CN113264954A - Fluorescent probe molecule for detecting hydrogen peroxide and preparation method thereof - Google Patents
Fluorescent probe molecule for detecting hydrogen peroxide and preparation method thereof Download PDFInfo
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- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 title claims abstract description 134
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title abstract description 9
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 39
- 238000006243 chemical reaction Methods 0.000 claims abstract description 18
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000012043 crude product Substances 0.000 claims abstract description 12
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims abstract description 12
- 239000000741 silica gel Substances 0.000 claims abstract description 12
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 12
- 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 claims abstract description 10
- 125000000051 benzyloxy group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])O* 0.000 claims abstract description 9
- 238000003756 stirring Methods 0.000 claims abstract description 9
- CBUOGMOTDGNEAW-UHFFFAOYSA-N 2-[4-(bromomethyl)phenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=C(CBr)C=C1 CBUOGMOTDGNEAW-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000010438 heat treatment Methods 0.000 claims abstract description 6
- 229910000027 potassium carbonate Inorganic materials 0.000 claims abstract description 6
- 239000007787 solid Substances 0.000 claims abstract description 6
- 150000001875 compounds Chemical class 0.000 claims abstract description 5
- 238000010992 reflux Methods 0.000 claims abstract description 5
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical group ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 8
- 229910052739 hydrogen Inorganic materials 0.000 claims description 7
- 239000001257 hydrogen Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 229910052736 halogen Chemical group 0.000 claims description 4
- 150000002367 halogens Chemical group 0.000 claims description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 4
- 239000012535 impurity Substances 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- 238000001514 detection method Methods 0.000 abstract description 29
- 239000000523 sample Substances 0.000 abstract description 15
- 230000035945 sensitivity Effects 0.000 abstract description 7
- 238000001816 cooling Methods 0.000 abstract description 3
- 239000002904 solvent Substances 0.000 abstract description 3
- 150000001642 boronic acid derivatives Chemical group 0.000 abstract description 2
- 238000002156 mixing Methods 0.000 abstract description 2
- 238000009833 condensation Methods 0.000 abstract 1
- 230000005494 condensation Effects 0.000 abstract 1
- 230000008859 change Effects 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- VFNKZQNIXUFLBC-UHFFFAOYSA-N 2',7'-dichlorofluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(O)C=C1OC1=C2C=C(Cl)C(O)=C1 VFNKZQNIXUFLBC-UHFFFAOYSA-N 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000007800 oxidant agent Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 description 2
- 239000002360 explosive Substances 0.000 description 2
- 238000001917 fluorescence detection Methods 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- VFIKPDSQDNROGM-UHFFFAOYSA-N 2-(4-methoxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane Chemical compound C1=CC(OC)=CC=C1B1OC(C)(C)C(C)(C)O1 VFIKPDSQDNROGM-UHFFFAOYSA-N 0.000 description 1
- ZZPNDIHOQDQVNU-UHFFFAOYSA-N 2-hydroxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane Chemical compound CC1(C)OB(O)OC1(C)C ZZPNDIHOQDQVNU-UHFFFAOYSA-N 0.000 description 1
- KKLCYBZPQDOFQK-UHFFFAOYSA-N 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborolane Chemical compound O1C(C)(C)C(C)(C)OB1C1=CC=CC=C1 KKLCYBZPQDOFQK-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 229910021580 Cobalt(II) chloride Inorganic materials 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 229910021578 Iron(III) chloride Inorganic materials 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 229910021586 Nickel(II) chloride Inorganic materials 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 229910000024 caesium carbonate Inorganic materials 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000009713 electroplating Methods 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- -1 hydrogen peroxide ions Chemical class 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- LWJROJCJINYWOX-UHFFFAOYSA-L mercury dichloride Chemical compound Cl[Hg]Cl LWJROJCJINYWOX-UHFFFAOYSA-L 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- QMMRZOWCJAIUJA-UHFFFAOYSA-L nickel dichloride Chemical compound Cl[Ni]Cl QMMRZOWCJAIUJA-UHFFFAOYSA-L 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- KMUONIBRACKNSN-UHFFFAOYSA-N potassium dichromate Chemical compound [K+].[K+].[O-][Cr](=O)(=O)O[Cr]([O-])(=O)=O KMUONIBRACKNSN-UHFFFAOYSA-N 0.000 description 1
- 239000012286 potassium permanganate Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- BAZAXWOYCMUHIX-UHFFFAOYSA-M sodium perchlorate Chemical compound [Na+].[O-]Cl(=O)(=O)=O BAZAXWOYCMUHIX-UHFFFAOYSA-M 0.000 description 1
- 229910001488 sodium perchlorate Inorganic materials 0.000 description 1
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
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- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/02—Boron compounds
- C07F5/025—Boronic and borinic acid compounds
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- 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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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Abstract
The invention discloses a fluorescent probe molecule for detecting hydrogen peroxide and a preparation method thereof, which comprises the steps of mixing a compound 2-R-based fluorescein, potassium carbonate, 4-bromomethyl phenylboronic acid pinacol ester and acetonitrile, heating and stirring, carrying out condensation reflux reaction, cooling the reaction to room temperature, and adding an acetonitrile solvent to obtain a crude product; purifying the obtained crude product by a forward silica gel chromatographic column to obtain a fluorescent probe molecule 2',7' -di R-3',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolane-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ] for detecting the hydrogen peroxide by white solid, detecting the content of the hydrogen peroxide by changing the intensity of a fluorescent signal, releasing 2-R-based fluorescein with high luminous efficiency by rearranging and eliminating borate groups caused by the chemical reaction of the hydrogen peroxide and the probe molecule, improving the specificity and the sensitivity of the detection, having simple preparation process, low cost, excellent specificity and sensitivity and being used for rapidly detecting the hydrogen peroxide on site, has wide application prospect.
