A colorimetric-fluorescent dual-mode detection reagent for on-site detection of tryptamine substances

Abstract

The present application relates to a kind of colorimetric-fluorescent dual-mode detection reagent for detecting color amine substance on site, which is composed of 4-mercapto-1,8 naphthalene dicarboxylic anhydride and organic solvent, and the reagent produces orange complex and quenches blue fluorescence by reacting 4-mercapto-1,8 naphthalene dicarboxylic anhydride with the substance to be measured, with a response time of less than 1 s, and the colorimetric-fluorescent detection limit can reach nM level.The method described in the present application can be used for on-site drug detection and analysis of color amine drugs in drug-related cases, with the characteristics of simple operation, strong anti-interference, fast response, sensitive reaction, easy visual observation, etc.The reagent provides an effective visual on-site detection technique for the detection of new and precise amine drugs, and has a broad application prospect, and provides a good research foundation for the field of drug detection, which can provide an effective technical means for the police department to quickly detect drugs on site.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of drug detection and analysis, and provides a colorimetric-fluorescent dual-mode detection reagent for on-site detection of tryptamine substances. The reagent has the characteristics of high sensitivity, anti-interference and fast response speed, and can meet the purpose of on-site, visual, rapid and low-cost detection of tryptamine substances. BACKGROUND

[0002] Tryptamine substances can be misused by illegal persons as hallucinogenic drugs, and belong to the third generation of drugs represented by new psychoactive substances. After smoking, it can affect the central nervous system, change the perception ability, make people feel happy, and produce illusion and hallucination. In severe cases, it can cause perception disorder and even mental disorder. Therefore, rapid detection of tryptamine substances has important practical significance.

[0003] At present, scholars at home and abroad have developed a series of methods for detecting tryptamine substances from the perspectives of biological health and food safety, mainly including chromatography, electrochemical method, etc. However, these analysis methods require expensive instruments and professional operators, and are difficult to be used for rapid on-site visual identification. Therefore, the development of fast, inexpensive, accurate and real-time detection means for tryptamine substances has become a research focus.

[0004] Compared with existing detection methods, optical sensing technology is widely used in on-site visual detection due to its high sensitivity, fast response speed, intuitive results and simple operation. This kind of technology relies on the construction of optically active probe molecules, and its basic structure includes a luminescent center and a key recognition group. Through the specific interaction between the recognition group and the active site of the target substance structure, the optical properties of the luminescent center change, and the specific detection of the target substance is realized.

[0005] The present application is aimed at the structural characteristics of tryptamine, i.e. having a primary amine fatty chain, and uses the end-chain amino group of the molecular structure as a recognition site. A probe is designed and prepared with naphthalene dicarboxylic anhydride as the luminescent center and sulfhydryl as the recognition group, and a colorimetric-fluorescent dual-mode detection reagent is developed for on-site detection of tryptamine substances. Based on the hydrogen bond interaction between the amino group in the tryptamine substance and 4-mercapto-1,8 naphthalene dicarboxylic anhydride, colorimetric and fluorescent dual-mode response phenomenon is generated, achieving the effect of detecting tryptamine substances, and realizing rapid, sensitive, specific and visual detection of tryptamine. The reagent is simple to prepare, portable and easy to operate, and has the potential for wide application. SUMMARY

[0006] The present application aims to provide a colorimetric-fluorescent dual-mode detection reagent for on-site detection of tryptamine substances, which is composed of 4-mercapto-1,8 naphthalene dicarboxylic anhydride and an organic solvent. The 4-mercapto-1,8 naphthalene dicarboxylic anhydride reacts with the to-be-detected substance to produce an orange complex and quench blue fluorescence, and the response time is less than 1 s, and the colorimetric-fluorescent detection limit can reach the nM level. The method can be used for on-site drug detection and rapid preliminary screening and analysis of tryptamine drugs in drug-related cases, and has the characteristics of simple operation, strong anti-interference, rapid response, sensitive reaction, convenient visual observation and the like. The reagent provides an effective visual on-site detection technical means for the detection of new and precise tryptamine drugs, has a wide application prospect, and at the same time provides a good research foundation for the field of drug detection, and can provide an effective technical means for the on-site rapid detection of drugs by the public security department.

