A heavy atom enhanced based chromophoric radiometric colorimetric sensor

Abstract

The application discloses a heavy atom enhanced calix cyanine dye radioactivity detection colorimetric sensor, the dye is prepared by introducing heavy atoms in the calix cyanine dye skeleton, and the radiation sensitivity is significantly improved; the dye reacts with hydroxyl radicals generated by water radiolysis under the action of radiation, conjugate structure is destroyed, and color change from green to yellow is generated; the application comprises a colorimetric test strip prepared from the dye, and a detection system integrated with a smartphone image recognition function, the test strip comprises a base material and an additive; the additive comprises a copolymer, and preparation reaction monomers of the copolymer comprise N,N-diethylaminoethyl acrylate and 3-methylpent-2-ene dimethyl acid; the application has the advantages of high sensitivity, low cost, simple operation, portability and the like, can be used for rapid, visual and quantitative detection of radionuclides in water solution, urine and the like samples, and has important application value in the field of nuclear medicine radiation safety monitoring.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of analytical chemistry detection, and particularly relates to a heavy atom enhanced phthalocyanine dye radioactivity detection colorimetric sensor. BACKGROUND

[0002] Radionuclides are increasingly widely used in the medical field, especially in diagnostic imaging and targeted radiotherapy. However, the use of radiopharmaceuticals also poses a serious challenge in radiation safety management.

[0003] Therefore, developing a method capable of quickly, accurately and low-cost evaluating the radioactivity in the patient's body has become a crucial and urgent challenge in the field of nuclear medicine. Currently, mainstream radiation detection equipment includes Geiger counters, scintillation detectors, semiconductor detectors and thermoluminescence dosimeters, etc. Among them, Geiger counters are commonly used in nuclear medicine departments due to their high sensitivity. However, such instruments usually have complex manufacturing processes, high equipment costs, high maintenance requirements and are not portable, making it difficult to popularize self-monitoring in the patient's home environment. Colorimetric analysis is an economical, rapid and portable detection method with a history of over a century. With the popularity of smartphones, their powerful image capture and data processing capabilities can work with colorimetric sensors to achieve qualitative, semi-quantitative and even quantitative analysis of target analytes by analyzing color changes, significantly improving the accuracy and fault tolerance of detection. Although some color developing reagents for uranium, plutonium and other nuclides have been developed, they produce color changes through complexation reactions and can be used to monitor industrial nuclear pollution wastewater, but these reagents are not suitable for 131 I、 18 F and other commonly used radionuclides in nuclear medicine.

[0004] Ionizing radiation can induce radiolysis of water, producing highly active species such as hydroxyl radicals ·OH that can degrade organic dye molecules and cause discoloration. Based on the above principle, the present application aims to develop a colorimetric sensing platform.

[0005] However, ordinary phthalocyanine dyes have insufficient sensitivity to radiation, making it difficult to achieve low-dose visual detection. Therefore, there is an urgent need for a strategy that can significantly enhance the radiation sensitivity of dyes to prepare high-performance colorimetric sensors for practical sample detection. SUMMARY

[0006] The present application aims to provide a heavy atom enhanced phthalocyanine dye radioactivity detection colorimetric sensor to solve the problems of insufficient radiation sensitivity of ordinary phthalocyanine dyes, high cost and poor portability of existing radiation detection equipment, and the inapplicability of traditional color developing reagents to the detection of commonly used radionuclides in nuclear medicine, achieving rapid visual qualitative and quantitative detection of radionuclides in aqueous solutions, urine and other samples.

[0007] The technical scheme adopted by the present application to achieve the above-mentioned purpose is:

[0008] A radioactivity detection colorimetric sensor, comprising a test strip.

[0009] Preferably, the test strip comprises a cyanine dye.

[0010] Preferably, the structure of the cyanine dye comprises one of formula (1), formula (2) and formula (3),

[0011] (1),

[0012] (2),

[0013] (3).

[0014] Preferably, the R group is a heavy atom.

[0015] Preferably, the heavy atom comprises one of a bromine, chlorine and iodine atom.

[0016] The present application significantly enhances the radiation sensitivity of the cyanine dye by introducing a chlorine, bromine or iodine heavy atom into the molecular structure of the cyanine dye, and can accurately match the demand for radioactivity detection in the field of nuclear medicine. The cyanine dye has a specific response to high activity species such as ·OH produced by ionizing radiation induced water radiolysis, and has no interference with common ions in biological fluids and metabolites such as glucose and uric acid in urine. The common ions in biological fluids include Na + , K + , Ca 2+ , Mg 2+ , Cl - . Therefore, the present application can be directly applied to the radioactivity detection of complex biological samples such as urine without complex sample pretreatment, ensuring the specificity and accuracy of the detection results.

[0017] Preferably, the preparation reaction monomer of the cyanine dye comprises a quaternary ammonium salt intermediate and a condensing agent.

[0018] Preferably, the quaternary ammonium salt intermediate comprises one of a quaternary ammonium onium salt and a heavy atom substituted quaternary ammonium salt.

[0019] Preferably, the heavy atom substituted quaternary ammonium salt comprises one of a bromine substituted quaternary ammonium salt, a chlorine substituted quaternary ammonium salt and an iodine substituted quaternary ammonium salt.

[0020] Preferably, the mass ratio of the quaternary ammonium salt intermediate to the condensing agent is 1:3-6.

[0021] Preferably, the preparation reaction monomer of the heavy atom substituted quaternary ammonium salt comprises iodinated n-butane and a heavy atom substituted indole derivative.

[0022] Preferably, the heavy atom-substituted indole derivative comprises one of a bromo-substituted indole derivative, a chloro-substituted indole derivative, and an iodo-substituted indole derivative.

[0023] Preferably, the mass ratio of the heavy atom-substituted indole derivative to the n-butyl iodide is 2:5-8.

[0024] Preferably, the preparation reaction monomer of the heavy atom-substituted indole derivative comprises one of a heavy atom reaction monomer and 3-methyl-2-butanone.

[0025] Preferably, the heavy atom reaction monomer comprises one of 4-bromo-phenylhydrazine, 4-chloro-phenylhydrazine, and 4-iodo-phenylhydrazine.

[0026] Preferably, the mass ratio of the heavy atom reaction monomer to the 3-methyl-2-butanone is 1:0.5-2.

[0027] Preferably, the test strip comprises a substrate.

[0028] Preferably, the substrate comprises one of filter paper, nitrocellulose membrane, glass fiber membrane, and polymer film.

[0029] Preferably, the test strip comprises an additive.

[0030] Preferably, the additive comprises one of glycerol and a copolymer.

[0031] Preferably, the preparation reaction monomer of the copolymer comprises N,N-diethylaminoethyl acrylate and dimethyl 3-methylpent-2-enedioate.

[0032] Preferably, the mass ratio of the N,N-diethylaminoethyl acrylate to the dimethyl 3-methylpent-2-enedioate is 1:1-3.

[0033] The copolymer formed by copolymerization of N,N-diethylaminoethyl acrylate and dimethyl 3-methylpent-2-enedioate can promote the uniform dispersion of the dye on the surface of the test strip substrate, avoid the shielding of the radiation response site caused by dye aggregation, thereby improving the contact efficiency of the dye with the high active species such as ·OH generated by ionizing radiation induced water radiolysis, and enhancing the response sensitivity of the test strip to radionuclides; on the other hand, the ester group in the copolymer can adjust the hydrophilic and hydrophobic properties of the copolymer, so that it can form good binding force with the substrate, reduce the loss of the dye during storage or detection, and at the same time, the ester group can wrap the merocyanine dye molecules through the steric hindrance effect, reduce the non-specific degradation rate of the dye under room temperature and light shielding conditions, and further ensure the specificity and stability of the dye to radiation response.

[0034] Preferably, the radioactivity detection colorimetric sensor comprises an image acquisition device.

[0035] Preferably, the image acquisition device comprises a smartphone camera.

[0036] Preferably, the radioactivity detection colorimetric sensor comprises a data processing device.

[0037] Preferably, the data processing device comprises an application program running on a smart phone or a cloud server.

[0038] Preferably, the application program is configured to extract color feature values of the image and convert the color feature values into radioactivity activity values according to a pre-stored standard curve.

[0039] Preferably, the color feature values comprise a hue value Hue in HSV color space.

