oxidation 3,3',5,5'-tetramethyl benzidine to produce blue oxidation state, and be fixed in situ by cellulose hydroxyl group immediately, avoid color overflow, make detection more accurate.The visual detection limit of the colorimetric test paper prepared by the application is 0.007mM, and hydrogen peroxide in the concentration range of 0.007-1.4mM can realize naked-eye colorimetric detection.Meanwhile, the colorimetric test paper after recognizing hydrogen peroxide can be reduced by ascorbic acid, so that the colorimetric test paper returns to light yellow, with the characteristics of repeated use.The colorimetric test paper of the application is simple to prepare, convenient to detect, sensitive to respond, and reusable, and has potential application prospect in the detection field of hydrogen peroxide.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical analysis and detection, and in particular to the preparation and application of a cellulose hydrogen peroxide colorimetric test paper. Background Art

[0002] Hydrogen peroxide is a major reactive oxygen species in living systems, with crucial applications in biological processes and the external environment. In the natural environment, hydrogen peroxide can be used to purify tap water. During biological activities, hydrogen peroxide participates in various oxidative stress responses and plays a vital role in sugar and protein metabolism and immune responses, serving as a signature product of enzyme degradation. Recent research has demonstrated that hydrogen peroxide is closely linked to cell proliferation, differentiation, and migration, regulating physiological and pathological processes as a signaling molecule. Furthermore, hydrogen peroxide plays a crucial role in numerous pathological processes, such as inflammation and immune defense. Many diseases, such as cancer and Alzheimer's disease, can also be caused by excessive hydrogen peroxide. Therefore, real-time, quantitative detection of hydrogen peroxide in living organisms and the external environment is of paramount importance.

[0003] A variety of methods are currently available for detecting hydrogen peroxide, including colorimetry, chromatography, titration, fluorescence, and electrochemistry. Notably, colorimetry has attracted increasing attention among researchers due to its convenience, sensitivity, and the absence of complex and expensive instrumentation. In recent years, the use of colorimetric test strips has garnered widespread attention in colorimetric hydrogen peroxide detection, as they allow for direct visual observation of the concentration in the test solution without the need for a photometer.

[0004] Chinese patent CN 112525896 A discloses a hydrogen peroxide test strip and a method for quantitatively detecting hydrogen peroxide. The test strip uses 3,3',5,5'-tetramethylbenzidine as a color developer. When Fe3O4 / Ti3C2 catalyzes the decomposition of hydrogen peroxide, 3,3',5,5'-tetramethylbenzidine is oxidized to produce a blue color. The test strip can be used to qualitatively determine the approximate hydrogen peroxide content by visually observing the color change of the test strip.

[0005] However, in the prior art, filter paper is often used as the base material for hydrogen peroxide test paper. Filter paper fibers are coarse, have poor binding properties with inorganic catalysts, are easily damaged after immersion testing, and are often only disposable, which also poses a problem for actual colorimetric detection. The selection of catalysts in test paper and the stability of the combination of catalysts and paper-based materials directly determine the sensitivity and stability of the test paper for hydrogen peroxide detection. Therefore, it is very necessary to provide a hydrogen peroxide test paper with high catalytic performance and good stability. Summary of the Invention

[0006] To address the above-mentioned problems, the present invention provides the preparation and application of a cellulose hydrogen peroxide colorimetric test paper. The test paper can perform colorimetric detection of hydrogen peroxide in a concentration range of 0.007 to 1.75 mM. The catalyst in the test paper is stably combined with the cellulose-based material. After color development detection, the test paper can be subjected to the action of a reducing reagent and reused multiple times. After up to six uses, the colorimetric results remain applicable.

[0007] In order to achieve the above object, the present invention provides a cellulose-based hydrogen peroxide colorimetric detection test paper, comprising gold nanoclusters and a substrate.

[0008] Furthermore, the particle size of the gold nanoclusters is 1 to 2 nm.

[0009] Furthermore, the substrate is one of a cellulose membrane or a filter paper.

