A colorimetric-fluorescent probe, test pen and method for detecting tryptamine

Abstract

The present invention relates to a colorimetric fluorescent probe for detecting tryptamine and a method for preparing the probe, a test pen and a method, wherein the probe is (E)-2-(3-cyano-4-R-5,5-dimethylfuran-2(5-hydrogen)-alkylene)malononitrile, wherein R is benzo[c][1,2,5]thiadiazole or 2,5-dibromobenzene. The probe is mixed with dimethyl sulfoxide, an organic solvent, and reacts with tryptamine. When R is benzo[c][1,2,5]thiadiazole, after reacting with tryptamine, the fluorescence color changes from red to colorless, and the colorimetric color changes from red to colorless; when R is 2,5-dibromobenzene, after reacting with tryptamine, the fluorescence color changes from orange to green, and the colorimetric color changes from orange to colorless. The test pen prepared by loading a sponge-based sensing material can conveniently and quickly realize the qualitative detection of tryptamine solid particles.

Description

Technical Field

[0001] The present invention belongs to the field of drug detection and analysis, and relates to a colorimetric-fluorescent probe, a test pen and a method for detecting tryptamine. The probe has a low detection limit and a fast response speed. Based on the constructed test pen, the purpose of low cost and real-time detection of tryptamine can be achieved. Background Art

[0002] Tryptamines are a type of new psychoactive substance, known for their hallucinogenic properties. Taking a small amount can have an impact on the mind, and taking more than 30 mg can cause anxiety, muscle tension, vomiting, and high fever. Therefore, it is of great practical significance to achieve rapid detection of tryptamines.

[0003] At present, scholars at home and abroad have developed a series of methods for the detection of tryptamine from the perspective of biological health and food safety, mainly including chromatography, Raman spectroscopy, electrochemical method, etc. Most of these methods require expensive instruments and professional operators. Therefore, the development of fast, accurate, real-time and low-cost detection methods for tryptamine 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 advantages such as high sensitivity, fast response speed, intuitive results, and simple operation. This type of technology relies on the construction of optically active probe molecules. Its basic structure includes a luminescent center and a bonded recognition group. Through the specific interaction between the recognition group and the active site of the target substance, the optical properties of the luminescent center change, thereby achieving specific detection of the target substance. However, most optical probes still have disadvantages such as poor detection limits and long response times.

[0005] The invention aims at the structural feature of tryptamine, that is, it has a fatty chain primary amine. Starting from the structure of the probe molecule itself, tricyanodihydrofuran (TCF) is used as an electron-withdrawing group. The aldehyde group at the para position of the conjugated bridge connected by C=C bond of TCF is used as the recognition site of tryptamine. The electron-withdrawing ability of the conjugated bridge is regulated by post-modification with a substituent benzo[c][1,2,5]thiadiazole or 2,5-dibromobenzene, and a colorimetric-fluorescence detection probe is designed. Moreover, a polyvinyl alcohol sponge is used as a probe substrate, and a test pen is designed and prepared to realize on-site, rapid, trace and visualized detection of tryptamine solid particles. Summary of the invention

[0006] The present invention aims to provide a colorimetric-fluorescent probe, a test pen and a method for detecting tryptamine, wherein the chemical name of the probe is (E)-2-(3-cyano-4-R-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile, wherein: R is benzo[c][1,2,5]thiadiazole or 2,5-dibromobenzaldehyde, and the detection method is to prepare the prepared probe and an organic solvent dimethyl sulfoxide, and after reacting with tryptamine, when R is benzo[c][1,2,5]thiadiazole, the fluorescence and color change from red to colorless, and the colorimetric color changes from red to colorless; when R is 2,5-dibromobenzene, the fluorescence color changes from orange to green, and the colorimetric color changes from orange to colorless. At the same time, a test pen is designed and prepared using polyvinyl alcohol sponge as a probe loading substrate, which can be used for rapid screening and analysis of tryptamine solid particles.

[0007] The colorimetric-fluorescent probe for detecting tryptamine described in the present invention has a chemical name: (E)-2-(3-cyano-4-R-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile, and a chemical structural formula (1):

[0008]

[0009] Wherein: R is benzo[c][1,2,5]thiadiazole or 2,5-dibromobenzene.

