CN110016008A - The fluorescence probe of specific recognition hydrogen polysulfide and biological thiol - Google Patents
The fluorescence probe of specific recognition hydrogen polysulfide and biological thiol Download PDFInfo
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Abstract
The invention discloses the fluorescence probe of a kind of specific recognition hydrogen polysulfide and biological thiol, the structural formula of fluorescence probe is as follows:A kind of preparation method of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol; include the following steps; step 1; the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- is dissolved in anhydrous acetonitrile; sequentially add 4-DMAP hydrochloride, triethylamine; inert gas shielding is spin-dried for solvent after back flow reaction, obtains compound A；Compound A and malononitrile are dissolved in anhydrous methylene chloride by step 2, are added triethylamine and are made alkali, inert gas shielding separates after ice bath reaction, obtains fluorescence probe.The present invention has the beneficial effect of specific recognition hydrogen polysulfide and biological thiol.
Description
Technical field
The present invention relates to fluorescence probe preparation technical fields.It is more particularly related to which a kind of specific recognition is more
The fluorescence probe of hydrogen sulfide and biological thiol.
Background technique
Biological thiol (cysteine (Cysteine, Cys), homocysteine (Homocysteine, Hcy), gluathione
Peptide (Glutataione, GSH)) be many protein and small molecule important component, have during cell activities
It plays an important role.Wherein, cysteine is not only the precursor of glutathione, acetylcoenzyme and taurine, while being also organism
The supplier of sulphur ligand in sulphur iron complex, cysteine is lacked in human body will lead to slow growth, trichochromes decoloration, water
The symptoms such as swollen, drowsiness, liver dysfunction, of flaccid muscles, in poor health.The concentration abnormality of cysteine may also can cause A Er
The hereby generation of Alzheimer disease, cardiovascular disease, cancer.Glutathione is intracellular the most abundant non-protein biology sulfydryl chemical combination
Many functions are closely related in object, with cell body:, the metabolism of xenobiotic active including cellular redox, into the cell
Signal transduction and gene regulation etc..Cysteine, glutathione can mutually convert under the action of biological enzyme again in vivo, they
Changes of contents in organism is maintained close ties with many diseases.Therefore, the detection for detecting their intracellular biological thiols has ten
Divide important meaning.
In adjusting various physiology courses, H2SnWith very high effect, including active ions channel, transcription factor and swollen
Tumor inhibiting factor；Biological thiol, hydrogen polysulfide how are detected, is concerned, in recent years, a large amount of fluorescence probe, including fluorescence
Albumen, fluorescent dye are used for the detection of certain object of organism, but present most probes do not have same time zone
Divide H2SnWith the ability of biological thiol this 2 kinds of substances, so develop a kind of probe that can distinguish them simultaneously have it is very heavy
The meaning wanted.
Summary of the invention
It is an object of the invention to solve at least the above problems, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide the fluorescence probe of a kind of specific recognition hydrogen polysulfide and biological thiol,
H can be distinguished simultaneously2SnAnd biological thiol.
It is a still further object of the present invention to provide the fluorescence probes of a kind of specific recognition hydrogen polysulfide and biological thiol
Preparation method, raw material are easy to get, and intermediate product preparation need not be isolated and purified, and lower step fluorescence probe system can be directly carried out
Standby, preparation method is simple.
In order to realize these purposes and other advantages according to the present invention, provide a kind of specific recognition hydrogen polysulfide and
The structural formula of the fluorescence probe of biological thiol, fluorescence probe is as follows:
Provide a kind of preparation method of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, including following step
It is rapid:
The chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- is dissolved in anhydrous acetonitrile, sequentially adds 4- diformazan by step 1
Amino-phenol hydrochloride, triethylamine, inert gas shielding are spin-dried for solvent after back flow reaction, purify to obtain compound A, compound A's
Structural formula are as follows:
Compound A and malononitrile are dissolved in anhydrous methylene chloride by step 2, are added triethylamine and are made alkali, inert gas
Protection separates after ice bath reaction, obtains fluorescence probe.
Preferably, the inert gas in step 1 and step 2 is argon gas.
