阅读说明：本技术 一种酰胺基芘衍生物荧光探针及其制备方法和应用 (Amido pyrene derivative fluorescent probe and preparation method and application thereof ) 是由 付颖 叶非 刘豫龙 杨柳 李璐 于 2021-10-13 设计创作，主要内容包括：本发明公开了一种酰胺基芘衍生物荧光探针及其制备方法和应用,属于荧光分析检测技术领域。本发明利用酰胺基为强氢键受体的结构特点,以芘衍生物为荧光团设计并合成了酰胺基芘衍生物荧光探针,并利用紫外可见光谱和荧光光谱等分别对目标化合物进行研究,证明本发明合成的酰胺基芘衍生物荧光探针为2,2-二氯-N-(芘-1-基)乙酰胺或N-(芘-1-基)呋喃-2-甲酰胺,并且2,2-二氯-N-(芘-1-基)乙酰胺可特异性识别阴离子SO-(4)～(2-)；N-(芘-1-基)呋喃-2-甲酰胺可特异性识别重金属离子Cu～(2+)。本发明合成的N-(芘-1-基)呋喃-2-甲酰胺分子探针属于“off-on”型探针,荧光开启为蓝色,Cu～(2+)检出限为4.73μM。(The invention discloses an amido pyrene derivative fluorescent probe and a preparation method and application thereof, and belongs to the technical field of fluorescence analysis and detection. The invention utilizes the structural characteristic that amido is a strong hydrogen bond receptor, designs and synthesizes amido pyrene derivative fluorescent probe by taking pyrene derivative as fluorophore, and utilizes ultraviolet visible spectrum, fluorescence spectrum and the like to respectively study target compounds, thereby proving that the amido pyrene derivative fluorescent probe synthesized by the invention is 2, 2-dichloro-N- (pyrene-1-yl) acetamide or N- (pyrene-1-yl) furan-2-formamide, and 2, 2-dichloro-N- (pyrene-1-yl) acetamide capable of specifically recognizing anion SO 4 2‑ (ii) a N- (pyrene-1-yl) furan-2-formamide capable of specifically identifying heavy metal ions Cu 2+ . The synthesized N- (pyrene-1-yl) furan-2-formamide molecular probe belongs to an off-on type probe, the fluorescence is turned on to be blue, and Cu is adopted 2+ The detection limit was 4.73. mu.M.)

1. An amido pyrene derivative fluorescent probe is characterized in that: the structural formula is shown as the following formula I, wherein: r is

2. The method for preparing an amidopyrene derivative fluorescent probe of claim 1, wherein: the method comprises the following steps:

dissolving a proper amount of 1-aminopyrene in dichloromethane, adding potassium carbonate as an acid-binding agent, and uniformly mixing to obtain a mixed solution; then slowly dripping acyl chloride into the mixed solution according to the proportion under the condition of ice-water bath, and after dripping is finished, placing the reaction system at normal temperature for reaction for 10-14 h; and after the reaction is finished, standing, filtering and evaporating to obtain a crude product, and recrystallizing the obtained crude product to obtain the target compound.

3. The method for preparing an amidopyrene derivative fluorescent probe according to claim 2, wherein: the mol ratio of the 1-aminopyrene to the acyl chloride is 1: (30-40).

4. The method for preparing an amidopyrene derivative fluorescent probe according to claim 2, wherein: the acyl chloride is dichloroacetyl chloride or 2-furoyl chloride.

5. The method for preparing an amidopyrene derivative fluorescent probe according to claim 2, wherein: the molar ratio of the 1-aminopyrene potassium carbonate is 1: (1.2-2).

6. The method for preparing an amidopyrene derivative fluorescent probe according to claim 2, wherein: the normal-temperature reflux reaction time is preferably 12 hours.

7. The application of the amido pyrene derivative fluorescent probe of claim 1 or the amido pyrene derivative fluorescent probe prepared by the method of any one of claims 2 to 6 is characterized in that: when R isWhen the amido pyrene derivative fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide is used for specifically identifying anion SO in sewage or wastewater₄ ^2-。

8. The application of the amido pyrene derivative fluorescent probe of claim 1 or the amido pyrene derivative fluorescent probe prepared by the method of any one of claims 2 to 6 is characterized in that: when R isWhen the amido pyrene derivative fluorescent probe N- (pyrene-1-yl) furan-2-formamide is used for specifically identifying heavy metal ions Cu in sewage, wastewater or organisms²⁺。

Technical Field

The invention belongs to the technical field of fluorescence analysis and detection, particularly relates to a fluorescent probe, and more particularly relates to an amido pyrene derivative fluorescent probe and a preparation method and application thereof.