[0010] The method for preparing a colorimetric-fluorescent probe for detecting tryptamine is carried out according to the following steps:

[0011] a. Mix malononitrile and 3-hydroxy-3-methyl-2-butanone in a molar ratio of 2.05:1, add to anhydrous ethanol, and add sodium ethoxide as a catalyst, set the temperature to 90° C., react for 5 hours, wash with ice anhydrous ethanol, filter and dry to obtain an intermediate product;

[0012] b. Mix the intermediate product obtained in step a with 2,1,3-benzothiadiazole-4,7-dicarboxaldehyde or 2,5-dibromoterephthalaldehyde in a molar ratio of 0.9:1, add to anhydrous ethanol, set the temperature to 90° C., react for 5 hours, wash with ice anhydrous ethanol and filter, dry, and purify by column chromatography to obtain probes (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5H)-alkylene)malononitrile and (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5H)-alkylene)malononitrile.

[0013] The test pen of the colorimetric-fluorescent probe for detecting tryptamine is composed of a pen tip (1), a hollow tube (2), a pen cover (3), a detection reagent (4), a spring (5) and a tip (6). The pen cover (3) is provided with a hollow tube (2), one end of the hollow tube (2) is connected to the pen tip (1), and the other end of the hollow tube (2) is connected to the tip (6) via a spring (5). The detection reagent (4) is placed in the hollow tube (2), and a groove is provided in the pen tip (1).

[0014] The colorimetric-fluorescent probe for detecting tryptamines is carried out according to the following steps:

[0015] Tryptamine solid detection:

[0016] a. Dissolve the obtained probe (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile or (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile in an organic solvent, dimethyl sulfoxide, to prepare a probe solution with a concentration of 500 μmol / L;

[0017] b. Testing with a test pen: In a pen cover (3) of 90.60 mm × 13.50 mm wide × 23.00 mm high, fill a polyvinyl alcohol sponge into the 5 mm × 5 mm × 1 mm groove of the pen tip (1), fill the probe solution prepared in step a into the hollow tube (2), and seal the probe solution with a sealing film;

[0018] c. Press the spring (5) connected to the hollow tube (2) in the test pen so that the tip (6) pierces the probe solution sealing film in step b. The probe solution is loaded into the polyvinyl alcohol sponge filled in the pen tip (1) based on the siphon effect. The pen tip (1) is used to gently wipe the surface of the object containing the tryptamine solid particles. Under the condition of an excitation wavelength of 365nm, the fluorescence color change is observed with the naked eye to perform a rapid qualitative analysis of the tryptamine in the sample to be tested.

[0019] Tryptamine solution test:

[0020] a. Dissolve the obtained probe (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile or (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile in an organic solvent, dimethyl sulfoxide, to prepare probe solutions with concentrations of 20 μmol / L and 100 μmol / L, to obtain a detection reagent;

[0021] b. Add the test solution to the detection reagent obtained in step a. After sufficient reaction, the fluorescence spectrum and absorption spectrum are measured using a fluorescence spectrometer and an ultraviolet spectrophotometer, and the changes in fluorescence and colorimetric signals are observed at 365 nm and under daylight illumination. For the probe (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile, the fluorescence color changes from red to colorless and the colorimetric color changes from red to colorless, indicating that tryptamine exists in the solution.

[0022] For the probe (E)-2-(3-cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile, the fluorescence color changes from orange to green and the colorimetric color changes from orange to colorless, indicating the presence of tryptamine in the solution.

[0023] The present invention discloses a colorimetric-fluorescent probe for detecting tryptamine. The detection principle of the probe is as follows: the amino group of the test object tryptamine attacks the aldehyde group of the probe molecule by nucleophilic attack, thereby generating an addition reaction, and then further generates an elimination reaction to remove water to form a Schiff base structure. The transformation of the probe to the product molecular structure corresponds to the change of fluorescence and colorimetric color.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The probe (E)-2-(3-cyano-4-R-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile (R is benzo[c][1,2,5]thiadiazole or 2,5-dibromobenzene) obtained by the method of the present invention can realize highly sensitive, rapid and visual detection of tryptamine solution. The test pen prepared by polyvinyl alcohol sponge can efficiently and conveniently realize sampling and detection of tryptamine solid particles without pre-treatment of the test object. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The fluorescence spectra and photos of the probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile before and after detecting tryptamine;

[0027] Figure 2 The ultraviolet absorption spectra and photos of the probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile before and after detecting tryptamine;

