Dual-function fluorescent molecular probe and preparation method thereof and application in detecting copper ions and glyphosate in Chinese herbal medicine

Abstract

The present invention relates to the technical field of copper ion and pesticide glyphosate detection, and discloses a bifunctional fluorescent molecular probe, a preparation method thereof, and an application in detecting copper ions and glyphosate in Chinese herbal medicine. The structural formula of the bifunctional fluorescent molecular probe is as follows. 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde is reacted with 2-pyridinecarboxylic acid hydrazide to obtain the bifunctional fluorescent molecular probe. The present invention also relates to the application of the bifunctional fluorescent molecular probe in copper ion sensing detection. The complex of the bifunctional fluorescent molecular probe and copper ions can be used for sensing detection of glyphosate. The synthesis of the bifunctional fluorescent molecular probe provided by the present invention only requires two steps or even one step, the raw materials are easy to obtain, and the operation is simple. The present invention realizes the quantitative detection of copper ions by the probe through the different functions displayed by ultraviolet and fluorescence spectra, has strong reversibility, can be recycled many times, has good selectivity, is not easily affected by other metal ions, and has a low detection limit.

Description

Technical Field

[0001] The invention relates to the technical field of copper ion and pesticide glyphosate detection, and in particular to a bifunctional fluorescent molecular probe and a preparation method thereof, and application thereof in detecting copper ions and glyphosate in Chinese herbal medicines. Background Art

[0002] Copper (II) ions are the most widely distributed form of copper in nature and are involved in many biochemical processes in organisms. With the advancement of global industrialization, many copper (II) ions are released into the environment, causing harm to the ecosystem. In addition, many diseases have been shown to be associated with copper imbalance in the body, including Menkes syndrome, Alzheimer's disease, Parkinson's disease, Wilson's disease, and coronary heart disease. Long-term excessive copper may cause a variety of symptoms such as cirrhosis, movement disorders, and neurodegeneration. Therefore, accurate and rapid detection of copper (II) ion content in different water environments, Chinese herbal medicines, living cells, and living zebrafish is of great significance to the environment and human health.

[0003] Existing technical means for measuring copper (II) ions, such as atomic fluorescence spectroscopy (AFS), inductively coupled plasma mass spectrometry (ICP-MS) and atomic absorption spectroscopy (AAS), can meet the general needs of copper (II) ion analysis. However, the high cost, complex and time-consuming pretreatment process, and high requirements for operators limit the use of the above methods. Compared with instrumental analysis, fluorescence sensing detection is low-cost, easy to operate, has no high requirements for operators, responds quickly, and can be used for biological imaging. Therefore, research on the fluorescence determination of copper (II) ions is urgently needed.

[0004] Glyphosate is an organophosphorus non-specific herbicide that is very effective against perennial weeds and is therefore widely used in various production sites for weed control. Glyphosate mainly inhibits the conversion of shikimate to phenylalanine, tyrosine and tryptophan by inhibiting enolpyruvyl shikimate phosphate synthase in plants, thereby interfering with protein synthesis and causing plant death.

[0005] Due to its poor selectivity and wide range of applications, improper use may affect production, and glyphosate residues have been found in some economic crops, especially in Chinese herbal medicines, which may affect its efficacy. Therefore, the development of glyphosate safeners is of great significance to improving production. However, there is currently no fast, efficient and convenient glyphosate detection technology. Therefore, there is an urgent need for a convenient technology to assist the development of glyphosate safeners. Summary of the invention

[0006] The purpose of the present invention is to overcome the problems of complex synthesis steps, poor selectivity and low sensitivity of probes for detecting copper ions and glyphosate in the prior art, and to provide a bifunctional fluorescent molecular probe and a preparation method thereof and an application in detecting copper ions and glyphosate in Chinese herbal medicines.

[0007] The invention provides a simple, rapid, highly specific and highly sensitive dual-function fluorescent detector of copper (II) ions and glyphosate, which has broad application prospects and is particularly suitable for the field of in vivo imaging.