Background

The metal ion fluorescent probe has important application in many fields of environmental protection, biomedicine, chemistry and the like. In terms of environmental protection, the pollution of heavy metal ions is becoming more serious in recent years, and therefore, the development of various heavy metal ion fluorescent probes for monitoring environmental pollution has important research value. In biomedicine, since many metal ions are involved in some important biochemical reactions in vivo or in the formation of coenzymes, since they affect normal physiological functions as signal molecules, the concentration form of some metal ions in vivo can reflect some chemical reaction processes and serve as a basis for disease diagnosis. Based on these important functions, the research of monitoring metal ions is of great value, and fluorescent molecular probes are presented in the field with excellent characteristics for monitoring the concentration of anions and cations in the environment.

Copper is a trace element required by human body and can assist various metal enzymes to play a role. In humans, copper ions are essential for the proper functioning and metabolic processes of the organ, and are an important component of many enzyme systems (e.g., oxidases and hydroxylases). However, if the copper ions contained in the human body are out of the normal range, the large amount of accumulated copper ions may also cause damage to human organs, and therefore, it is necessary to find a method for rapidly and effectively detecting the copper ions.

Although there are many reports in the prior art for detecting anionic and cationic fluorescent probes in the environment, most of these fluorescent probes have the problems of low yield, low detection sensitivity, low minimum detection limit, poor selectivity, sensitivity to pH, and the like. Therefore, it is still a challenge to develop a fluorescent molecular probe that has a novel structure, a simple preparation method, and high selectivity and sensitivity, and can detect anions and cations in the natural environment and in the living body.

For the above reasons, the present application has been made.

Disclosure of Invention

Aiming at the problems or defects in the prior art, the invention aims to provide an amido pyrene derivative fluorescent probe and a preparation method and application thereof. According to the invention, the structural characteristic that amido is taken as a strong hydrogen bond acceptor is utilized, the pyrene derivative is taken as a fluorophore to design and synthesize the amido pyrene derivative fluorescent probe, and the ultraviolet visible spectrum, the fluorescence spectrum and the like are utilized to respectively study a target compound, so that the amido pyrene derivative fluorescent probe provided by the invention can specifically identify heavy metal ions Cu²⁺Or anionic SO₄ ^2-And the detection limit is low.

In order to achieve the first object of the present invention, the present invention adopts the following technical solutions:

an amido pyrene derivative fluorescent probe has a structural formula shown as the following formula I, wherein: r is

Specifically, when R isWhen the target compound is 2, 2-dichloro-N- (pyrene-1-yl) acetamide, the structural formula is shown as the following formula II:

specifically, when R isWhen the target compound is N- (pyrene-1-yl) furan-2-formamide, the structural formula is shown as the following formula III:

the second purpose of the invention is to provide a preparation method of the amido pyrene derivative fluorescent probe, which comprises the following steps:

dissolving a proper amount of 1-aminopyrene in dichloromethane, adding potassium carbonate as an acid-binding agent, and uniformly mixing to obtain a mixed solution; then slowly dripping acyl chloride into the mixed solution according to the proportion under the condition of ice-water bath, and after dripping is finished, placing the reaction system at normal temperature for reaction for 10-14 h; and after the reaction is finished, standing, filtering and evaporating to obtain a crude product, and recrystallizing the obtained crude product to obtain the target compound.

Further, according to the technical scheme, the molar ratio of the 1-aminopyrene to the acyl chloride is 1: (30-40).

Further, in the technical scheme, the acyl chloride is dichloroacetyl chloride or 2-furoyl chloride.

Further, in the above technical solution, the amount of the dichloromethane used is not specifically limited as long as the 1-aminopyrene can be completely dissolved, for example, the amount of the 1-aminopyrene and the dichloromethane can be 0.01mol parts: (20 to 100) parts by volume, preferably 0.01 parts by mole: (20-40) parts by volume, wherein: the mol parts and the volume parts are as follows: mL was used as a reference.

