CN115304572A - Flavonoid fluorescent probe for detecting hydrazine and preparation method and application thereof - Google Patents
Flavonoid fluorescent probe for detecting hydrazine and preparation method and application thereof Download PDFInfo
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- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 title claims abstract description 156
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 92
- 229930003935 flavonoid Natural products 0.000 title claims abstract description 62
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 62
- 150000002215 flavonoids Chemical class 0.000 title claims abstract description 60
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000001514 detection method Methods 0.000 claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 20
- -1 flavonoid compound Chemical class 0.000 claims abstract description 18
- JECYUBVRTQDVAT-UHFFFAOYSA-N 2-acetylphenol Chemical compound CC(=O)C1=CC=CC=C1O JECYUBVRTQDVAT-UHFFFAOYSA-N 0.000 claims abstract description 14
- OTXINXDGSUFPNU-UHFFFAOYSA-N 4-tert-butylbenzaldehyde Chemical compound CC(C)(C)C1=CC=C(C=O)C=C1 OTXINXDGSUFPNU-UHFFFAOYSA-N 0.000 claims abstract description 14
- WETWJCDKMRHUPV-UHFFFAOYSA-N acetyl chloride Chemical compound CC(Cl)=O WETWJCDKMRHUPV-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000012346 acetyl chloride Substances 0.000 claims abstract description 7
- 239000000126 substance Substances 0.000 claims abstract description 4
- 238000005882 aldol condensation reaction Methods 0.000 claims abstract description 3
- 238000005886 esterification reaction Methods 0.000 claims abstract description 3
- 230000003647 oxidation Effects 0.000 claims abstract description 3
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 3
- 238000007363 ring formation reaction Methods 0.000 claims abstract description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 230000007613 environmental effect Effects 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 8
- 150000001875 compounds Chemical class 0.000 claims description 7
- HVQAJTFOCKOKIN-UHFFFAOYSA-N flavonol Natural products O1C2=CC=CC=C2C(=O)C(O)=C1C1=CC=CC=C1 HVQAJTFOCKOKIN-UHFFFAOYSA-N 0.000 claims description 7
- 235000011957 flavonols Nutrition 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 3
- 238000010438 heat treatment Methods 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 238000001917 fluorescence detection Methods 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 20
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 12
- 241000252212 Danio rerio Species 0.000 description 10
- 230000008859 change Effects 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- 239000012044 organic layer Substances 0.000 description 6
- 238000000799 fluorescence microscopy Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 3
- 239000012491 analyte Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- FULZLIGZKMKICU-UHFFFAOYSA-N N-phenylthiourea Chemical compound NC(=S)NC1=CC=CC=C1 FULZLIGZKMKICU-UHFFFAOYSA-N 0.000 description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- IOJUPLGTWVMSFF-UHFFFAOYSA-N benzothiazole Chemical compound C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
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- 229930003944 flavone Natural products 0.000 description 2
- 150000002212 flavone derivatives Chemical class 0.000 description 2
- 235000011949 flavones Nutrition 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
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- 238000010898 silica gel chromatography Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 2
- GOLORTLGFDVFDW-UHFFFAOYSA-N 3-(1h-benzimidazol-2-yl)-7-(diethylamino)chromen-2-one Chemical compound C1=CC=C2NC(C3=CC4=CC=C(C=C4OC3=O)N(CC)CC)=NC2=C1 GOLORTLGFDVFDW-UHFFFAOYSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- XJHABGPPCLHLLV-UHFFFAOYSA-N benzo[de]isoquinoline-1,3-dione Chemical compound C1=CC(C(=O)NC2=O)=C3C2=CC=CC3=C1 XJHABGPPCLHLLV-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000009616 inductively coupled plasma Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 230000009528 severe injury Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
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- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
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Abstract
The invention discloses a flavonoid fluorescent probe for detecting hydrazine and a preparation method and application thereof, wherein the fluorescent probe is a flavonoid compound obtained by performing aldol condensation and oxidation ring closure reaction on p-tert-butyl benzaldehyde and o-hydroxyacetophenone and then performing esterification reaction with acetyl chloride, and the chemical structural formula of the flavonoid compound is shown as a formula (I). The fluorescent probe has unique fluorescence selectivity, extremely high sensitivity and strong anti-interference capability to hydrazine in ethanol/water solution, and the detection limit can be as low as 0.116 mu M. The invention provides a simple and sensitive flavonoid fluorescent probe for detecting hydrazine, and the flavonoid fluorescent probe has a wide application prospect.