[0010] The present invention also provides a method for preparing the above-mentioned cellulose-based hydrogen peroxide colorimetric detection test paper, comprising the following preparation steps:

[0011] Preparation of cellulose membrane:

[0012] The dried cotton linters are placed in a NaOH / urea aqueous solution, stirred and dissolved to prepare a cellulose membrane, and the cellulose membrane is washed with water and dried to obtain a usable cellulose membrane;

[0013] Synthesis of gold nanocluster dispersion:

[0014] HAuCl4·4H2O was added to water, preheated, and then glutathione aqueous solution was added dropwise and stirred to react to obtain a gold nanocluster dispersion.

[0015] Preparation of test strips:

[0016] At room temperature, the cellulose membrane is soaked in the gold nanocluster dispersion, the soaked cellulose membrane is cleaned using ultrasonic waves, and dried to obtain a cellulose-based hydrogen peroxide colorimetric test paper.

[0017] Furthermore, the moisture content of the dried cotton linters is less than 5%, the NaOH / urea aqueous solution is prepared from 13-15 g of NaOH, 23-25 ​​g of urea and 160-164 g of deionized water, the mass ratio of the cotton linters to the NaOH / urea aqueous solution is (1-2):(23-26), the stirring and dissolving temperature is -8-13° C., the stirring rate is 2000-2300 rpm, and the drying temperature is 50-80° C.

[0018] Furthermore, the mass concentration of the HAuCl4·4H2O is 0.5-5%, the preheating temperature is 60-100°C, the mass concentration of the glutathione aqueous solution is 0.1-2%, the volume ratio of the HAuCl4·4H2O, water, and glutathione aqueous solution is (3-5):(80-100):(5-8), the stirring rate is 1500-1800 rpm, and the reaction time is 8-24 h.

[0019] Furthermore, the room temperature is 20-30°C, the ratio of the cellulose membrane to the gold nanocluster dispersion is (10-15g):(50-80mL), the soaking time is 1-10h, the ultrasonic cleaning time is 10-100s, and the drying temperature is 50-80°C.

[0020] The present invention also provides an application method of the above-mentioned hydrogen peroxide colorimetric detection test paper, comprising the following steps:

[0021] Prepare a hydrogen peroxide solution, immerse part or all of the hydrogen peroxide colorimetric test paper in the hydrogen peroxide solution, record the corresponding color after the color reaction, and obtain the relationship between the hydrogen peroxide concentration and the color change of the hydrogen peroxide colorimetric test paper;

[0022] Immerse part or all of the hydrogen peroxide colorimetric test paper in a hydrogen peroxide solution of unknown concentration, and calibrate the concentration of hydrogen peroxide in the unknown solution by the color displayed;

[0023] The tested test paper is immersed in a reducing agent solution, then taken out, washed with water, dried, and used again for the detection of hydrogen peroxide solution of unknown concentration.

[0024] Furthermore, the concentration of hydrogen peroxide in the hydrogen peroxide solution is 0.007 to 1.75 mM. The hydrogen peroxide solution is prepared by aqueous hydrogen peroxide solution and a dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine. The volume ratio of aqueous hydrogen peroxide solution to the dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine is 49:1, and the concentration of the dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine is 20 mM.

[0025] Furthermore, the concentration of hydrogen peroxide in the hydrogen peroxide solution is 0, 0.007, 0.021, 0.035, 0.07, 0.14, 0.28, 0.49, 0.70, 1.05, 1.40 and 1.75 mM.

[0026] Furthermore, the reducing agent solution is a 5 mM ascorbic acid solution, the soaking time is 2 to 4 hours, and the drying temperature is 50 to 80°C.

[0027] The present invention prepares a cellulose membrane from cotton linters, which is then immersed in a dispersion of gold nanoclusters. The gold nanoclusters self-assemble and immobilize on the cellulose membrane, thereby preparing a hydrogen peroxide colorimetric test strip. Based on the principle and preparation method of the colorimetric test strip of the present invention, the gold nanoclusters act as a pseudo-enzyme medium for hydrogen peroxide to promote the oxidation of 3,3',5,5'-tetramethylbenzidine by hydrogen peroxide, producing a blue oxidized state, which is instantly fixed in situ by the cellulose hydroxyl groups in the colorimetric test strip, avoiding color spillage and making the detection more accurate. In the hydrogen peroxide colorimetric test strip prepared by the present invention, the gold nanoclusters are stably bound to the cellulose membrane. After the hydrogen peroxide is detected, the oxidized state can be reduced using a reducing agent, enabling repeated detection of the hydrogen peroxide concentration.