[0040] The heavy atom enhanced-based cyanine dye radioactivity detection colorimetric sensor can exhibit a significant color change from green to yellow when interacting with radionuclides, which can be clearly recognized by the naked eye, enabling a preliminary qualitative judgment of the presence or absence of radioactivity. With the help of a smart phone to collect the image of the test strip, precise quantitative analysis can be further achieved by extracting the hue value in the HSV color space. The preparation process of the test strip is simple, the raw materials are easy to obtain, and the overall cost is much lower than that of traditional radiation detection equipment. The entire detection system does not need to rely on large and expensive professional instruments, and image acquisition and data reading can be completed through a smart phone only, which has low operation threshold and strong portability, and is especially suitable for on-site rapid screening and self-monitoring in patient home environment. The sensor can be effectively applied to the monitoring of residual radioactivity activity in patients after radiotherapy, and can also be used for the evaluation of the radioactivity level of medical waste water, and has a broad application prospect in the field of clinical nuclear medicine radiation safety management and radiation protection.

[0041] Preferably, a use of a radioactivity detection colorimetric sensor in radionuclide detection.

[0042] Preferably, the radionuclide comprises 131 I.

[0043] Preferably, the matrix for radionuclide detection comprises an aqueous solution or urine.

[0044] More preferably, the preparation copolymerization monomer also comprises isobutyl aminocrotonate, and the mass-volume ratio of isobutyl aminocrotonate to anhydrous ethanol is 0.5-2 g:10 mL. Isobutyl aminocrotonate, as a copolymerization monomer, participates in the synthesis of the copolymer together with N,N-diethylaminoethyl acrylate and dimethyl 3-methylpent-2-ene dioate, so as to optimize the performance of the sensor; the amino group in the molecule of isobutyl aminocrotonate can form a stronger hydrogen bond and a polar interaction between the molecules of the cyanine dye, so as to promote the dye to be distributed in a more uniform monodisperse state in the copolymer matrix, maximize the exposure of the dye to the response site of radiation, improve the contact efficiency with high active species such as ·OH generated by water radiolysis induced by ionizing radiation, enhance the response sensitivity of the test strip to low activity radionuclides, and show lower hue values and more significant color changes; meanwhile, the stability of the copolymer to the cyanine dye is improved, the non-specific degradation and loss of the dye during storage are reduced, and the storage stability of the test strip is improved.

[0045] The application further provides a preparation method of the quaternary ammonium onium salt, comprising:

[0046] Iodo-n-butane is dissolved in acetonitrile to obtain an iodo-n-butane solution; 2,3,3-trimethylindole is dissolved in acetonitrile, heated and refluxed at 85-95°C, the iodo-n-butane solution is added to the refluxing liquid, and the reaction is continued at 85-95°C for 35-45 h; after the reaction liquid is cooled to room temperature, filtration is performed to obtain a solid, the solid is washed with ethyl acetate for 2-4 times, and vacuum drying is performed to obtain the quaternary ammonium onium salt.

[0047] Preferably, in the iodo-n-butane solution, the mass-volume ratio of iodo-n-butane to acetonitrile is 8-10 g:5 mL.

[0048] Preferably, the mass-volume ratio of 2,3,3-trimethylindole to acetonitrile is 1-3 g:10 mL.

[0049] Preferably, the mass of the iodo-n-butane solution is measured by the mass of iodo-n-butane therein, and the mass ratio of 2,3,3-trimethylindole to iodo-n-butane is 2:8-10.

[0050] The application further provides a preparation method of the heavy atom-substituted indole derivative, comprising:

[0051] Preparation of the heavy atom-substituted indole derivative: heavy atom reaction monomers and 3-methyl-2-butanone are dissolved in glacial acetic acid, and reflux reaction is performed at 115-125°C for 7-9 h to obtain a reaction mixture; the reaction mixture is poured into ice water, extracted with ethyl acetate for 2-4 times, the organic phases are combined, washed with a saturated sodium chloride solution, dried with anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain the heavy atom-substituted indole derivative.

[0052] Preferably, the heavy atom reaction monomer comprises one of 4-bromophenylhydrazine, 4-chlorophenylhydrazine and 4-iodophenylhydrazine.

[0053] Preferably, the mass-volume ratio of the heavy atom reaction monomer to glacial acetic acid is 1g:2-5mL.

[0054] Preferably, the mass ratio of the heavy atom reaction monomer to 3-methyl-2-butanone is 1:0.5-2.

[0055] Preferably, the mass-volume ratio of the heavy atom reaction monomer to ice water is 1g:10-50mL.

[0056] Preferably, the volume ratio of ethyl acetate to ice water is 3:8-15.

[0057] Preferably, the heavy atom substituted indole derivative comprises one of a bromo-substituted indole derivative, a chloro-substituted indole derivative and an iodo-substituted indole derivative.

[0058] The present application also provides a preparation method of a heavy atom substituted quaternary ammonium salt, comprising:

[0059] Preparation of the heavy atom substituted quaternary ammonium salt: iodide n-butane is dissolved in acetonitrile to obtain an iodide n-butane solution; a heavy atom substituted indole derivative is dissolved in acetonitrile, heated to reflux at 85-95℃, the iodide n-butane solution is added to the refluxing liquid, and the reaction is continued at 85-95℃ for 35-45h; after the reaction liquid is cooled to room temperature, filtration is performed to obtain a solid, which is washed with ethyl acetate for 2-4 times, and vacuum dried to obtain the heavy atom substituted quaternary ammonium salt. Preferably, in the iodide n-butane solution, the mass-volume ratio of iodide n-butane to acetonitrile is 6.2g:5-10mL.

[0060] Preferably, the mass-volume ratio of the heavy atom substituted indole derivative to acetonitrile is 2g:5-15mL.

[0061] Preferably, the mass of the iodide n-butane solution is measured by the mass of iodide n-butane therein, and the mass ratio of the heavy atom substituted indole derivative to iodide n-butane is 2:5-8.

[0062] Preferably, the heavy atom substituted quaternary ammonium salt comprises one of a bromo-substituted quaternary ammonium salt, a chloro-substituted quaternary ammonium salt and an iodo-substituted quaternary ammonium salt.

[0063] The present application also provides a preparation method of a condensing agent, comprising:

[0064] Preparation of the condensing agent: under an ice water bath, phosphorus oxychloride is mixed with anhydrous dichloromethane, and cyclohexanone is added dropwise, and the reaction is refluxed at 35-45℃ for 2-4h to obtain a reaction liquid, the reaction liquid is added to ice water, and the precipitate is collected by vacuum filtration to obtain the condensing agent.

[0065] Preferably, the volume ratio of the phosphorus oxychloride to the anhydrous dichloromethane is 3:2-5.

[0066] Preferably, the mass volume ratio of the cyclohexanone to the anhydrous dichloromethane is 1g:2-5mL.

[0067] Preferably, the volume ratio of the ice water to the anhydrous dichloromethane is 10:2-5.

[0068] The application also provides a preparation method of the phycobilin dye, comprising:

[0069] The quaternary ammonium salt intermediate, sodium acetate and condensing agent are dissolved in acetic anhydride, stirred at 70-80℃ for 3-5h under nitrogen protection to obtain a reaction solution; the reaction solution is quenched with saturated sodium bicarbonate solution, extracted with dichloromethane for 2-4 times, the organic layers are combined, dried and concentrated to obtain a residue, and the residue is purified by column chromatography to obtain the phycobilin dye.

[0070] Preferably, the quaternary ammonium salt intermediate comprises one of quaternary ammonium onium salt and heavy atom substituted quaternary ammonium salt. Preferably, the mass volume ratio of the quaternary ammonium salt intermediate to acetic anhydride is 1g:5-15mL.

[0071] Preferably, the mass volume ratio of the sodium acetate to acetic anhydride is 0.93g:5-15mL.

[0072] Preferably, the mass volume ratio of the condensing agent to acetic anhydride is 4.2g:5-15mL.

[0073] Preferably, the volume ratio of the dichloromethane to acetic anhydride is 2:0.5-2.

[0074] The application also provides a preparation method of the copolymer, comprising:

[0075] The comonomer is dissolved in anhydrous ethanol, stirred at 150-250rpm for 5-15min, azobisisobutyronitrile is added, and the reaction is stirred at 65-75℃ and 200-400rpm for 7-9h under nitrogen protection to obtain a reaction solution; after the reaction solution is cooled to room temperature, petroleum ether is added, stirred at 150-250rpm for 5-15min, and the precipitate is collected by suction filtration after standing for 0.5-1.5h; the precipitate is washed with detergent for 2-4 times and vacuum dried at 35-45℃ for 10-15h to obtain the copolymer.

[0076] Preferably, the comonomer comprises N,N-diethylaminoethyl acrylate and 3-methylpent-2-ene dicarboxylic acid dimethyl ester.

[0077] Preferably, the mass volume ratio of the N,N-diethylaminoethyl acrylate to anhydrous ethanol is 1g:5-15mL.

