CN116102462A - Near infrared fluorescent probe responding to formaldehyde and preparation method and application thereof - Google Patents
Near infrared fluorescent probe responding to formaldehyde and preparation method and application thereof Download PDFInfo
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- CN116102462A CN116102462A CN202310126724.XA CN202310126724A CN116102462A CN 116102462 A CN116102462 A CN 116102462A CN 202310126724 A CN202310126724 A CN 202310126724A CN 116102462 A CN116102462 A CN 116102462A
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- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 title claims abstract description 186
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims description 35
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 21
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- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 18
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 12
- 238000001816 cooling Methods 0.000 claims description 9
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- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 8
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- 239000005695 Ammonium acetate Substances 0.000 claims description 7
- 235000019257 ammonium acetate Nutrition 0.000 claims description 7
- 229940043376 ammonium acetate Drugs 0.000 claims description 7
- CUONGYYJJVDODC-UHFFFAOYSA-N malononitrile Chemical compound N#CCC#N CUONGYYJJVDODC-UHFFFAOYSA-N 0.000 claims description 7
- HJOVHMDZYOCNQW-UHFFFAOYSA-N isophorone Chemical compound CC1=CC(=O)CC(C)(C)C1 HJOVHMDZYOCNQW-UHFFFAOYSA-N 0.000 claims description 5
- TXUICONDJPYNPY-UHFFFAOYSA-N (1,10,13-trimethyl-3-oxo-4,5,6,7,8,9,11,12,14,15,16,17-dodecahydrocyclopenta[a]phenanthren-17-yl) heptanoate Chemical compound C1CC2CC(=O)C=C(C)C2(C)C2C1C1CCC(OC(=O)CCCCCC)C1(C)CC2 TXUICONDJPYNPY-UHFFFAOYSA-N 0.000 claims description 4
- 229910021626 Tin(II) chloride Inorganic materials 0.000 claims description 4
- 235000010288 sodium nitrite Nutrition 0.000 claims description 4
- 239000001119 stannous chloride Substances 0.000 claims description 4
- 235000011150 stannous chloride Nutrition 0.000 claims description 4
- 238000004090 dissolution Methods 0.000 claims description 3
- 238000001556 precipitation Methods 0.000 claims description 3
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 abstract description 8
- 241000219195 Arabidopsis thaliana Species 0.000 abstract description 6
- 238000003384 imaging method Methods 0.000 abstract description 6
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- 241000196324 Embryophyta Species 0.000 description 13
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- 238000001514 detection method Methods 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
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- 239000000243 solution Substances 0.000 description 7
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 3
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- 238000000799 fluorescence microscopy Methods 0.000 description 3
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- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- MSTNYGQPCMXVAQ-RYUDHWBXSA-N (6S)-5,6,7,8-tetrahydrofolic acid Chemical compound C([C@H]1CNC=2N=C(NC(=O)C=2N1)N)NC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 MSTNYGQPCMXVAQ-RYUDHWBXSA-N 0.000 description 2
- 102100039702 Alcohol dehydrogenase class-3 Human genes 0.000 description 2
- AIJULSRZWUXGPQ-UHFFFAOYSA-N Methylglyoxal Chemical compound CC(=O)C=O AIJULSRZWUXGPQ-UHFFFAOYSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
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- 102000028528 s-formylglutathione hydrolase Human genes 0.000 description 2
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- 239000005460 tetrahydrofolate Substances 0.000 description 2
- QYNUQALWYRSVHF-OLZOCXBDSA-N (6R)-5,10-methylenetetrahydrofolic acid Chemical compound C([C@H]1CNC=2N=C(NC(=O)C=2N1C1)N)N1C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 QYNUQALWYRSVHF-OLZOCXBDSA-N 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
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- HHLFWLYXYJOTON-UHFFFAOYSA-N glyoxylic acid Chemical compound OC(=O)C=O HHLFWLYXYJOTON-UHFFFAOYSA-N 0.000 description 1
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- 229940120731 pyruvaldehyde Drugs 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C255/00—Carboxylic acid nitriles