Description
Technical Field
The invention belongs to the technical field of organic compound synthesis, fluorescent probes and fine chemical engineering, and particularly relates to a flavonoid fluorescent probe for detecting hydrazine and a preparation method and application thereof.
Background
Hydrazine (N) 2 H 4 ) Is a very important chemical substance because of its strong alkalinityThe product has good flammability and low toxicity, and can be widely used in the fields of industrial manufacture, agricultural production, aerospace and the like. However, N 2 H 4 Is considered to have a strong toxicity and a potential carcinogenic property. Due to its wide application and excellent water solubility, N 2 H 4 Can easily enter an environmental system, and causes serious environmental pollution. Furthermore, N 2 H 4 It is easily introduced through the food chain and accumulated in the human body, thereby causing severe damage to the respiratory tract, kidney, liver and central nervous system of the human body. Therefore, a rapid, effective, convenient and accurate N is developed and researched 2 H 4 The detection method has important significance for the health development of human beings.
Fluorescent probes have been identified as an effective N 2 H 4 Compared with other detection technologies such as absorption spectrum, atomic emission spectrum and inductively coupled plasma emission spectrum, the detection technology of the fluorescent probe has the advantages of simple operation, low cost, non-invasive detection, high spatial resolution and the like. To date, a considerable number of fluorescent probes based on different fluorophores have been developed to detect N 2 H 4 Fluorophores such as coumarin, 1,8-naphthalimide, fluorescein, and benzothiazole. However, these fluorescent probes often have the disadvantages of high detection limit, narrow pH range, poor light stability, and the like. Furthermore, most of these reported fluorescent probes are only applied to N in biological systems 2 H 4 Fluorescence imaging of (2), and detection of N usable in environmental systems 2 H 4 The fluorescent probes of (2) are still quite limited. Therefore, a novel method for detecting N with good selectivity and sensitivity in both biological and environmental systems is devised 2 H 4 The fluorescent probes of (2) still receive a great deal of attention from researchers.
In recent years, some fluorochromes having an Excited State Intramolecular Proton Transfer (ESIPT) characteristic have been widely used in the development of various fluorescent probes. The flavone and the derivatives thereof often have excellent photophysical characteristics and good biocompatibility, and can be used as a promising fluorophore to construct a plurality of fluorescent probes. Inspired by the above phenomenon, the invention prepares a novel flavone baseA fluorescent probe. The fluorescent probe can selectively identify N 2 H 4 After the hydrazine is reacted with hydrazine, the fluorescence emission intensity is obviously enhanced, and the detection limit is lower. The fluorescent probe can be applied to wide pH range and quantitative detection. In addition, the probe can also be applied to monitoring N in living zebra fish and environment in real time 2 H 4 The content of the fluorescent probe is significant for expanding the application range of the fluorescent probe.
Disclosure of Invention
Aiming at the defects of the prior art, the invention synthesizes the flavonoid fluorescent probe for detecting hydrazine, which has the advantages of good selectivity, strong anti-interference capability, wide pH value application range and low detection limit.
The invention also provides a preparation method of the flavonoid fluorescent probe for detecting hydrazine.
The invention also provides application of the flavonoid fluorescent probe for detecting hydrazine.