Preferably, in step 1, the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- and 4-DMAP hydrochloride
Molar ratio be 1:1-3, the molar ratio of the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- and triethylamine is 1:1-3,3- aldehyde radical -4-
The molal volume ratio of chloro- 7- lignocaine cumarin and anhydrous acetonitrile is 1mmol:5-15mL, wherein it is specific to carry out back flow reaction
For 80-100 DEG C of back flow reaction 4-5h.
Preferably, in step 1, the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- and 4-DMAP hydrochloride
Molar ratio be 1:1, the molar ratio of the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- and triethylamine is 1:1, and 3- aldehyde radical -4- is chloro-
The molal volume of 7- lignocaine cumarin and anhydrous acetonitrile ratio is 1mmol:5mL, wherein carrying out back flow reaction is specially 80 DEG C
Back flow reaction 4h.
Preferably, in step 2, the amount ratio of compound A and malononitrile is 1mmol:1-3mmol, compound A and three
The amount ratio of ethamine is 1mmol:0.36-1mmol, and the molal volume ratio of compound A and anhydrous methylene chloride is 1mmol:3-
9mL, wherein the ice bath reaction time is 50-120s.
Preferably, in step 2, the amount ratio of compound A and malononitrile is 1mmol:1mmol, compound A and three second
The amount ratio of amine is 1mmol:0.36mmol, and the molal volume ratio of compound A and anhydrous methylene chloride is 1mmol:3mL, wherein
The ice bath reaction time is 60s.
The present invention is include at least the following beneficial effects:
The first, blue light-emitting after fluorescence probe of the invention and biological thiol effect, with hydrogen polysulfide (H2Sn) turn to be yellow after effect
Light, being capable of specific recognition hydrogen polysulfide and biological thiol.
The second, raw material is easy to get, and intermediate product preparation need not be isolated and purified, and lower step fluorescence probe can be directly carried out
Preparation, preparation method are simple.
Further advantage, target and feature of the invention will be partially reflected by the following instructions, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Detailed description of the invention
Fig. 1 is the nucleus magnetic hydrogen spectrum figure of the fluorescence probe MCP1 of the invention；
Fig. 2 is the nuclear-magnetism carbon spectrogram of the fluorescence probe MCP1 of the invention；
Fig. 3 is the UV-Vis spectrum figure of the fluorescence probe MCP1 and biological thiol Cys interaction of the invention；
Fig. 4 is the UV-Vis spectrum figure of the fluorescence probe MCP1 and biological thiol GSH interaction of the invention；
Fig. 5 is the UV-Vis spectrum figure of the fluorescence probe MCP1 and hydrogen polysulfide interaction of the invention；
Fig. 6 is the fluorescent emission figure of the fluorescence probe MCP1 and biological thiol Cys interaction of the invention；
Fig. 7 is the fluorescent emission figure of the fluorescence probe MCP1 and biological thiol Cys interaction of the invention；
Fig. 8 is the fluorescent emission figure of the fluorescence probe MCP1 and biological thiol GSH interaction of the invention；
Fig. 9 is the fluorescent emission figure of the fluorescence probe MCP1 and biological thiol GSH interaction of the invention；
Figure 10 is the fluorescent emission figure of the fluorescence probe MCP1 and hydrogen polysulfide interaction of the invention；
Figure 11 is the fluorescent emission figure of the fluorescence probe MCP1 and hydrogen polysulfide interaction of the invention；
Figure 12 is the fluorescent emission of fluorescence probe MCP1 selectivity test under 386nm shooting condition of the invention
Figure；
Figure 13 is the fluorescent emission of fluorescence probe MCP1 selectivity test under 511nm shooting condition of the invention
Figure；
Figure 14 is the kinetic measurement schematic diagram of the fluorescence probe MCP1 and biological thiol Cys of the invention；
Figure 15 is the kinetic measurement schematic diagram of the fluorescence probe MCP1 and biological thiol GSH of the invention；
Figure 16 is the kinetic measurement schematic diagram of the fluorescence probe MCP1 and hydrogen polysulfide of the invention.
Specific embodiment
The present invention will be further described in detail below with reference to the embodiments, to enable those skilled in the art referring to specification
Text can be implemented accordingly.