- C07C255/63—Carboxylic acid nitriles containing cyano groups and nitrogen atoms further bound to other hetero atoms, other than oxygen atoms of nitro or nitroso groups, bound to the same carbon skeleton
- C07C255/65—Carboxylic acid nitriles containing cyano groups and nitrogen atoms further bound to other hetero atoms, other than oxygen atoms of nitro or nitroso groups, bound to the same carbon skeleton with the nitrogen atoms further bound to nitrogen atoms
- C07C255/66—Carboxylic acid nitriles containing cyano groups and nitrogen atoms further bound to other hetero atoms, other than oxygen atoms of nitro or nitroso groups, bound to the same carbon skeleton with the nitrogen atoms further bound to nitrogen atoms having cyano groups and nitrogen atoms being part of hydrazine or hydrazone groups bound to the same carbon skeleton
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- G01—MEASURING; TESTING
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- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a fluorescent opening near infrared fluorescent probe responding to formaldehyde, and a preparation method and application thereof, wherein the structure is as shown in a structural formula (I):
the prepared near infrared fluorescent probe has good sensitivity, specific selectivity and rapid response speed, is stable in a wider pH range and is not interfered by other related ions, and is a good fluorescent molecular probe for detecting formaldehyde. The target product obtained by the method can be used for imaging formaldehyde in the root tip tissue of the arabidopsis thaliana.
Description
Technical Field
The invention relates to preparation and application of a fluorescent opening near infrared fluorescent probe responding to formaldehyde, and belongs to the technical field of biochemistry.
Background
Formaldehyde (FA) is one of the simplest aldehydes and is also known as a reactive carbonyl species due to its high reactivity. Formaldehyde, in addition to being widely found in our living environment, can also be produced by organisms. In normal physiological processes formaldehyde can be formed endogenously by oxidation of methanol, dissociation of 5, 10-methylene-THF and decarboxylation of glyoxylate and can be detoxified by binding to Tetrahydrofolate (THFA) in the C1 metabolic system or catalyzed by Glutathione (GSH) -dependent formaldehyde dehydrogenase (FALDH) and S-formyl glutathione hydrolase (S-FGH), which can bring the formaldehyde concentration in a dynamic equilibrium state. On the other hand, excessive formaldehyde expression induced by excessive exposure may cause various diseases such as memory deterioration and various neurodegenerative diseases due to high activity of formaldehyde. Based on the physiological and metabolic characteristics of plants and environmental adaptability, phytoremediation is an energy-saving, environment-friendly and effective method for removing formaldehyde in air. However, in situ observation of the formaldehyde metabolism mechanism in plants remains challenging due to the lack of suitable tools. Therefore, in order to understand the physiological function of formaldehyde in plants, it is important to develop methods that can be used for real-time detection and imaging of formaldehyde in plants.
With the development and application of the small molecular fluorescent probe, researchers provide a new idea for detecting formaldehyde in organisms. Formaldehyde detection in organisms can be achieved by combining fluorescent probes with fluorescent microscopy. The method has the advantages of high space-time resolution, high sensitivity and high selectivity, and therefore, the method is attracting attention. Most fluorescent probes reported to date can image formaldehyde in living animal cells. Only a few cases are used for imaging formaldehyde in plants. During the detection of formaldehyde in plants, the autofluorescence of the plants and tissue light scattering may interfere with the detection accuracy.
Therefore, in order to achieve higher resolution and more accurate detection, there is an urgent need to develop near infrared fluorescent probes with stronger tissue penetration for imaging formaldehyde in plant tissue.
Disclosure of Invention
The invention breaks through the limitation of the problems in the prior art, and provides a preparation method and application of a fluorescent-opening near infrared fluorescent probe responding to formaldehyde. Fluorescent probes capable of rapidly responding to formaldehyde are prepared and applied to imaging formaldehyde in plants.