The technical scheme is as follows: in order to achieve the purpose of the invention, the technical scheme of the invention is as follows: a flavonoid fluorescent probe for detecting hydrazine has a chemical structure shown in formula (I):
the preparation method of the flavonoid fluorescent probe for detecting hydrazine is characterized in that the flavonoid fluorescent probe is synthesized according to the following experimental steps;
(1) Performing aldol condensation and oxidation ring-closing reaction on o-hydroxyacetophenone and p-tert-butyl benzaldehyde to obtain a flavonol compound (II);
(2) Under the alkaline condition, performing esterification reaction on acetyl chloride and the compound (II) obtained in the step (1) to obtain a flavonoid fluorescent probe compound (I);
the specific synthetic route of the flavonoid fluorescent probe for detecting hydrazine is as follows:
the steps are completed by adopting the following method: dissolving p-tert-butyl benzaldehyde, 5.5mmol of o-hydroxyacetophenone and sodium hydroxide solid in ethanol, stirring at room temperature for reaction for 30min, heating and refluxing for 3H, cooling to room temperature after the reaction, and adding H into the mixture 2 O 2 Stirring and reacting for 2 hours, after the reaction is finished, evaporating partial solvent of the mixture obtained by the reaction in vacuum, extracting the mixture for multiple times by using dichloromethane, combining organic layers, washing the organic layers by using saturated saline solution, drying the organic layers by using anhydrous sodium sulfate, filtering the organic layers, and then separating and purifying the organic layers by using silica gel column chromatography to obtain a flavonol compound (II); dissolving flavonol compound (II) and triethylamine in anhydrous dichloromethane, and stirring at room temperature for 30min. Subsequently, an anhydrous dichloromethane solution containing acetyl chloride was slowly added to the above mixture, stirred at room temperature for 12 hours, and after completion of the reaction, the reaction solution was poured into water and extracted with dichloromethane. And (4) combining organic phases, washing with water, evaporating the organic solvent under reduced pressure, and purifying by column chromatography to obtain the flavonoid fluorescent probe (I).
The flavonoid fluorescent probe for detecting hydrazine has unique fluorescence response to hydrazine in ethanol/water solution.
Preparing ethanol solution of flavonoid fluorescent probe (I), and adding various analyte solutions containing quantitative analyte such as Ag + 、Ca 2+ 、Cd 2+ 、Cu 2+ 、Mg 2+ 、Ni 2+ 、CO 3 2- 、HCO 3- 、HPO 4 2- 、HSO 3 - 、I - 、SO 3 2- 、SO 4 2- 、H 2 O 2 GSH and Hcy aqueous solutions, selective recognition effects on different analytes are researched through fluorescence spectrum tests, the results are shown in figure 1, and the intensity change of fluorescence emission spectra shows that the flavonoid fluorescent probe (I) for detecting hydrazine has unique fluorescence responsiveness to hydrazine, and the fluorescence intensity is remarkably enhanced after the flavonoid fluorescent probe (I) reacts with hydrazine; in addition, a certain amount of the fluorescent probe solution was taken, and hydrazine was gradually added to 1 equivalent, and the fluorescence intensity of the fluorescent probe (I) at 540nm increased with the increase of the hydrazine concentration, and the results are shown in the figure2 is shown in the specification; drawing the process of dripping hydrazine in the solution containing the fluorescent probe (I), selecting the fluorescence intensity as a vertical coordinate, dripping equivalent as a horizontal coordinate, and performing linear fitting to obtain a linear regression curve Y =1014.9940X +9496.0068 with a fitting coefficient R 2 Is 0.9947, and the result is shown in figure 3, which shows that the linear correlation is high, so the fluorescent probe can be used for quantitative analysis and detection of hydrazine molecules.
The flavonoid fluorescent probe for detecting hydrazine has excellent anti-interference performance on different analytes when detecting hydrazine, and the result is shown in figure 4, and the fluorescence intensity of the fluorescent probe (I) is hardly influenced by adding other analytes, so that the fluorescent probe (I) has unique fluorescence selectivity and strong anti-interference capability on hydrazine in an ethanol/water solution.
The flavonoid fluorescent probe for detecting hydrazine has the characteristic of wide pH value application range, as shown in figure 5, the probe still has stable fluorescence emission for identifying hydrazine when the pH value is within the range of 5.0-9.0, and the wider pH value application range is beneficial to improving the actual detection performance of the fluorescent probe.
The invention also comprises the research of applying the flavonoid fluorescent probe to the fluorescence imaging of the living zebra fish, as shown in figure 6, which shows that the flavonoid fluorescent probe can be successfully used for detecting hydrazine molecules in a biological system.
The invention also comprises the research of applying the flavonoid fluorescent probe to the detection of the hydrazine content in samples of distilled water, tap water, river water and lake water, and the research shows that the flavonoid fluorescent probe can be successfully applied to the detection of hydrazine in an environmental system as shown in Table 1.