[0007] The present application aims to provide a colorimetric-fluorescent dual-mode detection reagent for on-site detection of tryptamine substances, which is composed of 4-mercapto-1,8 naphthalene dicarboxylic anhydride and an organic solvent. The 4-mercapto-1,8 naphthalene dicarboxylic anhydride reacts with the to-be-detected substance to produce an orange complex and quench blue fluorescence, and the response time is less than 1 s, and the colorimetric-fluorescent detection limit can reach the nM level. The method can be used for on-site drug detection and rapid preliminary screening and analysis of tryptamine drugs in drug-related cases, and has the characteristics of simple operation, strong anti-interference, rapid response, sensitive reaction, convenient visual observation and the like. The reagent provides an effective visual on-site detection technical means for the detection of new and precise tryptamine drugs, has a wide application prospect, and at the same time provides a good research foundation for the field of drug detection, and can provide an effective technical means for the on-site rapid detection of drugs by the public security department.

[0008] Preparation of the detection reagent:

[0009] a. 4-bromo-1,8 naphthalene dicarboxylic anhydride and sodium sulfide nonahydrate are respectively dissolved in an organic solvent N,N-dimethylformamide according to a molar ratio of 1:2, and reacted at room temperature for 6 h;

[0010] b. After the reaction is completed, the reaction product in step a is placed in 500 mL ice water, and dilute hydrochloric acid is added dropwise to adjust the pH to 1-2. Then the product is suction filtered, washed with a large amount of water until the pH is neutral, and dried to obtain a crude product;

[0011] c. The crude product obtained in step b is recrystallized with 1,4-dioxane, and then suction filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8 naphthalene dicarboxylic anhydride;

[0012] d. 4-mercapto-1,8 naphthalene dicarboxylic anhydride is weighed and dissolved in a solvent such as ethyl acetate, acetone or toluene, and after ultrasonic dissolution, a 4-mercapto-1,8 naphthalene dicarboxylic anhydride solution with a concentration of 1-100 μM is obtained, which is the colorimetric-fluorescent dual-mode detection reagent;

[0013] Preparation of the to-be-detected substance solution:

[0014] e. Tryptamine substances are weighed and dissolved in a solvent such as ethyl acetate, acetone or toluene, and after ultrasonic dissolution, transferred to a volumetric flask and diluted to 1000 mL to obtain a detection tryptamine standard solution with a concentration of 100 μM;

[0015] Colorimetric-fluorescent dual-mode detection of tryptamine substances:

[0016] f. Add 1 mL of the test reagent obtained in step d to the test bottle, add 1 mL of the analyte solution from step e, and then measure it with a UV spectrophotometer and a fluorescence spectrometer. Observe the colorimetric and fluorescence signal changes under sunlight and 365 nm light conditions. Confirm the presence of tryptamine substances based on the signal changes. The tryptamine substances include tryptamine and tryptamine derivatives that are abused as drugs.

[0017] When tryptamine-like substances are added to the reagent, a colorimetric phenomenon of color change from colorless to orange and a blue fluorescence quenching phenomenon are immediately produced, with a response time of <1s.

[0018] The present invention discloses a colorimetric-fluorescence dual-mode detection reagent for on-site detection of tryptamines. The probe described in this method is chemically named 4-mercapto-1,8-naphthalenedicarboxylic anhydride, obtained by reacting 4-bromo-1,8-naphthalenedicarboxylic anhydride with sodium sulfide nonahydrate. Its chemical structural formula (I) is:

[0019]

[0020] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0021] The present invention discloses a colorimetric-fluorescence dual-mode detection reagent for on-site detection of tryptamines. This method is simple to prepare, low in cost, mild in reaction, fast in response, sensitive in reaction, stable and clear in color development, can be directly observed with the naked eye, and has strong anti-interference ability. It provides an excellent technical means for on-site detection of tryptamines and provides a good research foundation for the field of detection of novel psychoactive substances. Attached Figure Description

[0022] Figure 1 These are images showing the colorimetric response of the detection reagent of this invention to a series of concentrations of tryptophan.

[0023] Figure 2 The UV absorption spectra of the detection reagent of this invention for a series of concentrations of tryptophan are shown.