[0008] In order to achieve the above object, the first aspect of the present invention provides a bifunctional fluorescent molecular probe, wherein the structural formula of the bifunctional fluorescent molecular probe is as follows:

[0009]

[0010] Existing fluorescent probes only use fluorescence spectra to quantitatively detect metal ions. The bifunctional fluorescent molecular probe of the present invention can provide a variety of different data by combining ultraviolet ratio quantitative detection with fluorescence spectrum quantitative detection, increasing the accuracy and reliability of metal ions, namely copper (II) ions. The ultraviolet spectrum can provide higher contrast and sensitivity. The most intuitive phenomenon is that the bifunctional fluorescent molecular probe of the present invention has a chromogenic sensing function, which not only causes a change in the color of the solution under sunlight, but also produces a change in the fluorescence color under 365nm ultraviolet light. This dual observation mode makes it a chromogenic sensing fluorescent probe suitable for different lighting conditions.

[0011] The second aspect of the present invention provides a method for preparing the bifunctional fluorescent molecular probe described in the first aspect, wherein the method comprises: reacting 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde with 2-pyridinecarboxyhydrazide to obtain the bifunctional fluorescent molecular probe.

[0012] The third aspect of the present invention provides the use of the bifunctional fluorescent molecular probe described in the first aspect or the bifunctional fluorescent molecular probe obtained by the preparation method described in the second aspect in copper ion sensing detection.

[0013] The fourth aspect of the present invention provides a complex of the bifunctional fluorescent molecular probe described in the first aspect or the bifunctional fluorescent molecular probe obtained by the preparation method described in the second aspect and copper ions. As a cheap, easy-to-operate, highly selective detection technology, the bifunctional fluorescent molecular probe can greatly assist the research on glyphosate safeners.

[0014] The fifth aspect of the present invention provides the use of the complex described in the fourth aspect in glyphosate sensing detection.

[0015] Through the above technical solution, the beneficial technical effects achieved by the present invention are as follows:

[0016] (1) Based on specific recognition, the bifunctional fluorescent molecular probe of the present invention can be used as a specific indicator for the presence of copper (II) ions in different water environments, Chinese herbal medicines, living cells and living zebrafish. It can provide real-time qualitative and quantitative information through fluorescence detection and produce color changes, making it a highly specific indicator suitable for visual colorimetric detection. At the same time, the probe has high reversibility and repeatability for the detection of copper ions, and can also be used as a sensor for detecting copper (II) ions in the environment and Chinese herbal medicines, living cells and living zebrafish, and can perform repeated detections without damaging its performance.

[0017] (2) The synthesis of the bifunctional fluorescent molecular probe provided by the present invention only requires two steps or even one step, the raw materials are easily available, and the operation is simple.

[0018] (3) The present invention realizes the quantitative detection of copper ions and glyphosate by the probe through the different functions displayed by ultraviolet and fluorescence spectra. It has strong reversibility, can be recycled multiple times, has good selectivity, is not easily affected by other ions, has a low detection limit, and can be widely used to detect copper (II) ions and glyphosate in complex environments.

[0019] Copper ion: UV detection limit is 4.59×10 -7 M, the fluorescence detection limit is 1.15×10 -6 M;

[0020] Glyphosate: UV detection limit is 1.33×10 -6 M, the fluorescence detection limit is 5.88×10 -7 M. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a high-resolution mass spectrum of the bifunctional fluorescent molecular probe J-3 synthesized in Example 1 of the present invention.

[0022] Figure 2 The nuclear magnetic resonance of the bifunctional fluorescent molecular probe J-3 synthesized in Example 1 of the present invention is 1 HNMR spectrum.

[0023] Figure 3 The nuclear magnetic resonance of the bifunctional fluorescent molecular probe J-3 synthesized in Example 1 of the present invention is 13 CNMR image.

[0024] Figure 4 (A) is J-3+Cu 2+ Or other metal ion fluorescence spectra, (B) is [J-3-Cu 2+ ]+Fluorescence spectra of pesticides, amino acids or anions.

[0025] Figure 5 (A) J-3 and J-3+Cu 2+ UV absorption spectrum, (B) is [J-3-Cu 2+ ] and [J-3-Cu 2+ ]+UV absorption spectrum of glyphosate.