In order to solve the technical problems, the invention provides a near infrared fluorescent probe responding to formaldehyde, which has a structure shown in a structural formula (I):
the invention also provides a preparation method of the near infrared fluorescent probe responding to formaldehyde, which comprises the following steps:
(1) Adding 3, 5-trimethyl-2-cyclohexene-1-one into a reactor, adding toluene for dissolution, then adding malononitrile, ammonium acetate and acetic acid, heating for reaction, and cooling for precipitation after the reaction is finished to obtain a product 1, wherein the molar ratio of 3, 5-trimethyl-2-cyclohexene-1-one, malononitrile, ammonium acetate and acetic acid is 1:1:0.15:0.30;
(2) Adding a product 1 into a reactor, adding 4-acetamidobenzaldehyde, dissolving with acetonitrile, then adding piperidine, heating to react, and cooling to separate out after the reaction is finished to obtain a product 2, wherein the molar ratio of the product 1 to the 4-acetamidobenzaldehyde is 1:1;
(3) Adding the product 2 into a reactor, dissolving with isopropanol and hydrochloric acid, heating for reaction, and cooling to separate out after the reaction is finished to obtain a product 3;
(4) Adding the product 3 into a reactor, dissolving with hydrochloric acid, adding sodium nitrite and stannous chloride, and obtaining the product DISE after the reaction is finished.
The reaction process is as follows:
preferably, the temperature of the elevated reaction in the step (1) is 100 to 140 ℃.
Preferably, the temperature of the elevated reaction in the step (2) is 0 to 100 ℃.
Preferably, the temperature of the elevated reaction in the step (3) is 80 to 120 ℃.
Meanwhile, the invention also provides an application of the near infrared fluorescent probe responding to formaldehyde or the near infrared fluorescent probe responding to formaldehyde prepared by the preparation method of the near infrared fluorescent probe responding to formaldehyde in plants.
The invention has the beneficial effects that:
1. the fluorescent probe obtained by the method has the characteristics of high selectivity and high sensitivity, and the detection limit reaches 0.048 mu M. The probe has good stability under physiological pH, and is beneficial to real-time in-situ detection of formaldehyde; the emission wavelength of the probe reaches the near infrared region, which can effectively avoid the interference of autofluorescence of plant tissues so as to be used for formaldehyde imaging of the plant tissues.
2. The raw materials used in the synthesis methods reported in the present invention are all commercially available. The synthesis method has mild synthesis conditions and high yield, which is beneficial to saving the cost and ensuring the yield of the target product.
3. The formaldehyde fluorescent probe reported by the invention can be applied to formaldehyde detection in Arabidopsis root tip tissues. The probe can emit fluorescence in 600-800nm wave band, and successfully avoids the autofluorescence of the Arabidopsis root tip tissue in 400-500nm wave band, thereby obtaining clear and reliable fluorescence imaging pictures.
Drawings
FIG. 1 is a nuclear magnetic resonance spectrum of a probe DISE according to the invention;
FIG. 2 is a nuclear magnetic resonance spectrum of the probe DISE according to the invention;
FIG. 3 is a graph showing the ultraviolet-visible absorption spectrum and fluorescence intensity change before and after the probe DISE reacts with formaldehyde in the present invention;
fig. 4 is a graph showing the fluorescence intensity of the probe dis reacted with other analytes and the change of fluorescence intensity with time after the probe dis reacted with formaldehyde at different concentrations in the present invention.
FIG. 5 is a photograph of fluorescence imaging of the probe DISE of the invention in Arabidopsis thaliana root tip tissue.
Detailed description of the preferred embodiments
The invention is further described below. The following examples are only for more clearly illustrating the technical aspects of the present invention, and are not intended to limit the scope of the present invention.