[0078] Preferably, the mass volume ratio of dimethyl 3-methylpent-2-enoate to anhydrous ethanol is 1-3 g:10 mL.

[0079] Preferably, the mass volume ratio of azobisisobutyronitrile to anhydrous ethanol is 32 mg:5-15 mL.

[0080] Preferably, the volume ratio of petroleum ether to anhydrous ethanol is 2-8:1.

[0081] Preferably, the detergent is petroleum ether.

[0082] Preferably, the volume ratio of the detergent to anhydrous ethanol is 1:0.5-2.

[0083] More preferably, the co-monomer further comprises isobutyl aminocrotonate.

[0084] More preferably, the mass volume ratio of isobutyl aminocrotonate to anhydrous ethanol is 0.5-2 g:10 mL.

[0085] The present application also provides a preparation method of a test strip, comprising:

[0086] The cyanine dye is dissolved in anhydrous ethanol, stirred for 5-15 min to obtain a cyanine dye stock solution; the cyanine dye stock solution is added to an additive, stirred at 300-500 rpm for 15-25 min to obtain a cyanine dye working solution; the substrate is cut into a length of 8-15 cm and a width of 1-5 cm, dried in an oven at 65-75℃ for 1-3 h to obtain a pretreated substrate; the pretreated substrate is immersed in the cyanine dye working solution at room temperature for 25-35 min, and after being taken out, the excess solution on the surface is drained, and dried at room temperature and in a ventilated and light-proof condition for 2-4 h, cut into a length of 0.5-2 cm and a width of 0.5-2 cm to obtain a test strip.

[0087] Preferably, the molar volume ratio of the cyanine dye to anhydrous ethanol is 10 mmol:0.5-2 L.

[0088] Preferably, the additive comprises one of glycerol and a copolymer.

[0089] Preferably, the volume ratio of the additive to the cyanine dye stock solution is 1:20.

[0090] Preferably, the substrate comprises one of filter paper, nitrocellulose membrane, glass fiber membrane and polymer film.

[0091] More preferably, the substrate is a Whatman filter paper strip.

[0092] The present application also provides a preparation method of a radioactivity detection system, comprising:

[0093] 10-50 μL of the test solution is added to the center of the test strip, and a test strip with deionized water is added as a blank control; after the test strip is incubated at room temperature for 1-3 h, a photo of all the test strips is taken vertically above the test strip at a distance of 30-70 cm under standard light conditions of white light 3900-4100 K using a smart phone camera; the hue value in the HSV color space is automatically extracted from the selected detection area, and the hue value is used as a parameter for quantitative analysis of the radioactivity because the hue value is sensitive to color types and is relatively less affected by changes in light intensity, and a radioactivity detection system integrating the test strip, smart phone image acquisition and data processing is constructed.

[0094] The present application has the following beneficial effects: first, the heavy atom enhancement significantly improves the radiation sensitivity of the dye, which meets the needs of nuclear medicine detection; second, the dye specifically responds to ·OH generated by radiation and is not affected by common ions and metabolites in biological fluids, so it can directly detect complex samples such as urine; third, the test strip preparation process is simple and low-cost, and the detection does not require large instruments, and only a smart phone is needed for qualitative observation and quantitative analysis, which is highly portable and has a low operation threshold. Therefore, the present application is a heavy atom enhanced phycobilin dye radioactivity detection colorimetric sensor with excellent performance, low cost and convenient operation, which can meet the needs of nuclear medicine radiation safety monitoring, patient residual radioactivity assessment and on-site rapid screening. BRIEF DESCRIPTION OF DRAWINGS

[0095] Figure 1 The synthesis route diagram of the phycobilin dye prepared in Example 1 is shown.

[0096] Figure 2 The synthesis route diagram of the phycobilin dye prepared in Example 2 is shown.

[0097] Figure 3 The synthesis route diagram of the phycobilin dye prepared in Example 3 is shown.

[0098] Figure 4 The synthesis route diagram of the phycobilin dye prepared in Comparative Example 1 is shown.

[0099] Figure 5 The synthesis route diagram of the phycobilin dye prepared in Comparative Example 2 is shown.

[0100] Figure 6 The phycobilin dye prepared in Example 1 is used to detect Na 131Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 1 versus Na

[0101] Figure 7 Response color change plot for the merocyanine dye prepared for Example 1 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 2 versus Na

[0102] Figure 8 Response color change plot for the merocyanine dye prepared for Example 2 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0103] Figure 9 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0104] Figure 10 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0105] Figure 11 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0106] Figure 12 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0107] Figure 13 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0108] Figure 14 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0109] Figure 15 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0110] Figure 16 Response color change plot for the merocyanine dye prepared for Example 3 versus Na 131 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0111] Figure 17 Radiation response mechanism test results plot for the merocyanine dye.

[0112] Figure 18 Response UV-Vis spectral change plot for the merocyanine dye prepared for Example 3 versus Na

[0113] Figure 19 A schematic diagram of the color change response of the cyanine dye prepared in Example 3 to the interference test.

[0114] Figure 20 A schematic diagram of the UV-Vis spectrum change response of the cyanine dye prepared in Example 3 to the stability test.

[0115] Figure 21 A schematic diagram of the color change response of the cyanine dye prepared in Example 3 to the stability test.

[0116] Figure 22 A schematic diagram of the validation test results of the radioactivity detection system.

[0117] Figure 23 A schematic diagram of the evaluation test results of the urine matrix effect of the radioactivity detection system.

[0118] Figure 24 A schematic diagram of the workflow of the radioactivity detection system. DETAILED DESCRIPTION

[0119] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of protection of the present application.

[0120] The concepts involved in the present application will be described below with reference to the drawings. It should be noted that the following descriptions of the concepts are only to make the content of the present application easier to understand, and do not limit the scope of protection of the present application. Meanwhile, the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below with reference to the drawings and in combination with the embodiments.

[0121] Example 1:

[0122] The synthetic route of the cyanine dye prepared in this embodiment is shown in Figure 1 Compound 1 in the figure is a quaternary ammonium onium salt, and compound 7 is a condensing agent.

[0123] The detection process in the radioactivity detection system in this embodiment is shown in Figure 24

[0124] ​Preparation of quaternary ammonium salt: iodide-n-butane was dissolved in acetonitrile to obtain a solution of iodide-n-butane; 2,3,3-trimethylindole was dissolved in acetonitrile, and the solution was heated to reflux at 90°C, and the solution of iodide-n-butane was added to the refluxing solution, and the reaction was continued at 90°C for 40h, and the reaction solution was cooled to room temperature, and then filtered to obtain a solid, which was washed with ethyl acetate for 3 times and dried under vacuum to obtain the quaternary ammonium salt. The mass / volume ratio of iodide-n-butane to acetonitrile in the solution of iodide-n-butane was 9.3g:5mL; the mass / volume ratio of 2,3,3-trimethylindole to acetonitrile was 2g:10mL; the mass of iodide-n-butane in the solution of iodide-n-butane was used as the measurement, and the mass ratio of 2,3,3-trimethylindole to iodide-n-butane was 2:9.3.

[0125] Preparation of condensing agent: under ice water bath, phosphorus oxychloride was mixed with anhydrous dichloromethane, and cyclohexanone was added dropwise, and the reaction was continued at 40°C for 3h to obtain a reaction solution, and the reaction solution was added to ice water, and the precipitate was collected by vacuum filtration to obtain the condensing agent. The volume ratio of phosphorus oxychloride to anhydrous dichloromethane was 3:4, the mass / volume ratio of cyclohexanone to anhydrous dichloromethane was 1g:4mL, and the volume ratio of ice water to anhydrous dichloromethane was 10:1.

[0126] Preparation of cyanine dye: the quaternary ammonium salt intermediate, sodium acetate and the condensing agent were dissolved in acetic anhydride, and the reaction was stirred at 75°C for 4h under nitrogen protection to obtain a reaction solution; the reaction solution was quenched with saturated sodium bicarbonate solution, extracted with dichloromethane for 3 times, and the combined organic layer was dried and concentrated to obtain a residue, which was purified by column chromatography to obtain IR-780 dye, i.e. cyanine dye. The quaternary ammonium salt intermediate was quaternary ammonium salt, the mass / volume ratio of the quaternary ammonium salt intermediate to acetic anhydride was 1g:10mL, the mass / volume ratio of sodium acetate to acetic anhydride was 0.93g:10mL, the mass / volume ratio of the condensing agent to acetic anhydride was 4.2g:10mL, and the volume ratio of dichloromethane to acetic anhydride was 2:1.

