Aggregation-induced emission compound with dual fluorescence emission characteristics and application thereof

Abstract

The application discloses an aggregation-induced emission compound with double fluorescence emission characteristics and a preparation method and application thereof, and the proportional fluorescence probe has a structure of general formula I. The probe molecule has a polar response group tetraphenyl ethylene as a fluorescence report unit, introduces a hydrophilic oxygen atom, connects an alpha carbon of an acetamide, and connects a polar detection sensitization group Tag chain on an amino group of the acetamide. The probe molecule improves the conjugation degree of oxygen atom lone pair electrons and a tetraphenyl ethylene pi electron system, so that the probe molecule simultaneously produces n→pi* and pi→pi* molecular transitions, and a proportional fluorescence probe which can sensitively respond to polarity change is constructed. The fluorescence probe designed in the application has the advantages that the fluorescence probe is not easily interfered by probe concentration, a detection environment and light bleaching, and experiments prove that the proportional fluorescence intensity of the fluorescence probe can sensitively respond to polarity, and the proportional fluorescence intensity and water content are linearly fitted, so that the purposes of detecting water content in an organic reagent and alcohol content in wine are achieved.

Description

Technical Field

[0001] This invention belongs to the field of fluorescent probe technology, specifically relating to the efficient, accurate and rapid detection of water content in organic solvents and alcohol content in wine. Background Technology

[0002] Water is the most common impurity in organic solvents, and its content in organic solvents is subject to strict requirements. For example, the synthesis of Grignard reagents requires that the reaction system be free of water. In the past, the Karl Fischer method was used to quantitatively determine the water content in organic solvents, but due to its drawbacks such as reagent instability, limitations on batch analysis, and the need for specialized equipment, there is a need to develop efficient and accurate methods to determine the water content in organic solvents. Ethanol is the main component of alcoholic beverages and an important indicator of their quality. The determination of the ethanol volume percentage is crucial for quality control and monitoring of the brewing process in the beverage industry. Currently, many methods for determining ethanol have been developed, such as gas chromatography and high-performance liquid chromatography, but these methods have relatively strict operating conditions and high instrument requirements, thus hindering the real-time on-site determination of alcohol content in industrial fermentation. Therefore, it is necessary to develop rapid and simple methods to determine the ethanol content in alcoholic beverages.

[0003] Fluorescent probes are molecular systems that, with the aid of analytical tools, can convert many microscopic chemical reactions and physiological processes into emitted fluorescent signals. Among the fluorescent probes studied, small organic molecule fluorescent probes have become an important research topic due to their simple design and ease of use. Currently, many fluorescent probes have been applied to the detection of polarity and significant progress has been made. However, single-signal fluorescent probes are easily affected by various factors, while proportional fluorescent probes can greatly enhance their sensitivity and quantification capabilities. However, most conventional fluorescent probes are hydrophobic. These probes exhibit fluorescence quenching (ACQ) in concentrated solutions or in aggregated states. Compared to ordinary ACQ dyes, AIE dyes (aggregation-induced emission dyes) produce higher fluorescence emission. Aggregation-induced emission (AIE) refers to the phenomenon where fluorescent molecules exhibit weak fluorescence in dilute solutions, but produce strong fluorescence when the molecules are aggregated or in a solid state. Numerous studies have found that fluorescent molecules with the AIE effect typically show little or no fluorescence in pure organic solvents. The higher the proportion of water or poor solvents, the worse the solubility of the fluorescent molecules, the more molecules aggregate, and the more significantly the fluorescence of the molecules is enhanced. AIE probes have advantages such as high resolution, resistance to photobleaching, and large Stokes shift, which are essentially superior to traditional ACQ dyes. Tetraphenylene has excellent AIE luminescence properties and has become the most widely studied AIE luminescence material.

[0004] Therefore, it is necessary to construct an AIE proportional fluorescent probe capable of detecting both water and alcohol content. Summary of the Invention

[0005] The purpose of this invention is to provide an efficient, accurate and rapid method for detecting the water content in organic solvents and the alcohol content in wine.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] This invention provides a compound for detecting the water content in organic solvents. The compound has the structure of general formula I as follows:

[0008]

[0009] In general formula I:

[0010] R1, R2, and R3 are each individually selected from one or more of H, hydroxyl, phenyl, C1-C18 alkyl, C1-C18 alkylhydroxy, C2-C18 alkylcarboxyl, C1-C18 alkylamide, C1-C18 alkylazido, C2-C18 alkenyl, C2-C18 alkynyl, C1-C18 alkylamino, C1-C18 alkylsulfonic acid, C1-C18 alkoxy, or halogen (one or more of F, Cl, Br, and I), and the number of them is 1 to 5, preferably 1 or 2;

[0011] R4 is selected from H, halogens (one or more of F, Cl, Br, I), hydroxyl, amino, C1-C18 alkoxy, C1-C18 alkyl mercapto, C1-C18 alkyl dithio, C1-C18 alkylamino, C1-C18 alkylimino, C1-C18 alkylhydrazine, C1-C18 haloalkyl, C1-C18 alkylazido, O 6 - Benzylguanine, isooxocyanate, isothiocyanate, C2-C18 alkynyl, tetrazine group, morpholine, triphenylphosphine or p-toluenesulfonamide, one or more of unsubstituted or substituted C1-C18 alkyl or C6-C18 aromatic groups, wherein the substituents on the substituted C1-C18 alkyl or C6-C18 aromatic groups are one or more of hydroxyl, aldehyde, sulfonic acid, phosphoric acid, C1-C18 carboxylic acid ester, C1-C18 sulfonate, C1-C18 phosphate ester, amide, sulfonyl chloride or sulfonamide groups.

[0012] L1 is the connecting chain -(CH2CH2O) m - Here, m is an integer between 0 and 20;

[0013] L2 is the connecting chain, -(CH2) n- Here, n is an integer from 1 to 20; and, when R1, R2 and R3 = H, m = 0, and n = 2, R4 does not have a hydroxyl group.

