CN104031637A - Azo fluorescent probe for detecting biological hydrogen sulfide and application thereof - Google Patents
Azo fluorescent probe for detecting biological hydrogen sulfide and application thereof Download PDFInfo
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- CN104031637A CN104031637A CN201410285548.5A CN201410285548A CN104031637A CN 104031637 A CN104031637 A CN 104031637A CN 201410285548 A CN201410285548 A CN 201410285548A CN 104031637 A CN104031637 A CN 104031637A
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Abstract
The invention provides a fluorescent probe for detecting biological hydrogen sulfide, of which the fluorescence emission wavelength is greater than 500nm. The fluorescent probe has high stability, and thus, can be stored and used for a long time. The fluorescent probe can effectively avoid interference from the biomacromolecule background fluorescence. The fluorescent probe does not have fluorescence itself, has fluorescence only after reaction with H2S, and has high signal to noise ratio for detection and favorable sensitivity. The fluorescent probe has excellent selectivity, and can specifically detect H2S in a complex biological sample. The fluorescent probe has favorable biomembrane permeability, and thus, can be used for specific detection of biological hydrogen sulfide in living cells. The structural formula of the fluorescent probe is disclosed in the specification.
Description
Technical field
The invention belongs to field of biological detection, relate to a kind of fluorescent probe and application thereof of detection of biological hydrogen sulfide.
Background technology
Hydrogen sulfide (H
2s), be a kind of colourless, inflammable, gas with rotten egg smell, be considered to for a long time toxic substance.But recent research shows, Mammals can be at H
2under the katalysis of S synthetic enzyme, take homocysteine or halfcystine as the synthetic a small amount of H of raw material
2s, and profit uses it as the physiological processs such as the diastole of signaling molecule modulating vascular, inflammatory reaction, glucose metabolism.H
2s is the 3rd found gas courier after NO, CO.It can be by opening ATP sensitive potassium-channel in vascular smooth muscle and vasodilation, thereby reach hypotensive effect.H
2s also has the cardiovascular protection effects such as Angiogensis, vascular remodeling, atherosclerosis, anti-platelet aggregation, and the Ischemia-reperfusion Injury damage of heart, kidney and other organs is had to provide protection.In inflammatory reaction, H
2s can lower by suppressing the activation of NF-κ B the level of many short inflammation gene expressions (as iNOS, COX-2 etc.) and inflammatory cell molecule, adhesion molecule.H in mammalian body
2the dyssynthesis of S will cause the diseases such as Alzheimer disease, hypertension, diabetes.At present, to H
2during the research of S physiological function and mechanism of action is further being carried out, and H
2s releasing agent is also used as the medicine with potential result for the treatment of and is subject to extensive concern.Thereby, develop quick, sensitive, easy detection of biological H
2the method of S is not only to H
2the functional study of S has huge pushing effect, is also the H that screening has potential treatment meaning simultaneously
2the necessary technique means of S donor molecule.
Traditional detection H
2the method of S comprises spectrophotometry and electrochemical process.But the detection error of these two kinds of methods is larger, and can only measure the H in blood plasma or tissue homogenate
2s, can not be for measuring the even H of biological tissue of cell
2s.Fluorescent probe detection method is the novel method growing up over the past two years.The method is utilized H
2the reductibility of S or nucleophilicity, design can with H
2the fluorescent probe of S specific reaction, these fluorescent probes itself are without fluorescence, with H
2after S reaction, can generate the product with hyperfluorescenceZeng Yongminggaoyingguang, thereby can be used for biological H
2the Sensitive Detection of S.Because small molecules fluorescent probe has advantages of that volume is little, microbial film permeability is good, thereby fluorescent probe method is except being applicable to detect the H in blood plasma, tissue homogenate
2s, is also applicable to H in viable cell or even animal tissues
2the detection of S.