Further, according to the technical scheme, the molar ratio of the 1-aminopyrene potassium carbonate is 1: (1.2-2), preferably 1: (1.5-2).

Specifically, in the above technical scheme, the normal temperature refers to a natural room temperature condition in four seasons, no additional cooling or heating treatment is performed, and the normal temperature is generally controlled to be 10-30 ℃, preferably 15-25 ℃.

Further, in the above technical scheme, the normal-temperature reflux reaction time is preferably 12 hours.

Specifically, the synthetic route of the amido pyrene derivative fluorescent probe is shown as the following formula IV, wherein: r is

The third purpose of the invention is to provide the application of the amido pyrene derivative fluorescent probe.

In one aspect, the invention provides the use of the 2, 2-dichloro-N- (pyrene-1-yl) acetamide for specifically identifying anions in sewage, wastewater or organisms and the like.

Further, in the above technical scheme, the anion is SO₄ ^2-。

On the other hand, the invention also provides application of the N- (pyrene-1-yl) furan-2-formamide in specific recognition of heavy metal ions in sewage or wastewater, organisms and the like.

Further, in the above technical scheme, the heavy metal ions are Cu²⁺。

Compared with the prior art, the invention has the following beneficial effects:

(1) according to the invention, by utilizing the structural characteristic that amido is a strong hydrogen bond receptor, an amido pyrene derivative fluorescent probe is designed and synthesized by taking a pyrene derivative as a fluorophore, and the ultraviolet visible spectrum, the fluorescence spectrum and the like are utilized to respectively research a target compound, SO that the amido pyrene derivative fluorescent probe synthesized by the invention is proved to be 2, 2-dichloro-N- (pyrene-1-yl) acetamide or N- (pyrene-1-yl) furan-2-formamide, and the 2, 2-dichloro-N- (pyrene-1-yl) acetamide can specifically identify anions SO₄ ^2-(ii) a The N- (pyrene-1-yl) furan-2-formamide belongs to an off-on probe and can specifically identify heavy metal ions Cu²⁺。

(2) The synthesized N- (pyrene-1-yl) furan-2-formamide molecular probe belongs to an off-on type probe, and fluorescence is used for detectingTurn off as blue, Cu²⁺The detection limit was 4.73. mu.M.

Drawings

FIG. 1 is a graph showing the effect of different cations, different anions and different pesticides on the UV spectrum of 2, 2-dichloro-N- (pyrene-1-yl) acetamide, a compound prepared in example 1, in comparison with the effect of (a), (b) and (c) in sequence; (d) is a graph comparing the effect of different cations, anions, or pesticides on the fluorescence spectrum of 2, 2-dichloro-N- (pyrene-1-yl) acetamide, a compound prepared in example 1;

FIG. 2 shows the addition of varying amounts of SO₄ ^2-Comparative graph of absorption titration spectrum of 2, 2-dichloro-N- (pyrene-1-yl) acetamide as the fluorescent probe prepared in the following example 1;

FIG. 3(a) and (b) are graphs showing the effect of different cations on fluorescence and UV spectra of the compound N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2;

FIG. 4 shows the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide + Cu prepared in example 2²⁺A plot of fluorescence intensity versus presence of various competing cations;

in FIG. 5, (a) and (b) are sequentially added with different amounts of Cu²⁺A fluorescence titration spectrum contrast chart and an absorption titration spectrum contrast chart of the fluorescent probe N- (pyrene-1-yl) furan-2-formamide prepared in the following example 2;

in FIG. 6, (a) and (b) are the fluorescent probes N- (pyrene-1-yl) furan-2-carboxamide and Cu prepared in example 2 in this order²⁺Graph of reversibility test results and Job's graph;

FIG. 7 is a graph showing the effect of time on the fluorescence intensity of N- (pyrene-1-yl) furan-2-carboxamide, a fluorescent probe prepared in example 2.

Detailed Description

The present invention will be described in further detail below with reference to examples. The present invention is implemented on the premise of the technology of the present invention, and the detailed embodiments and specific procedures are given to illustrate the inventive aspects of the present invention, but the scope of the present invention is not limited to the following embodiments.