A fluorescent opening near infrared fluorescent probe responding to formaldehyde has a structure shown in a structural formula (I):
a preparation method of a fluorescent-opening near infrared fluorescent probe responding to formaldehyde comprises the following steps:
(1) Adding 3, 5-trimethyl-2-cyclohexanecene-1-one into a reactor, adding toluene for dissolution, then adding malononitrile, ammonium acetate and acetic acid, heating for reaction, and cooling for precipitation after the reaction is finished to obtain a product 1, wherein the molar ratio of 3, 5-trimethyl-2-cyclohexanecene-1-one, malononitrile, ammonium acetate and acetic acid is 1:1:0.15:0.30;
(2) Adding a product 1 into a reactor, adding 4-acetamidobenzaldehyde, dissolving with acetonitrile, then adding piperidine, heating to react, and cooling to separate out after the reaction is finished to obtain a product 2, wherein the molar ratio of the product 1 to the 4-acetamidobenzaldehyde is 1:1;
(3) Adding the product 2 into a reactor, dissolving with isopropanol and hydrochloric acid, heating for reaction, and cooling to separate out after the reaction is finished to obtain a product 3;
(4) Adding the product 3 into a reactor, dissolving with hydrochloric acid, adding sodium nitrite and stannous chloride, and obtaining the product DISE after the reaction is finished.
The reaction process is as follows:
the near infrared fluorescent probe responding to formaldehyde can be applied to formaldehyde detection in Arabidopsis thaliana root tip tissues.
Examples:
3, 5-trimethyl-2-cyclohexen-1-one (3.0 g,22.0mmol,1 equiv.) and anhydrous toluene were added to a two-necked flask at room temperature under nitrogen atmosphere, and malononitrile (1.51 g,22.0mmol,1 equiv.) and ammonium acetate (251 mg,3.3mmol,0.15 equiv.) were added to the solution under stirring. And acetic acid (0.41 ml,6.5mmol,0.3 equiv.) was added dropwise to the solution at room temperature. Then the temperature is raised to 125 ℃ and stirred for 20h. After the completion of the reaction, extraction was performed with methylene chloride, and drying was performed with anhydrous magnesium sulfate. The solvent was distilled off to give brown crystals. Finally, compound 1 was obtained as a white solid in 70.5% yield by flash column chromatography on a silica gel column (petroleum ether/dichloromethane=1/1, v/v).
Compound 1 (1 g,5.4mmol,1 equiv.) and 4-acetamidobenzaldehyde (878 mg,5.4mmol,1 equiv.) are added to a 100mL two-neck flask equipped with a condenser at room temperature under nitrogen atmosphere, dissolved with anhydrous acetonitrile and stirred. Simultaneously, piperidine (0.1 mL) was added to the solution. Heating to 80 ℃ and refluxing for 2 hours. After the reaction was completed, the solid was washed with cold acetonitrile to obtain compound 2 as an orange solid in 83.3% yield.
Compound 2 (1.0 g,3.0 mmol) was added to a 250mL two-necked flask equipped with a condenser, and the compound was dissolved with 30mL of isopropanol and 10mL of hydrochloric acid (2M), followed by refluxing at 100℃for 1.5h. After the reaction, ethyl acetate was used for extraction, and anhydrous magnesium sulfate was used for drying. After evaporation of the solvent, flash column chromatography on a silica gel column (dichloromethane/petroleum ether=9/1, v/v) gave compound 3 as a red solid in 73.0% yield.
Compound 3 (300 mg,1.04mmol,1 equiv.) is charged to a single neck flask with 100mL, dissolved with 10mL hydrochloric acid (6M) and the temperature reduced to 0 ℃. Aqueous sodium nitrite (193 mg,2.8mmol,2.7 equiv.) was added dropwise under ice bath and stirring continued for 1.5h. A hydrochloric acid solution of stannous chloride (788.81 mg,4.16mmol,4 equiv.) was then gradually added dropwise to the system under ice-bath and stirring continued for 2h. After the completion of the reaction, the pH of the reaction solution was adjusted to neutral with sodium hydroxide solution. The reaction solution was extracted with dichloromethane and dried over anhydrous magnesium sulfate. After evaporation of the solvent, the compound DISE was obtained in a yield of 78.8% and was used as such after washing with cold diethyl ether.