[0024] Figure 3 The results are colorimetric tests of the specificity and resistance to interference of the detection reagent of this invention for para-tryptamine, common drugs, structural analogs and potential coexisting substances.

[0025] Figure 4 This is a visualization of the fluorescence response of the detection reagent of the present invention to a series of concentrations of tryptophan;

[0026] Figure 5 The fluorescence spectra of the detection reagent of this invention for a series of concentrations of tryptophan are shown.

[0027] Figure 6The results show the fluorescence test results of the detection reagent of this invention for the specificity and resistance to interference of tryptamine, common drugs, structural analogs and potential coexisting substances. Detailed Implementation

[0028] The present invention will be further described below through specific embodiments, but the invention is not limited to these embodiments.

[0029] Example 1

[0030] Preparation of detection reagents:

[0031] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0032] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0033] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0034] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate the solution for 10 min until homogeneous. Use this solution as the probe stock solution. Dilute 1 mL of the solution with ethyl acetate to 1000 mL to obtain a 10 μM colorimetric-fluorescent dual-mode reagent for 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0035] Preparation of the analyte solution:

[0036] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of ethyl acetate. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain the tryptamine standard solution with a concentration of 100μM.

[0037] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0038] f. Add 1 mL of the 10 μM test reagent obtained in step d to the test vial, and add 1 mL of the 100 μM analyte solution from step e to obtain a mixed solution with a tryptamine concentration of 50 μM. Within 1 second, the solution changes from colorless to orange-red. Simultaneously, under 365 nm UV light irradiation, the blue fluorescence signal of the test reagent exhibits quenching. The rapid response and significant color / fluorescence change indicate that the detection system contains tryptamine-like substances. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm, and fluorescence spectroscopy, with an excitation wavelength of 380 nm, showed a decrease in fluorescence emission peak intensity at 450 nm.

[0039] Example 2

[0040] Preparation of detection reagents:

[0041] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0042] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0043] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0044] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthoic anhydride and dissolve it in 100 mL of acetone. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 1 mL of the solution and dilute it with acetone to 1000 mL to obtain a colorimetric-fluorescent dual-mode reagent with a concentration of 10 μM 4-mercapto-1,8-naphthoic anhydride.

[0045] Preparation of the analyte solution:

[0046] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of acetone. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain the tryptamine standard solution with a concentration of 100μM.

[0047] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0048] f. Add 1 mL of the 10 μM test reagent obtained in step d to the test vial, and add 1 mL of 100 μM tryptophan-acetone solution to obtain a mixed solution with a tryptophan concentration of 50 μM. Within 1 second, the solution changes from colorless to orange-red. Simultaneously, under 365 nm UV light irradiation, the blue fluorescence signal of the test reagent exhibits quenching. The rapid response and significant color / fluorescence change indicate that the detection system contains tryptophan-like substances. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm, and fluorescence spectroscopy, with an excitation wavelength of 380 nm, showed a decrease in fluorescence emission peak intensity at 450 nm.

[0049] Example 3

[0050] Preparation of detection reagents:

[0051] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0052] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0053] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0054] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of toluene. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 1 mL of the solution and dilute it with toluene to 1000 mL to obtain a 10 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0055] Preparation of the analyte solution:

[0056] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of toluene. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain a 100μM standard solution for tryptamine detection.

[0057] f. Add 1 mL of the 10 μM test reagent obtained in step d to the test vial, and add 1 mL of 100 μM tryptamine toluene solution to obtain a mixed solution with a tryptamine concentration of 50 μM. Within 1 second, the solution changes from colorless to orange-red. Simultaneously, under 365 nm UV light irradiation, the blue fluorescence signal of the test reagent exhibits quenching. The rapid response and significant color / fluorescence change indicate that the detection system contains tryptamine-like substances. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm, and fluorescence spectroscopy, with an excitation wavelength of 380 nm, showed a decrease in fluorescence emission peak intensity at 450 nm.

[0058] Example 4

[0059] Preparation of detection reagents:

[0060] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0061] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0062] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0063] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 0.1 mL of the solution and dilute it with ethyl acetate to 1000 mL to obtain a 1 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0064] Preparation of the analyte solution:

[0065] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of ethyl acetate. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain the tryptamine standard solution with a concentration of 100μM.