For a better understanding of the invention, and not as a limitation on the scope thereof, all numbers expressing quantities, percentages, and other numerical values used in this application are to be understood as being modified in all instances by the term "about". At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

The structural characterization method of the synthesized compound is as follows:

(1) digital melting point tester

The dissolution range of the compound is measured by adopting a WRS-3 type digital melting point instrument of Shanghai apparatus electro-physical optical instruments Limited. Melting point measurement range: 23-320 ℃, and the minimum temperature display value is 0.1 ℃.

(2) Infrared spectrum (IR)

The infrared spectrum of the compound is measured by an IRTracer-100 type Fourier transform infrared spectrometer (Shimadzu corporation, Japan), the potassium bromide tablet is pressed, and the spectrum range is 400-4000 cm^-1。

(3) Nuclear magnetic resonance spectroscopy (NMR)

The NMR spectrum of the compound was measured by an Av-400 NMR spectrometer (Bruker, Germany) using deuterated chloroform (CDCl)₃) TMS (tetramethylsilane) as internal standard with frequency of 400MHz (respectively¹H NMR) and 100 MHz: (¹³C NMR)。

(4) High Resolution Mass Spectrum (HRMS)

The mass spectra of the compounds were determined using a high resolution mass spectrometer, model microOTOF-Q II 10410, Bruker (Bruker) Germany.

The detection limits referred to in the following examples were calculated as follows:

the detection Limit (LOD) is calculated as follows:

LOD＝3s/k_a

wherein s is the standard deviation of the maximum fluorescence intensity or absorbance of the same probe sample determined 10 times; k is a radical of_aIs the slope of the linear equation in the UV-Vis or fluorescence concentration titration experiment.

The test methods used in the following examples are all conventional methods unless otherwise specified; the raw materials and reagents used are, unless otherwise specified, those commercially available from ordinary commercial sources. For example, the 1-aminopyrenes involved in the following examples of the invention are commercially available with Cas numbers: 1606-67-3.

Example 1

The amido pyrene derivative fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide of the embodiment is prepared by the following method, and comprises the following steps:

1-Aminopyrene (2.2g, 0.01mol) was dissolved in 30mL of dichloromethane, and K was added₂CO₃(2.5g) as an acid-binding agent, and uniformly mixing to obtain a mixed solution; slowly dripping dichloroacetyl chloride (3.5g, 0.35mol) into the mixed solution by using a constant-pressure dropping funnel at the temperature of 0 ℃ in an ice water bath, and reacting for 12 hours at normal temperature after finishing dripping; standing after the reaction is finished, and filtering K₂CO₃Removing the solvent of the filtrate by rotary evaporation to obtain a crude product; and recrystallizing the obtained crude product by using dichloromethane/petroleum ether as a solvent to obtain a white powdery target compound, wherein the yield is 61 percent, and the melting point is 224.7-225.2 ℃.

The synthetic route of the amido pyrene derivative fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide is shown as the following formula VI:

the structural characterization test results are as follows:

the infrared spectrum peak data of the target compound prepared in this example is as follows:

3234-3115(C-N)；3076-3008(C-H)；1668(C＝O)。

the nmr hydrogen spectra data of the target compound prepared in this example are as follows:

11.13(s,1H),8.46–7.97(m,9H),6.90(s,1H).

the nmr carbon spectrum data of the target compound prepared in this example are as follows:

163.95,131.36,131.05,128.61,127.75,127.48,126.93,126.87,125.60,125.55,125.35,124.91,124.40,123.55,123.55,122.80,69.29.

from the high-resolution mass spectrum of the target compound prepared in this example, the molecular weight/molecular ion peak/theoretical molecular weight was 327.0218/326.0147(-H)/326.0140(-H) in this order.

From the above data, it can be seen that the target compound prepared in this example is 2, 2-dichloro-N- (pyrene-1-yl) acetamide.

Example 2

The amido pyrene derivative fluorescent probe N- (pyrene-1-yl) furan-2-formamide of the embodiment is prepared by the following method, and comprises the following steps:

1-Aminopyrene (2.2g, 0.01mol) was dissolved in 30mL of dichloromethane, and K was added₂CO₃(2.5g) as an acid-binding agent, and uniformly mixing to obtain a mixed solution; slowly dripping 2-furoyl chloride (3.5g, 0.35mol) into the mixed solution by using a constant-pressure dropping funnel at the temperature of 0 ℃ in an ice water bath, and reacting for 12 hours at normal temperature after finishing dripping; standing after the reaction is finished, and filtering K₂CO₃Removing the solvent of the filtrate by rotary evaporation to obtain a crude product; and recrystallizing the obtained crude product by using dichloromethane/petroleum ether as a solvent to obtain a target compound which is a green flaky crystal, wherein the yield is 74%, and the melting point is 249.0-249.1 ℃.