[0014] Preferably, in general formula I, R1, R2 and R3 are each individually selected from one or more of H, hydroxyl, phenyl, C1-C10 alkyl, C1-C10 alkylhydroxy, C2-C10 alkylcarboxyl, C1-C10 alkylamido, C1-C10 alkylazido, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkylamino, C1-C10 alkylsulfonic acid, C1-C10 alkoxy or halogen;

[0015] R4 is selected from H, halogen, hydroxyl, amino, C1-C10 alkoxy, C1-C10 alkyl mercapto, C1-C10 alkyl dithio, C1-C10 alkylamino, C1-C10 alkylimino, C1-C10 alkylhydrazine, C1-C10 haloalkyl, C1-C10 alkylazido, O 6 - Benzylguanine, isooxocyanate, isothiocyanate, C2-C18 alkynyl, tetrazine group, morpholine, triphenylphosphine or p-toluenesulfonamide, one or more of unsubstituted or substituted C1-C10 alkyl or C6-C10 aromatic groups, wherein the substituents on the substituted C1-C10 alkyl or C6-C10 aromatic groups are one or more of hydroxyl, aldehyde, sulfonic acid, phosphoric acid, C1-C10 carboxylic acid ester, C1-C10 sulfonate, C1-C10 phosphate ester, amide, sulfonyl chloride or sulfonamide.

[0016] L1 is -(CH2CH2O) m -;

[0017] L2 is -(CH2) n - Here, m is an integer from 0 to 20, and n is an integer from 1 to 20; and when R1, R2, and R3 = H, m = 0, and n = 2, R4 does not have a hydroxyl group.

[0018] Further preferably, in general formula I, R1, R2 and R3 are each individually selected from one of H, hydroxyl, phenyl, C1-C10 alkyl, C1-C10 alkylhydroxy, C2-C10 alkylcarboxyl, C1-C10 alkylamide, C1-C10 alkylazido, C2-C10 alkenyl, C2-C10 alkynyl, C1-C10 alkylamino, C1-C10 alkylsulfonic acid, and C1-C10 alkoxy.

[0019] R4 is selected from H, halogen, hydroxyl, amino, C1-C10 alkoxy, C1-C10 alkyl mercapto, C1-C10 alkyl dithio, C1-C10 alkylamino, C1-C10 alkylimino, C1-C10 alkylhydrazine, C1-C10 haloalkyl, C1-C10 alkylazido, O6 - Benzylguanine, isooxocyanate, isothiocyanate, C2-C18 alkynyl, tetrazine group, morpholine, triphenylphosphine or p-toluenesulfonamide, unsubstituted or substituted C1-C10 alkyl or C6-C10 aromatic group, wherein the substituent group on the substituted C1-C10 alkyl or C6-C10 aromatic group is one or more of hydroxyl, aldehyde, sulfonic acid, phosphate, C1-C10 carboxylic acid ester, C1-C10 sulfonate, C1-C10 phosphate ester, amide, sulfonyl chloride or sulfonamide.

[0020] L1 is -(CH2CH2O) m -;

[0021] L2 is -(CH2) n - Here, m is an integer from 0 to 20, and n is an integer from 1 to 20; and when R1, R2, and R3 = H, m = 0, and n = 2, R4 does not have a hydroxyl group.

[0022] In a further preferred embodiment, in general formula I, R1, R2 and R3 are each individually selected from one of H, hydroxyl, methane and methoxy.

[0023] R4 is selected from one of H, halogen, hydroxyl, amino, and hydroxymethyl.

[0024] L1 is -(CH2CH2O) m -;

[0025] L2 is -(CH2) n - Here, m is an integer from 0 to 20, and n is an integer from 1 to 20; and when R1, R2, and R3 = H, m = 0, and n = 2, R4 does not have a hydroxyl group.

[0026] An application of the compound described above for detecting water content in organic solvents, wherein the compound represented by general formula I is used as a proportional fluorescent probe.

[0027] A proportional fluorescent probe for detecting water content in organic solvents, comprising the compound of claim 1 or a derivative thereof.

[0028] The proportional fluorescent probe is used for detecting water content in organic solvents and alcohol content in wine.

[0029] Advantages of this invention:

[0030] The fluorescent probe molecule of the present invention has the property of aggregation-induced fluorescence emission, which can produce high fluorescence emission, and also has the advantages of high fluorescence resolution, photobleaching resistance, and large Stokes shift.

[0031] The reason why the fluorescent probe molecules of this invention respond to polarity is as follows: the environmental polarity changes the degree of aggregation of tetraphenylethylene molecules with AIE properties to regulate the fluorescence intensity of tetraphenylethylene, thereby realizing the response of tetraphenylethylene probe molecules to polarity.

[0032] The probe molecule of this invention uses a polar responsive group, tetraphenylethylene, as the fluorescent reporter unit. A hydrophilic oxygen atom is introduced and then linked to the α-carbon of acetamide, which increases the conjugation degree between the lone pair electrons of the oxygen atom and the tetraphenylethylene π system. This allows the probe molecule to simultaneously generate n→π* and π→π* molecular transitions, thereby producing proportional fluorescence. Then, a polar detection-sensitizing group, Tag chain, is linked to the amino group of acetamide to construct a proportional fluorescent probe that can sensitively respond to changes in polarity.

[0033] The fluorescent probe molecule of the present invention can perform proportional fluorescence detection of polarity, and by linearly fitting the proportional fluorescence intensity with the water content, it can achieve the purpose of detecting the water content in organic reagents and the alcohol content in wine.

[0034] The benzene ring portion on the tetraphenylethylene group in the fluorescent probe molecule of the present invention can also be linked to functional groups targeting organelles (such as triphenylphosphine) and drugs targeting bacteria (such as vancomycin) to achieve the detection of the polarity of organelles and the bacterial internal environment.