[0028] Based on the recognition principle, the present invention rationally designs a cellulose-based hydrogen peroxide colorimetric test strip for detecting hydrogen peroxide. The response parameters of the test strip were tested, including a detection time of approximately 20 minutes, a visual detection limit of 0.007 mM, colorimetric detection within a concentration range of 0.007 to 1.4 mM, and a number of recycling times of approximately six times.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] (1) The cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention is simple to prepare and convenient to detect. Compared with the fluorescent probe method that requires complex instruments, it has practical application value.

[0031] (2) The cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention has a sensitive response, a low visual detection limit, good selectivity, is reusable, and can be detected by the naked eye. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The present invention provides a color reaction diagram of cellulose-based hydrogen peroxide colorimetric test paper and filter paper-based hydrogen peroxide colorimetric test paper to hydrogen peroxide.

[0033] Figure 2 The cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention is immersed in a colorimetric corresponding diagram for 20 minutes;

[0034] Figure 3 Schematic diagram of the mechanism of hydrogen peroxide identification by the cellulose-based hydrogen peroxide colorimetric test paper prepared in the present invention;

[0035] Figure 4 This is a scanning electron microscope image of the cellulose film prepared in the present invention;

[0036] Figure 5 This is a scanning electron microscope image of the cellulose-based hydrogen peroxide colorimetric test paper prepared in the present invention;

[0037] Figure 6This is a scanning electron microscope image of the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention after color development to detect hydrogen peroxide;

[0038] Figure 7 This is a transmission electron microscopy image of the gold nanoclusters synthesized by the present invention;

[0039] Figure 8 A visual observation diagram of the colorimetric reaction of the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention with hydrogen peroxide at concentrations of 0, 0.007, 0.021, 0.035, 0.07, 0.14, 0.28, 0.49, 0.70, 1.05, 1.40 and 1.75 mM;

[0040] Figure 9 The colorimetric reaction of the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention with hydrogen peroxide at concentrations of 0, 0.007, 0.021, 0.035, 0.07, 0.14, 0.28, 0.49, 0.70, 1.05, 1.40 and 1.75 mM, and the corresponding relationship between color intensity and hydrogen peroxide concentration;

[0041] Figure 10 The cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention reacts colorimetrically with hydrogen peroxide at concentrations of 0, 0.007, 0.021, 0.035, 0.07, 0.14, 0.28, 0.49, 0.70, 1.05, 1.40 and 1.75 mM, and a fitting relationship curve between color intensity and hydrogen peroxide concentration;

[0042] Figure 11 The cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention and the colorimetric test paper of hydrogen peroxide and K+, Na+, Zn 2 +、Ca 2 +, urea, ascorbic acid and glucose (all concentrations are normal urine content);

[0043] Figure 12 This is a comparison chart of the reuse of the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention. DETAILED DESCRIPTION

[0044] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the specific embodiments of the present invention and the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0045] Unless otherwise specified, in the embodiments of the present invention, the hydrogen peroxide solution is prepared from an aqueous hydrogen peroxide solution and a dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine, the volume ratio of the aqueous hydrogen peroxide solution to the dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine is 49:1, and the concentration of the dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine is 20 mM.

[0046] Example 1

[0047] A method for preparing cellulose-based hydrogen peroxide colorimetric test paper, comprising the following steps:

[0048] Preparation of cellulose membrane:

[0049] At -12.5°C, 8 g of cotton linters with a water content of 2% were placed in 200 g of a NaOH / urea aqueous solution prepared from 14 g of NaOH, 24 g of urea, and 162 g of deionized water. The solution was stirred and dissolved at 2300 rpm. A cellulose film with a thickness of 0.5 mm was prepared by a casting method. The cellulose film was rinsed with water and dried at 60°C for 3 h to obtain a usable cellulose film.