[0028] Figure 3The fluorescence spectra and photos of the probe molecule (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile before and after detecting tryptamine;

[0029] Figure 4 The ultraviolet absorption spectra and photos of the probe molecule (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile before and after detecting tryptamine;

[0030] Figure 5 The structure diagram of the test pen prepared by the present invention is shown in Figure a, which is a cross-sectional view and Figure b is an isometric view. Notes: 1. pen tip; 2. hollow tube; 3. pen cover; 4. detection reagent; 5. spring; 6. tip;

[0031] Figure 6 The photographs are of the test pen prepared by the present invention detecting tryptamine particles, wherein ac is a fluorescence photograph before detection; ad is a fluorescence photograph after detection; bc is a colorimetric photograph before detection; and bd is a colorimetric photograph after detection. DETAILED DESCRIPTION

[0032] The present invention is further described below through specific implementations, but the invention is not limited to these embodiments.

[0033] Example 1

[0034] Preparation of probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile:

[0035] a. Mix 10.036mmol, 0.66g of malononitrile and 4.896mmol, 0.5g of 3-hydroxy-3-methyl-2-butanone in a molar ratio of 2.05:1, add to 20mL of anhydrous ethanol, and add 0.675mmol, 0.046g of sodium ethoxide as a catalyst, set the temperature to 90°C, react for 5h, wash with ice anhydrous ethanol, filter and dry to obtain an intermediate product;

[0036] b. Mix 0.124 mmol, 24.8 mg of the intermediate product obtained in step a with 0.156 mmol, 30 mg of 2,1,3-benzothiadiazole-4,7-dicarboxaldehyde in a molar ratio of 0.9:1, add to 20 mL of anhydrous ethanol, set the temperature to 90°C, react for 5 hours, wash with ice anhydrous ethanol and filter, dry, and purify by column chromatography to obtain the probe (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile.

[0037] Example 2

[0038] Preparation of Probe Molecule (E)-2-(3-cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile

[0039] a. Mix 10.036mmol, 0.66g of malononitrile and 3-hydroxy-3-methyl-2-butanone (4.896mmol, 0.5g) in a molar ratio of 2.05:1, add to 20mL of anhydrous ethanol solvent, and add sodium ethoxide (0.675mmol, 0.046g) as a catalyst, set the temperature to 90°C, react for 5h, wash with ice anhydrous ethanol, filter and dry to obtain an intermediate product;

[0040] b. Mix 1.8 mmol, 0.358 g of the intermediate product obtained in step a with 2 mmol, 0.584 g of 2,5-dibromoterephthalaldehyde in a molar ratio of 0.9:1, add to 20 mL of anhydrous ethanol, set the temperature to 90°C, react for 5 hours, wash with ice anhydrous ethanol and filter, dry, and purify by column chromatography to obtain the probe (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile.

[0041] Example 3

[0042] Fluorescence Detection of Tryptamine Solution Using Probe Molecule (E)-2-(3-Cyano-4-(Benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-Hydrogen)-Alkylene)malononitrile

[0043] The probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 1 was dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the probe molecule was prepared to be 20 μM; 1.8 mL of the probe solution was taken in a cuvette, and 0.2 mL of a tryptamine solution was added. After the reaction, a fluorescence spectrum test was performed, and photos before and after the reaction were taken with a digital camera. As a result, it can be seen from the fluorescence spectrum that with the addition of the tryptamine solution, the emission peak at 641 nm decreases until it disappears. It can be seen from the photographs that the fluorescence color changes from red before the reaction to colorless, indicating that tryptamine exists in the solution. Figure 1 shown.

[0044] Example 4

[0045] Colorimetric Detection of Tryptamine Solution Using Probe Molecule (E)-2-(3-Cyano-4-(Benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-H)-alkylidene)malononitrile

[0046] The probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 1 was dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the probe molecule was 20 μM; 1.8 mL of the probe solution was taken in a cuvette, and 0.2 mL of a tryptamine solution was added. After the reaction, an ultraviolet absorption spectrum test was performed, and photos before and after the reaction were taken with a digital camera. As a result, it can be seen from the ultraviolet absorption spectrum that with the addition of the tryptamine solution, the ultraviolet absorption at 355 nm gradually increased while the ultraviolet absorption at 520 nm gradually decreased. It can be seen from the photographs that the colorimetric color changed from red before the reaction to colorless, indicating that tryptamine exists in the solution. Figure 2 shown.