[0026] Figure 6 (A) is the competitive detection diagram of probe J-3 for different metal cations in DMF / water (8:2, v / v, 0.01 M Hepes, pH = 7.4) system, (B) is the competitive detection diagram of [J-3-Cu 2+ ]Competitive detection diagram of different anions and pesticides in DMF / water (8:2, v / v, 0.01 M Hepes, pH=7.4) system.

[0027] Figure 7 (A) is J-3+Cu 2+ The fluorescence titration curve of [J-3-Cu 2+ ]+fluorescence titration curve of glyphosate.

[0028] Figure 8 (A) is J-3+Cu 2+ The UV titration curve of [J-3-Cu 2+ ]+UV titration curve of glyphosate.

[0029] Fig. 9 (A) is J-3+Cu 2+ Reversibility detection diagram, (B) is [J-3-Cu 2+ ]+Reversibility detection diagram of glyphosate.

[0030] Fig.10 Probe J-3, J-3+Cu 2+ and [J-3-Cu 2+ ]+The effect of glyphosate in sunlight.

[0031] Fig.11 Probe J-3, J-3+Cu 2+ and [J-3-Cu 2+ ]+The effect of glyphosate under 365nm ultraviolet light. DETAILED DESCRIPTION

[0032] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0033] The first aspect of the present invention provides a bifunctional fluorescent molecular probe, wherein the structural formula of the bifunctional fluorescent molecular probe is as follows:

[0034]

[0035] The second aspect of the present invention provides a method for preparing the bifunctional fluorescent molecular probe described in the first aspect, wherein the method comprises: reacting 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde with 2-pyridinecarboxyhydrazide to obtain the bifunctional fluorescent molecular probe.

[0036] In some embodiments of the invention, the method comprises the following steps:

[0037] (1) dissolving 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde in an organic solvent to obtain a solution;

[0038] (2) adding 2-pyridinecarboxylic acid hydrazide to the solution obtained in step (1), heating under reflux to react and obtaining the bifunctional fluorescent molecular probe.

[0039] During the preparation of the bifunctional fluorescent molecular probe of the present invention, the reaction that occurs is:

[0040]

[0041] The 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde is obtained by reacting triphenylamine boric acid and 5-bromothiophene-2-carboxaldehyde, and can also be obtained from a reagent company.

[0042] In some embodiments of the present invention, the organic solvent in step (1) is dry anhydrous ethanol.

[0043] In some embodiments of the present invention, the molar ratio of 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde to 2-pyridinecarboxylic acid hydrazide is 14:15.

[0044] In some embodiments of the present invention, the reaction time is 4 hours.

[0045] In some embodiments of the present invention, the progress of the reaction is monitored by TLC.

[0046] In some embodiments of the present invention, the method further comprises: after stopping the reaction, cooling the reaction liquid to room temperature to precipitate, filtering, washing and drying the precipitate to obtain the bifunctional fluorescent molecular probe.

[0047] The third aspect of the present invention provides the use of the bifunctional fluorescent molecular probe described in the first aspect or the bifunctional fluorescent molecular probe obtained by the preparation method described in the second aspect in copper ion sensing detection.

[0048] In some embodiments of the present invention, the bifunctional fluorescent molecular probe is used for sensing and detecting copper ions in water environments, Chinese herbal medicines, living cells, and living zebrafish.

[0049] The fourth aspect of the present invention provides a complex of the bifunctional fluorescent molecular probe described in the first aspect or the bifunctional fluorescent molecular probe obtained by the preparation method described in the second aspect and copper ions.

[0050] In the present invention, the preparation method of the complex may include: mixing the bifunctional fluorescent molecular probe and an equimolar amount of copper ions in a mixed solution of DMF and water to obtain a complex of the bifunctional fluorescent molecular probe and copper ions.

[0051] The fifth aspect of the present invention provides the use of the complex described in the fourth aspect in glyphosate sensing detection.

[0052] In some embodiments of the present invention, the complex is used for sensing and detecting glyphosate in water environments, Chinese herbal medicines, living cells, and living zebrafish.

[0053] In some embodiments of the present invention, the sensing assay is a selective sensing assay and a competitive sensing assay.

[0054] In some embodiments of the present invention, the sensing detection is visual qualitative detection, ultraviolet light detection, fluorescence detection and reversible detection.