[0127] Preparation of test paper strip: the cyanine dye was dissolved in anhydrous ethanol and stirred for 10 min to obtain a cyanine dye stock solution; the cyanine dye stock solution was added to an additive and stirred at 400 rpm for 20 min to obtain a cyanine dye working solution; the substrate was cut into a length of 10 cm and a width of 2 cm, dried in a 70°C oven for 2 h to obtain a pretreated substrate; the pretreated substrate was immersed in the cyanine dye working solution at room temperature for 30 min, then the excess solution on the surface was drained after being taken out, and dried at room temperature and in the dark for 3 h, cut into a length of 1 cm and a width of 1 cm to obtain the test paper strip. In the cyanine dye stock solution, the cyanine dye was Cl-IR780 dye, and the molar volume ratio of the cyanine dye to anhydrous ethanol was 10 mmol:1 L; in the cyanine dye working solution, the additive was glycerol, and the volume ratio of the additive to the cyanine dye stock solution was 1:20; the substrate was a Whatman filter paper strip, model 1001-085.

[0128] Preparation of a radioactivity detection system: 20 μL of the sample to be tested was added to the center of the test paper strip, and a test paper strip with added deionized water was used as a blank control; after incubation of the test paper strips at room temperature for 2 h, the photos of all the test paper strips were taken vertically at a distance of 50 cm above the test paper strips under standard light conditions of white light 4000 K using a smartphone camera; the hue value in the HSV color space was automatically extracted from the selected detection area, and was used as a parameter for quantitative analysis of radioactivity because the hue value is sensitive to color types and is relatively less affected by changes in light intensity, thereby completing the construction of a radioactivity detection system integrating the test paper strip, image acquisition and data processing of a smartphone.

[0129] Example 2: Compared with Example 1, the difference lies in the preparation of the cyanine dye.

[0130] The synthetic route chart for preparing the cyanine dye in this example is shown in Figure 2 The compound 2 is a bromo-substituted indole derivative, the compound 3 is a bromo-substituted quaternary ammonium salt, and the compound 7 is a condensing agent.

[0131] Preparation of a bromo-substituted indole derivative: 4-bromo phenylhydrazine and 3-methyl-2-butanone were dissolved in glacial acetic acid and refluxed at 120°C for 8 h to obtain a reaction mixture; the reaction mixture was poured into ice water, extracted with ethyl acetate for 3 times, the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain the bromo-substituted indole derivative. The mass-volume ratio of 4-bromo phenylhydrazine to glacial acetic acid was 1 g:4 mL, the mass ratio of 4-bromo phenylhydrazine to 3-methyl-2-butanone was 1:0.7, and the mass-volume ratio of 4-bromo phenylhydrazine to ice water was 1 g:20 mL. The volume ratio of ethyl acetate to ice water was 3:10.

[0132] Preparation of bromo-substituted quaternary ammonium salt: iodide-n-butane was dissolved in acetonitrile to obtain a solution of iodide-n-butane; bromo-substituted indole derivative was dissolved in acetonitrile, heated to reflux at 90℃, and the solution of iodide-n-butane was added to the refluxing solution, and the reaction was continued at 90℃ for 40h. After the reaction solution was cooled to room temperature, the solid was collected by filtration, washed with ethyl acetate for 3 times, and dried under vacuum to obtain the bromo-substituted quaternary ammonium salt. The mass / volume ratio of iodide-n-butane to acetonitrile in the solution of iodide-n-butane was 6.2g:5mL; the mass / volume ratio of bromo-substituted indole derivative to acetonitrile was 2g:10mL; the mass of iodide-n-butane in the solution of iodide-n-butane was used as the measurement; and the mass ratio of indole derivative to iodide-n-butane was 2:6.2.

[0133] Preparation of condensing agent: under ice water bath, phosphorus oxychloride was mixed with anhydrous dichloromethane, and cyclohexanone was added dropwise, and the reaction was continued at reflux for 3h to obtain a reaction solution. The reaction solution was added to ice water, and the precipitate was collected by vacuum filtration to obtain the condensing agent. The volume ratio of phosphorus oxychloride to anhydrous dichloromethane was 3:4; the mass / volume ratio of cyclohexanone to anhydrous dichloromethane was 1g:4mL; and the volume ratio of ice water to anhydrous dichloromethane was 10:1.

[0134] Preparation of cyanine dye: quaternary ammonium salt intermediate, sodium acetate, and condensing agent were dissolved in acetic anhydride, and stirred at 75℃ under nitrogen protection for 4h to obtain a reaction solution. The reaction solution was quenched with saturated sodium bicarbonate solution, extracted with dichloromethane for 3 times, and the combined organic layers were dried and concentrated to obtain a residue. The residue was purified by column chromatography to obtain Br-IR780 dye, i.e., cyanine dye. The mass / volume ratio of quaternary ammonium salt intermediate to acetic anhydride was 1g:10mL; the mass / volume ratio of sodium acetate to acetic anhydride was 0.93g:10mL; the mass / volume ratio of condensing agent to acetic anhydride was 4.2g:10mL; and the volume ratio of dichloromethane to acetic anhydride was 2:1.

[0135] Example 3: Compared with Example 1, the difference is in the preparation of cyanine dye.

[0136] The synthetic route chart for preparing cyanine dye in this example is shown in Figure 3 Figure, wherein compound 4 is chloro-substituted indole derivative, compound 5 is chloro-substituted quaternary ammonium salt, and compound 7 is condensing agent.

[0137] Preparation of chloro-substituted indole derivative: 4-chlorophenylhydrazine was dissolved in glacial acetic acid with 3-methyl-2-butanone, and the reaction mixture was refluxed at 120 °C for 8 h. The reaction mixture was poured into ice water, and the organic phase was extracted with ethyl acetate three times, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain the chloro-substituted indole derivative. The mass / volume ratio of 4-chlorophenylhydrazine to glacial acetic acid was 1 g:4 mL, the mass ratio of 4-chlorophenylhydrazine to 3-methyl-2-butanone was 1:0.7, and the mass / volume ratio of 4-chlorophenylhydrazine to ice water was 1 g:20 mL. The volume ratio of ethyl acetate to ice water was 3:10.

[0138] Preparation of chloro-substituted quaternary ammonium salt: n-butyliodide was dissolved in acetonitrile to obtain an n-butyliodide solution. The chloro-substituted indole derivative was dissolved in acetonitrile and heated to reflux at 90 °C. The n-butyliodide solution was added to the refluxing solution, and the reaction was continued at 90 °C for 40 h. After the reaction solution was cooled to room temperature, the solid was filtered, washed with ethyl acetate three times, and dried under vacuum to obtain the chloro-substituted quaternary ammonium salt. The mass / volume ratio of n-butyliodide to acetonitrile in the n-butyliodide solution was 6.2 g:5 mL. The mass / volume ratio of the chloro-substituted indole derivative to acetonitrile was 2 g:10 mL. The mass of the n-butyliodide solution was measured based on the mass of the n-butyliodide therein. The mass ratio of the indole derivative to n-butyliodide was 2:6.2.

[0139] Preparation of condensing agent: Phosphorus oxychloride was mixed with anhydrous dichloromethane under an ice water bath, and cyclohexanone was added dropwise. The reaction was refluxed for 3 h to obtain a reaction solution. The reaction solution was added to ice water, and the precipitate was collected by vacuum filtration to obtain the condensing agent. The volume ratio of phosphorus oxychloride to anhydrous dichloromethane was 3:4. The mass / volume ratio of cyclohexanone to anhydrous dichloromethane was 1 g:4 mL. The volume ratio of ice water to anhydrous dichloromethane was 10:1.

[0140] Preparation of cyanine dye: The quaternary ammonium salt intermediate, sodium acetate, and the condensing agent were dissolved in acetic anhydride under nitrogen protection, and the reaction was stirred at 75 °C for 4 h to obtain a reaction solution. The reaction solution was quenched with saturated sodium bicarbonate solution, extracted with dichloromethane three times, and the organic layers were combined, dried, and concentrated to obtain a residue. The residue was purified by column chromatography to obtain Cl-IR780 dye, i.e., the cyanine dye. The quaternary ammonium salt intermediate was the chloro-substituted quaternary ammonium salt. The mass / volume ratio of the quaternary ammonium salt intermediate to acetic anhydride was 1 g:10 mL. The mass / volume ratio of sodium acetate to acetic anhydride was 0.93 g:10 mL. The mass / volume ratio of the condensing agent to acetic anhydride was 4.2 g:10 mL. The volume ratio of dichloromethane to acetic anhydride was 2:1.

[0141] Example 4: The difference between this example and Example 3 is in the preparation of the test strip.