[0050] Synthesis of gold nanocluster dispersion:

[0051] 4 mL of 2% HAuCl4·4H2O was added to 90 mL of water, preheated to 80°C, and 6 mL of 1.5% glutathione was added dropwise. The mixture was stirred and reacted for 8 h to obtain a gold nanocluster dispersion.

[0052] Preparation of test strips:

[0053] At 25° C., 10 g of the cellulose membrane was soaked in 50 mL of the gold nanocluster dispersion for 3 h. The soaked cellulose membrane was cleaned using ultrasonic waves for 30 s and dried at 60° C. for 3 h to obtain a cellulose-based hydrogen peroxide colorimetric test paper.

[0054] Example 2

[0055] A method for preparing filter paper-based hydrogen peroxide colorimetric test paper, comprising the following steps:

[0056] Synthesis of gold nanocluster dispersion:

[0057] 4 mL of 2% HAuCl4·4H2O was added to 90 mL of water, preheated to 80°C, and 6 mL of 1.5% glutathione was added dropwise. The mixture was stirred and reacted for 8 h to obtain a gold nanocluster dispersion.

[0058] Preparation of test strips:

[0059] At 25° C., 10 g of filter paper was soaked in 50 mL of the gold nanocluster dispersion for 3 h. The soaked filter paper was cleaned using ultrasonic waves for 30 s and dried at 60° C. for 3 h to obtain filter paper-based hydrogen peroxide colorimetric test paper.

[0060] Application Example 1

[0061] The original color of the colorimetric test paper may have a significant impact on the color change of the developer, such as Figure 1 As shown, the original color of the cellulose-based hydrogen peroxide colorimetric test paper prepared in Example 1 of the present invention is significantly lighter than that of the filter paper-based hydrogen peroxide colorimetric test paper prepared in Example 2, showing a light yellow color and having less interference with color development. Furthermore, the cellulose-based hydrogen peroxide colorimetric test paper and the filter paper-based hydrogen peroxide colorimetric test paper prepared in the present invention were immersed in a 1 mM concentration hydrogen peroxide solution to observe the color development effect of the test paper. Figure 1 As can be seen, the color development effect of the cellulose-based hydrogen peroxide colorimetric test paper is more obvious than that of the filter paper-based hydrogen peroxide colorimetric test paper. These results show that the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention has a stronger colorimetric detection ability than the conventional filter paper-based hydrogen peroxide colorimetric test paper.

[0062] Application Example 2

[0063] In order to study the detection potential of the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention, the test paper was placed in a hydrogen peroxide solution to observe the color development performance. The specific operation is to place the cellulose-based hydrogen peroxide colorimetric test paper prepared in Example 1 of the present invention in 5mL of a hydrogen peroxide solution with a concentration of 1mM, let it stand for 30min, observe and record the change in the color of the colorimetric test paper over time, the time is 0, 1, 2, 3, 4, 6, 8, 10, 12, 15, 18, 20, 25 and 30min, and the blank control group is set up with ultrapure water instead of the hydrogen peroxide aqueous solution. The results show that in the blank control group without hydrogen peroxide, the color of the test paper does not change, and still presents the initial light yellow color. As shown Figure 2 As shown, in the group containing hydrogen peroxide, the color development gradually deepened at 1, 2, 3, 4, 6, 8, 10, 12, 15, 18, and 20 minutes. The light yellow colorimetric test paper at 0 minutes gradually changed from light yellow to blue, and the color became increasingly darker. The color development at 20, 25, and 30 minutes was not clearly distinguishable to the naked eye. These results indicate that the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention has the potential to visually detect hydrogen peroxide.

[0064] Based on the color development reaction of the test paper, the present invention proposes a mechanism diagram of cellulose-based hydrogen peroxide colorimetric test paper for identifying hydrogen peroxide, such as Figure 3The interaction between gold nanoclusters and cellulose molecules improves overall stability, catalyzing the oxidation of 3,3',5,5'-tetramethylbenzidine to form a color. The gold nanoclusters are then fixed in situ by the hydroxyl groups of cellulose and attached to the cellulose membrane, preventing the color from spilling out.