[0035] Therefore, this invention can detect the water content in organic solvents and the alcohol content in wine, and is expected to detect the polarity of specific regions of organs, tissues, cells and organelles. Attached Figure Description

[0036] Figure 1 The intermediate compound 1 described in this invention 1 H NMR spectrum.

[0037] Figure 2 The intermediate compound 2 described in this invention 1 H NMR spectrum.

[0038] Figure 3 The intermediate compound 3 of the present invention 1 H NMR spectrum.

[0039] Figure 4 The intermediate H-TAG described in this invention 1 H NMR spectrum.

[0040] Figure 5 The proportional fluorescent probe T-TAG for polarity detection described in this invention 1 H NMR spectrum.

[0041] Figure 6The proportional fluorescent probe T-TAG for polarity detection described in this invention 13 C NMR spectrum.

[0042] Figure 7 This is the absorption spectrum of the proportional fluorescent probe T-TAG for polarity detection described in this invention in aqueous solution.

[0043] Figure 8 This is the absorption spectrum of the proportional fluorescent probe T-TAG for polarity detection described in this invention in acetonitrile solution.

[0044] Figure 9 The absorption and fluorescence emission spectra of the proportional fluorescent probe T-TAG for polarity detection described in this invention in different solvents are shown.

[0045] Figure 10 The absorption and fluorescence emission spectra of the proportional fluorescent probe T-TAG for polarity detection described in this invention are shown in two-phase mixed solvents of water and 1,4-dioxane with different volume ratios.

[0046] Figure 11 The fluorescence intensity ratio of the proportional fluorescent probe T-TAG for polarity detection described in this invention is I. 473nm / I 371nm The Boltzmann function fit relationship with solvent polarity Δf.

[0047] Figure 12 This is a graph showing the change in fluorescence intensity (λ = 320 nm) over time of the proportional fluorescent probe T-TAG for polarity detection described in this invention in a pure water system.

[0048] Figure 13 This is a transmission electron microscope (TEM) image of the proportional fluorescent probe T-TAG for polarity detection described in this invention in a pure water system.

[0049] Figure 14 The fluorescence intensity ratio of the proportional fluorescent probe T-TAG for polarity detection described in this invention is I. 473nm / I 371nm Fitted curves showing the relationship between the water content and the water content in a mixed solvent of water and 1,4-dioxane.

[0050] Figure 15 The fluorescence emission spectra of the proportional fluorescent probe T-TAG for polarity detection described in this invention are in two-phase mixed solvents of water and acetonitrile with different volume ratios.

[0051] Figure 16 The fluorescence intensity ratio of the proportional fluorescent probe T-TAG for polarity detection described in this invention is I. 468nm / I 406nmFitted curves showing the relationship between the water content in the mixed solvent of water and acetonitrile.

[0052] Figure 17 The fluorescence emission spectra of the proportional fluorescent probe T-TAG for polarity detection described in this invention are shown in two-phase mixed solvents of water and N,N-dimethylformamide with different volume ratios.

[0053] Figure 18 The fluorescence intensity ratio of the proportional fluorescent probe T-TAG for polarity detection described in this invention is I. 468nm / I 441nm Fitted curves showing the relationship between the water content and the water content in a mixed solvent of water and N,N-dimethylformamide.

[0054] Figure 19 The fluorescence emission spectra of the proportional fluorescent probe T-TAG for polarity detection described in this invention are in two-phase mixed solvents of water and ethanol with different volume ratios.

[0055] Figure 20 The fluorescence intensity ratio of the proportional fluorescent probe T-TAG for polarity detection described in this invention is I. 468nm / I 365nm Fitted curves showing the relationship between the water content in a mixed solvent of water and ethanol. Detailed Implementation Plan

[0056] The present invention will be further described in detail below with reference to specific embodiments.

[0057] The fluorescent probe used to detect water content in organic solvents is named T-TAG, and its structural formula is as follows:

[0058]

[0059] The method for preparing the fluorescent probe T-TAG, which can be used to detect water content in organic solvents, includes the following steps:

[0060]

[0061] (1)(a) Zinc powder was uniformly suspended in anhydrous tetrahydrofuran under N2 atmosphere. Titanium tetrachloride was slowly added at -5°C and the reaction solution was refluxed at room temperature for 2.5 hours. Then, pyridine was added at -5°C. After stirring for 10 minutes, benzophenone and 4-hydroxybenzophenone dissolved in anhydrous tetrahydrofuran were added and the reaction solution was refluxed for 17 hours. After quenching with 10% potassium carbonate aqueous solution, the mixture was filtered. The filtrate was extracted three times with dichloromethane and concentrated under reduced pressure. After column purification, compound 1 was obtained; (b) Compound 1 and potassium carbonate were reacted under N2 atmosphere. The mixture was placed in a round-bottom flask and reacted for 30 minutes. Then, acetonitrile and ethyl bromoacetate were added and refluxed for 16 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure and purified by column chromatography to obtain compound 2. (c) Compound 2 was dissolved in tetrahydrofuran at room temperature and the reaction solution was added to an alkaline solution and stirred for 18 hours. Then, the reaction solution was poured into deionized water and the pH was adjusted with hydrochloric acid. The resulting white solid was filtered, and the filtrate was extracted with dichloromethane. The filtered white solid was dissolved in dichloromethane and combined with the extracted organic phase. Then, the mixture was dried with anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 3.

[0062] (2)(d) Sodium hydride was dissolved in dry N,N-dimethylformamide under N2 atmosphere, 2-(2-aminoethoxy)ethanol was added under ice bath conditions and stirred for 45 minutes, then 1-chloro-6-iodohexane was added and stirred for 4 hours. The reaction solution was then dissolved in dichloromethane and washed twice with saturated sodium chloride solution, then washed twice with water, and concentrated under reduced pressure to obtain H-TAG.