The invention has the beneficial effects that: (1) The fluorescent probe has the advantages of simple synthetic route, mild reaction conditions and simple and convenient purification method; (2) The invention realizes the selective and rapid detection of hydrazine, and has the advantages of large Stokes displacement, high fluorescence intensity, good selectivity, strong anti-interference capability and detection limit as low as 0.116 mu M. Therefore, the invention is a rapid and sensitive hydrazine detection reagent, and has wide application prospects in the fields of analytical chemistry and environmental detection.
Drawings
FIG. 1 shows a flavonoid fluorescent probe (I) (1X 10) -5 M) in ethanol, with 1 equivalent of different analytes added;
FIG. 2 shows a flavonoid fluorescent probe (I) (1X 10) -6 M) in ethanol/water solution for hydrazine fluorescence titration. In the figure, FL intensity is fluorescence emission intensity, wavelet is Wavelength, and excitation Wavelength is 365nm;
FIG. 3 is a linear fitting graph of the flavonoid fluorescent probe (I) with the concentration of the selected hydrazine of different equivalent weights as the abscissa and the fluorescence intensity as the ordinate; the abscissa is the concentration of the dropwise added hydrazine, and the unit is mu M;
FIG. 4 shows a flavonoid fluorescent probe (I) (1X 10) -6 M) histogram of the change in fluorescence intensity after addition of 10 equivalents of the other analyte to an aqueous solution in which an equivalent amount of hydrazine is present;
FIG. 5 is a graph showing the effect of different pH values on the fluorescence intensity of the flavonoid fluorescent probe (I) for identifying hydrazine;
FIG. 6 is fluorescence imaging of flavonoid fluorescent probe (I) applied to live zebrafish containing hydrazine at different concentrations.
Table 1 shows the results of measurement of the concentration of hydrazine in an actual environmental water sample by using the flavonoid fluorescent probe (I).
Detailed Description
The present invention will be described in further detail with reference to the following examples and the accompanying drawings.
Example one
Preparation of flavonol compound (II)
Dissolving 5mmol of p-tert-butyl benzaldehyde, 5.5mmol of o-hydroxyacetophenone and 12.5mmol of sodium hydroxide solid in 25mL of ethanol, stirring at room temperature for reaction for 30min, heating and refluxing for 3H, cooling to room temperature after the reaction, and adding 1.5mL of H into the mixture 2 O 2 Stirring for 2 hr, distilling partial solvent under reduced pressure, extracting with 25mL dichloromethane for three times, mixing organic layers, washing with saturated saline solution, separating with separating funnel, drying with anhydrous sodium sulfate, filtering, and separating and purifying with silica gel column chromatography to obtain flavonol compound (II) in light yellowSolid, yield: 65 percent. 1 H NMR(400MHz，CDCl 3 )δ：8.24(m，1H)，8.18(d，J＝6.4Hz，2H)，7.68(m，1H)，7.57(d，J＝6.6Hz，1H)，7.55(d，J＝6.4Hz，2H)，7.40(m，1H)，6.96(s，1H)，1.38(s，9H)；
Example two
Preparation of flavonoid fluorescent probe (I)
Dissolving 0.5mmol of flavonol compound (II) and 1.0mmol of triethylamine in 15mL of anhydrous dichloromethane, and stirring at room temperature for 30min; then, 5mL of anhydrous dichloromethane containing 1.0mmol of acetyl chloride is slowly added into the mixture, the mixture is stirred at room temperature for 12 hours, and after the reaction is finished, the reaction solution is poured into water and extracted by dichloromethane; and (3) combining organic phases, washing with water, distilling under reduced pressure to remove the organic solvent, and purifying the crude product by column chromatography to obtain the flavonoid fluorescent probe (I) which is a light yellow solid with the yield: 46 percent. 1 H NMR(600MHz，CDCl 3 )δ：8.25(d，J＝6Hz，1H)，7.83(d，J＝12Hz，2H)，7.69(t，J＝12Hz，1H)，7.54(d，J＝12Hz，3H)，7.41(t，J＝12Hz，1H)，2.37(s，3H)，1.37(s，9H)； 13 C NMR(150MHz，CDCl 3 )δ：172.27，168.20，156.46，155.70，154.99，133.96，133.61，128.17，127.22，126.16，125.81，125.20，123.70，118.15，35.11，31.21，20.76；HRMS(m/z)：calcd for C 21 H 21 O 4 [M+H] + ：337.1440；found：337.1472。
EXAMPLE III
Selective research of flavonoid fluorescent probe on different analytes
Preparation of fluorescent Probe (I) (1X 10) -5 M) in ethanol, 1 equivalent of a standard solution of the different analytes, such as Ag, being added separately + 、Ca 2+ 、Cd 2+ 、Cu 2+ 、Mg 2+ 、Ni 2+ 、CO 3 2- 、HCO 3- 、HPO 4 2- 、HSO 3 - 、I - 、SO 3 2- 、SO 4 2- 、H 2 O 2 GSH and Hcy in water, as can be seen from FIG. 1, fluorescence when hydrazine is addedThe solution of the probe (I) emits bright green fluorescence, and meanwhile, the fluorescence enhancement phenomenon is obvious at 540 nm; and after other analytes are added, no fluorescence change is caused, which shows that the fluorescent probe (I) has good specificity recognition effect on hydrazine and can be used as a specific fluorescent probe for detecting hydrazine.