Description
Technical Field
The invention relates to a fluorescent probe molecule for detecting hydrogen peroxide and a preparation method thereof, belonging to the fields of organic chemistry and analysis and detection.
Background
Hydrogen peroxide, also known as hydrogen peroxide, is a strong oxidant and is widely applied to the fields of industrial bleaching, electroplating, surgical disinfection, military rocket propellant and the like. Hydrogen peroxide participates in a variety of important physiological processes in the human body [ Journal of the American Chemical Society,2012,134, 1305-. Abnormal concentrations of hydrogen peroxide in humans have been implicated in a number of diseases such as Alzheimer's disease, cancer, diabetes, DNA damage, cardiovascular disease and neurodegenerative disease [ Nature,2004,430, 686-. However, hydrogen peroxide is highly corrosive to the human respiratory tract and can cause blindness upon direct contact with the eyes. More importantly, hydrogen peroxide is a typical hazardous chemical oxidant and also a non-standard explosive. Because of its super strong oxidability and combustion-supporting property, it is simple to make explosive, and possesses large harmfulness. Therefore, the method has important practical significance for the ultra-sensitive, rapid and specific accurate identification and detection of the hydrogen peroxide.
To date, detection methods for hydrogen peroxide have mainly included: colorimetric methods [ Dyes and Pigments,2021,186,108954 and Sensors and activators B: Chemical,2018,271, 336-. However, these methods either require expensive instruments, cumbersome detection procedures and semi-quantitative capability. In comparison, fluorescence spectroscopy has received much attention because of its advantages of fast response, high sensitivity, good selectivity, and low cost [ Biosensors and Bioelectronics,2016,79, 237-. Therefore, fluorescence detection based on hydrogen peroxide is a preferred method.
At present, no fluorescent probe molecule with high detection speed, high sensitivity, strong anti-interference capability, stable performance and low price exists for the fluorescent detection of hydrogen peroxide. Therefore, there is an urgent need to develop a fluorescent probe molecule with the above advantages and simple preparation to meet the requirement of hydrogen peroxide detection, so as to solve the problem of insufficient on-site and rapid hydrogen peroxide detection. The method for detecting hydrogen peroxide by using the fluorescent probe with excellent preparation performance is a preferred method for rapid and on-site detection in the fields of food safety, environmental protection, public health and the like at present and in the future, and provides powerful guarantee and technical support for the safety of people's lives.
Disclosure of Invention
The invention aims to provide a fluorescent probe molecule for detecting hydrogen peroxide and a preparation method thereof, which comprises the steps of mixing a compound 2-R-based fluorescein, potassium carbonate, 4-bromomethyl phenylboronic acid pinacol ester and acetonitrile, heating for reaction, cooling the reaction to room temperature, and adding an acetonitrile solvent to obtain a crude product; and purifying the obtained crude product by a forward silica gel chromatographic column to obtain a white solid fluorescent probe molecule 2',7' -di R-3',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolane-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ] for detecting hydrogen peroxide.
The chemical name of the fluorescent probe for detecting hydrogen peroxide is 2',7' -di R-3',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolan-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ], and the structural formula of the fluorescent probe is as follows:
wherein: r is hydrogen or halogen.