[0063] (3)(e) Compound 3 was dissolved in N,N-dimethylformamide under N2 atmosphere and stirred in an ice bath for 20 minutes. Triethylamine was then added and stirred for 20 minutes. HBTU was then added and stirred for 2.5 hours. H-TAG was then added and stirred at 30°C for 5 hours. The reaction solution was poured into water and extracted with dichloromethane. The organic phase was dried with anhydrous sodium sulfate and concentrated under reduced pressure. After column purification, compound T-TAG was obtained.

[0064] Preferably, in step (1)(a), the molar ratio of zinc powder, titanium tetrachloride, pyridine, benzophenone, and 4-hydroxybenzophenone is 4:2:1:(0.4~0.5):0.4; the molar concentration of zinc powder suspended in anhydrous tetrahydrofuran is 0.55~0.6mol / L; the molar concentration of benzophenone dissolved in anhydrous tetrahydrofuran is 0.15~0.18mol / L; and the volume ratio of ethyl acetate to petroleum ether in column purification is 1:10; in step (b), compound 1, ethyl bromoacetate, and carbon... The molar ratio of potassium bromoacetate is 1:(1.2–1.3):2.1; the molar concentration of ethyl bromoacetate dissolved in acetonitrile solution is 0.24–0.25 mol / L; the volume ratio of ethyl acetate to petroleum ether in column purification is 1:10; the molar ratio of compound 2 to solid sodium hydroxide in (c) is 1:(5.9–6); the molar concentration of compound 2 dissolved in tetrahydrofuran is 0.06–0.08 mol / L; the molar concentration of solid sodium hydroxide dissolved in water is 3–3.5 mol / L.

[0065] Preferably, in steps (2) and (d), the molar ratio of 2-(2-aminoethoxy)ethanol, 1-chloro-6-iodohexane, and sodium hydride is in the range of 1:1.1:1.2; and the molar concentration of sodium hydride dissolved in N,N-dimethylformamide is in the range of 1 to 1.2 mol / L.

[0066] Preferably, in step (3)(e), the molar ratio of compound 3, H-TAG, triethylamine and HBTU is 1:(1.18~1.2):1.5:3; the molar concentration of compound 3 dissolved in N,N-dimethylformamide is 0.1~0.12mol / L; and the volume ratio of methanol to dichloromethane in column purification is 1:20.

[0067] Example 1

[0068] A method for preparing a proportional fluorescent probe for detecting water content, comprising the following steps:

[0069] 1) Synthesis of Compound 1:

[0070]

[0071] Zinc powder (5.38 g, 82.32 mmol) was uniformly suspended in anhydrous tetrahydrofuran (140 mL) under a nitrogen atmosphere. Titanium tetrachloride (41.16 mL, 41.16 mmol) was slowly added at -5 °C, and the reaction mixture was refluxed at 25 °C for 2.5 h. Then, pyridine (1.70 mL, 20.58 mmol) was added at -5 °C, and the mixture was stirred for 10 min. Benzophenone (1.50 g, 8.24 mmol) and 4-hydroxybenzophenone (1.96 g, 9.96 mmol) dissolved in anhydrous tetrahydrofuran (50 mL) were added, and the reaction mixture was refluxed for 17 h. After quenching with 10% potassium carbonate aqueous solution, the mixture was filtered, extracted three times with dichloromethane, concentrated under reduced pressure in the dichloromethane phase, and purified by column chromatography (ethyl acetate / petroleum ether = 1 / 10, v / v) to give compound 1 (white solid) in 30% yield.

[0072] 1 H NMR (400MHz, DMSO-d6), δ: 9.33 (s, 1H), 7.10 (s, 15H), 6.75 (s, 2H), 6.51 (s, 2H).

[0073] Synthesis of compound 2:

[0074]

[0075] Compound 1 (0.62 g, 1.80 mmol) and potassium carbonate (0.52 g, 3.78 mmol) were reacted in a round-bottom flask under N2 atmosphere for 30 min. Then, acetonitrile (9.0 mL) and ethyl bromoacetate (0.38 g, 2.24 mmol) were added and refluxed for 16 h. After cooling to room temperature, the mixture was concentrated under reduced pressure and purified by column chromatography (ethyl acetate / petroleum ether = 1 / 10, v / v) to give compound 2 (white solid) in 65% yield.

[0076] 1 H NMR (400MHz, CDCl3), δ: 7.09(s,9H),7.02(s,6H),6.93(s,2H),6.65(s,2H),4.54(s,2H),4.26(s,2H),1.28(s,3H).

[0077] Synthesis of compound 3:

[0078]

[0079] Compound 2 (0.20 g, 0.46 mmol) was dissolved in tetrahydrofuran (7.6 mL) at room temperature, and the reaction solution was added to an alkaline solution (1.16 g, 2.74 mmol of sodium hydroxide solid dissolved in 0.8 mL of water) and stirred for 18 hours. Then, the reaction solution was poured into 20 mL of deionized water and the pH was adjusted to 2 with 4 mol / L hydrochloric acid. The resulting white solid was filtered, and the filtrate was extracted with dichloromethane. The filtered white solid was dissolved in dichloromethane and combined with the extracted organic phase. The mixture was then dried under reduced pressure with anhydrous sodium sulfate to obtain compound 3 (white solid) in 96% yield.

[0080] 1 H NMR (400MHz, DMSO-d6), δ: 12.92(s,1H),7.11(s,9H),6.97(s,6H),6.85(s,2H),6.68(s,2H),4.57(s,2H).

[0081] 2) Synthesis of compound H-TAG:

[0082]

[0083] Sodium hydride (0.26 g, 6.2 mmol) was dissolved in dry N,N-dimethylformamide (6 mL) under N2 atmosphere. 2-(2-aminoethoxy)ethanol (0.52 mL, 5.16 mmol) was added under ice bath conditions and stirred for 45 minutes. Then 1-chloro-6-iodohexane (1.40 g, 5.68 mmol) was added and stirred for 4 hours. The reaction solution was then dissolved in dichloromethane and washed twice with saturated sodium chloride solution, followed by two washes with water. The solution was concentrated under reduced pressure to obtain H-TAG (pale yellow viscous liquid) in 54% yield.