[0066] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0067] f. Add 1 mL of the 1 μM test reagent obtained in step d to the test bottle, and then add 1 mL of the 100 μM ethyl tryptophan solution obtained in step e, to obtain a mixed solution with a tryptophan concentration of 50 μM. Within 1 second, the solution changes from colorless to orange-red. Simultaneously, under 365 nm UV light irradiation, the blue fluorescence signal of the test reagent exhibits quenching. These phenomena demonstrate rapid response and significant color / fluorescence changes, indicating that the detection system contains tryptophan-like substances. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm, and fluorescence spectroscopy, with an excitation wavelength of 380 nm, showed a decrease in fluorescence emission peak intensity at 450 nm.

[0068] Example 5

[0069] Preparation of detection reagents:

[0070] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0071] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0072] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0073] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Dilute 2 mL of the solution with ethyl acetate to 1000 mL to obtain a 20 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0074] Preparation of the analyte solution:

[0075] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of ethyl acetate. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain the tryptamine standard solution with a concentration of 100μM.

[0076] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0077] f. Add 1 mL of the 20 μM test reagent obtained in step d to the test vial, and add 1 mL of the 100 μM ethyl tryptophan solution from step e to obtain a mixed solution with a tryptophan concentration of 50 μM. Within 1 second, the solution changes from colorless to orange-red. Simultaneously, under 365 nm UV light irradiation, the blue fluorescence signal of the test reagent exhibits quenching. The rapid response and significant color / fluorescence changes indicate that the detection system contains tryptophan-like substances. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm, and fluorescence spectroscopy, with an excitation wavelength of 380 nm, showed a decrease in fluorescence emission peak intensity at 450 nm.

[0078] Example 6

[0079] Preparation of detection reagents:

[0080] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0081] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0082] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0083] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 5 mL of the solution and dilute it with ethyl acetate to 1000 mL to obtain a 50 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0084] Preparation of the analyte solution:

[0085] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of ethyl acetate. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain the tryptamine standard solution with a concentration of 100μM.

[0086] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0087] f. Add 1 mL of the 50 μM test reagent obtained in step d to the test bottle, and add 1 mL of the 100 μM ethyl tryptophan solution from step e to obtain a mixed solution with a tryptophan concentration of 50 μM. Within 1 second, the solution can be observed to change from colorless to orange-red. At the same time, under 365 nm ultraviolet light irradiation, the blue fluorescence signal of the test reagent is found to quench. The above phenomena are rapid and the color / fluorescence changes are obvious, indicating that the detection system contains tryptophan. In addition, ultraviolet spectroscopy shows that a new absorption peak appears at 490 nm. Fluorescence spectroscopy, with an excitation wavelength of 380 nm, shows that the intensity of the fluorescence emission peak decreases at 450 nm.

[0088] Example 7

[0089] Preparation of detection reagents:

[0090] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0091] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0092] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0093] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 10 mL of the solution and dilute it with ethyl acetate to 1000 mL to obtain a 100 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0094] Preparation of the analyte solution:

[0095] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of ethyl acetate. Sonicate for 10min until homogeneous, then transfer to a volumetric flask and dilute to 1000mL to obtain the tryptamine standard solution with a concentration of 100μM.

[0096] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0097] f. Add 1 mL of the 100 μM test reagent obtained in step d to the test bottle, and add 1 mL of the 100 μM ethyl tryptophan solution from step e to obtain a mixed solution with a tryptophan concentration of 50 μM. Within 1 second, the solution can be observed to change from colorless to orange-red. At the same time, under 365 nm ultraviolet light irradiation, the blue fluorescence signal of the test reagent shows a quenching phenomenon. The above phenomena are rapid and the color / fluorescence changes are obvious, indicating that the detection system contains tryptophan-like substances. In addition, ultraviolet spectroscopy revealed a new absorption peak at 490 nm. Fluorescence spectroscopy, with an excitation wavelength of 380 nm, showed a decrease in the intensity of the fluorescence emission peak at 450 nm.