[0047] Example 5

[0048] Fluorescence Detection of Tryptamine Solution Using Probe Molecule (E)-2-(3-Cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile

[0049] The probe molecule (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 2 was dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the probe molecule was prepared to be 100 μM; 1.8 mL of the probe solution was taken in a cuvette, and 0.2 mL of a tryptamine solution was added respectively. After the reaction, a fluorescence spectrum test was performed, and photos before and after the reaction were taken with a digital camera. As a result, it can be seen from the fluorescence spectrum that with the addition of the tryptamine solution, the emission peak at 535 nm gradually increased while the emission peak at 613 nm gradually decreased. It can be seen from the photographs that the fluorescence color changed from orange to green, proving the presence of tryptamine in the solution. Figure 3 shown.

[0050] Example 6

[0051] Colorimetric Detection of Tryptamine Solution Using Probe Molecule (E)-2-(3-Cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-Hydrogen)-Alkylene) Malononitrile

[0052] The probe molecule (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 2 was dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the probe molecule was 100 μM; 1.8 mL of the probe solution was taken in a cuvette, and 0.2 mL of a tryptamine solution was added respectively. After the reaction, an ultraviolet absorption spectrum test was performed, and photos before and after the reaction were taken with a digital camera. As a result, it can be seen from the ultraviolet absorption spectrum that with the addition of the tryptamine solution, the ultraviolet absorption at 342 nm gradually increased while the ultraviolet absorption at 453 nm gradually decreased. It can be seen from the photographs that the colorimetric color changed from orange to colorless, indicating that tryptamine exists in the solution, such as Figure 4 shown.

[0053] Example 7

[0054] Detection of solid particles of tryptamine by the prepared test pen

[0055] The test pen is composed of a pen tip 1, a hollow tube 2, a pen cover 3, a detection reagent 4, a spring 5 and a tip 6. The pen cover 3 is provided with a hollow tube 2, one end of the hollow tube 2 is connected to the pen tip 1, and the other end of the hollow tube 2 is connected to the tip 6 through the spring 5. The detection reagent 4 is placed in the hollow tube 2, and a groove is provided in the pen tip 1. The specific operation is performed according to the following steps:

[0056] Tryptamine solid detection:

[0057] a. The probe (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile obtained in Example 1 was dissolved in an organic solvent, dimethyl sulfoxide, to prepare a probe solution with a concentration of 500 μmol / L;

[0058] b. Testing with a test pen: In a pen cap 3 of 90.60 mm × 13.50 mm wide × 23.00 mm high, fill the polyvinyl alcohol sponge in the groove of the pen tip 1 of 5 mm × 5 mm × 1 mm, fill the probe solution prepared in step a in the hollow tube 2, and seal the probe solution with a sealing film;

[0059] c. Press the spring 5 connected to the hollow tube 2 in the test pen so that the tip 6 pierces the probe solution sealing membrane in step b. The probe solution is loaded into the polyvinyl alcohol sponge filled in the pen tip 1 based on the siphon effect. Use the pen tip 1 to gently wipe the surface of the object containing the solid particles of tryptamine. Under the condition of an excitation wavelength of 365nm, observe the change in fluorescence color with the naked eye for rapid qualitative analysis of tryptamine in the sample to be tested. If the fluorescence color of the contact part between the particles and the sponge changes from red to colorless, and the colorimetric color changes from red to colorless, it indicates the presence of tryptamine particles.

[0060] Example 8

[0061] The prepared test pen detects the solid particles of tryptamine:

[0062] The test pen is carried out according to Example 7:

[0063] Tryptamine solid detection:

[0064] a. The probe molecule (E)-2-(3-cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 2 was dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the probe molecule was 500 μM;

[0065] b. Testing with a test pen: In a pen cap 3 of 90.60 mm × 13.50 mm wide × 23.00 mm high, fill the polyvinyl alcohol sponge in the groove of the pen tip 1 of 5 mm × 5 mm × 1 mm, fill the probe solution prepared in step a in the hollow tube 2, and seal the probe solution with a sealing film;

[0066] c. Press the spring 5 connected to the hollow tube 2 in the test pen so that the tip 6 pierces the probe solution sealing membrane in step b. The probe solution is loaded into the polyvinyl alcohol sponge filled in the pen tip 1 based on the siphon effect. Use the pen tip 1 to gently wipe the surface of the object containing the solid particles of tryptamine. Under the condition of an excitation wavelength of 365nm, observe the change in fluorescence color with the naked eye for rapid qualitative analysis of tryptamine in the sample to be tested. If the fluorescence color of the contact part between the particles and the sponge changes from orange to green, and the colorimetric color changes from orange to colorless, it indicates the presence of tryptamine particles.