[0055] In some embodiments of the present invention, the sensing detection is quantitative detection under ultraviolet spectroscopy and fluorescence spectroscopy.

[0056] The present invention will be described in detail below through examples.

[0057] In the following examples and comparative examples, if no specific conditions are specified, the experiments were carried out under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used, if no manufacturer is specified, are all conventional products that can be obtained through commercial channels.

[0058] Example 1

[0059] This example is used to illustrate the preparation of the target compound J-3

[0060] (1) Synthesis of intermediate M (5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde):

[0061]

[0062] Triphenylamine boric acid (3.47g, 12mmol), 5-bromothiophene-2-carboxaldehyde (1.91g, 10mmol), potassium carbonate (2.49g, 18mmol) and catalyst tetrakis(triphenylphosphine)palladium (0.139g, 0.12mmol) were added to a 30mL THF / water (1 / 1, v / v) mixed solution, and refluxed at 70°C under the protection of nitrogen until the raw material point disappeared as monitored by TLC. After stopping the reaction, the mixture was cooled to room temperature, extracted three times with dichloromethane (30mL×3 times), and the organic layer was collected. The organic layer was extracted three times with a saturated sodium chloride solution (15mL×3 times), the organic layer was separated, anhydrous sodium sulfate was added to dry, and the solvent was spin-dried to obtain a viscous black crude product. Finally, separation and purification were performed by column chromatography to obtain 1.93g of a green solid.

[0063] (2) Synthesis of target compound

[0064]

[0065] The intermediate M (100 mg, 0.28 mmol) was dissolved in 20 mL of anhydrous ethanol, and then 0.30 mmol of another reactant 2-pyridine carboxylic acid hydrazide was added, and the reaction was heated under reflux for about 4 h until the reaction was complete as monitored by thin layer chromatography (TLC). After stopping the reaction, the reaction liquid was cooled to room temperature, at room temperature, a large amount of precipitate was precipitated, filtered, and the filter cake was rinsed three times with 20 mL of ice ethanol to finally obtain the target compound J-3.

[0066] The target compound J-3 obtained in this example is a new compound that has not been reported before. After purification, high-resolution mass spectrometry (HRMS), nuclear magnetic resonance spectroscopy ( 1 H NMR, 13 C NMR) to characterize it. The high-resolution mass spectrum of probe J-3 is shown in Figure 1 As shown, the NMR of probe J-3 1 H NMR Figure 2 As shown, the NMR of probe J-3 13 C NMR Figure 3 shown. 1 H NMR(400MHz, DMSO-d6)δppm)12.17(s,1H),8.80(s,1H),8.71(d,J=4.6Hz,1H),8.12(d,J=7.7Hz,1H),8.05(td,J=7.7,1.5Hz,1H),7. 72–7.61(m,3H),7.43(d,J=3.8Hz,1H),7.41–7.38(m,1H),7.35(t,J=7.8Hz,4H),7.10(dd,J=13.2,7.5Hz,6H),6.98(d,J=8.6Hz,2H).13 C NMR(101MHz,DMSO-d6)δ160.63,150.07,148.95,147.93,147.17,146.20,144.45,138.49, 137.86,132.83,130.17,127.43,127.35,127.22,125.04,124.16,123.66,123.13,123.02.

[0067] Example 2

[0068] This example is used to illustrate the complex of the target compound J-3 and copper ions [J-3-Cu 2+ ] preparation.

[0069] The target compound J-3 (1×10 -5 mol / L) and copper ions (1×10 -5 mol / L) DMF / water (8:2, v / v, 0.01 M Hepes, pH = 7.4) buffer solution system, the prepared complex [J-3-Cu 2+ ]The concentration is 1×10 - 5 mol / L.

[0070] The target compound J-3 synthesized in Example 1 was dissolved in DMF to a concentration of 1×10 -5 mol / L DMF / water (8:2, v / v, 0.01 M Hepes, pH=7.4) system, denoted as solution A.

[0071] The target compound J-3 (1×10 -5 mol / L) and copper ions (1×10 -5 mol / L) were mixed in equal amounts to prepare the complex [J-3-Cu 2+ ]The concentration is 1×10 -5 mol / L DMF / water (8:2, v / v, 0.01M Hepes, pH=7.4) system, denoted as solution B.