Example four
Change of fluorescence intensity of flavonoid fluorescent probe along with increase of hydrazine addition amount
Taking 1mL of the stock solution prepared in the third embodiment out of the stock solution prepared in the third embodiment, adding the stock solution into a 10mL volumetric flask, adding hydrazine molecule standard solutions with different equivalent weights, carrying out constant volume to 10mL, and measuring the change of fluorescence intensity of an emission wavelength at 540nm, wherein the result is shown in FIG. 2, and the fluorescence of the flavonoid fluorescent probe solution is gradually enhanced along with the increase of the addition amount of hydrazine, which indicates that the flavonoid fluorescent probe has highly sensitive fluorescence response to the change of the hydrazine concentration. The results of linear fitting with the hydrazine concentration of different selected equivalent as the abscissa and the fluorescence intensity as the ordinate are shown in fig. 3, and the fluorescence intensity has a good linear relationship with the hydrazine concentration change.
EXAMPLE five
Anti-interference performance of flavonoid fluorescent probe to different analytes in hydrazine molecule detection
1mL of the stock solution of the fluorescent probe in example III was taken out and added to a 10mL volumetric flask, and an equivalent amount of the hydrazine standard solution was added, and then 10 equivalents of the other analytes were added, respectively, to make the volume to 10mL. Competing analytes include Ag + 、Ca 2+ 、Cd 2+ 、Cu 2+ 、Mg 2+ 、Ni 2+ 、CO 3 2- 、HCO 3- 、HPO 4 2- 、HSO 3 - 、I - 、SO 3 2- 、SO 4 2- 、H 2 O 2 And observing the change of fluorescence intensity of fluorescence emission at 540nm by GSH and Hcy, wherein the result is shown in figure 4, and the result shows that when other analytes are added, the fluorescence intensity of the flavonoid fluorescent probe is hardly influenced, which indicates that the flavonoid fluorescent probe has good anti-interference performance on other analytes when detecting hydrazine molecules.
EXAMPLE six
The influence of different pH values on the identification of hydrazine by the flavonoid fluorescent probe is as follows:
in order to obtain the optimal pH value range for detecting hydrazine, the change of the fluorescence intensity of the flavonoid fluorescent probe (I) under different pH values in the presence and absence of hydrazine is studied, and as can be seen from figure 5, the fluorescence intensity of the fluorescent probe (I) without hydrazine is still very weak in the pH value range of 3-9; in contrast, the fluorescence intensity of the fluorescent probe (I) is remarkably enhanced and kept stable in the presence of hydrazine within the pH value range of 5-9. The pH value range of the flavonoid fluorescent probe (I) is wide, and the actual detection performance of the flavonoid fluorescent probe (I) is improved.
EXAMPLE seven
The flavonoid fluorescent probe is applied to living zebra fish hydrazine identification fluorescence imaging.
Zebra fish were grown in embryo culture medium containing 1-phenyl-2-thiourea (PTU) and cultured at 28 ℃ for 72h. Treating zebrafish with 10 μ M flavonoid fluorescent probe (I) for 0.5h; after three washes with PBS buffer, zebrafish were further incubated with different concentrations of hydrazine (50 μ M and 100 μ M). As shown in FIG. 6, zebrafish stained with 10. Mu.M flavonoid fluorescent probe (I) showed no significant fluorescent signal; after 50 mu M hydrazine is added, the fluorescence emission of the living zebra fish pretreated by the flavonoid fluorescent probe (I) is obviously increased and is continuously enhanced along with the increase of the concentration of the hydrazine; the results show that the probe has good biocompatibility and is suitable for detecting the content of hydrazine in live zebra fish in real time.