Summary of the invention
An object of the present invention is to provide a kind of azo fluorescent probe of detection of biological hydrogen sulfide, is the biological hydrogen sulfide (H of a kind of novel specific detection
2s) small molecules fluorescent probe, this fluorescent probe this in physiological environment without fluorescence, but can with H
2the rapid reaction of S specificity, generates the product with hyperfluorescenceZeng Yongminggaoyingguang, thereby realizes H
2the specificity inspection of S.The fluorescent emission wavelength >500nm of described fluorescent probe, has the structure shown in formula (I):
Wherein, R
1for polysubstituted aromatic nucleus, substituting group is electron withdrawing group, can be fluorine, chlorine, bromine, iodine, nitro, trifluoromethyl, cyano group etc.; R
2for fluorescence dye, can be rhodamine, anthocyanidin series dyes, tonka bean camphor, fluorine boron two pyrroles (BODIPY) etc.
Another object of the present invention is to provide the fluorescent probe shown in formula I at detection of biological H
2application in S, can realize by following steps: in system to be detected (blood plasma, homogenate, cell culture medium), add the fluorescent probe shown in formula I, making its final concentration is 20 μ M.37
ounder C, hatch observe system fluorescence intensity 1 hour.When system to be checked is blood plasma or tissue homogenate, can record fluorescence intensity by spectrophotofluorometer; When system to be detected is viable cell, can record fluorescence intensity by flow cytometer or fluorescent microscope.
The fluorescent probe the present invention relates to has following beneficial effect: (1) good stability, can preserve for a long time use; (2) there is long fluorescent emission wavelength (>500nm), can effectively avoid the interference from biomacromolecule background fluorescence; (3) because probe itself is without fluorescence, only with H
2after S reaction, just have fluorescence, therefore, detection signal-to-noise ratio is high, and sensitivity is good; (4) there is outstanding selectivity, can in complex biological sample, detect specifically H
2s; (5) there is good microbial film permeability, thereby can be used for viable cell H
2the detection of S.
Accompanying drawing explanation
Fig. 1 is fluorescent probe molecule II and H
2change in fluorescence before and after S reaction.
Fig. 2 is fluorescent probe molecule III and H
2change in fluorescence before and after S reaction.
Fig. 3 is that fluorescent probe molecule II is to H
2the selectivity of S.
Fig. 4 is that fluorescent probe molecule III is to H
2the selectivity of S.
Fig. 5 is that fluorescent probe molecule II presents NaHS concentration dependent fluorescence is strengthened in bovine serum.
Embodiment
The present invention is further described with the following examples by reference to the accompanying drawings, rather than limits the scope of the invention.
embodiment 1: the probe molecule shown in Formula Il and H
2change in fluorescence before and after S reaction
Probe molecule, with a small amount of dissolve with ethanol, is added respectively to the PBS solution of PBS damping fluid or NaHS, and the final concentration that makes probe molecule is 20 μ M, and NaHS final concentration is 500 μ M.React and in fluorescence spectrophotometer, measure after 1.0 hours, and then definite probe molecule and H
2after S reaction, fluorescence intensity significantly strengthens, and result is referring to Fig. 1.
embodiment 2: the probe molecule shown in Formula Il I and H
2change in fluorescence before and after S reaction
Probe molecule, with a small amount of dissolve with ethanol, is added respectively to the PBS solution of PBS damping fluid or NaHS, and the final concentration that makes probe molecule is 20 μ M, and NaHS final concentration is 500 μ M.React and in fluorescence spectrophotometer, measure after 1.0 hours, and then definite probe molecule and H
2after S reaction, fluorescence intensity significantly strengthens, and result is referring to Fig. 2.
embodiment 3: probe molecule II is to H
2the selectivity of S
Probe molecule, with a small amount of dissolve with ethanol, then is configured to solution with PBS damping fluid.Add respectively the testing sample dissolving with PBS damping fluid, the concentration that makes final probe molecule is 20 μ M, and the concentration of testing sample is 500 μ M.React and in fluorescence spectrophotometer, measure after 1.0 hours, and then definite probe molecule is to H
2the selectivity of S.Result is referring to Fig. 3, and probe molecule is to H
2s has very high selectivity.