[0072] Application Example 1

[0073] Add different metal cations (Al 3+ , K + , Fe 3+ , Fe 2+ 、Na + Cr 3+ , Ca 2+ 、Zn 2+ Mg 2+, Mn 2+ , Ba 2+ , Hg 2+ , Pb 2+ 、Cd 2+ 、Co 2+ 、Ni 2+ 、Ag + and Cu 2+ ), fluorescence spectra such as Figure 4 (A) is shown. Figure 4 As can be seen in (A), the addition of Cu 2+ After addition, the fluorescence intensity decreased significantly. The addition of other metal ions did not cause significant changes in fluorescence intensity. 2+ It can quench the fluorescence of probe J-3, which is unique compared with other ions, so probe J-3 can specifically recognize Cu 2+ .

[0074] Add pesticides, amino acids or anions (glyphosate, parathion, DDVP, dimethoate, parathion methyl, malathion, glutamic acid, alanine, phenylalanine, NO3 - 、S2O3 2- 、HSO3 - 、HCO3 - 、ClO4 - , Cl - 、HPO42 - 、BF4 - 、F - 、CrO4 2- ,I - ), fluorescence spectra such as Figure 4 (B) As shown. It can be seen that the addition of glyphosate makes [J-3-Cu 2+ ] system's fluorescence was significantly restored, while the addition of other pesticides, amino acids or anions had little effect on the system's fluorescence. The above results indicate that glyphosate can restore the fluorescence of [J-3-Cu 2+ ] complex has a unique fluorescence enhancement compared with other pesticides and anions, so the probe [J-3-Cu 2+ ] can specifically recognize glyphosate.

[0075] Figure 5 (A) is solution A and Cu is added thereto 2+ The UV spectrum after adding Cu 2+ After that, the 405nm ultraviolet absorption peak decreases and red-shifts to 410nm. Figure 5 (B) is the UV spectrum of solution B and after glyphosate is added thereto. It can be seen that after adding glyphosate, the absorbance of the absorption peak at 410 nm increases and blue-shifts to 405 nm.

[0076] The above results show that Cu 2+ It can quench the fluorescence of probe J-3, which is unique compared with other ions. Probe J-3 can specifically recognize Cu 2+ Glyphosate can make the probe [J-3-Cu 2+ ] fluorescence recovery is unique compared with other ions, and the probe [J-3-Cu 2+ ] can specifically recognize glyphosate.

[0077] Application Example 2

[0078] Detection of Cu using Solution A 2+ For different metal cations Al 3+ , K + , Fe 3+ , Fe 2+ 、Na + Cr 3+ , Ca 2+ 、Zn 2+ Mg 2+ , Mn 2+ , Ba 2+ , Hg 2+ , Pb 2+ 、Cd 2+ 、Co 2+ 、Ni 2+ 、Ag + The fluorescence intensity of solution A and after adding different metal cations to it at 538nm was measured, and the fluorescence intensity of solution A after adding other metal cations and then adding an equal amount of Cu 2+ The fluorescence intensity of the system at 538 nm is as follows: Figure 6 As shown in (A). It can be seen that probe J-3 has a strong affinity for Cu 2+ The recognition has good anti-interference ability, and the presence of other metal ions in the system will not affect its recognition effect.

[0079] Solution B was used to detect the effects of glyphosate on different pesticides, amino acids or anions (glyphosate, parathion, DDVP, dimethoate, parathion methyl, malathion, glutamic acid, alanine, phenylalanine, NO3 - 、S2O3 2- 、HSO3 - 、HCO3 - 、ClO4 - , Cl - 、HPO4 2- 、BF4 - 、F - 、CrO4 2- ,I - ) competitiveness, such as Figure 6(B) As shown. It can be seen that the probe [J-3-Cu 2+ ] The complex has a good anti-interference ability for the recognition of glyphosate, and the presence of other pesticides, amino acids or anions in the system will not affect its recognition effect.

[0080] Application Example 3

[0081] Different concentrations of copper ions / glyphosate were added to solution A / solution B, and the changes in their fluorescence spectra were detected, such as Figure 7 shown.