The preparation method of the fluorescent probe molecule for detecting hydrogen peroxide comprises the following steps:
a. adding a compound 2-R-based fluorescein, potassium carbonate, 4-bromomethyl phenylboronic acid pinacol ester and acetonitrile into a 100mL round-bottom flask according to a molar ratio of 1:2:3:500, heating and stirring at 90 ℃, condensing and refluxing for 20 hours, and stopping the reaction, wherein: r is hydrogen or halogen;
b. after the reaction is cooled to room temperature, adding 50mL of acetonitrile, fully stirring, filtering to remove impurities, and drying to obtain a crude product;
c. purifying the crude product obtained in the step b by a forward silica gel chromatographic column, wherein the eluent is dichloromethane, and drying the eluted product to obtain the white solid fluorescent probe molecule for detecting the hydrogen peroxide
The fluorescent probe molecule for detecting hydrogen peroxide is 2',7' -di R-3',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolane-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ], has a fluorescein derivative as a fluorescent group and 4-methoxyphenylboronic acid pinacol ester as an identification group, and comprises the following specific synthesis steps:
the detection principle of the fluorescent probe molecule for detecting hydrogen peroxide is as follows: under an alkaline condition, hydrogen peroxide oxidizes borate groups on probe molecules to cause rearrangement elimination reaction, and finally 2-R-based fluorescein, boric acid pinacol ester and 4-methylenecyclohexa-2, 5-diene-1-ketone are generated, so that a fluorophore (2-R-based fluorescein) in the probe is released, and the fluorescent lighting detection of hydrogen peroxide is realized; the change of the fluorescence intensity detected before and after the reaction and the concentration of the hydrogen peroxide form a linear relation, so that the quantitative detection of the hydrogen peroxide can be realized according to the change of the fluorescence intensity.
The fluorescent probe molecule for detecting hydrogen peroxide can realize rapid and specific detection of hydrogen peroxide; the purpose of detecting the hydrogen peroxide is realized. The fluorescent probe molecule comprises the following steps of:
(1) sodium carbonate (0.6g, 5.7mmol) and sodium bicarbonate (3.7g, 4.4mmol) were mixed and dissolved in acetonitrile solvent (100mL) to obtain an alkaline buffer solution;
(2) dissolving 41.6mg of probe molecules in 50mL of the alkaline buffer solution obtained in the step (1) to obtain a 1mM probe molecule solution;
(3) reacting the hydrogen peroxide standard solution with the fluorescent probe obtained in the step (2), and then measuring the fluorescence intensity, wherein the fluorescence intensity corresponds to the hydrogen peroxide content and is used as a calibration curve for quantitative analysis;
(4) uniformly distributing the probe molecule solution obtained in the step (2) on a silica gel plate to obtain a silica gel plate sensor;
(5) and (4) calculating according to the correction curve obtained in the step (3) to obtain the hydrogen peroxide content in the sample to be detected.
The fluorescent probe molecule for detecting hydrogen peroxide has the detection limit of 13.6 nmol/L. The fluorescence intensity of the detected molecules and the concentration of the hydrogen peroxide form a good linear relation under the condition that the concentration of the hydrogen peroxide standard solution is 0-80 mu mol/L, and the detection time is less than 20 s.
Compared with the prior hydrogen peroxide detection technology, the invention has the beneficial effects that: the fluorescent probe molecule disclosed by the invention can be used for identifying hydrogen peroxide with high sensitivity, high speed and specificity under an alkaline condition to generate a 2-R-based fluorescein substance with high fluorescence efficiency, so that the fluorescence detection of the hydrogen peroxide is realized. The kit has the advantages of high detection sensitivity, good specificity, high response speed and the like. The fluorescent probe molecule for detecting hydrogen peroxide has the advantages of quick response, high sensitivity, strong specificity, low cost and the like in the detection process, so that the fluorescent probe molecule is extremely easy to be practically applied and popularized in the field of on-site and timely detection.
Drawings
FIG. 1 is a one-dimensional hydrogen nuclear magnetic spectrum of a fluorescent probe molecule in example 1 of the present invention, wherein the abscissa is chemical shift and the ordinate is signal intensity;
FIG. 2 is a diagram showing the selective detection and identification of hydrogen peroxide by fluorescent probe molecules in example 3 of the present invention, wherein the abscissa represents interferents such as different compounds, and the ordinate represents fluorescence intensity;
FIG. 3 is a linear graph of the fluorescence intensity of the fluorescent probe according to example 4 of the present invention and the concentration of hydrogen peroxide, in which the abscissa is the concentration of hydrogen peroxide and the ordinate is the fluorescence intensity;
fig. 4 is a graph of fluorescence intensity and hydrogen peroxide concentration of a silica gel plate-based probe molecule in example 5 of the present invention, where the fluorescence picture is a fluorescence picture of different amounts of hydrogen peroxide reacted with the probe molecule.