[0142] Preparation of the copolymer: N,N-diethylaminoethyl acrylate and dimethyl 3-methylpent-2-enedioate were dissolved in anhydrous ethanol, stirred at 200 rpm for 10 min, and azobisisobutyronitrile was added. The reaction was stirred at 70°C and 300 rpm for 8 h under nitrogen protection to obtain a reaction solution. After the reaction solution was cooled to room temperature, petroleum ether was added, stirred at 200 rpm for 10 min, and the precipitate was collected by suction filtration after standing for 1 h. The precipitate was washed with detergent three times and dried at 40°C under vacuum for 12 h to obtain the copolymer. The mass-volume ratio of N,N-diethylaminoethyl acrylate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of dimethyl 3-methylpent-2-enedioate to anhydrous ethanol was 1.24 g:10 mL, the mass-volume ratio of azobisisobutyronitrile to anhydrous ethanol was 32 mg:10 mL, and the volume ratio of petroleum ether to anhydrous ethanol was 4:1. The detergent was petroleum ether, and the volume ratio of the detergent to anhydrous ethanol was 1:1.

[0143] Preparation of the test strip: The copolymer was dissolved in anhydrous ethanol to obtain a copolymer solution. The phycobilin dye was dissolved in anhydrous ethanol and stirred for 10 min to obtain a phycobilin dye stock solution. The phycobilin dye stock solution was added to an additive and stirred at 400 rpm for 20 min to obtain a phycobilin dye working solution. The substrate was cut into a length of 10 cm and a width of 2 cm, dried in an oven at 70°C for 2 h to obtain a pretreated substrate. The pretreated substrate was immersed in the phycobilin dye working solution at room temperature for 30 min, and then the excess solution on the surface was drained. The substrate was dried at room temperature and in the dark under ventilation for 3 h, cut into a length of 1 cm and a width of 1 cm, and then obtained as a test strip. In the copolymer solution, the mass-volume ratio of the copolymer to anhydrous ethanol was 1 g:20 mL. In the phycobilin dye stock solution, the phycobilin dye was Cl-IR780 dye, and the molar-volume ratio of the phycobilin dye to anhydrous ethanol was 10 mmol:1 L. In the phycobilin dye working solution, the additive was the copolymer solution, and the volume ratio of the additive to the phycobilin dye stock solution was 1:20. The substrate was a Whatman filter paper strip with a model number of 1001-085.

[0144] Example 5: This example is different from Example 4 in that the copolymer is prepared.

[0145] Preparation of the copolymer: N,N-diethylaminoethyl acrylate, dimethyl 3-methylpent-2-enedioate and isobutyl aminocrotonate were dissolved in anhydrous ethanol, stirred at 200 rpm for 10 min, azobisisobutyronitrile was added, and the reaction was carried out at 70°C and 300 rpm for 8 h under nitrogen protection to obtain a reaction solution; after the reaction solution was cooled to room temperature, petroleum ether was added, stirred at 200 rpm for 10 min, and the precipitate was collected by suction filtration after standing for 1 h; the precipitate was washed with detergent 3 times and vacuum dried at 40°C for 12 h to obtain the copolymer. The mass-volume ratio of N,N-diethylaminoethyl acrylate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of dimethyl 3-methylpent-2-enedioate to anhydrous ethanol was 1.5 g:10 mL, the mass-volume ratio of isobutyl aminocrotonate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of azobisisobutyronitrile to anhydrous ethanol was 32 mg:10 mL, and the volume ratio of petroleum ether to anhydrous ethanol was 4:1; the detergent was petroleum ether, and the volume ratio of the detergent to anhydrous ethanol was 1:1.

[0146] Example 6: Compared with Example 4, the difference lies in the preparation of the copolymer.

[0147] Preparation of the copolymer: N,N-diethylaminoethyl acrylate, dimethyl 3-methylpent-2-enedioate and isobutyl aminocrotonate were dissolved in anhydrous ethanol, stirred at 200 rpm for 10 min, azobisisobutyronitrile was added, and the reaction was carried out at 70°C and 300 rpm for 8 h under nitrogen protection to obtain a reaction solution; after the reaction solution was cooled to room temperature, petroleum ether was added, stirred at 200 rpm for 10 min, and the precipitate was collected by suction filtration after standing for 1 h; the precipitate was washed with detergent 3 times and vacuum dried at 40°C for 12 h to obtain the copolymer. The mass-volume ratio of N,N-diethylaminoethyl acrylate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of dimethyl 3-methylpent-2-enedioate to anhydrous ethanol was 1.5 g:10 mL, the mass-volume ratio of isobutyl aminocrotonate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of azobisisobutyronitrile to anhydrous ethanol was 32 mg:10 mL, and the volume ratio of petroleum ether to anhydrous ethanol was 4:1; the detergent was petroleum ether, and the volume ratio of the detergent to anhydrous ethanol was 1:1.

[0148] Example 7: Compared with Example 4, the difference lies in the preparation of the copolymer.

[0149] Preparation of the copolymer: N,N-diethylaminoethyl acrylate, dimethyl 3-methylpent-2-enedioate and isobutyl aminocrotonate were dissolved in anhydrous ethanol, stirred at 200 rpm for 10 min, azobisisobutyronitrile was added, and the reaction was carried out at 70 °C and 300 rpm for 8 h under nitrogen protection to obtain a reaction solution; the reaction solution was cooled to room temperature, petroleum ether was added, stirred at 200 rpm for 10 min, and the precipitate was collected by suction filtration after standing for 1 h; the precipitate was washed with detergent 3 times and dried at 40 °C under vacuum for 12 h to obtain the copolymer. The mass-volume ratio of N,N-diethylaminoethyl acrylate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of dimethyl 3-methylpent-2-enedioate to anhydrous ethanol was 1.24 g:10 mL, the mass-volume ratio of isobutyl aminocrotonate to anhydrous ethanol was 1.5 g:10 mL, the mass-volume ratio of azobisisobutyronitrile to anhydrous ethanol was 32 mg:10 mL, and the volume ratio of petroleum ether to anhydrous ethanol was 4:1; the detergent was petroleum ether, and the volume ratio of the detergent to anhydrous ethanol was 1:1.

[0150] Comparative Example 1: The comparative example is different from Example 1 in that a cyanine dye was prepared.

[0151] The synthetic route diagram for preparing the cyanine dye in the comparative example is shown in Figure 4 , in which compound 1 is a quaternary onium salt.

[0152] Preparation of the quaternary onium salt: n-butyl iodide was dissolved in acetonitrile to obtain a n-butyl iodide solution; 2,3,3-trimethylindole was dissolved in acetonitrile, heated to reflux at 90 °C, and the n-butyl iodide solution was added to the refluxing liquid, which was then continuously refluxed at 90 °C for 40 h; the reaction solution was cooled to room temperature, filtered to obtain a solid, which was washed with ethyl acetate 3 times and dried under vacuum to obtain the quaternary onium salt. The mass-volume ratio of n-butyl iodide to acetonitrile in the n-butyl iodide solution was 9.3 g:5 mL; the mass-volume ratio of 2,3,3-trimethylindole to acetonitrile was 2 g:10 mL; the mass of the n-butyl iodide solution was measured based on the mass of n-butyl iodide therein, and the mass ratio of 2,3,3-trimethylindole to n-butyl iodide was 2:9.3.

[0153] Preparation of the cyanine dye: the quaternary onium salt and triethyl orthoformate were dissolved in anhydrous pyridine, and the reaction was carried out at 120 °C under argon protection for 16 h to obtain a reaction solution, which was cooled to room temperature, and the solvent was removed by distillation under reduced pressure; the reaction solution was purified by column chromatography to obtain Cy3 dye, i.e., the cyanine dye. The mass-volume ratio of the quaternary onium salt to anhydrous pyridine was 216 mg:1 mL, and the mass-volume ratio of triethyl orthoformate to anhydrous pyridine was 296 mg:1 mL; the stationary phase of the column chromatography was silica gel, and the eluent included dichloromethane and methanol, and the volume ratio of dichloromethane to methanol was 10:1.

[0154] Comparative Example 2: This comparative example is compared with Example 1, except that the preparation of the cyanine dye is different.