[0067] Example 9

[0068] The probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylene)malononitrile was used to detect the unknown solution:

[0069] The probe molecule (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 1 is dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the prepared probe molecule is 20 μM; 1.8 mL of the probe solution is taken in a cuvette, and 0.2 mL of an unknown solution is added. After the reaction, fluorescence spectrum and ultraviolet absorption spectrum tests are performed, and photos before and after the reaction are taken with a digital camera. If after the unknown solution is added, the emission peak at 641 nm in the fluorescence spectrum gradually decreases or disappears, the ultraviolet absorption at 355 nm in the ultraviolet absorption spectrum gradually increases while the ultraviolet absorption at 520 nm gradually decreases, it can be seen from the photographs that the fluorescence color changes from red to colorless, and the colorimetric color changes from red to colorless, indicating that tryptamine exists in the unknown solution.

[0070] Example 10

[0071] Detection of unknown solution by probe molecule (E)-2-(3-cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile

[0072] The probe molecule (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 2 is dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the prepared probe molecule is 100 μM; 1.8 mL of the probe solution is taken in a cuvette, and 0.2 mL of an unknown solution is added. After the reaction, fluorescence spectrum and ultraviolet absorption spectrum tests are performed, and photos before and after the reaction are taken with a digital camera. If after the unknown solution is added, the emission peak at 535 nm in the fluorescence spectrum gradually increases while the emission peak at 613 nm gradually decreases, and the ultraviolet absorption at 342 nm in the ultraviolet absorption spectrum gradually increases while the ultraviolet absorption at 453 nm gradually decreases, it can be seen from the taken photos that the fluorescence color changes from orange to green, and the colorimetric color changes from orange to colorless, then tryptamine is present in the unknown solution.

[0073] Embodiment 11

[0074] Detection of unknown solid particles by the prepared test pen:

[0075] a. The probe (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile obtained in Example 1 was dissolved in an organic solvent, dimethyl sulfoxide, to prepare a probe solution with a concentration of 500 μmol / L;

[0076] b. Testing with a test pen: In a pen cap 3 of 90.60 mm × 13.50 mm wide × 23.00 mm high, fill the polyvinyl alcohol sponge in the groove of the pen tip 1 of 5 mm × 5 mm × 1 mm, fill the probe solution prepared in step a in the hollow tube 2, and seal the probe solution with a sealing film;

[0077] c. Press the spring 5 connected to the hollow tube 2 in the test pen so that the tip 6 pierces the sealing membrane of the probe solution in step b. The probe solution is loaded into the polyvinyl alcohol sponge filled in the pen tip 1 based on the siphon effect. Use the pen tip 1 to gently wipe the surface of the object containing unknown solid particles. Under the condition of an excitation wavelength of 365nm, observe the change in fluorescence color with the naked eye. If the fluorescence color of the contact part between the particles and the sponge changes from red to colorless, and the colorimetric color changes from orange to colorless, it indicates the presence of tryptamine particles.

[0078] Example 12

[0079] Detection of unknown solid particles by the prepared test pen:

[0080] a. The probe molecule (E)-2-(3-cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile prepared in Example 2 was dissolved in an organic solvent, dimethyl sulfoxide, and the concentration of the probe molecule was 500 μM;

[0081] b. Testing with a test pen: In a pen cover 3 of 90.60 mm × 13.50 mm wide × 23.00 mm high, fill the polyvinyl alcohol sponge into the groove of the pen tip of 15 mm × 5 mm × 1 mm, fill the probe solution prepared in step a into the hollow tube 2, and seal the probe solution with a sealing film;

[0082] c. Press the spring 5 connected to the hollow tube 2 in the test pen so that the tip 6 pierces the probe solution sealing membrane in step b. The probe solution is loaded into the polyvinyl alcohol sponge filled in the pen tip 1 based on the siphon effect. Use the pen tip 1 to gently wipe the surface of the object containing unknown solid particles. Under the condition of an excitation wavelength of 365nm, observe the change of fluorescence color with the naked eye. If the fluorescence color of the contact part between the particles and the sponge changes from orange to green, and the colorimetric color changes from orange to colorless, it proves that tryptamine particles exist.