Detailed Description
The following is a description of specific embodiments of the present invention.
Example 1
Synthesis of fluorescent probe molecule 2',7' -dichloro-3 ',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolan-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ]:
a. adding 1.5mmol of dichlorofluorescein, 0.6g of dichlorofluorescein, 4mmol of potassium carbonate, 0.56g of dichlorofluorescein, 6mmol of 4-bromomethylbenzeneboronic acid pinacol ester, 1.8g of dichlorofluorescein, and 50mL of acetonitrile into a 100mL round-bottom flask, heating and stirring at the temperature of 90 ℃ to dissolve, condensing and refluxing for overnight reaction for 20 hours, and stopping the reaction after the reaction is completed;
b. cooling the reaction to room temperature, adding 50mL of acetonitrile, fully stirring, and filtering to remove impurities to obtain a crude product;
c. and (c) purifying the crude product obtained in the step (b) by a forward silica gel chromatographic column, wherein an eluent is dichloromethane, and drying the eluted product to obtain a white solid, namely the fluorescent probe molecule 2',7' -dichloro-3 ',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolane-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ] for detecting hydrogen peroxide.
FIG. 1 shows nuclear magnetic data of hydrogen spectra of probe molecule 2',7' -dichloro-3 ',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolan-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ]:
1H NMR(400MHz,DMSO-d6)δ8.04(d,J＝7.5Hz,1H),7.86-7.60(m,6H),7.47(t,J＝10.3Hz,4H),7.35(d,J＝7.5Hz,1H),7.19(s,2H),6.83(s,2H),5.51-5.22(m,4H),1.30(s,25H)。
example 2
Synthesis of fluorescent probe molecule 2',7' -dihydro-3 ',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolan-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ]:
a. adding fluorescein 1.5mmol, 0.5g, potassium carbonate 4mmol, 0.56g, 4-bromomethylbenzeneboronic acid pinacol ester 6mmol, 1.8g and 50mL of acetonitrile into a 100mL round-bottom flask, heating and stirring at the temperature of 90 ℃ to dissolve, condensing and refluxing for overnight reaction for 20 hours, and stopping the reaction after the reaction is completed;
b. after the reaction is cooled to room temperature, adding 50mL of acetonitrile, fully stirring, and filtering to remove impurities to obtain a crude product;
c. purifying the crude product obtained in the step b by a forward silica gel chromatographic column, wherein an eluent is dichloromethane, and drying the eluted product to obtain a white solid fluorescent probe molecule for detecting hydrogen peroxide;
hydrogen spectrum nuclear magnetic data:1H NMR(400MHz,DMSO-d6)δ11.04(s,2H),7.98(d,J＝7.6Hz,1H),7.76(dt,J＝29.5,7.4Hz,2H),7.30(d,J＝7.6Hz,1H),6.88(s,2H),6.62(s,2H)。
example 3
Selective detection of hydrogen peroxide by fluorescent probe molecules:
the fluorescent probe molecules prepared in example 1 are used to evaluate the specificity detection and recognition of the probe molecules on hydrogen peroxide, and FIG. 2 shows that the fluorescence intensity of the detection test paper changes when various common interferents or strong oxidants are added to the fluorescent probe, for example, 1mmol/L of interferent is (Bi (NO) is added3)3、CoCl2、Cs2CO3、FeCl3、FeSO4、HgCl2、K2Cr2O7、KCl、KI、KMnO4、MgCl2、Na2SO3、NaClO4、NaNO2、NH4NO3、NiCl2、Pb(CH3CO2)2、CH4N2S、CH4ON、CH4N2O or Zn (CH)3CO2)2) Probe needleThe excitation wavelength is 365 nm, and the emission wavelength is 526 nm; when hydrogen peroxide is added into the fluorescent probe solution, the fluorescence intensity of the fluorescent probe solution is increased dramatically, and when various common interfering species are added, the fluorescence intensity of the fluorescent probe solution is hardly changed obviously, so that the fluorescent probe molecule has a good function of selectively identifying the hydrogen peroxide.
Example 4
Linear relationship between change in fluorescent probe molecule intensity and hydrogen peroxide concentration:
using the fluorescent probe molecules of example 1 as a linear relationship for evaluating the fluorescence intensity of such probe molecules with the concentration of hydrogen peroxide ions, fig. 3 is a graph of the change of the fluorescence intensity of the fluorescent probe molecules at the emission center wavelength of 526 nm with the concentration of hydrogen peroxide, and the results of fig. 3 show that: the fluorescence intensity change of the fluorescent probe molecule solution and the concentration change of the hydrogen peroxide show good linear relation in the range of 0-80 umol/L, so that the probe molecules have excellent hydrogen peroxide detection capability and show good detection performance.