<embodiment 1>
The structural formula of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, fluorescence probe is as follows:
The preparation method of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, comprising the following steps:
It is anhydrous to be dissolved in 5mL by step 1 for the chloro- 7- lignocaine cumarin (marked as 1,1mmol, 279mg) of 3- aldehyde radical -4-
In acetonitrile (acetonitrile), 4-DMAP hydrochloride (1mmol, 173mg) is added, triethylamine is eventually adding
(NEt3) (1mmol, 139 μ L) as alkali, argon gas is protected, and after 80 DEG C of back flow reaction 4h, is spin-dried for solvent, is obtained mixture, it mixes
Object purifies to obtain compound A, wherein mixture is red brown solid, purifies to obtain dark-brown compound A, amounts to 304mg, yield
It is 80%；
Principal product in mixture is compound A；
Reaction equation are as follows:
Compound A (304mg, 0.8mmol) and malononitrile (marked as 3,0.8mmol, 52.8 μ L) are dissolved in by step 2
In 2.4mL anhydrous methylene chloride (DCM), adds triethylamine (0.288mmol, 40 μ L) and make alkali, in ice bath (Ice Bath), argon
Under gas shielded, 1min is reacted, separation obtains fluorescence probe (marked as MCP1), wherein fluorescence probe is red solid, is amounted to
171.2mg yield 50%；
Reaction equation are as follows:
Can be seen that raw material fundamental reaction is complete in step 1 according to contact plate, mixture without purification, can directly into
The reaction of row step 2 influences to can be ignored substantially on the purification of step 2, when mixture is anti-without purifying directly progress step 2
At once, the amount of mixture by weight of yield carries out for 80%.
<embodiment 2>
The structural formula of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, fluorescence probe is as follows:
The preparation method of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, comprising the following steps:
Step 1, by the chloro- 7- lignocaine cumarin (marked as 1,1mmol, 279mg) of 3- aldehyde radical -4- be dissolved in 10mL without
In water-acetonitrile (acetonitrile), 4-DMAP hydrochloride (2mmol, 346mg) is added, triethylamine is eventually adding
(NEt3) (2mmol, 278 μ L) as alkali, argon gas is protected, and after 100 DEG C of back flow reaction 4.5h, is spin-dried for solvent, is obtained mixture, it mixes
Object is closed to purify to obtain compound A, wherein mixture is red brown solid, purifies to obtain dark-brown compound A, amounts to 291.8mg,
Yield is 76.8%；
Principal product in mixture is compound A；
Reaction equation are as follows:
Step 2, by compound A (291.8mg, 0.77mmol) and malononitrile (marked as 3,1.54mmol, 101.6 μ L),
It is dissolved in 4.62mL anhydrous methylene chloride (DCM), adds triethylamine (0.55mmol, 76.4 μ L) and make alkali, in ice bath (Ice
Bath), under argon gas protection, 50min is reacted, separation obtains fluorescence probe (marked as MCP1), wherein fluorescence probe is red
Solid amounts to 161mg, yield 49%；
Reaction equation are as follows:
<embodiment 3>
The structural formula of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, fluorescence probe is as follows:
The preparation method of the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, comprising the following steps:
Step 1, by the chloro- 7- lignocaine cumarin (marked as 1,1mmol, 279mg) of 3- aldehyde radical -4- be dissolved in 15mL without
In water-acetonitrile (acetonitrile), 4-DMAP hydrochloride (3mmol, 519mg) is added, triethylamine is eventually adding
(NEt3) (3mmol, 417 μ L) as alkali, argon gas is protected, and after 90 DEG C of back flow reaction 5h, is spin-dried for solvent, is obtained mixture, it mixes
Object purifies to obtain compound A, wherein mixture is red brown solid, purifies to obtain dark-brown compound A, amounts to 297.2mg, is produced
Rate is 78.2%；
Principal product in mixture is compound A；
Reaction equation are as follows:
Step 2, by compound A (297.2304mg, 0.78mmol) and malononitrile (marked as 3,2.350.8mmol,
155.1 μ L), it is dissolved in 7.02mL anhydrous methylene chloride (DCM), adds triethylamine (0.78288mmol, 108.3 μ L) and make alkali,
At ice bath (Ice Bath), argon gas protection, 120s is reacted, separation obtains fluorescence probe (marked as MCP1), wherein fluorescence
Probe is red solid, amounts to 160.7mg, yield 48%；
Reaction equation are as follows:
1, probe hydrogen spectrum measurement
As shown in Figure 1, the nucleus magnetic hydrogen spectrum figure of fluorescence probe MCP1,1H NMR(300MHz,CDCl3)δ7.67(s,1H),
7.35 (d, J=9.9Hz, 1H), 6.93 (d, J=9.3Hz, 2H), 6.71 (d, J=9.0Hz, 2H), 6.50 (d, J=2.1Hz,
1H), 6.47 (s, 1H), 3.46 (q, J=7.2Hz, 4H), 1.32-1.17 (m, 6H).