[0065] In order to study the changes in the structural morphology of the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention after color development (detection of a hydrogen peroxide solution with a concentration of 1 mM), the cellulose membrane, the cellulose-based hydrogen peroxide colorimetric test paper, and the color-developed cellulose-based hydrogen peroxide colorimetric test paper were characterized by scanning electron microscopy. The results are shown in FIG. Figures 4 to 7 As shown. Figure 4 It can be seen that the cellulose membrane prepared in the present invention has a loose and porous structure. Figure 5 The results show that the structure of cellulose-based hydrogen peroxide colorimetric test paper (i.e. loaded with gold nanoclusters) does not change significantly compared with cellulose membrane. Figure 6 As shown, it can be seen that the structure of the test paper has not changed significantly compared to before color development. These results indicate that the gold nanoclusters in the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention are stably bound to cellulose, and the loose and porous structure of cellulose allows the color development reaction to proceed on the cellulose surface. These effects may improve the detection performance of the test paper and enable it to have the ability to perform multiple cycles of detection. Figure 7 This is a transmission electron microscope image of gold nanoparticles in the gold nanocluster dispersion in an embodiment of the present invention. It can be seen that the particle size of the gold nanoparticles is concentrated in the range of 1 to 2 nm.

[0066] The cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention was immersed in hydrogen peroxide solutions with concentrations of 0, 0.007, 0.021, 0.035, 0.07, 0.14, 0.28, 0.49, 0.70, 1.05, 1.40 and 1.75 mM, respectively, and incubated for 20 min. The color of the colorimetric test paper was observed and recorded. The results are shown in FIG. Figure 8 As shown. Figure 8 It can be seen that as the concentration of hydrogen peroxide increases, the color intensity increases. Figure 9 The relationship between hydrogen peroxide concentration and color intensity shows that at low hydrogen peroxide concentrations, the color intensity increases linearly. When the hydrogen peroxide concentration is greater than 1.40 mM, the color intensity remains almost unchanged. In addition, the color intensity of the test paper after hydrogen peroxide response is converted into a digital signal through Photoshop software. The intensity signal is plotted against the logarithm of the hydrogen peroxide concentration to study the relationship between the color intensity of the colorimetric test paper and the hydrogen peroxide concentration. The fitting result is y = (84.65 ± 1.96) x + 193.71 ± 1.92, and the square of the correlation coefficient R 2=0.9952, where y represents the color intensity and x represents the logarithm of the concentration. The results show that the colorimetric test paper can achieve colorimetric detection in the concentration range of 0.007 to 1.75 mM.

[0067] In actual detection, the analyte usually contains not only hydrogen peroxide, but also a large number of impurity ions or molecules. Selective detection of hydrogen peroxide is one of the important indicators for evaluating the performance of test paper. + 、Na + 、Zn 2+ , Ca 2+ , urea, ascorbic acid and glucose (all concentrations are normal human urine concentrations) were incubated for 20 minutes respectively, and the specific detection ability of the test paper for hydrogen peroxide (concentration is 1mM) was compared. The colorimetric results are as follows Figure 11 As shown. Figure 11 It can be seen that the colorimetric test paper responds best to hydrogen peroxide, and the test paper turns a distinct blue, while the color of the test paper remains almost unchanged after co-incubation with other substances, indicating that the colorimetric test paper has relatively good selectivity for hydrogen peroxide.

[0068] Cyclic detection capability is also one of the important indicators for evaluating the performance of test paper. The colorimetric test paper is incubated with 1mM hydrogen peroxide solution for 20 minutes, and the test paper turns blue. Then the blue test paper is immersed in a 5mM ascorbic acid aqueous solution for 3 hours. The hydrogen peroxide colorimetric test paper that has turned back to light yellow is taken out and washed with deionized water to remove ascorbic acid and the reduced 3,3',5,5'-tetramethylbenzidine. It is then dried at 60°C for 3 hours to restore the detection capability of the test paper. Repeat the above operation 6 times, and record the color after each color development and reduction. The results are as follows: Figure 12 As shown. Figure 12 It can be seen from the results that after 6 cycles of color development, the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention still has a good effect on the color development detection of hydrogen peroxide. This result shows that the cellulose-based hydrogen peroxide colorimetric test paper prepared by the present invention has good reusability.