Example 5
FIG. 4 is a graph showing the actual detection performance and effect of the silica gel plate-based supported fluorescent probe molecules on hydrogen peroxide:
uniformly dispersing the fluorescent probe molecules in the embodiment 1 on a silica gel plate to obtain a detection sensor for detecting hydrogen peroxide; by dropping a hydrogen peroxide solution and observing the change of the fluorescence intensity of the probe molecules on the sensor under the irradiation of an ultraviolet lamp, the test result shown in fig. 4 shows that: the silica gel plate loaded with the fluorescent probe molecules can efficiently detect the hydrogen peroxide, shows obvious fluorescence intensity change and shows good hydrogen peroxide detection performance.
It should be further noted that the above embodiment is only a derivative using dichlorofluorescein as a fluorescent group and phenylboronic acid pinacol ester as a recognition group, which describes a preferred embodiment of the present invention, and all the similar fluorescent derivatives that have not been described in the embodiment of the present invention can achieve the same purpose and achieve the same detection effect and technical effect as the embodiment.
The scope of the present invention is not limited thereto, and any changes and modifications made to the technical solution of the present invention by those skilled in the art and related thereto without departing from the spirit of the present invention shall fall within the scope of the present invention defined in the claims.
Claims (2)
1. A fluorescent probe molecule for detecting hydrogen peroxide, characterized in that the chemical name of the fluorescent probe is 2',7' -di R-3',6' -bis ((4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxolan-2-yl) benzyloxy) -3H-spiro [ isobenzofuran-1, 9' -xanthene ], and the structural formula is:
wherein: r is hydrogen or halogen.
2. The method for preparing a fluorescent probe molecule for detecting hydrogen peroxide according to claim 1, comprising the steps of:
a. adding a compound 2-R-based fluorescein, potassium carbonate, 4-bromomethyl phenylboronic acid pinacol ester and acetonitrile into a round-bottom flask according to a molar ratio of 1:2:3:500, heating and stirring at the temperature of 90 ℃, condensing and refluxing for 20 hours, and stopping the reaction, wherein: r is hydrogen or halogen;
b. after the reaction is cooled to room temperature, adding 50mL of acetonitrile, fully stirring, filtering to remove impurities, and drying to obtain a crude product;
c. and (c) purifying the crude product obtained in the step (b) by using a forward silica gel chromatographic column, wherein an eluent is dichloromethane, and drying an eluted product to obtain the white solid fluorescent probe molecule for detecting hydrogen peroxide.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113896739A (en) * | 2021-08-26 | 2022-01-07 | 福建师范大学 | Reactive rhodamine B derivative fluorescent probe and preparation method and application thereof |
| CN114657780A (en) * | 2022-03-21 | 2022-06-24 | 武汉纺织大学 | Preparation method of nanofiber membrane for hydrogen peroxide fluorescence detection |
| KR102648134B1 (en) * | 2022-09-13 | 2024-03-14 | 경북대학교 산학협력단 | Novel fluorescein derivative and use thereof for detecting hydrogen peroxide |
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| US20140248218A1 (en) * | 2013-02-08 | 2014-09-04 | The Regents Of The University Of California | Ros-sensitive fluorescent probes |
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| US20140248218A1 (en) * | 2013-02-08 | 2014-09-04 | The Regents Of The University Of California | Ros-sensitive fluorescent probes |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113896739A (en) * | 2021-08-26 | 2022-01-07 | 福建师范大学 | Reactive rhodamine B derivative fluorescent probe and preparation method and application thereof |
| CN113896739B (en) * | 2021-08-26 | 2023-09-22 | 福建师范大学 | Reactive rhodamine B derivative fluorescent probe and preparation method and application thereof |
| CN114657780A (en) * | 2022-03-21 | 2022-06-24 | 武汉纺织大学 | Preparation method of nanofiber membrane for hydrogen peroxide fluorescence detection |
| CN114657780B (en) * | 2022-03-21 | 2023-11-28 | 武汉纺织大学 | Preparation method of nanofiber membrane for hydrogen peroxide fluorescence detection |
| KR102648134B1 (en) * | 2022-09-13 | 2024-03-14 | 경북대학교 산학협력단 | Novel fluorescein derivative and use thereof for detecting hydrogen peroxide |
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