2, probe carbon spectrum measurement
As shown in Fig. 2, the nucleus magnetic hydrogen spectrum figure of fluorescence probe MCP1,13C NMR(75MHz,CDCl3)δ166.8,159.9,
157.6,153.3,150.2,127.7,117.9,115.2,113.7,112.8,109.9,103.3,101.9,97.1,83.3,
45.2,41.0,12.5。
3, probe MCP1 and H2SnWith the UV-Vis spectrum detection of RSH interaction
3.1 solution are prepared
Blank solution: DMSO, mixing are added into PBS solution, wherein the volume ratio of DMSO and PBS solution is 2:
8；
Probe-PBS solution containing object to be detected: DMSO, fluorescence probe MCP1, to be detected is sequentially added into PBS solution
Object mixes, wherein the volume ratio of DMSO and PBS solution is 2:8, and the concentration of fluorescence probe MCP1 is 10 μM, object to be detected with it is glimmering
The mass ratio of light probe MCP1 is 100:1, and object to be detected is one of biological thiol Cys, biological thiol GSH, hydrogen polysulfide.
3.2 test
Use the baseline of blank solution adjustment UV-Vis spectrum visible spectrophotometer；
The UV-Vis spectrum curve that probe-PBS solution containing object to be detected respectively changes over time is measured respectively, wherein is contained
UV-Vis spectrum curve that probe-PBS solution of biological thiol Cys changes over time as shown in figure 3, the GSH containing biological thiol spy
The UV-Vis spectrum curve that needle-PBS solution changes over time is as shown in figure 4, probe-PBS solution containing hydrogen polysulfide becomes at any time
The UV-Vis spectrum curve of change is as shown in Figure 5.
3.3 result
By Fig. 3-4 it is found that the fluorescence exciting wavelength of biological thiol is 386nm.
As shown in Figure 4, the fluorescence exciting wavelength of hydrogen polysulfide is 511nm.
4, probe MCP1 and H2SnWith the fluorescent emission detection of RSH interaction
4.1 solution are prepared
Blank solution, with the preparation of blank solution in " preparation of 3.1 solution "；
Probe-PBS solution containing object to be detected: with probe-PBS solution containing object to be detected in " preparation of 3.1 solution "
Configuration；
4.2 test
Use the baseline of blank solution adjustment UV-Vis spectrum visible spectrophotometer；
Probe-PBS solution containing object to be detected is surveyed respectively respectively to change over time under 386nm and 511nm shooting condition
Fluorescence emission curves, wherein
Fig. 6: the fluorescence emission curves that probe-PBS solution containing Cys changes over time under the excitation of 386nm excitation wavelength；
Fig. 7: the fluorescence emission curves that probe-PBS solution containing Cys changes over time under the excitation of 511nm excitation wavelength；
Fig. 8: the fluorescence emission curves that probe-PBS solution containing GSH changes over time under the excitation of 386nm excitation wavelength；
Fig. 9: the fluorescence emission curves that probe-PBS solution containing GSH changes over time under the excitation of 511nm excitation wavelength；
Figure 10: contain H2SnProbe-PBS solution under the excitation of 386nm excitation wavelength the fluorescent emission that changes over time it is bent
Line；
Figure 11: contain H2SnProbe-PBS solution under the excitation of 511nm excitation wavelength the fluorescent emission that changes over time it is bent
Line；
4.3 result
The fluorescence emission wavelengths of biological thiol Cys and biological thiol GSH known to Fig. 6-9 are 486nm.
The H known to Figure 10-112SnFluorescence emission wavelengths be 576nm.