[0155] The synthetic route chart for the preparation of the cyanine dye in this comparative example is shown in FIG. 1, wherein compound 1 is the quaternary onium salt and compound 6 is the condensing agent. Figure 5

[0156] Preparation of the quaternary onium salt: iodide-n-butane was dissolved in acetonitrile to obtain an iodide-n-butane solution; 2,3,3-trimethylindole was dissolved in acetonitrile, heated to reflux at 90°C, and the iodide-n-butane solution was added to the refluxing solution, which was then refluxed at 90°C for 40 h. After the reaction solution was cooled to room temperature, the solid was filtered, washed with ethyl acetate for 3 times, and dried under vacuum to obtain the quaternary onium salt. The mass / volume ratio of iodide-n-butane to acetonitrile in the iodide-n-butane solution was 9.3 g:5 mL; the mass / volume ratio of 2,3,3-trimethylindole to acetonitrile was 2 g:10 mL; the mass of the iodide-n-butane solution was measured based on the mass of iodide-n-butane therein; and the mass ratio of 2,3,3-trimethylindole to iodide-n-butane was 2:9.3.

[0157] Preparation of the condensing agent: 1,1,3,3-tetramethoxypropane and aniline were dissolved in ethanol under argon protection, cooled to 0°C in an ice bath, and concentrated hydrochloric acid was added dropwise. After stirring at room temperature for 3 h, the solvent was removed, the precipitate was washed with ice water once, and dried under high vacuum to obtain the condensing agent. The volume ratio of 1,1,3,3-tetramethoxypropane to ethanol was 1:15, the volume ratio of aniline to ethanol was 1.1:15, and the volume ratio of concentrated hydrochloric acid to ethanol was 1.2:15.

[0158] Preparation of the cyanine dye: the quaternary onium salt and the condensing agent were dissolved in anhydrous pyridine under argon protection, refluxed at 120°C for 16 h to obtain a reaction solution, which was cooled to room temperature, and the solvent was removed by distillation under reduced pressure. The cyanine dye, Cy5 dye, was obtained by column chromatography purification. The mass / volume ratio of the quaternary onium salt to anhydrous pyridine was 216 mg:1 mL, and the mass / volume ratio of the condensing agent to anhydrous pyridine was 296 mg:1 mL. The column chromatography used silica gel as the stationary phase, and the eluent included dichloromethane and methanol, with a volume ratio of 10:1.

[0159] Comparative Example 3: This comparative example is compared with Example 4, except that the preparation of the copolymer is different.

[0160] ​Preparation of the copolymer: N,N-diethylaminoethyl acrylate was dissolved in anhydrous ethanol, stirred at 200 rpm for 10 min, and azobisisobutyronitrile was added. The reaction was carried out at 70°C and 300 rpm for 8 h under nitrogen protection to obtain a reaction solution. After the reaction solution was cooled to room temperature, petroleum ether was added, and the mixture was stirred at 200 rpm for 10 min. The precipitate was collected by suction filtration after standing for 1 h. The precipitate was washed with detergent three times and dried at 40°C under vacuum for 12 h to obtain the copolymer. The mass-volume ratio of N,N-diethylaminoethyl acrylate to anhydrous ethanol was 1 g:10 mL, the mass-volume ratio of azobisisobutyronitrile to anhydrous ethanol was 32 mg:10 mL, and the volume ratio of petroleum ether to anhydrous ethanol was 4:1. The detergent was petroleum ether, and the volume ratio of the detergent to anhydrous ethanol was 1:1.

[0161] Comparative Example 4: This comparative example is different from Example 4 in the preparation of the copolymer.

[0162] Preparation of the copolymer: N,N-diethylaminoethyl acrylate was dissolved in anhydrous ethanol, stirred at 200 rpm for 10 min, and azobisisobutyronitrile was added. The reaction was carried out at 70°C and 300 rpm for 8 h under nitrogen protection to obtain a reaction solution. After the reaction solution was cooled to room temperature, petroleum ether was added, and the mixture was stirred at 200 rpm for 10 min. The precipitate was collected by suction filtration after standing for 1 h. The precipitate was washed with detergent three times and dried at 40°C under vacuum for 12 h to obtain the copolymer. The mass-volume ratio of N,N-diethylaminoethyl acrylate to anhydrous ethanol was 1.24 g:10 mL, the mass-volume ratio of azobisisobutyronitrile to anhydrous ethanol was 32 mg:10 mL, and the volume ratio of petroleum ether to anhydrous ethanol was 4:1. The detergent was petroleum ether, and the volume ratio of the detergent to anhydrous ethanol was 1:1.

[0163] Test Example 1: Test of the radiosensitivity of the cyanine dyes.

[0164] Test sample: cyanine dyes prepared in Examples 1-3 and Comparative Examples 1-2.

[0165] Test method: The cyanine dyes prepared in Examples 1-3 and Comparative Examples 1-2 were dispersed in deionized water to obtain IR780 dye solution, Br-IR780 dye solution, Cl-IR780 dye solution, Cy3 dye solution, and Cy5 dye solution, i.e., cyanine dye solution, at 10 μmol / L. Na 131 I radioactive solution was diluted to 0 μCi, 1 μCi, 5 μCi, 10 μCi, 20 μCi, and 40 μCi to obtain Na 131 I radioactive solutions with different activities. A plurality of 1.5 mL EP tubes were taken, and 500 μL of the Na 131I radioactive solution, another set of 500 μL of blank deionized water is added as a blank group, 50 μL of the cyanine dye solution is added respectively, incubated at room temperature in the dark environment for 2 h, then the color change of each group of solution is observed directly by naked eye and recorded, then the ultraviolet-visible absorption spectrum change of each group of solution is recorded by using ultraviolet-visible spectrophotometer; the IR780 dye solution is at 800 nm, the Br-IR780 dye solution is at 780 nm, and the Cl-IR780 dye solution is at 800 nm.

[0166] The radiation sensitivity test of the cyanine dye prepared in the application is as shown in the figure, wherein the cyanine dye prepared in example 1 is as shown in the figure. Figures 6-16 The response UV-Vis spectrum change graph of the cyanine dye prepared in example 1 to Na 131 I is as shown in the figure. Figure 6 The color change graph of the cyanine dye prepared in example 1 to Na 131 I is as shown in the figure. Figure 7 The response UV-Vis spectrum change graph of the cyanine dye prepared in example 2 to Na 131 I is as shown in the figure. Figure 8 The color change graph of the cyanine dye prepared in example 2 to Na 131 I is as shown in the figure. Figure 9 The response UV-Vis spectrum change graph of the cyanine dye prepared in example 3 to Na 131 I is as shown in the figure. Figure 10 The color change graph of the cyanine dye prepared in example 3 to Na 131 I is as shown in the figure. Figure 11 The relationship graph of the absorption value of the cyanine dye prepared in example 3 at 800 nm and the radioactivity of Na 131 I is as shown in the figure. Figure 12 The response UV-Vis spectrum change graph of the cyanine dye prepared in comparative example 1 to Na 131 I is as shown in the figure. Figure 13 The color change graph of the cyanine dye prepared in comparative example 1 to Na 131 I is as shown in the figure. Figure 14 The response UV-Vis spectrum change graph of the cyanine dye prepared in comparative example 2 to Na 131 I is as shown in the figure. Figure 15 The color change graph of the cyanine dye prepared in comparative example 2 to Na 131 I is as shown in the figure. Figure 16 The blank group and each cyanine dye and 0 μCi Na 131The solution color of each group remained the initial color, the cyanine dye solution prepared in Comparative Example 1 was initially red, the cyanine dye solution prepared in Comparative Example 2 was initially blue, the cyanine dye solution prepared in Example 1 was initially green, the cyanine dye solution prepared in Example 2 was initially green, and the cyanine dye solution prepared in Example 3 was initially green. The Na 131 After the irradiation, the cyanine dye solutions prepared in Comparative Example 1 and Comparative Example 2 did not show obvious color changes at the highest activity of 40 μCi, and only the absorbance at the characteristic wavelength slightly decreased; the cyanine dye solution prepared in Example 1 did not show significant color changes and changes in the absorption spectrum at all test activities; the cyanine dye solutions prepared in Example 2 and Example 3 showed obvious dose-dependent color changes, and as the Na 131 As the radioactivity increased from 1 μCi to 40 μCi, both dye solutions gradually changed from the initial green to yellow, and the higher the activity, the more obvious the color change, wherein the cyanine dye solution prepared in Example 3 showed a discernible color change at an activity of 1 μCi, and the change in absorbance at the characteristic absorption wavelength of 800 nm was linear with the Na 131 The radioactivity showed a good linear relationship in the test range, and the determination coefficient R2 was 0.946. The color change and the change in absorbance of the cyanine dye solution prepared in Example 2 were slightly weaker than those of the cyanine dye solution prepared in Example 3, which comprehensively indicated that the radiation sensitivity of the cyanine dye prepared in Example 3 was optimal.

[0167] Test Example 2: Test of the radiation response mechanism of the cyanine dye.