The synthetic route of the amido pyrene derivative fluorescent probe N- (pyrene-1-yl) furan-2-formamide is shown as the following formula seven:

the structural characterization test results are as follows:

the infrared spectrum peak data of the target compound prepared in this example is as follows:

3361-3302(C-N)；2976-2877(C-H)；1653(C＝O)。

the nmr hydrogen spectra data of the target compound prepared in this example are as follows:

10.76(s,1H),8.42–7.99(m,10H),7.51(dd,J＝3.5,0.8Hz,1H),6.79(dd,J＝3.5,1.8Hz,1H).

the nmr carbon spectrum data of the target compound prepared in this example are as follows:

157.81,148.10,146.34,131.38,131.23,130.94,129.52,127.75,127.71,127.50,126.95,126.16,125.87,125.64,125.60,125.38,124.84,124.25,123.32,115.35,112.67.

from the high-resolution mass spectrum of the target compound prepared in this example, the molecular weight/molecular ion peak/theoretical molecular weight was 311.0946/310.0876(-H)/310.0868(-H) in this order.

From the above data, it can be seen that the target compound prepared in this example is N- (pyrene-1-yl) furan-2-carboxamide.

And (3) performance testing:

(first) the selectivity test of the fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide prepared in example 1 on different metal ions, anions or pesticides:

50mL of 10 concentration ultrapure water was used^-2mol/L metal ion standard solution (Na)⁺、K⁺、Ca²⁺、Zn²⁺、Mg²⁺、Fe^{3 +}、Al³⁺、Ni²⁺、Sn²⁺、Pb²⁺、Hg²⁺、Cr²⁺、Ag⁺、Co²⁺、Mn²⁺、Cu²⁺、Ba²⁺、Fe²⁺、Cu⁺) Anion standard solution (F)^-、Cl^-、I^-、ClO₄ ^-、NO₃ ^-、HSO₄ ^-、CH₃COO^-、CN^-、SCN^-、NO₂ ^-、SO₄ ^2-、H₂PO₄ ^-) Pesticide standard solution (cypermethrin, cyhalothrin, oxyfluorfen, mesotrione, cyfluthrin, teflubenzuron, flusilazole, flucycloxuron, isoxaflutole, tembotrione, sulcotrione, NTBC, glyphosate and clethodim) and concentration of 1 x 10^－5mol/L of the fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide prepared in example 1 in dimethyl sulfoxide as a pure solutionAnd (4) preparing standard solution for later use.

The standard solution of the fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide prepared in example 1 and standard solutions of different metal ions, anions and pesticides are prepared into a plurality of 10mL samples to be detected according to the volume ratio of 1:5, a fluorescence spectrophotometer is used for carrying out fluorescence spectrum measurement on the samples, and an ultraviolet visible spectrophotometer is used for carrying out ultraviolet spectrum measurement on the samples, so that a target object which can be identified by the probe is determined. As shown in FIG. 1, when various cations, anions and pesticides were added, almost no change was observed in the fluorescence spectrum and ultraviolet spectrum, but only SO₄ ^2-The UV spectrum of (1) has slight change, and a new peak appears at 445 nm.

(II) titration detection of different concentrations of sulfate ions by the fluorescent probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide prepared in example 1:

50mL of the solution with the concentration of 10 is prepared^-2Ultra-pure aqueous solution of sulfate ions in mol/L and concentration of 1X 10^－5A pure dimethyl sulfoxide solution of 2, 2-dichloro-N- (pyrene-1-yl) acetamide, a fluorescent probe prepared in example 1, in mol/L was used as a standard solution. Adding sulfate ion standard solutions with different volumes (0-10 equivalent) into a standard solution of a probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide to prepare solutions to be detected with different concentration ratios; and (3) respectively carrying out fluorescence spectrum and ultraviolet spectrum measurement on the solution so as to determine the identification capability of the probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide on sulfate ions with different concentrations. As shown in fig. 2, with SO₄ ^2-When the ratio of (A) to (B) is increased from 0 to 10 equivalents, the probe 2, 2-dichloro-N- (pyrene-1-yl) acetamide shows a new peak at 445nm and becomes more and more pronounced, and the intensity reaches a maximum at 10 equivalents.