5, the selectivity test of probe MCP1
5.1, solution allocation
Mother liquor: DMSO, fluorescence probe MCP1 are sequentially added into PBS solution, wherein the volume ratio of DMSO and PBS solution
For 2:8, the concentration of fluorescence probe MCP1 is 10 μM；
Probe-PBS solution containing object to be detected: sequentially added into PBS solution DMSO, fluorescence probe MCP1, it is related to
Detectable substance mixes, wherein the volume ratio of DMSO and PBS solution is 2:8, and the concentration of fluorescence probe MCP1 is 10 μM, object to be detected
Mass ratio with fluorescence probe MCP1 is 100:1, and object to be detected includes: various common zwitterions, amino acid, active oxygen nitrogen class
Substance, i.e. H2Sn、H2O2、S2O3 2-、SO3 2-、SO4 2-、HSO3 -、ClO-、O2-、NO、NO2 -、NO3 -、Ca2+、K+、Mg2+、Zn2+, half Guang
Propylhomoserin Cys, glutathione GSH, alanine Ala, arginine Arg, aspartic acid Asp, lysine Lys, methionine Met, network
Propylhomoserin Tyl, glycine Gly, glutamic acid Glu, histidine, isoleucine Iso, phenylalanine Phe, proline Pro, silk ammonia
Sour Ser, threonine Thr, valine Val.
5.2, it tests
On the downside of 386nm and 511nm shooting condition, by Fluorescence Spectrometer test, respectively probe-the PBS containing object to be detected is molten
The fluorescence emission curves that liquid changes over time, wherein Figure 12 is indicated on the downside of excitation wavelength 386nm shooting condition respectively containing to be detected
Probe-PBS solution fluorescence emission spectrum of object；Figure 13 indicates the probe-on the downside of 511nm shooting condition respectively containing object to be detected
The fluorescence emission spectrum of PBS solution；
5.3, result
As shown in figure 12, under 386nm shooting condition, when object to be detected is H2O2、S2O3 2-、SO3 2-、SO4 2-、HSO3 -、
ClO-、O2-、NO、NO2 -、NO3 -、Ca2+、K+、Mg2+、Zn2+, alanine Ala, arginine Arg, aspartic acid Asp, lysine
Lys, methionine Met, tyrosine Tyl, glycine Gly, glutamic acid Glu, histidine, isoleucine Iso, phenylalanine
When Phe, proline Pro, serine Ser, threonine Thr, valine Val, system fluorescence has no significant change, and is specifically shown in Figure 12
Middle 0thers indicates that biological thiol cysteine Cys, biological thiol glutathione GSH cause fluorescence at 486nm (blue light region)
The different degrees of variation of intensity, and H2SnCause fluorescence intensity change at 576nm (yellow light area)；
As shown in figure 13, under 386nm shooting condition, when object to be detected is H2O2、S2O3 2-、SO3 2-、SO4 2-、HSO3 -、
ClO-、O2-、NO、NO2 -、NO3 -、Ca2+、K+、Mg2+、Zn2+, alanine Ala, arginine Arg, aspartic acid Asp, lysine
Lys, methionine Met, tyrosine Tyl, glycine Gly, glutamic acid Glu, histidine, isoleucine Iso, phenylalanine
Phe, proline Pro, serine Ser, threonine Thr, valine Val, biological thiol cysteine Cys, biological thiol paddy Guang
When sweet peptide GSH, system fluorescence has no significant change, and being specifically shown in 0thers in Figure 12 indicates, only H2SnIn 576nm (yellow light area)
Place causes fluorescence intensity change；
That is, probe MCP1 being capable of specific recognition hydrogen polysulfide (H2Sn) and biological thiol.
6, probe MCP1 and H2SnWith the kinetic measurement of RSH effect
Probe MCP1 itself is without fluorescence, according to Fig. 6, Fig. 8 and Figure 11 it is found that biological thiol cysteine is added
Cys, biological thiol glutathione GSH, H2SnThe fluorescence intensity of system gradually increases over time afterwards, wherein according to figure
14-16 is it is found that the equilibration time of biological thiol cysteine Cys, biological thiol glutathione GSH are 15min, H2SnBalance
Time is 10min, shows probe MCP1 to H2SnThere is faster response with RSH.
7, probe mechanism study
Research shows that probe is as follows to biological thiol (biothiols), the Response Mechanism of hydrogen polysulfide:
Using hydrogen polysulfide, biological thiol in terms of structure and chemical property after existing some differences, with probe effect
The compound for generating different structure, issues different fluorescence, reaches specific recognition hydrogen polysulfide and biological thiol.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details and legend shown and described herein.