[0168] Test sample: cyanine dye prepared in Example 2-3.

[0169] Test method: The cyanine dye prepared in Example 2-3 was dispersed in deionized water to obtain a 10 μmol / L Br-IR780 dye solution and a Cl-IR780 dye solution, i.e., a cyanine dye solution; the Na 131 The radioactive solution was diluted to 0 μCi, 20 μCi, 50 μCi, and 100 μCi to obtain Na 131 The radioactive solution; 1 mL of the cyanine dye solution was taken and divided into two groups, one group was added with 1 mL of deionized water as a water system group, and the other group was added with 1 mL of methanol as a methanol system group; 200 μL of the Na 131 The radioactive solution was diluted to 0 μCi, 20 μCi, 50 μCi, and 100 μCi to obtain Na

[0170] The test results of the radiation response mechanism of the anthocyanin dye prepared in this invention are as follows: Figure 17 As shown, in the water system group, with Na 131 As the activity of I increased from 0 μCi to 100 μCi, both Br-IR780 and Cl-IR780 solutions gradually changed from their initial green color to yellow, and the degree of color change became more pronounced with increasing activity. The absorbance values ​​at the corresponding characteristic absorption wavelengths also decreased significantly. The absorbance value of the Br-IR780 dye prepared in Example 2 decreased by 42.3% in the 100 μCi activity group compared to the 0 μCi group, and the absorbance value of the Cl-IR780 dye prepared in Example 3 decreased by 51.7% in the 100 μCi activity group compared to the 0 μCi group. In the methanol system group, regardless of Na... 131 Regardless of the I activity, both Br-IR780 and Cl-IR780 solutions maintained their initial green color, and the absorbance values ​​at the characteristic absorption wavelengths were less than 3% different from the 0 μCi group, showing no significant change. These results indicate that methanol can inhibit the radiation response of the dyes by scavenging ·OH, demonstrating that ·OH generated by water radiolysis is the key reactive species causing the destruction of the conjugated structure of Br-IR780 and Cl-IR780 dyes, resulting in changes in color and absorbance. Furthermore, Cl-IR780 exhibits slightly higher reaction efficiency with ·OH than Br-IR780, which is related to the stronger H- aggregation tendency of Br-IR780, reducing its contact efficiency with ·OH.

[0171] Experimental Example 3: Interference resistance test of cyanine dyes.

[0172] Test sample: cyanine dye prepared in Example 3.

[0173] Test method: Prepare buffer solutions of the cyanine dye prepared in Example 3, and add high concentrations of possible interfering substances to each solution, with a final concentration of 50 mmol / L, including Na⁺ (NaCl), K⁺ (KCl), Ca²⁺ (CaCl2), Mg²⁺ (MgCl2), Cl⁻ (NaCl), glucose, uric acid, and normal mouse urine. After incubating in a dark environment at room temperature for 2 hours, observe and record the color changes of each solution directly, and then use a UV-Vis spectrophotometer to measure the absorbance value at 800 nm.

[0174] The UV-Vis spectrum change of the anthocyanin dye prepared in Example 3 in response to interference testing is shown in the figure below. Figure 18 As shown, the color change diagram of the cyanine dye prepared in Example 3 in response to interference testing is as follows. Figure 19As shown, the buffer solution of the blank group is stable green, the solutions of each group added with sodium chloride, potassium chloride, calcium chloride, magnesium chloride, glucose, uric acid solution and the group added with normal mouse urine are consistent with the blank group in solution color and have no obvious change. The absorbance values of each group at 800 nm have very small difference compared with the blank group and have no significant downward trend, indicating that the ions and metabolites in common biological fluids do not interfere with the color and absorbance of the cyanine dye prepared in Example 3, and the cyanine dye prepared in Example 3 has excellent anti-interference ability.

[0175] Test Example 4: Stability test of cyanine dye.

[0176] Test sample: cyanine dye prepared in Example 3.

[0177] Test method: The cyanine dye prepared in Example 3 was dispersed in deionized water and stored in a sealed container at room temperature in the dark. At 0 h, 12 h, 24 h, 3 d and 7 d of storage, part of the solution was taken out to observe the color change with the naked eye, and the absorbance value of the solution at 800 nm was measured using a UV-Vis spectrophotometer. The observation results and absorbance data at each time node were recorded.

[0178] The UV-Vis spectrum change chart of the response of the cyanine dye prepared in Example 3 to the stability test is shown in Figure 20 As shown, the color change chart of the response of the cyanine dye prepared in Example 3 to the stability test is shown in Figure 21 As shown, the Cl-IR780 aqueous solution remained stable green at 0 h, 12 h, 24 h and 3 d of storage at room temperature in the dark, and the characteristic absorption peak at 800 nm had no obvious shift and the absorbance value was basically consistent. At 7 d of storage, the solution color changed obviously yellow, and the absorbance value at 800 nm decreased significantly, indicating that the Cl-IR780 aqueous solution has good stability within 3 d, which can meet the requirements of test strip preparation and short-term storage. For long-term storage, it is recommended to use solid form or dissolved in organic solvent.

[0179] Test Example 5: In vitro detection verification test of radioactivity detection system.

[0180] Test sample: radioactivity detection system prepared in Example 3.

[0181] Test method: Na 131I was diluted into a series of different activities, including 0, 2, 5, 10, 20, 40 μCi, and 20 μL of each activity solution was added to the center of the test strip prepared in Example 2. The test strip with added drop buffer was used as a blank control. After 2 h, the photos of all test strips were taken vertically 50 cm above the strips under standard light conditions of white light, 4000 K, using the camera of a smart phone. The hue value of the center of each test strip photo was analyzed using the phone App, and the results were expressed as Na 131 I activity as the abscissa and the corresponding hue value as the ordinate to draw the standard curve.

[0182] The results of the validation test of the radioactivity detection system are shown in Figure 22 The color of the test strip gradually changed from green to yellow as the activity increased. The hue value decreased linearly with the increase of activity, showing a good linear relationship, indicating that the standard curve could be used for quantitative analysis of unknown samples.

[0183] Test Example 6: Evaluation test of urine matrix effect of the radioactivity detection system.

[0184] Test sample: The radioactivity detection system prepared in Example 3.

[0185] Test method: The urine of normal untreated mice was directly added to the test strip and compared with the test strip with added deionized water. Healthy Balb / c mice were injected with 100 μCi of Na 131 I solution through the tail vein, and the urine of the mice was collected at 30 min, 1 h, and 2 h after injection, respectively. The collected urine samples were added to the test strip prepared in Example 3, and the blank urine before injection was set as a control. After drying, the photos were taken using a smart phone and the hue value was analyzed.

[0186] The results of the evaluation test of the urine matrix effect of the radioactivity detection system are shown in Figure 23 There was no significant difference in the color of the test strip and the measured hue value between the urine sample and the deionized water sample, indicating that the components in the urine did not cause non-specific color change of the cyanine dye prepared in Example 3, proving the feasibility of the method for urine sample detection. Compared with the blank control, all urine samples collected after Na 131 I injection caused obvious color change of the test strip, from green to yellow-green or yellow. The color change of the urine sample collected at 30 min was the most significant, indicating that the radioactivity in the urine at this time was the highest, which was consistent with the physiological process of rapid clearance of Na 131 I through the kidneys. The degree of color change of the samples at 1 h and 2 h was weakened in turn, indicating that the radioactivity gradually decreased over time.

[0187] Test Example 7: Radiation response sensitivity test of the test strip.

[0188] Test samples: test strips prepared in Examples 3-7.

[0189] Test method: Na 131 I was diluted with deionized water to a series of activity solutions of 0 μCi, 0.2 μCi, 0.3 μCi, 0.5 μCi, 0.8 μCi, 1.0 μCi, 1.5 μCi, 2.0 μCi, 2.2 μCi; 20 μL of Na 131 I solution corresponding to the activity was added to the center of each test strip prepared in Examples 3-7, and a test strip to which 20 μL of deionized water was added was used as a blank control; after incubation at room temperature in the dark for 2 h, the image was taken vertically 50 cm above the test strip using the camera of a smart phone under standard light conditions of white light 4000 K, and the HSV hue value of the central region of each test strip was extracted.

[0190] The results of the radiation response sensitivity test of the test strips are shown in Table 1.