[0084] 1 H NMR(400MHz, CDCl3), δ:3.67–3.62(m,2H),3.62–3.58(m,2H),3.55(dd,J=9.1,4.2Hz,4H),3.50(dd,J=12.8,6.2Hz,2 H),3.13–2.78(m,2H),1.85–1.75(m,2H),1.68–1.58(m,2H),1.52–1.43(m,2H),1.43–1.35(m,2H),1.01–0.64(m,2H).

[0085] 3) Synthesis of compound T-TAG:

[0086]

[0087] Compound 3 (0.26 g, 0.64 mmol) was dissolved in N,N-dimethylformamide (6 mL) under N2 atmosphere and stirred in an ice bath for 20 minutes. Triethylamine (0.194 g, 0.96 mmol) was then added and stirred for 20 minutes. HBTU (0.36 g, 1.92 mmol, O-benzotriazole-tetramethylurea hexafluorophosphate) was then added and stirred for 2.5 hours. H-TAG (0.18 g, 0.76 mmol) was then added and stirred at 30 °C for 5 hours. The reaction solution was poured into 20 mL of water and extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. Column purification (methanol / dichloromethane = 1 / 20, v / v) yielded compound T-TAG (a light yellow gelatinous solid) in 46% yield.

[0088] 1 H NMR(400MHz, CDCl3), δ:7.70(s,1H),7.54(s,1H),7.09(s,7H),7.02(s,6H),6.95(s,2H),6.66(s,2H),4 .42(s,2H),4.22(s,1H),3.59(s,3H),3.57(s,5H),3.51(s,2H),3.45(s,2H),1.76(s,4H),1.59(s,4H).

[0089] 13 C NMR (101MHz, CDCl3), δ: 168.36, 155.91, 144.00, 143.91, 140.89, 140.28, 137.82, 132.92, 131.49, 131.47, 131.04, 128.99, 127.94, 127.86, 127 .81,126.66,126.59,126.55,114.12,71.48,70.62,70.25,69.95,68.36 ,67.49,45.16,38.97,32.71,30.58,29.88,29.64,29.13,26.87,25.60.

[0090] Example 2: Water solubility test of fluorescent probe T-TAG

[0091] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO (dimethyl sulfoxide).

[0092] First, the UV-Vis absorption spectrometer (200nm–800nm) was calibrated using a blank solvent (membrane-filtered deionized water). Different concentrations of probe T-TAG test solutions were prepared by sequentially adding probe T-TAG stock solution to quartz cuvettes containing 3mL of ultrapure water. After mixing, the quartz cuvettes containing the test solutions were placed in the test chamber of the UV-Vis absorption spectrometer (200nm–800nm) for absorption spectrum scanning (see [link]). Figure 7 ).

[0093] Test results as follows Figure 7 As shown, within the concentration range of 5–30 μmol / L, the absorbance corresponding to 5 μmol / L is 0.23007, 10 μmol / L is 0.32342, 15 μmol / L is 0.44744, 20 μmol / L is 0.56021, 25 μmol / L is 0.269699, and 30 μmol / L is 0.78406. Under these six concentration conditions, the absorbance of the fluorescent probe T-TAG in water was fitted, and the absorbance showed a linear relationship with the concentration, conforming to Beer-Lambert law. This demonstrates that the probe T-TAG has good water solubility within a certain concentration range.

[0094] Example 3: Lipid solubility test of fluorescent probe T-TAG

[0095] First, the UV-Vis absorption spectrometer (200nm–800nm) was calibrated using a blank solvent (membrane-filtered deionized water). The probe T-TAG stock solution prepared in Example 2 was added sequentially to a quartz cuvette containing 3mL of acetonitrile solution. Then, probe T-TAG test solutions of different concentrations were prepared and mixed thoroughly. The quartz cuvette containing the test solutions was placed in the test chamber of the UV-Vis absorption spectrometer (200nm–800nm) for absorption spectrum scanning (see [link]). Figure 8 The test results are as follows: Figure 8 As shown, within the concentration range of 5–30 μmol / L, the absorbance corresponding to 5 μmol / L is 0.46603, 10 μmol / L is 0.95137, 15 μmol / L is 1.45885, 20 μmol / L is 1.98297, 25 μmol / L is 2.50862, and 30 μmol / L is 3.14818. Under these six concentration conditions, the absorbance of the fluorescent probe T-TAG in acetonitrile solution was fitted, and the absorbance showed a linear relationship with concentration, conforming to Beer-Lambert law. This demonstrates that the probe T-TAG has good lipophilicity within a certain concentration range.

[0096] Example 4: Absorption and emission spectra of the fluorescent probe T-TAG in different solvents

[0097] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. The UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent (membrane-filtered deionized water). Then, an equal volume of the 20 mmol / L probe T-TAG stock solution was mixed with different solvents (toluene, ethyl acetate, tetrahydrofuran, dichloromethane, dimethyl sulfoxide, acetonitrile, methanol, and water) in quartz cuvettes. After thorough mixing, a test solution (20 mmol / L, 2 mL) was prepared. The quartz cuvette containing the test solution was placed in the test chamber of the UV-Vis absorption spectrometer (200 nm–800 nm) for absorption spectroscopy scanning. Subsequently, the test solution was excited using a fluorescence spectrometer (350 nm–600 nm) to obtain the fluorescence emission spectrum of the probe. Table 1 shows the variation of absorption and emission wavelengths of the fluorescent probe T-TAG in the corresponding solvents with polarity. The short and long absorption wavelengths are represented by λabs1 and λabs2, respectively, and the emission wavelength is represented by λem. (See Table 1 for more details.) Figure 9 (and Table 1).

[0098] Test results as follows Figure 9 Table 1 shows that the fluorescent probe T-TAG exhibits dual absorption peaks and dual fluorescence emission peaks in eight solvents, including toluene. Furthermore, the fluorescence emission peak of the T-TAG probe shows a significant red shift: with increasing solution polarity, the fluorescence emission spectrum of the probe red-shifts from 389 nm with low-polarity toluene as the test solvent to 478 nm with high-polarity water as the test solvent. Detection Results Figure 9 Table 1 demonstrates that the absorption and emission wavelengths of the fluorescent probe T-TAG are both polar-sensitive.