[0098] Example 8

[0099] Preparation of detection reagents:

[0100] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0101] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0102] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0103] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 1 mL of the solution and dilute it with ethyl acetate to 1000 mL to obtain a 10 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0104] Preparation of the analyte solution:

[0105] e. Weigh 0.016g of tryptamine and dissolve it in 100mL of ethyl acetate. Dilute the solution to prepare tryptamine standard solutions of 0.1, 1, 50, 100, 200, 300, 400, 600, 800, 1000, 2000, 3000, 4000 and 5000μM respectively.

[0106] f. Add 1 mL of the 100 μM test reagent obtained in step d to the test vial, and add 1 mL of the ethyl tryptophan solution prepared in step e. Within 1 second, the solution color can be observed to gradually change from colorless to orange-red as the tryptophan concentration increases. Simultaneously, under 365 nm UV irradiation, the blue fluorescence of the test reagent gradually quenches with increasing tryptophan concentration until it is completely quenched. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm, with the absorbance showing a gradient increase. Fluorescence spectroscopy, using an excitation wavelength of 380 nm, showed a gradual decrease in fluorescence emission peak intensity at 450 nm. Figure 1 , 4 As shown, the spectrum is as follows Figure 2 , 5 As shown.

[0107] Example 9

[0108] Preparation of detection reagents:

[0109] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0110] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0111] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0112] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 1 mL of the solution and dilute it with ethyl acetate to 1000 mL to obtain a 10 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0113] Preparation of the analyte solution:

[0114] e. Weigh 0.008g of tryptamine and dissolve it in 10mL of ethyl acetate. Sonicate for 10min until homogeneous to obtain the tryptamine standard solution with a concentration of 5mM.

[0115] Preparation of solutions for other substances: The analytes included tryptamine analogues (indole, 4-fluoroaniline, p-toluidine, N,N-dimethylformamide), other psychoactive substances (barbiturates, methamphetamine, cannabis, heroin, cocaine, 4,5-methylenedioxyamphetamine, morphine, methamphetamine, methadone, buprenorphine), and capsule materials (starch and ethyl cellulose, used to prepare the controlled drug "Zero Capsule" with tryptamine as the main component), all of which were prepared as 50 mM ethyl acetate solutions;

[0116] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0117] f. Add 1 mL of the 10 μM test reagent obtained in step d to the test vial, along with 1 mL of the 5 mM tryptophan solution from step e and 1 mL of the remaining 50 mM solution. Within 1 second, it can be observed that when only tryptophan is present, the test reagent solution changes from colorless to orange-red; when other substances are present, the color of the test reagent solution remains unchanged. Simultaneously, under 365 nm UV light irradiation, when only tryptophan is present, the blue fluorescence of the test reagent quenches; when other substances are present, the blue fluorescence of the test reagent solution remains unchanged. Furthermore, UV spectroscopy revealed a new absorption peak at 490 nm when only tryptophan is present, while no new absorption peak appears at 490 nm when other substances are present. Fluorescence spectroscopy revealed a decrease in the fluorescence emission peak intensity at 450 nm when only tryptophan is present, while the fluorescence emission peak intensity at 450 nm remains unchanged when other substances are present. A line graph was plotted based on the absorption peak intensity at 490 nm in the UV spectrum and the emission peak intensity at 450 nm in the fluorescence spectrum. Figure 3 , 6 ).

[0118] The above results demonstrate that this test reagent has excellent visual differentiation and detection capabilities for tryptamine, common drugs, structural analogs, and potential coexisting substances.

[0119] Example 10

[0120] Preparation of detection reagents:

[0121] a. 4-Bromo-1,8-naphthoic anhydride and sodium sulfide nonahydrate were dissolved in the organic solvent N,N-dimethylformamide at a molar ratio of 1:2 and reacted at room temperature for 6 hours.

[0122] b. After the reaction is complete, place the reaction product from step a in 500 mL of ice water, add dilute hydrochloric acid to adjust the pH to 1-2, filter the product, wash with plenty of water until the pH is neutral, and dry to obtain the crude product.

[0123] c. The crude product obtained in step b was recrystallized with 1,4-dioxane, then filtered at low temperature and dried to obtain a yellow solid probe 4-mercapto-1,8-naphthalenedicarboxylic anhydride.