As shown in figure 1 of the drawings, 1 H NMR(600MHz，CDCl3)：δ7.43(d，J＝8.4Hz，2H)，7.03(d，J＝15.6Hz，1H)，6.87-6.82(m，3H)，6.77(s，1H)，5.52(s，1H)，3.68(s，2H)，2.58(s，2H)，2.46(s，2H)，1.08(s，7H).
as shown in the figure 2 of the drawings, 13 C NMR(150MHz，CDCl 3 )：δ169.3，154.9，152.5，137.6，129.3，126.6，125.4，122.0，114.0，113.2，112.0，43.0，39.2，32.0，28.0.
probe dis was dissolved in acetonitrile to prepare a 10mM stock solution. The test solution was acetonitrile/Phosphate Buffer (PBS) (1:1, v: v, ph=7.4, containing 0.4% tween 80) and the probe was diluted to the desired concentration by the test solution. As shown in FIG. 3, the absorption peak of the probe DISE is about 460nm, the weaker fluorescence emission peak is about 670nm, and the emission peak of the probe DISE after reaction with formaldehyde is obviously enhanced at 670 nm. As shown in FIG. 3-b, probe response to increasing concentrations of formaldehyde. As the concentration of formaldehyde increases, the fluorescence intensity of the probe DISE increases, and as the concentration increases to 500. Mu.M, the response of the probe DISE to formaldehyde reaches equilibrium (FIG. 3-c). As shown in FIG. 3-d, the fluorescence intensity ratio of the probe DISE in the range of 20-80. Mu.M formaldehyde concentration showed a linear correlation, and the detection limit was calculated to be 0.048. Mu.M based on L=3σ/k. The result shows that the fluorescent probe has high sensitivity for detecting formaldehyde.
To verify the specificity of the probe DISE for Formaldehyde (FA) detection, the present application examined the probe for various potential interfering substances (1. Blank;2.GSH;3.Cys;4.S 2- ；5.HS - ；6.ONOO - ；7.H 2 O 2 ；8.GlO - ；9. · OH；10.ROO · ；11.Ca 2+ ；12.Na + ；13.K + ；14.Mg 2+ ；15.Fe 2+ ；16.Fe 3+ ；17.Mn 2+ ；18.Zn 2+ ；19.CO 3 2- ；20.SO 3 2- ；21.SO 4 2- ；22.HSO 3 - The method comprises the steps of carrying out a first treatment on the surface of the Acetone; glyoxal; methyl glyoxy; fa) fluorescence response in the presence of fa. As shown in FIG. 4-a, it can be seen that the addition of glyoxal or pyruvaldehyde has an effect on the fluorescence intensity of the probe, but far from that of formaldehydeThe light intensity is obviously increased, other species have no obvious fluorescent response to formaldehyde, and the results show that the probe has high selectivity to formaldehyde and can detect formaldehyde in a physiological environment. As shown in fig. 4-b, the change of the fluorescence intensity of the probe with time after adding formaldehyde with different concentrations is also tested, and experimental results show that the reaction can be completed within 10-20min after adding formaldehyde, the fluorescence intensity is obviously enhanced, the probe can rapidly respond to formaldehyde, and the requirement of detecting the dynamic change of formaldehyde in real time can be met.
Application examples:
probe dis fluorescence imaging of formaldehyde in arabidopsis root tip tissue.
Arabidopsis thaliana is used as a model plant for biological experiments due to its characteristics of simple structure, short growth cycle and strong reproductive capacity. We used a laser confocal microscope to detect formaldehyde in Arabidopsis seedling root tip tissue. As shown in FIG. 5, the experimental results showed that weak fluorescence was observed in the root tip tissue of Arabidopsis thaliana, indicating that a small amount of formaldehyde was contained therein. A significant increase in fluorescence intensity can be observed when exogenous formaldehyde is added or endogenous formaldehyde is produced by stimulation with methanol. And when NaHSO is used 3 After removal of endogenous formaldehyde generated by methanol stimulation, fluorescence intensity of Arabidopsis root tip tissue is reduced. Experimental results show that the probe DISE can be applied to detection of formaldehyde content in Arabidopsis thaliana root tip tissues.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that modifications and variations could be made by those skilled in the art without departing from the technical principles of the present invention, and such modifications and variations should also be regarded as being within the scope of the invention.