(III) the selectivity test of the fluorescent probe N- (pyrene-1-yl) furan-2-formamide prepared in the example 2 on different metal ions:

50mL of 10 concentration ultrapure water was used^-2mol/L metal ion standard solution (Na)⁺、K⁺、Ca²⁺、Zn²⁺、Mg²⁺、Fe^{3 +}、Al³⁺、Ni²⁺、Sn²⁺、Pb²⁺、Hg²⁺、Cr²⁺、Ag⁺、Co²⁺、Mn²⁺、Cu²⁺、Ba²⁺、Fe²⁺、Cu⁺) And a concentration of 1X 10^－5A pure acetonitrile solution of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 in mol/L was used as a standard solution.

Preparing a plurality of samples to be detected from the standard solution of the probe N- (pyrene-1-yl) furan-2-formamide and the standard solutions of different metal ions according to the volume ratio of 1:5, performing fluorescence spectrum measurement on the samples by using a fluorescence spectrophotometer, and performing ultraviolet spectrum measurement on the samples by using an ultraviolet visible spectrophotometer so as to determine the target object which can be identified by the probe. As shown in FIG. 3, it can be seen from FIG. 3(a) that the addition of copper ions turns on the fluorescence, which is significantly enhanced but has different enhancing effect, while the fluorescence of other ions under the same conditions is negligible. As can be seen from FIG. 3(b), when various metal ions were added to the system, the absorption spectrum of the probe N- (pyrene-1-yl) furan-2-carboxamide was found in Cu²⁺In the presence of this, a blue shift occurs and the absorbance decreases significantly.

(IV) the interference detection of the specific recognition of copper ions by the fluorescent probe N- (pyrene-1-yl) furan-2-formamide prepared in the example 2:

50mL of 10 concentration ultrapure water was used^-2mol/L metal ion standard solution (Na)⁺、K⁺、Ca²⁺、Zn²⁺、Mg²⁺、Fe^{3 +}、Al³⁺、Ni²⁺、Sn²⁺、Pb²⁺、Hg²⁺、Cr²⁺、Ag⁺、Co²⁺、Mn²⁺、Cu²⁺、Ba²⁺、Fe²⁺、Cu⁺) And a concentration of 1X 10^－5A pure acetonitrile solution of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 in mol/L was used as a standard solution. The concentration is 1 x 10^－5Acetonitrile pure solution of mol/L fluorescent probe N- (pyrene-1-yl) furan-2-formamide and concentration of acetonitrile pure solution is 10^{- 2}Preparing a plurality of same samples to be detected by mol/L copper ion standard solution according to the volume ratio of 1:5, and respectively introducing other samples to be detectedThe anti-interference capability of the probe N- (pyrene-1-yl) furan-2-formamide for identifying copper ions in the presence of different metal ions is detected by the same amount of metal ion standard solution. As can be seen from FIG. 4, the recognition capability of the probe N- (pyrene-1-yl) furan-2-formamide for copper ions is not influenced by other metal ions, and the probe has good anti-interference capability.

(V) titration detection of different concentrations of copper ions by the fluorescent probe N- (pyrene-1-yl) furan-2-formamide prepared in example 2:

50mL of the solution with the concentration of 10 is prepared^-2Pure solution of ultrapure water containing copper ions in mol/L and having a concentration of 1X 10^－5A pure acetonitrile solution of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 in mol/L was used as a standard solution. Adding copper ion standard solutions with different volumes (0-3 equivalent) into a standard solution of a probe N- (pyrene-1-yl) furan-2-formamide to prepare solutions to be detected with different concentration ratios; and respectively carrying out fluorescence spectrum and ultraviolet spectrum measurement on the solution so as to determine the recognition capability of the probe N- (pyrene-1-yl) furan-2-formamide on copper ions with different concentrations.