[0099] Example 5: Absorption and fluorescence emission spectra of the fluorescent probe T-TAG in mixed solvents of different polarities.

[0100] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, the UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent (membrane-filtered deionized water). Then, equal volumes of the probe stock solution were diluted to different concentrations (0%–100% water content) using water and 1,4-dioxane, respectively, to create test systems (2 mL) with different polarities. These polarity test systems were added to quartz cuvettes and mixed thoroughly to prepare the test solutions. The quartz cuvettes containing the test solutions were placed in the test chamber of the UV-Vis absorption spectrometer (200 nm–800 nm) for absorption spectroscopy scanning. Subsequently, the test solutions were excited using a fluorescence spectrometer (350 nm–600 nm) to obtain the fluorescence emission spectrum of the probe. By mixing different volumes of water and dioxane, test systems with polarities Δf = 0.086–0.316 were constructed, where the polarity of the mixed system increased accordingly with increasing water content. The polarity of the mixed solvent can be calculated according to Formula 1, where the mixed dielectric constant (ε) mix ) and mixed refractive index The calculations can be performed using formulas 2 and 3. The absorption wavelengths of the fluorescent probe T-TAG at short and long wavelengths are represented by λabs1 and λabs2, respectively, and the emission wavelength is represented by λem. (See...) Figure 10 , Figure 11 (and Table 2).

[0101]

[0102] ε mix =f a ε a +f b ε b Formula 2

[0103]

[0104] Where: Δf: polarity of the mixed solvent; ε: dielectric constant of the solvent; n: refractive index of the solvent; ε mix : is the dielectric constant of the mixed solvent; f a : represents the polarity of solution a; f b : represents the polarity of solution b; ε a ε is the dielectric constant of solvent a; b : is the dielectric constant of solvent b; n mix : is the refractive index of the mixed solvent; n a : where a is the refractive index of solvent; n b : is the refractive index of solvent b.

[0105] Test results as follows Figure 10 , Figure 11As shown in Table 2, the absorbance of the probe T-TAG did not change significantly with the change in the volume ratio of water and 1,4-dioxane, proving that the solubility of the fluorescent probe T-TAG in the water and 1,4-dioxane mixture did not change significantly (Δf = 0.086–0.320). This ensures that the changes in the UV-Vis absorption and fluorescence emission spectra of the 20 mmol / L fluorescent probe in the water and 1,4-dioxane mixture are only affected by polarity. The fluorescent probe T-TAG exhibits a double absorption peak in a mixture of water and 1,4-dioxane. The polarity response of the fluorescence emission peak of T-TAG is mainly reflected in the increase of water volume fraction and polarity in the mixture. A significant redshift of approximately 102 nm occurs at the emission peak position: when Δf = 0.086–0.306, the emission peak of T-TAG is in the short wavelength range (approximately 371 nm); when Δf = 0.311–0.320, the emission peak is in the long wavelength range (approximately 473 nm). When the polarity of the water-1,4-dioxane mixture is Δf = 0.086–0.320, the fluorescence intensity ratio (Ig) at wavelengths of 473 nm and 371 nm is... 473nm / I 371nm The solution polarity can be fitted using the Boltzmann function (see [link]). Figure 11 As the volume fraction of water in the mixture increases, the polarity increases, and the proportional fluorescence emission intensity also shows a significant increase. This is presumably because with the increase in water volume fraction, the molecular configuration changes, forming aggregates, and the aggregation-induced fluorescence emission from these aggregates is enhanced. Function analysis shows that the proportional fluorescence parameter I... 473nm / I 371nm The polarity of the fluorescence changes with the polarity of the mixture, showing a significant change around Δf = 0.30. Analysis of the absorption and emission peak positions reveals that when Δf = 0.086–0.306, the peak positions are all in the short wavelength range, and when Δf = 0.311–0.320, the peak positions are all in the long wavelength range, consistent with the proportional fluorescence polarity response results (showing a significant change around Δf = 0.30). Considering the double absorption and fluorescence emission peaks of the fluorescent probe T-TAG, and the proportional fluorescence intensity (Ig)... 473nm / I 371nm The polarity has a Boltzmann function fitting relationship, and the fluorescent probe T-TAG can be used as a proportional fluorescent probe for detecting polarity.

[0106] Example 6: Time-scan fluorescence emission spectroscopy of fluorescent probe T-TAG in a pure water system

[0107] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, 2 mL of the test stock solution was added to 2 mL of pure water to prepare a 20 mmol / L test solution. After mixing, the solution was placed in a quartz cuvette and subjected to a fluorescence spectrometer (350 nm–600 nm) for 2.5 h of spectral scanning to obtain the fluorescence time-scan spectrum of the probe. Subsequently, a transmission electron microscope (TEM) was used to scan the stable aggregates.

[0108] Test results as follows Figure 12 , Figure 13 As shown, after preparing the test solution and mixing for 2.5 hours, the fluorescence intensity remained essentially unchanged, and the probe T-TAG mainly existed as irregularly shaped sheet-like aggregates in the pure water solvent, indicating that the fluorescent probe aggregated in the pure water system. These experimental results demonstrate that the polarity-responsiveness of the probe T-TAG originates from the aggregation altering the morphology of the AIE molecule and regulating its fluorescence properties. They also prove that the significant increase in proportional fluorescence emission intensity is due to the aggregation forming and regulating the AIE molecule morphology as polarity increases.

[0109] Example 7: Fluorescence emission spectrum test of fluorescent probe T-TAG in mixed solutions of 1,4-dioxane with different water contents (0%–100%).

[0110] Please refer to Example 5 for test procedures (see Example 5). Figure 14 ).