Claims (6)
1. A near infrared fluorescent probe responding to formaldehyde has a structure shown in a structural formula (I):
2. the near infrared fluorescent probe responsive to formaldehyde according to claim 1, wherein the preparation method comprises the following steps:
(1) Adding 3, 5-trimethyl-2-cyclohexene-1-one into a reactor, adding toluene for dissolution, then adding malononitrile, ammonium acetate and acetic acid, heating for reaction, and cooling for precipitation after the reaction is finished to obtain a product 1, wherein the molar ratio of 3, 5-trimethyl-2-cyclohexene-1-one, malononitrile, ammonium acetate and acetic acid is 1:1:0.15:0.30;
(2) Adding a product 1 into a reactor, adding 4-acetamidobenzaldehyde, dissolving with acetonitrile, then adding piperidine, heating to react, and cooling to separate out after the reaction is finished to obtain a product 2, wherein the molar ratio of the product 1 to the 4-acetamidobenzaldehyde is 1:1;
(3) Adding the product 2 into a reactor, dissolving with isopropanol and hydrochloric acid, heating for reaction, and cooling to separate out after the reaction is finished to obtain a product 3;
(4) Adding the product 3 into a reactor, dissolving with hydrochloric acid, adding sodium nitrite and stannous chloride, and obtaining the product DISE after the reaction is finished.
The reaction process is as follows:
3. the method for preparing a near infrared fluorescent probe responsive to formaldehyde according to claim 2, wherein the temperature of the heating reaction in the step (1) is 100 to 140 ℃.
4. The method for preparing a near infrared fluorescent probe responsive to formaldehyde according to claim 2 or 3, wherein the temperature of the heating reaction in the step (2) is 0 to 100 ℃.
5. The method for preparing a near infrared fluorescent probe responsive to formaldehyde according to any one of claims 2 to 4, wherein the elevated reaction temperature in the step (3) is 80 to 120 ℃.
6. Use of the near infrared fluorescent probe responsive to formaldehyde according to claim 1 or prepared by the preparation method of the near infrared fluorescent probe responsive to formaldehyde according to any one of claims 2 to 5 in plants.
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| CN113896708A (en) * | 2021-10-29 | 2022-01-07 | 南京碳硅人工智能生物医药技术研究院有限公司 | Design, synthesis and activity research of formaldehyde fluorescent probe |
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Non-Patent Citations (3)
| Title |
|---|
| 《ZHANGTAO MIN》: "A near-infrared fluorescence-on fluorescent probe for formaldehyde imaging in Arabidopsis thaliana", 《DYES AND PIGMENTS》, vol. 218, no. 111446, 1 June 2023 (2023-06-01), pages 1 - 6 * |
| LIN, NAI-JIE等: "A ratiometric fluorescent probe for fast detection and bioimaging of formaldehyde", 《ORGANIC & BIOMOLECULAR CHEMISTRY 》, vol. 21, no. 10, 14 February 2023 (2023-02-14), pages 2167 - 2171 * |
| YONGHE TANG等: "Development of a Two-Photon Fluorescent Probe for Imaging of Endogenous Formaldehyde in Living Tissues", 《ANGEWANDTE CHEMIE INTERNATIONAL EDITION》, vol. 55, no. 10, 1 March 2016 (2016-03-01), pages 3356 - 3359, XP055500907, DOI: 10.1002/anie.201510373 * |
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| CN113896708A (en) * | 2021-10-29 | 2022-01-07 | 南京碳硅人工智能生物医药技术研究院有限公司 | Design, synthesis and activity research of formaldehyde fluorescent probe |
| CN113896708B (en) * | 2021-10-29 | 2024-03-12 | 南京碳硅人工智能生物医药技术研究院有限公司 | Design, synthesis and activity research of formaldehyde fluorescent probe |
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