[0191] Table 1 Results of the radiation response sensitivity test of the test strips

[0192]

[0193] The test strip prepared in Example 3 used glycerol as an additive, and the hue value was relatively high; the test strip prepared in Example 4 used a copolymer prepared from N,N-diethylaminoethyl acrylate and 3-methylpent-2-ene dimethyl malonate as an additive, and the hue value was significantly lower than that of Example 3, indicating that the addition of the base copolymer can promote the response of the dye to radiation, help the dye to disperse uniformly on the substrate surface, and make ·OH more easily contact the dye molecules and initiate a reaction; Example 5 further optimized the interaction between the copolymer and the dye by increasing the proportion of 3-methylpent-2-ene dimethyl malonate in the copolymer, resulting in a lower hue value than Example 4 and further improved radiation response ability; Example 6 introduced isobutyl aminocrotonate into the copolymer system, which further improved the dispersibility and radiation sensitivity of the dye on the substrate, and the hue value was lower than that of Example 5, and the radiation response effect was better; Example 7 achieved the best effect of the copolymer and the dye by optimizing the proportion of isobutyl aminocrotonate in the copolymer, and the hue value was the lowest among all the test samples, and the radiation response ability was the best; Comparative Example 3 used only N,N-diethylaminoethyl acrylate as a single monomer to prepare a copolymer, and the copolymer had limited auxiliary effect on the dye, and the radiation response effect was weaker than that of Example 4; Comparative Example 4 used 3-methylpent-2-ene dimethyl malonate as a single monomer to prepare a copolymer, and the radiation response effect was also weaker than that of Example 4.

[0194] Test Example 8: Storage stability test of test strips.

[0195] Test samples: test strips prepared in Examples 3-7.

[0196] Test method: the test strips prepared in Examples 3-7 were respectively placed in sealed light-proof containers, and stored at room temperature and 50% relative humidity, and 3 pieces of each sample test strip were taken out at 0 d, 3 d, 7 d and 14 d of storage, respectively; 20 μL of 0 μCi deionized water was added to each test strip, and after incubation at room temperature for 30 min, the absorbance of the test strip at 800 nm was determined by ultraviolet-visible spectrophotometry, and the determination was performed in triplicate to obtain the average value; the retention rate of the absorbance at each time point relative to the initial absorbance was calculated, and the absorbance retention rate (%) = (absorbance at a certain time point / absorbance at 0 d) x 100%.

[0197] The results of the storage stability test of the test strips are shown in Table 2.

[0198] Table 2 Results of the storage stability test of the test strips

[0199]

[0200] The test strip of Example 3 has glycerol as an additive, and the absorbance retention rate decreases obviously over time during storage, and the retention rate is low, indicating that although glycerol can play a certain moisturizing role, it cannot effectively inhibit the non-specific degradation of dye molecules and also cannot enhance the binding force of the dye and the substrate, resulting in easy migration or structural damage of the dye during storage; Example 4 has a copolymer formed by copolymerization of N,N-diethylaminoethyl acrylate and 3-methylpent-2-ene dimethyl malonate, and the absorbance retention rate is higher than that of Example 3, the N,N-diethylamino group of N,N-diethylaminoethyl acrylate can form a hydrogen bond with the cyanine dye, reducing the aggregation of dye molecules, and the ester group of 3-methylpent-2-ene dimethyl malonate can adjust the hydrophilicity and hydrophobicity of the copolymer, thereby improving the binding capacity of the copolymer and the substrate and reducing the loss of the dye, thereby significantly improving the storage stability compared with Example 3; Example 5 further improves the absorbance retention rate by adjusting the proportion of 3-methylpent-2-ene dimethyl malonate in the copolymer; Example 6 adds isobutyl aminocrotonate to the copolymer, the amino group of isobutyl aminocrotonate can form a stronger hydrogen bond with the cyanine dye, and the steric hindance of the isobutyl ester group can wrap the dye molecules, thereby inhibiting the damage of the external environment to the structure of the dye and enhancing the storage stability of the test strip; Example 7 optimizes the proportion of isobutyl aminocrotonate, so that the hydrogen bond and steric hindance effect in the copolymer are in an optimal balance, and the storage stability is optimal; Comparative Example 3 only uses N,N-diethylaminoethyl acrylate as a single monomer to prepare a copolymer, and lacks the ester group of 3-methylpent-2-ene dimethyl malonate to adjust the hydrophilicity and hydrophobicity, so that the wrapping capacity of the copolymer for the dye is insufficient, and the dye is prone to aggregation and degradation during storage, so the absorbance retention rate is lower than that of Example 4, and the storage stability is poor; Comparative Example 4 only uses 3-methylpent-2-ene dimethyl malonate as a single monomer to prepare a copolymer, and lacks the amino group of N,N-diethylaminoethyl acrylate to form a hydrogen bond with the dye, so that the binding force between the dye and the copolymer is weak and the dye is prone to loss, and therefore the absorbance retention rate is also lower than that of Example 4, and the storage stability is poor.

[0201] The above-described examples and / or embodiments are only used to illustrate the preferred embodiments and / or implementations of the present technology, and do not limit the implementation of the present technology in any form, and any person skilled in the art can make some changes or modifications to other equivalent embodiments without departing from the scope of the technology disclosed in the present disclosure, but should be considered as the same technology or embodiment as the present application.

[0202] The principles and implementation manners of the present application are described herein by using specific examples, and the above example descriptions are only used to help understand the method of the present application and its core idea. The above descriptions are only preferred embodiments of the present application, and it should be pointed out that, due to the limited nature of the language expression, there are objectively infinite specific structures, and for ordinary skilled persons in the technical field, some improvements, refinements or changes can be made without departing from the principles of the present application, and the above technical features can also be combined in an appropriate manner; these improvements, refinements, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, shall be regarded as the protection scope of the present application.

Claims

1. A radioassay colorimetric sensor characterized by: The radioactivity detection colorimetric sensor comprises a test strip, the test strip comprises a cyanine dye, the structure of the cyanine dye is formula (3), (3)， The R group is a heavy atom, which includes a bromine atom or a chlorine atom; The matrix for radioactivity detection includes an aqueous solution or urine.

2. A radioassay colorimetric sensor according to claim 1, characterized in that: The preparation reaction monomers of the cyanine dye include a quaternary ammonium salt intermediate and a condensing agent, the quaternary ammonium salt intermediate includes one of a quaternary ammonium onium salt and a heavy atom substituted quaternary ammonium salt, the heavy atom substituted quaternary ammonium salt includes a bromine substituted quaternary ammonium salt or a chlorine substituted quaternary ammonium salt, and the mass ratio of the quaternary ammonium salt intermediate to the condensing agent is 1:3-6.

3. A radioassay colorimetric sensor according to claim 2, characterized in that: The preparation reaction monomers of the heavy atom substituted quaternary ammonium salt include iodinated n-butane and a heavy atom substituted indole derivative, the heavy atom substituted indole derivative includes a bromine substituted indole derivative or a chlorine substituted indole derivative, and the mass ratio of the heavy atom substituted indole derivative to iodinated n-butane is 2:5-8.

4. A radioassay colorimetric sensor according to claim 3, characterized in that: The preparation reaction monomers of the heavy atom substituted indole derivative include a heavy atom reaction monomer and 3-methyl-2-butanone, the heavy atom reaction monomer includes 4-bromophenylhydrazine or 4-chlorophenylhydrazine, and the mass ratio of the heavy atom reaction monomer to 3-methyl-2-butanone is 1:0.5-2.

5. A radioassay colorimetric sensor according to claim 1, characterized in that: The test strip includes a substrate, the substrate includes one of filter paper, glass fiber membrane and polymer film.

6. A radioassay colorimetric sensor according to claim 1, characterized in that: The test strip includes an additive, the additive includes one of glycerol and a copolymer, the preparation reaction monomers of the copolymer include N,N-diethylaminoethyl acrylate and 3-methylpent-2-enyl dimethyl malonic acid.

7. A radioassay colorimetric sensor according to claim 6, characterized in that: The test strip includes an additive, and the mass ratio of N,N-diethylaminoethyl acrylate to 3-methylpent-2-enyl dimethyl malonic acid is 1:1-3.

8. A radioassay colorimetric sensor according to claim 1, characterized in that: The radioactivity detection colorimetric sensor includes an image acquisition device, and the image acquisition device includes a smartphone camera.

9. A radioassay colorimetric sensor according to claim 1, characterized in that: The radioactivity detection colorimetric sensor includes a data processing device, the data processing device includes an application program running on a smartphone or a cloud server, the application program is configured to extract color feature values of an image, and convert the color feature values into a radioactivity activity value according to a pre-stored standard curve, and the color feature values include a hue value Hue in an HSV color space.

10. Use of a radioactivity detecting colorimetric sensor according to any one of claims 1-9 for the detection of radionuclides, characterized in that: The radionuclide includes 131 I, the matrix for the radionuclide detection includes an aqueous solution or urine.

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