[0082] Figure 7 (A) shows the fluorescence spectrum change of the system and Cu 2+ The relationship between the concentration of copper ions and the fluorescence intensity is shown in Table 1. With the increase of copper ion concentration, the fluorescence intensity gradually weakens, and the change of fluorescence intensity is obvious, which proves that probe J-3 can be used for Cu 2+ The quantitative detection is clear and easy to identify.

[0083] like Figure 7 As shown in (B), with the increase of glyphosate concentration, the fluorescence intensity gradually increased, and the change of fluorescence intensity was obvious, proving that [J-3-Cu 2+ ] The complex can be used for quantitative detection of glyphosate, and the phenomenon is obvious and easy to identify.

[0084] Different concentrations of copper ions / glyphosate were added to solution A / solution B, and the changes in their UV spectra were detected, such as Figure 8 shown.

[0085] Figure 8 (A) shows the change of the UV spectrum of the system and Cu 2+ With the increase of copper ion concentration, the UV absorbance at 405nm gradually weakened and red-shifted to 410nm. The UV spectrum changed significantly, proving that probe J-3 can be used for Cu 2+ The quantitative detection is clear and easy to identify.

[0086] like Figure 8 As shown in (B), with the increase of glyphosate concentration, the 410nm UV absorbance gradually increased and blue-shifted to 405nm. The UV spectrum changed significantly, proving that [J-3-Cu 2+ ] The complex can be used for quantitative detection of glyphosate, and the phenomenon is obvious and easy to identify.

[0087] Application Example 4

[0088] like Fig. 9 As shown in (A), EDTA (concentration of 1×10 -5 mol / L) and Cu 2+ (Concentration is 1×10-5 mol / L) and then detect the reversible change of its fluorescence intensity at 538 nm.

[0089] like Fig. 9 As shown in (B), glyphosate (2×10 -5 mol / L) and Cu 2+ (Concentration is 1×10 -5 mol / L) and then detected the reversible change of its fluorescence intensity at 538 nm.

[0090] The results showed that probe J-3 was sensitive to Cu 2+ The recognition of [J-3-Cu 2+ ]The recognition of the complex glyphosate has good reversibility and can be used repeatedly.

[0091] Application Example 5

[0092] Fig.10 To add 1×10 -5 mol / L copper ions and then add 1×10 -5 mol / L glyphosate solution color changes under sunlight. It can be seen that it changes from light yellow on the left to dark yellow in the middle and then to light yellow on the right.

[0093] Fig.11 To add 1×10 -5 mol / L copper ions and then add 1×10 -5 The color change of mol / L glyphosate solution under 365nm UV light. It can be seen that the color changes from bright yellow on the left to light yellow in the middle and then to bright yellow on the right.

[0094] Application Example 6

[0095] Tap water, wolfberry extract, and honeysuckle extract were selected to study the probe J-3 / [J-3-Cu 2+ ]The detection ability of the complex in actual water samples and Chinese medicine extracts.

[0096] The Chinese medicine extract was filtered to remove insoluble impurities, and then the filtered extract was mixed with the probe J-3 / [J-3-Cu 2+ The complex was prepared into a test solution to test the fluorescence, and the fluorescence showed almost no change, indicating that the Cu 2+ / The glyphosate content is very low and does not reach J-3 / [J-3-Cu 2+ ] complex detection limit. Then tap water and Chinese medicine extract were used to prepare the concentrations of 2.5, 5, and 10×10 -5 mol / L Cu 2+ The solution was prepared with tap water and Chinese herbal medicine extract at concentrations of 5, 10, and 15×10-5 mol / L glyphosate solution was used for spike recovery experiment.

[0097] Table 1 shows the fluorescence molecular probe J-3 for the determination of Cu in tap water, wolfberry extract, and honeysuckle extract 2+ The experimental results.

[0098] Table 1

[0099]

[0100] Note: n is the number of test repetitions.

[0101] Table 2 is a list of fluorescent molecular probes [J-3-Cu 2+ ] Experimental results of complex determination of glyphosate in tap water, wolfberry extract and honeysuckle extract.