[0111] Test results as follows Figure 14As shown, the fluorescence intensity ratio at wavelengths of 473 nm and 371 nm exhibits a Boltzmann fit relationship with water content ranging from 0% to 100%. Linear analysis of the experimental results for water content ranging from 0% to 10% reveals the following: at 1% water content, the fluorescence intensity ratio at wavelengths of 473 nm and 371 nm is 0.12313; at 3% water content, it is 0.15048; at 5% water content, it is 0.15705; and at 7% water content, the ratio is... The fluorescence intensity ratio was 0.14811. At 9% water content, the fluorescence intensity ratio at 473 nm to 371 nm was 0.16643; at 10% water content, the ratio was 0.18628. The calibration curve was y = 0.516x + 0.129 (R² = 0.8817), where y represents the fluorescence intensity ratio at 473 nm and 371 nm, and x represents the water content from 0% to 10%. Based on the 3σ / n rule (S / N = 3), the limit of detection (LOD) was calculated to be 0.0111%, demonstrating that the fluorescent probe T-TAG can sensitively detect slight changes in water content in the organic solvent 1,4-dioxane.

[0112] Example 8: Fluorescence emission spectrum test of fluorescent probe T-TAG in acetonitrile mixed solutions with different water contents (0%–100%)

[0113] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, the UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent (membrane-filtered deionized water). Then, equal volumes of the probe stock solution were diluted to different concentrations (0%–100% water content) using water and acetonitrile respectively to create test systems (2 mL) with different polarities. These test systems were added to quartz cuvettes and mixed thoroughly to prepare the test solutions. The quartz cuvettes containing the test solutions were placed in the test chamber of the UV-Vis absorption spectrometer (200 nm–800 nm) for absorption spectroscopy scanning. Subsequently, the test solutions were excited using a fluorescence spectrometer (350 nm–600 nm) to obtain the fluorescence emission spectrum of the probe (see [link to relevant documentation]). Figure 15 and Figure 16 ).

[0114] Test results as follows Figure 15 , Figure 16As shown, two fluorescence emission peaks are observed at wavelengths of 406 nm and 468 nm. The fluorescence emission peak at 368 nm decreases with increasing wavelength, i.e., a redshift occurs. The trend of the fluorescence emission peak at 454 nm is similar to that in the water and dioxane mixed solution system. When the water content increases to 70%, its intensity increases, redshifting to wavelength 468 nm; when the water content exceeds 80%, the new peak at wavelength 468 nm gradually strengthens. Linear analysis of the experimental results with water content of 0%–10% shows that: at 0% water content, the fluorescence intensity ratio at wavelengths of 468 nm and 406 nm is 0.22786; at 1% water content, the ratio is 0.23904; at 3% water content, the ratio is 0.24881; and at 5% water content, the ratio is 0.24881. The fluorescence intensity ratio at 468 nm to 406 nm is 0.26091; at 7% water content, the ratio is 0.26886; at 9% water content, the ratio is 0.26836; and at 10% water content, the ratio is 0.27357. The calibration curve is y = 0.429x + 0.234(R). 2 =0.9302), where y represents the ratio of fluorescence intensity at wavelengths of 468 nm and 406 nm, and x represents the water content from 0% to 10%. Based on the 3σ / n rule (S / N = 3), the limit of detection (LOD) is calculated to be 0.0146%, demonstrating that the fluorescent probe T-TAG can sensitively detect slight changes in water content in the organic solvent acetonitrile.

[0115] Example 9: Fluorescence emission spectrum test of fluorescent probe T-TAG in mixed solutions of N,N-dimethylformamide with different water contents (0%–100%).

[0116] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, the UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent (membrane-filtered deionized water). Then, equal volumes of the probe stock solution were diluted to different concentrations using water and N,N-dimethylformamide, respectively, to create test systems (2 mL) with different polarities. These test systems were added to quartz cuvettes and mixed thoroughly to prepare the test solutions. The quartz cuvettes containing the test solutions were placed in the test chamber of the UV-Vis absorption spectrometer (200 nm–800 nm) for absorption spectroscopy scanning. Subsequently, the test solutions were excited using a fluorescence spectrometer (350 nm–600 nm) to obtain the fluorescence emission spectrum of the probe (see [link to relevant documentation]). Figure 17 and Figure 18 ).

[0117] Test results as follows Figure 17 , Figure 18 As shown, there is a fluorescence emission peak at a wavelength of 411 nm, but as the water content increases, the fluorescence emission peak red-shifts to a wavelength of 468 nm, resulting in two stages of fluorescence change. First, when the water content increases to 30%, the fluorescence intensity of T-TAG at 411 nm gradually decreases; when the water content increases to 60%, it red-shifts to 422 nm; and when the water content increases to 70%–100%, its fluorescence intensity peak red-shifts to 468 nm. Linear analysis of the experimental results with water content ranging from 0% to 10% showed that: at 0% water content, the fluorescence intensity ratio at wavelengths of 468 nm and 411 nm was 0.21774; at 1% water content, the ratio was 0.22651; at 3% water content, the ratio was 0.22859; and at 5% water content, the ratio was... The fluorescence intensity ratio at 1 nm is 0.23070; at 7% water content, the fluorescence intensity ratio at 468 nm to 411 nm is 0.23519; at 9% water content, the ratio is 0.23885; and at 10% water content, the ratio is 0.23937. The calibration curve is y = 0.190x + 0.222(R). 2 =0.9223), where y represents the ratio of fluorescence intensity at wavelengths of 468 nm and 411 nm, and x represents the water content from 0% to 10%. Based on the 3σ / n rule (S / N = 3), the limit of detection (LOD) is calculated to be 0.0215%, demonstrating that the fluorescent probe T-TAG can sensitively detect slight changes in water content in the organic solvent N,N-dimethylformamide.

[0118] Example 10: Detection of water content (0%–100%) in organic solvents using the fluorescent probe T-TAG.