[0124] d. At room temperature, weigh 0.23 g of 4-mercapto-1,8-naphthalenedicarboxylic anhydride and dissolve it in 100 mL of ethyl acetate. Sonicate for 10 min until homogeneous. Use this as the probe stock solution. Take 1 mL of the solution and dilute it with ethyl acetate to 1000 mL to obtain a 10 μM 4-mercapto-1,8-naphthalenedicarboxylic anhydride colorimetric-fluorescence dual-mode reagent.

[0125] Preparation of the analyte solution:

[0126] e. Weigh 0.008g of tryptamine and dissolve it in 10mL of ethyl acetate. Sonicate for 10min until homogeneous to obtain the tryptamine standard solution with a concentration of 5mM.

[0127] Preparation of solutions for other substances: The analytes selected include tryptamine analogues (indole, 4-fluoroaniline, p-toluidine, N,N-dimethylformamide), other psychoactive substances (barbiturates, methamphetamine, cannabis, heroin, cocaine, 4,5-methylenedioxyamphetamine, morphine, methamphetamine, methadone, buprenorphine), and capsule materials (starch and ethyl cellulose), all of which are prepared as 100 mM ethyl acetate solutions;

[0128] Colorimetric-fluorescence dual-mode detection of tryptamines:

[0129] f. Add 1 mL of the 10 μM detection reagent obtained in step d to the test bottle, and then add 1 mL of the mixed solution prepared from tryptophan and the remaining substances in step e (v:v = 1:1). Within 1 second, all system solutions can be observed to change from colorless to orange-red. Simultaneously, under 365 nm UV light irradiation, the blue fluorescence of all system solutions is quenched. Furthermore, UV spectroscopy reveals that when only tryptophan is present, a new absorption peak appears at 490 nm; fluorescence spectroscopy shows that the fluorescence emission peak intensity at 450 nm is reduced. A line graph is plotted based on the absorption peak intensity at 490 nm in the UV spectrum and the emission peak intensity at 450 nm in the fluorescence spectrum. Figure 3 ,6).

[0130] The results show that the detection of tryptamine by this reagent is not affected by common drugs, structural analogs, and potential coexisting substances.

Claims

1. A method for the on-site detection of a tryptamine species using a colorimetric-fluorometric dual-mode test reagent, characterized in that The method is carried out according to the following steps: Preparation of detection reagent: a. 4-bromo-1,8 naphthalene dianhydride and sodium sulfide nine hydrate are respectively dissolved in organic solvent N,N-dimethylformamide according to the molar ratio of 1:2, and reacted at room temperature for 6 h; b. After the reaction is completed, the reaction product in step a is placed in 500 mL ice water, and dilute hydrochloric acid is added dropwise to adjust the pH to 1-2. Then the product is suction filtered, washed with a large amount of water until the pH is neutral, and dried to obtain a crude product; c. The crude product obtained in step b is recrystallized with 1,4-dioxane, followed by low-temperature suction filtration and drying to obtain a yellow solid probe 4-mercapto-1,8 naphthalene dianhydride; d. 4-mercapto-1,8 naphthalene dianhydride is weighed and dissolved in solvent ethyl acetate, acetone or toluene. After ultrasonic dissolution, a 4-mercapto-1,8 naphthalene dianhydride solution with a concentration of 1-100 μM is obtained, which is a colorimetric-fluorescent dual-mode detection reagent; Preparation of test substance solution: e. Tryptamine substances are weighed and dissolved in solvent ethyl acetate, acetone or toluene. After ultrasonic dissolution, the solution is transferred to a volumetric flask and diluted to 1000 mL to obtain a detection tryptamine standard solution with a concentration of 100 μM; Colorimetric-fluorescent dual-mode detection of tryptamine substances: f. 1 mL of the detection reagent obtained in step d is added to a test bottle, 1 mL of the test substance solution in step e is added, and then the ultraviolet spectrophotometer and fluorescence spectrometer are used for determination. The colorimetric and fluorescent signal changes under daylight and 365 nm light are observed, and the presence of tryptamine substances, including tryptamine and tryptamine derivatives abused as drugs, is confirmed according to the signal changes.

2. A method for the on-site detection of a tryptamine species using the colorimetric-fluorometric dual-mode test reagent of claim 1, wherein, After the addition of tryptamine substances to the reagent, the colorimetric phenomenon of changing from colorless to orange and the blue fluorescence quenching phenomenon occur immediately, and the response time is <1 s.

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