[0083] The results showed that the prepared test pen had good potential for qualitative analysis of tryptamines.

Claims

1. A colorimetric-fluorescent probe for detecting tryptamine, characterized in that: The chemical structure of the probe is (1): Wherein: R is benzo[c][1,2,5]thiadiazole or 2,5-dibromobenzene, and its structural formula is:

2. A method for preparing a colorimetric-fluorescent probe as claimed in claim 1, characterized in that Follow these steps: a. Mix malononitrile and 3-hydroxy-3-methyl-2-butanone in a molar ratio of 2.05:1, add to anhydrous ethanol, and add sodium ethoxide as a catalyst, set the temperature to 90° C., react for 5 hours, wash with ice anhydrous ethanol, filter and dry to obtain an intermediate product; b. Mix the intermediate product obtained in step a with 2,1,3-benzothiadiazole-4,7-dicarboxaldehyde or 2,5-dibromoterephthalaldehyde in a molar ratio of 0.9:1, add to anhydrous ethanol, set the temperature to 90° C., react for 5 hours, wash with ice anhydrous ethanol and filter, dry, and purify by column chromatography to obtain probes (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5H)-alkylene)malononitrile and (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5H)-alkylene)malononitrile.

3. A test pen containing the colorimetric-fluorescent probe as claimed in claim 1, characterized in that: The test pen is composed of a pen tip (1), a hollow tube (2), a pen cover (3), a detection reagent (4), a spring (5) and a tip (6). The pen cover (3) is provided with a hollow tube (2), one end of the hollow tube (2) is connected to the pen tip (1), and the other end of the hollow tube (2) is connected to the tip (6) via a spring (5). The detection reagent (4) is placed in the hollow tube (2), a groove is provided in the pen tip (1), and the detection reagent (4) contains a colorimetric fluorescent probe.

4. A method for detecting a colorimetric-fluorescent probe for detecting tryptamine according to claim 1, characterized in that: Follow these steps: Tryptamine solid detection: a. Dissolve the obtained probe (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile or (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile in an organic solvent, dimethyl sulfoxide, to prepare a probe solution with a concentration of 500 μmol / L; b. Testing with a test pen: In a pen cover (3) of 90.60 mm × 13.50 mm wide × 23.00 mm high, fill a polyvinyl alcohol sponge into the 5 mm × 5 mm × 1 mm groove of the pen tip (1), fill the probe solution prepared in step a into the hollow tube (2), and seal the probe solution with a sealing film; c. Press the spring (5) connected to the hollow tube (2) in the test pen so that the tip (6) pierces the probe solution sealing film in step b. The probe solution is loaded into the polyvinyl alcohol sponge filled in the pen tip (1) based on the siphon effect. The pen tip (1) is used to gently wipe the surface of the object containing the tryptamine solid particles. Under the condition of an excitation wavelength of 365nm, the fluorescence color change is observed with the naked eye to perform a rapid qualitative analysis of the tryptamine in the sample to be tested. Tryptamine solution test: a. Dissolve the obtained probe (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile or (E)-2-(3-cyano-4-(2,5-dibromobenzene)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile in an organic solvent, dimethyl sulfoxide, to prepare probe solutions with concentrations of 20 μmol / L and 100 μmol / L, to obtain a detection reagent; b. Add the test solution to the detection reagent obtained in step a. After sufficient reaction, the fluorescence spectrum and absorption spectrum are measured using a fluorescence spectrometer and an ultraviolet spectrophotometer, and the changes in fluorescence and colorimetric signals are observed at 365 nm and under daylight illumination. For the probe (E)-2-(3-cyano-4-(benzo[c][1,2,5]thiadiazole)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile, the fluorescence color changes from red to colorless and the colorimetric color changes from red to colorless, indicating that tryptamine exists in the solution. For the probe (E)-2-(3-cyano-4-(2,5-dibromophenyl)-5,5-dimethylfuran-2(5-hydrogen)-alkylidene)malononitrile, the fluorescence color changes from orange to green and the colorimetric color changes from orange to colorless, indicating the presence of tryptamine in the solution.

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