[0119] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, the UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent (membrane-filtered deionized water). Then, equal volumes of the probe stock solution were diluted with water and organic solvents (1,4-dioxane, acetonitrile, and N,N-dimethylformamide) to different concentrations (0%–100% water content) to create test systems (2 mL) with different polarities. These test systems were added to quartz cuvettes and mixed thoroughly to prepare the test solution. The fluorescence intensity ratio at the two peak wavelengths was measured. Based on the standard curve of fluorescence intensity ratio versus water content, the water content in the actual sample was calculated and compared with the accurate value on the reagent label (see Table 3).

[0120] The test results are shown in Table 3: The large difference between the water content value of 1,4-dioxane and the label value may be due to the solution being left for too long; the increased water content of acetonitrile solution suggests that the solvent is a commonly used laboratory reagent and may have been contaminated; while the water content of N,N-dimethylformamide is not much different from the label value because the reagent bottle is sealed with a ring and the storage conditions meet the laboratory requirements.

[0121] Example 11: Fluorescence emission spectrum test of fluorescent probe T-TAG in ethanol mixed solutions with different water contents (0%–100%)

[0122] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, the UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent. Then, equal volumes of the probe stock solution were diluted with water and ethanol to different concentrations (0%–100% water content) to create test systems (2 mL) with different polarities. These test systems were added to quartz cuvettes and mixed thoroughly to prepare the test solutions. The quartz cuvettes containing the test solutions were placed in the test chamber of the UV-Vis absorption spectrometer (200 nm–800 nm) for absorption spectroscopy scanning. Subsequently, the test solutions were excited using a fluorescence spectrometer (350 nm–600 nm) to obtain the fluorescence emission spectrum of the probe (see [link to relevant documentation]). Figure 19 and Figure 20 ).

[0123] Test results as follows Figure 19 , Figure 20As shown, a significant blue shift in the spectral shape occurred with increasing ethanol volume ratio. Linear analysis of the experimental results for water contents ranging from 0% to 10% revealed the following: at 0% water content, the fluorescence intensity ratio at wavelength 468 nm to 365 nm was 42.78366; at 10% water content, the ratio was 41.0711; at 20% water content, the ratio was 30.50826; and at 30% water content… At a specific wavelength, the fluorescence intensity ratio at 468 nm to 365 nm was 14.16903; at 40% water content, the ratio was 1.00058. The calibration curve was y = 11.468x + 48.000 (R² = 0.9430), where y represents the fluorescence intensity ratio at 468 nm and 365 nm, and x represents the ethanol content from 0% to 40%. According to the 3σ / n rule (S / N = 3), the limit of detection (LOD) for ethanol was 0.0269%, demonstrating that the fluorescent probe T-TAG can easily and quantitatively detect ethanol content.

[0124] Example 12: Detection of alcohol content (0%–100% ethanol) in real samples using the fluorescent probe T-TAG.

[0125] The fluorescent probe T-TAG was prepared into a 20 mmol / L test stock solution using anhydrous DMSO. First, the UV-Vis absorption spectrometer (200 nm–800 nm) was calibrated using a blank solvent (membrane-filtered deionized water). Then, the same volume of the probe stock solution was added to commercial spirits of different alcohol contents (0%–100% ethanol) to create test systems of different polarities (2 mL). These polarity-different test systems were added to quartz cuvettes and mixed thoroughly to prepare the test solution. The fluorescence intensity ratio at the two peak wavelengths was measured. Based on the standard curve of fluorescence intensity ratio versus ethanol content, the alcohol content of the actual sample was calculated and compared with the accurate value on the reagent label (see Table 4).

[0126] The test results are shown in Table 4: the difference between the detected alcohol content and the alcohol content on the label is within 2%, which proves that the fluorescent probe T-TAG has a certain degree of reliability in detecting the ethanol content in the actual sample.

[0127] The instruments used were a UV-Vis spectrophotometer, model: Perkin Elmer Lambda 35UV / VIS; and a fluorescence spectrophotometer, model: F-4600, Hitachi High Technology Corporation.

[0128] Table 1

[0129]

[0130] Table 1 summarizes the absorption and emission wavelength data of the proportional fluorescent probe T-TAG for polarity detection described in this invention in different polar solvents.

[0131] Table 2

[0132]

[0133] Table 2 summarizes the photophysical data of the proportional fluorescent probe T-TAG for polarity detection described in this invention in different polarity mixed solvents.

[0134] Table 3

[0135]

[0136] Table 3 summarizes the determination data of water content in different organic solvents for the proportional fluorescent probe T-TAG used for polarity detection described in this invention.

[0137] Table 4

[0138]

[0139] Table 4 summarizes the data on the determination of alcohol content in different actual samples using the proportional fluorescent probe T-TAG for polarity detection described in this invention.

[0140] Although the present invention has been described in detail through the above preferred examples, it should be understood that the above description should not be considered as a limitation of the present invention.

[0141] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of the present invention.

Claims

1. A class of aggregation-induced emission compounds with dual fluorescence emission properties, characterized in that: The structure of the aggregation-induced emission compound of Formula I with dual fluorescence emission characteristics described above is as follows: 。 2. The application of an aggregation-induced emission compound with dual fluorescence emission characteristics as described in claim 1 in the detection of water content in organic solvents or alcohol content in wine.

3. The application according to claim 2, characterized in that, The compound shown in Formula I is used as an example of an aggregation-induced emission compound with dual fluorescence emission characteristics in the detection of water content in organic solvents or alcohol content in wine.

4. An aggregation-induced emission composition with dual fluorescence emission characteristics for detecting the water content in organic solvents or the alcohol content in wine, characterized in that: The compound containing the aggregation-induced emission properties of a class of compounds with dual fluorescence emission characteristics as described in claim 1.

5. The application of the aggregation-induced emission composition with dual fluorescence emission characteristics for water content detection according to claim 4, characterized in that: The aforementioned aggregation-induced emission composition with dual fluorescence emission characteristics can be used to detect the water content in organic solvents or the alcohol content in wine.

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