[0069] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a cellulose-based hydrogen peroxide colorimetric test paper, characterized in that: The cellulose-based hydrogen peroxide colorimetric test paper comprises gold nanoclusters and a substrate, and the preparation method comprises the following steps: Preparation of cellulose membrane: The dried cotton linters are placed in a NaOH / urea aqueous solution, stirred and dissolved to prepare a cellulose membrane, and the cellulose membrane is washed with water and dried to obtain a usable cellulose membrane; Synthesis of gold nanocluster dispersion: HAuCl4·4H2O was added to water, preheated, and then glutathione aqueous solution was added dropwise and stirred to react to obtain a gold nanocluster dispersion. Preparation of test strips: At room temperature, the cellulose membrane is soaked in the gold nanocluster dispersion, the soaked cellulose membrane is cleaned using ultrasonic waves, and dried to obtain a cellulose-based hydrogen peroxide colorimetric test paper; The moisture content of the dried cotton linters is less than 5%. The NaOH / urea aqueous solution is prepared from 13-15 g of NaOH, 23-25 ​​g of urea, and 160-164 g of deionized water. The mass ratio of the cotton linters to the NaOH / urea aqueous solution is (1-2):(23-26). The stirring and dissolving temperature is -8-13° C., the stirring rate is 2000-2300 rpm, and the drying temperature is 50-80° C.

2. The method according to claim 1, characterized in that The particle size of the gold nanoclusters is 1-2 nm.

3. The method according to claim 1, characterized in that The substrate is one of a cellulose membrane or a filter paper.

4. The method according to claim 1, wherein The mass concentration of the HAuCl4·4H2O is 0.5-5%, the preheating temperature is 60-100°C, the mass concentration of the glutathione aqueous solution is 0.1-2%, the volume ratio of the HAuCl4·4H2O, water, and the glutathione aqueous solution is (3-5):(80-100):(5-8), the stirring rate is 1500-1800 rpm, and the reaction time is 8-24 h.

5. The method according to claim 1, wherein The room temperature is 20-30° C., the ratio of the cellulose membrane to the gold nanocluster dispersion is (10-15 g): (50-80 mL), the soaking time is 1-10 h, the ultrasonic cleaning time is 10-100 s, and the drying temperature is 50-80° C.

6. Use of the hydrogen peroxide colorimetric test paper prepared by the method according to any one of claims 1 to 5, characterized in that: The colorimetric test paper is used for qualitative and quantitative detection of hydrogen peroxide.

7. The use according to claim 6, characterized in that The following steps are involved: Prepare a hydrogen peroxide solution, immerse part or all of the hydrogen peroxide colorimetric test paper in the hydrogen peroxide solution, record the corresponding color after the color reaction, and obtain the relationship between the hydrogen peroxide concentration and the color change of the hydrogen peroxide colorimetric test paper; Immerse part or all of the hydrogen peroxide colorimetric test paper in a hydrogen peroxide solution of unknown concentration, and calibrate the concentration of hydrogen peroxide in the unknown solution by the color displayed; The tested test paper is immersed in a reducing agent solution, then taken out, washed with water, dried, and used again for the detection of hydrogen peroxide solution of unknown concentration.

8. The use according to claim 7, characterized in that The concentration of hydrogen peroxide in the hydrogen peroxide solution is 0.007 to 1.75 mM. The hydrogen peroxide solution is prepared from an aqueous hydrogen peroxide solution and a dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine. The volume ratio of the aqueous hydrogen peroxide solution to the dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine is 49:1, and the concentration of the dimethyl sulfoxide solution of 3,3',5,5'-tetramethylbenzidine is 20 mM.

9. The use according to claim 7, characterized in that The concentrations of hydrogen peroxide in the hydrogen peroxide solution are 0, 0.007, 0.021, 0.035, 0.07, 0.14, 0.28, 0.49, 0.70, 1.05, 1.40 and 1.75 mM.

10. The use according to claim 7, characterized in that The reducing agent solution is a 5 mM ascorbic acid solution, the soaking time is 2 to 4 hours, and the drying temperature is 50 to 80°C.

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