embodiment 4: probe molecule III is to H
2the selectivity of S
Probe molecule, with a small amount of dissolve with ethanol, then is configured to solution with PBS damping fluid.Add respectively the testing sample dissolving with PBS damping fluid, the concentration that makes final probe molecule is 20 μ M, and the concentration of testing sample is 500 μ M.React and in fluorescence spectrophotometer, measure after 1.0 hours, and then definite probe molecule is to H
2the selectivity of S.Result is referring to Fig. 4, and probe molecule is to H
2s has very high selectivity.
embodiment 5: probe molecule II presents NaHS concentration dependent fluorescence is strengthened for bovine serum
Probe molecule, with a small amount of dissolve with ethanol, then is configured to solution with bovine serum and a small amount of ethanol.Add respectively the testing sample dissolving with PBS damping fluid, the concentration that makes final probe molecule is 20 μ M, adds respectively successively the H that volume is identical
2o
2the PBS solution of (final concentration 1 mM), NaHS (final concentration is 10 μ M, 20 μ M, 30 μ M, 40 μ M, 50 μ M) reaction was measured after 1.0 hours in fluorescence spectrophotometer, and then definite probe molecule presents NaHS concentration dependent fluorescence is strengthened in bovine serum.Result is referring to Fig. 5.
embodiment 6: probe molecule II is for rat plasma endogenous H
2s's is quantitative
Adopt marker method to detect endogenous H in fresh rat blood plasma
2s concentration.Fresh rat blood plasma adds the long-pending ethanol of monoploid, and precipitation is except Deproteinization.Except the blood plasma after albumen is divided into 5 parts, add respectively successively the ZnCl that volume is identical
2the PBS solution of (1 mM), deionized water, NaHS (final concentration is 10 μ M, 20 μ M, 30 μ M).The ethanolic soln (final concentration is 20 μ M) that adds respectively isopyknic probe I I in above-mentioned plasma solutions, reacts and with 468 nm, excites after 1 hour, records 517 nm place fluorescence intensities in microplate reader.It is 18 μ M that calculating records endogenous H2S concentration.
Claims (2)
1. an azo fluorescent probe for detection of biological hydrogen sulfide, is characterized in that, the fluorescent emission wavelength >500nm of this probe, and chemical structural formula is:
R wherein
1for polysubstituted aromatic nucleus, substituting group is electron withdrawing group, selects fluorine, chlorine, bromine, iodine, nitro, trifluoromethyl or cyano group; R
2for fluorescence dye, select rhodamine, anthocyanidin series dyes, tonka bean camphor or fluorine boron two pyrroles.
2. the application of azo fluorescent probe according to claim 1 in detection of biological hydrogen sulfide.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104403663A (en) * | 2014-12-11 | 2015-03-11 | 华东理工大学 | Fluorescent probe for detecting endogenous H2S, as well as preparation method and application of fluorescent probe |
| CN106083888A (en) * | 2016-07-12 | 2016-11-09 | 济南大学 | A kind of detect the fluorescent probe of hydrogen sulfide in cancerous cell |
| CN106967038A (en) * | 2017-04-10 | 2017-07-21 | 台州学院 | A kind of synthesis and application of the fluorescence probe of Dual channel detection hydrogen sulfide |
| CN111518546A (en) * | 2020-05-19 | 2020-08-11 | 遵义医科大学 | Hypoxic microenvironment response fluorescent probe and preparation method and application thereof |
| CN112745303A (en) * | 2019-10-30 | 2021-05-04 | 南京大学 | Hypoxic fluorescent probe and application thereof |
| CN113189093A (en) * | 2021-04-28 | 2021-07-30 | 西南石油大学 | Method for detecting and monitoring hydrogen sulfide gas |