As shown in FIG. 5(a), the abscissa is the wavelength (nm), the ordinate is the fluorescence intensity, Cu²⁺The ratio of (A) to (B) is increased from 0 to 3 equivalents, and the fluorescence of the probe N- (pyrene-1-yl) furan-2-carboxamide is gradually increased and reaches the highest at 3 equivalents. In FIG. 5(b), Cu at various concentrations is shown²⁺The absorption spectrum of N- (pyrene-1-yl) furan-2-carboxamide changes with Cu²⁺The amount increased, the absorption peak of the probe at 500nm gradually blue-shifted, and the absorbance slowly decreased and stabilized at 3 equivalents. Fluorescence intensity and Cu²⁺The concentration has good linear relation, the detection limit is as low as 4.73 mu M, and the aim of Cu treatment is realized²⁺And (4) carrying out quantitative detection.

(VI) reversible detection of copper ions by the fluorescent probe N- (pyrene-1-yl) furan-2-formamide prepared in example 2

50mL of the solution with the concentration of 10 is prepared^-2Pure solution of ultrapure water containing copper ions in mol/L and having a concentration of 1X 10^－5mol/L pure acetonitrile solution of the fluorescent probe N- (pyrene-1-yl) furan-2-formamide prepared in example 2 was used as a standard solution for future use, and electrons were removedEDTA was weighed by an analytical balance and prepared to a concentration of 10 with ultrapure water^-2Pure solution of mol/L is ready for use. In probe-Cu²⁺Adding a proper amount of EDTA solution into the ladder system, and carrying out fluorescence spectrum and ultraviolet spectrum measurement on the EDTA solution; introducing equal amount of copper ions into the same system again, and measuring the fluorescence spectrum and the ultraviolet spectrum again; and repeating for multiple times to determine the reversible recognition capability of the probe for the copper ions.

FIG. 6(a) shows the probe N- (pyrene-1-yl) furan-2-carboxamide vs Cu after EDTA addition²⁺In response to the change, Cu is added into a probe N- (pyrene-1-yl) furan-2-formamide system²⁺Turning on fluorescence; after the equivalent amount of EDTA is added, the fluorescence is closed; then adding equal amount of Cu²⁺Then, the fluorescence is turned on again; the fluorescence spectrum was obtained by repeating the above steps several times, as shown in FIG. 6 (a). This is because of Cu²⁺The chelating ability with EDTA is larger than the coordination ability with probe N- (pyrene-1-yl) furan-2-formamide, which indicates that the probe N- (pyrene-1-yl) furan-2-formamide can be used for detecting Cu²⁺The chemically reversible probe of (1).

(hepta) test of the complexation ratio of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 to copper ions:

50mL of the solution with the concentration of 10 is prepared^-2Pure solution of ultrapure water containing copper ions in mol/L and having a concentration of 1X 10^－5A pure acetonitrile solution of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 in mol/L was used as a standard solution. Preparing solutions with different proportions from the probe standard solution and the copper ion standard solution according to the volume ratio of 1:0 to 1: 10; and (3) respectively measuring the solution by a fluorescence spectrum and an ultraviolet spectrum so as to determine the coordination ratio of the probe to the copper ions. The results in FIG. 6(b) show that [ N- (pyrene-1-yl) furan-2-carboxamide]/([ N- (pyrene-1-yl) furan-2-carboxamide)]+[Cu²⁺]) Is 0.5. Thus, it was demonstrated that, in the initial stage of the reaction, the probe N- (pyrene-1-yl) furan-2-carboxamide was reacted with Cu²⁺The binding ratio of (A) to (B) is 1: 1.

(eight) measurement of influence of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 on copper ion with time:

50mL of the solution with the concentration of 10 is prepared^-2Ultra pure aqueous solution of copper ions in mol/L and concentration of 1X 10^－5A pure acetonitrile solution of the fluorescent probe N- (pyrene-1-yl) furan-2-carboxamide prepared in example 2 in mol/L was used as a standard solution. Preparing the two into a sample to be measured according to a certain proportion, fixing time intervals, and measuring the sample by fluorescence spectrum and ultraviolet spectrum at different time intervals to determine the time pair probe-Cu²⁺Influence of the system. The results in FIG. 7 show that the probe N- (pyrene-1-yl) furan-2-carboxamide itself can be in a high steady state within 100 min; adding Cu²⁺Thereafter, the fluorescence intensity of the probe N- (pyrene-1-yl) furan-2-carboxamide reached a maximum level within 30 minutes and remained stable for a long time.