Example eight
And (3) detecting the concentration of hydrazine in the environmental water sample by using the flavonoid fluorescent probe.
The concentration of hydrazine in tap water, river water and lake water environment water samples is detected by adopting a flavonoid fluorescent probe (I). As shown in table 1, the concentration of hydrazine detected by probe (I) was very close to the actual concentration added to the water sample (table 1); the results show that the flavonoid fluorescent probe (I) can be used for accurately determining the concentration of hydrazine in an environmental water sample.
TABLE 1
Claims (8)
1. A flavonoid fluorescent probe for detecting hydrazine is characterized in that the chemical structural formula is shown as formula (I),
2. the method for preparing a flavonoid fluorescent probe for detecting hydrazine according to claim 1, which is synthesized according to the following experimental steps;
(1) The flavonol compound (II) is obtained by the aldol condensation and the oxidation ring-closing reaction of o-hydroxyacetophenone and p-tert-butyl benzaldehyde, and the structural formula is as follows:
(2) Under the alkaline condition, acetyl chloride and the compound (II) obtained in the step (1) are subjected to esterification reaction to obtain the flavonoid fluorescent probe compound (I) for detecting hydrazine, and the structural formula is as follows:
the reaction formula of the preparation process is as follows:
3. the method for preparing flavonoid fluorescent probe for detecting hydrazine according to claim 2, wherein said method comprisesThe step (1) is completed by adopting the following method: dissolving p-tert-butyl benzaldehyde, o-hydroxyacetophenone and sodium hydroxide solid in ethanol, stirring at room temperature for reaction for 30min, heating and refluxing for 3H, cooling to room temperature after reaction, and adding H into the mixture 2 O 2 The reaction was stirred for 2 hours to obtain compound (II).
4. The method for preparing a flavonoid fluorescent probe used for detecting hydrazine according to claim 3, wherein the molar ratio of p-tert-butyl benzaldehyde, o-hydroxyacetophenone and sodium hydroxide is 1: 1.1: 2.5.
5. The method for preparing a flavonoid fluorescent probe for detecting hydrazine according to claim 2, wherein the step (2) is carried out by adopting the following method: dissolving the compound (II) and triethylamine in anhydrous dichloromethane, and stirring at room temperature for 30min; subsequently, an anhydrous dichloromethane solution containing acetyl chloride was slowly added to the above mixture, and the reaction was stirred at room temperature for 12 hours to obtain compound (I).
6. The method for preparing a flavonoid fluorescent probe used for detecting hydrazine according to claim 5, wherein the molar ratio of triethylamine to acetyl chloride is 1: 1.
7. The use of the flavonoid fluorescent probe of claim 1 for detecting hydrazine.
8. The use according to claim 7, characterized in that the detection of the content of hydrazine molecules is carried out in the environment; the detection is fluorescence detection; the fluorescent probe has excellent detection performance on hydrazine molecules, the minimum detection limit can be as low as 0.116 mu M, and the like, and can be applied to detection of hydrazine molecules in biological and environmental systems.
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| CN103865962A (en) * | 2014-01-24 | 2014-06-18 | 潍坊医学院 | Enzymatic preparation method and application of quercetin-3-O-fatty acid ester |
| CN111484470A (en) * | 2020-03-28 | 2020-08-04 | 齐鲁工业大学 | Fluorescent probe for detecting hydrazine, preparation method and application thereof |
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| CN103865962A (en) * | 2014-01-24 | 2014-06-18 | 潍坊医学院 | Enzymatic preparation method and application of quercetin-3-O-fatty acid ester |
| CN111484470A (en) * | 2020-03-28 | 2020-08-04 | 齐鲁工业大学 | Fluorescent probe for detecting hydrazine, preparation method and application thereof |
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| BIN LIU等: "Fluorescence monitor of hydrazine in vivo by selective deprotection of flavonoid", SENSORS AND ACTUATORS B: CHEMICAL, vol. 202, pages 194 * |
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