| CN113358289A (en) * | 2021-04-28 | 2021-09-07 | 西南石油大学 | Characteristic response material for monitoring and detecting hydrogen sulfide gas leakage and preparation method thereof |
| CN113493821A (en) * | 2020-04-03 | 2021-10-12 | 北京康普森农业科技有限公司 | Method for detecting fishy smell gene of egg |
| CN114354593A (en) * | 2022-01-07 | 2022-04-15 | 西南石油大学 | Composite test paper for detecting hydrogen sulfide gas leakage |
| CN114380856A (en) * | 2021-12-31 | 2022-04-22 | 北京化工大学 | Silorhodamine derivative for detecting brain hydrogen sulfide and preparation method and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111233885B (en) * | 2020-03-20 | 2022-04-22 | 福建师范大学 | Fluorescent probe for detecting methanol and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102417229A (en) * | 2011-10-24 | 2012-04-18 | 沈阳建筑大学 | Method for removing azo dye waste water chroma by using hydrogen sulfide waste gas |
-
2014
- 2014-06-24 CN CN201410285548.5A patent/CN104031637B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102417229A (en) * | 2011-10-24 | 2012-04-18 | 沈阳建筑大学 | Method for removing azo dye waste water chroma by using hydrogen sulfide waste gas |
Non-Patent Citations (2)
| Title |
|---|
| SHUN-HUA LI 等: "A cyclometalated palladium–azo complex as a differential chromogenic probe for amino acids in aqueous solution", 《CHEM. COMMUN.》, 2 December 2004 (2004-12-02) * |
| 高敏 等: "检测硫化氢分子的荧光探针", 《化学进展》, vol. 26, no. 6, 28 May 2014 (2014-05-28) * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104403663A (en) * | 2014-12-11 | 2015-03-11 | 华东理工大学 | Fluorescent probe for detecting endogenous H2S, as well as preparation method and application of fluorescent probe |
| CN106083888A (en) * | 2016-07-12 | 2016-11-09 | 济南大学 | A kind of detect the fluorescent probe of hydrogen sulfide in cancerous cell |
| CN106967038A (en) * | 2017-04-10 | 2017-07-21 | 台州学院 | A kind of synthesis and application of the fluorescence probe of Dual channel detection hydrogen sulfide |
| CN106967038B (en) * | 2017-04-10 | 2019-06-11 | 台州学院 | A kind of synthesis and application of the fluorescence probe of Dual channel detection hydrogen sulfide |
| CN112745303A (en) * | 2019-10-30 | 2021-05-04 | 南京大学 | Hypoxic fluorescent probe and application thereof |
| CN112745303B (en) * | 2019-10-30 | 2022-04-22 | 南京大学 | Hypoxic fluorescent probe and application thereof |
| CN113493821A (en) * | 2020-04-03 | 2021-10-12 | 北京康普森农业科技有限公司 | Method for detecting fishy smell gene of egg |
| CN111518546A (en) * | 2020-05-19 | 2020-08-11 | 遵义医科大学 | Hypoxic microenvironment response fluorescent probe and preparation method and application thereof |
| CN113189093A (en) * | 2021-04-28 | 2021-07-30 | 西南石油大学 | Method for detecting and monitoring hydrogen sulfide gas |
| CN113358289B (en) * | 2021-04-28 | 2022-04-15 | 西南石油大学 | Characteristic response material for monitoring and detecting hydrogen sulfide gas leakage and preparation method thereof |
| CN113358289A (en) * | 2021-04-28 | 2021-09-07 | 西南石油大学 | Characteristic response material for monitoring and detecting hydrogen sulfide gas leakage and preparation method thereof |
| CN114380856A (en) * | 2021-12-31 | 2022-04-22 | 北京化工大学 | Silorhodamine derivative for detecting brain hydrogen sulfide and preparation method and application thereof |
| CN114380856B (en) * | 2021-12-31 | 2023-03-03 | 北京化工大学 | Silorhodamine derivative for detecting brain hydrogen sulfide and preparation method and application thereof |
| CN114354593A (en) * | 2022-01-07 | 2022-04-15 | 西南石油大学 | Composite test paper for detecting hydrogen sulfide gas leakage |
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