[0102] Table 2

[0103]

[0104] As shown in Table 1, the recovery rates of spiked water samples were 94.2-107.2%, and the relative standard deviations were all less than 10%; as shown in Table 2, the recovery rates of spiked water samples were 91.1-134.8%, and the relative standard deviations were all less than 5%. 2+ The complex can be used for quantitative detection of Cu in tap water and Chinese medicine extracts 2+ / Glyphosate content.

[0105] The present invention designs and synthesizes a copper (II) ion and glyphosate dual-functional specific recognition fluorescent molecular probe. Probe J-3 has a strong fluorescence intensity in the absence of copper (II) ions, but has obvious fluorescence quenching at 538nm after adding copper (II) ions, and has high selectivity and anti-interference. In addition, the detection limit of probe J-3 for copper (II) ions is low, and the ultraviolet detection limit is 4.59×10 -7 M, the fluorescence detection limit is 1.15×10 -6 M. [J-3-Cu 2+ The detection limit of the complex for glyphosate is low, and the UV detection limit is 1.33×10 -6 M, the fluorescence detection limit is 5.88×10 -7 M. The ability to effectively detect glyphosate present in copper ions in water environments, Chinese herbal medicines, living cells and living zebrafish with high sensitivity has certain guiding significance for production activities.

[0106] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. A bifunctional fluorescent molecular probe, characterized in that: The structural formula of the bifunctional fluorescent molecular probe is as follows:

2. A method for preparing a bifunctional fluorescent molecular probe according to claim 1, characterized in that: The method comprises: reacting 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde with 2-pyridinecarboxylic acid hydrazide to obtain the bifunctional fluorescent molecular probe.

3. The preparation method according to claim 2, wherein The method comprises the following steps: (1) dissolving 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde in an organic solvent to obtain a solution; (2) adding 2-pyridinecarboxylic acid hydrazide to the solution obtained in step (1), heating under reflux to react and obtaining the bifunctional fluorescent molecular probe.

4. The preparation method according to claim 3, wherein The 5-(4-(diphenylamino)phenyl)thiophene-2-carboxaldehyde is obtained by reacting triphenylamineboronic acid and 5-bromothiophene-2-carboxaldehyde.

5. The preparation method according to claim 3, wherein: The organic solvent in step (1) is dry anhydrous ethanol.

6. The preparation method according to any one of claims 2 to 5, wherein The molar ratio of 5-(4-(diphenylamino)phenyl)thiophene-2-carbaldehyde to 2-pyridinecarboxylic acid hydrazide is 14:

15.

7. The preparation method according to any one of claims 2 to 5, wherein The reaction time is 4 hours.

8. The preparation method according to any one of claims 2 to 5, wherein The progress of the reaction was monitored by TLC.

9. The preparation method according to any one of claims 2 to 5, wherein The method further comprises: after stopping the reaction, cooling the reaction liquid to room temperature to precipitate, filtering, washing and drying the precipitate to obtain the dual-function fluorescent molecular probe.

10. Use of the bifunctional fluorescent molecular probe according to claim 1 or the bifunctional fluorescent molecular probe obtained by the preparation method according to any one of claims 2 to 9 in copper ion sensing detection.

11. The use according to claim 10, wherein: The dual-function fluorescent molecular probe is used for sensing and detecting copper ions in water environments, Chinese herbal medicines, living cells and living zebrafish.

12. A complex of the bifunctional fluorescent molecular probe according to claim 1 or the bifunctional fluorescent molecular probe obtained by the preparation method according to any one of claims 2 to 9 and copper ions.

13. Use of the complex of the bifunctional fluorescent molecular probe and copper ions according to claim 12 in glyphosate sensing detection.

14. The use according to claim 13, wherein: The complex is used for sensing and detecting glyphosate in water environment, Chinese herbal medicine, living cells and living zebrafish.

15. The use according to claim 10 or 13, wherein: The sensing assay is a selective sensing assay and a competitive sensing assay.

16. The use according to claim 10 or 13, wherein: The sensing detection includes visual qualitative detection, ultraviolet light detection, fluorescence detection and reversible detection.

17. The use according to claim 10 or 13, wherein: The sensing detection is a quantitative detection under ultraviolet spectrum and fluorescence spectrum.

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