Claims (7)
1. the fluorescence probe of specific recognition hydrogen polysulfide and biological thiol, which is characterized in that the structural formula of fluorescence probe is as follows:
2. the preparation method of the fluorescence probe of specific recognition hydrogen polysulfide as described in claim 1 and biological thiol, special
Sign is, comprising the following steps:
The chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- is dissolved in anhydrous acetonitrile, sequentially adds 4- dimethylamino by step 1
Phenol hydrochloride, triethylamine, inert gas shielding are spin-dried for solvent after back flow reaction, purify to obtain compound A, the structure of compound A
Formula are as follows:
Compound A and malononitrile are dissolved in anhydrous methylene chloride by step 2, are added triethylamine and are made alkali, inert gas shielding,
It is separated after ice bath reaction, obtains fluorescence probe.
3. the preparation method of the fluorescence probe of specific recognition hydrogen polysulfide as claimed in claim 2 and biological thiol, special
Sign is that the inert gas in step 1 and step 2 is argon gas.
4. the preparation method of the fluorescence probe of specific recognition hydrogen polysulfide as claimed in claim 2 and biological thiol, special
Sign is, in step 1, the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- and the molar ratio of 4-DMAP hydrochloride are
The molar ratio of the chloro- 7- lignocaine cumarin of 1:1-3,3- aldehyde radical -4- and triethylamine is 1:1-3, the chloro- 7- diethyl of 3- aldehyde radical -4-
The molal volume of aminocoumarin and anhydrous acetonitrile ratio is 1mmol:5-15mL, wherein carrying out back flow reaction is specially 80-100
DEG C back flow reaction 4-5h.
5. the preparation method of the fluorescence probe of specific recognition hydrogen polysulfide as claimed in claim 4 and biological thiol, special
Sign is, in step 1, the chloro- 7- lignocaine cumarin of 3- aldehyde radical -4- and the molar ratio of 4-DMAP hydrochloride are
The molar ratio of the chloro- 7- lignocaine cumarin of 1:1,3- aldehyde radical -4- and triethylamine is 1:1, the chloro- 7- lignocaine of 3- aldehyde radical -4-
The molal volume of cumarin and anhydrous acetonitrile ratio is 1mmol:5mL, wherein carrying out back flow reaction is specially 80 DEG C of back flow reactions
4h。
6. the preparation method of the fluorescence probe of specific recognition hydrogen polysulfide as claimed in claim 2 and biological thiol, special
Sign is, in step 2, the amount ratio of compound A and malononitrile is 1mmol:1-3mmol, the dosage of compound A and triethylamine
Than for 1mmol:0.36-1mmol, the molal volume ratio of compound A and anhydrous methylene chloride is 1mmol:3-9mL, wherein ice bath
Reaction time is 50-120s.
7. the preparation method of the fluorescence probe of specific recognition hydrogen polysulfide as claimed in claim 6 and biological thiol, special
Sign is, in step 2, the amount ratio of compound A and malononitrile is 1mmol:1mmol, the amount ratio of compound A and triethylamine
For 1mmol:0.36mmol, the molal volume ratio of compound A and anhydrous methylene chloride is 1mmol:3mL, wherein when ice bath reacts
Between be 60s.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110669043A (en) * | 2019-10-12 | 2020-01-10 | 南宁师范大学 | Fluorescent probe for identifying cysteine, homocysteine, glutathione and hydrogen sulfide and preparation method thereof |
| CN111718319A (en) * | 2020-07-27 | 2020-09-29 | 山西大学 | Fluorescent probe for distinguishing and detecting mercaptan and monitoring Cys/GSH metabolism and preparation method thereof |
| CN112939918A (en) * | 2021-02-05 | 2021-06-11 | 山西大学 | Coumarin derivative CTT and synthesis method and application thereof |
| CN115490664A (en) * | 2022-09-29 | 2022-12-20 | 湖南师范大学 | Synthesis and application of fluorescent probe for selectively detecting homocysteine |
| WO2023097820A1 (en) * | 2021-12-03 | 2023-06-08 | 德州学院 | Flavonol compound, and preparation method therefor and use thereof in detection of biological mercaptan |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104418874A (en) * | 2013-08-28 | 2015-03-18 | 苏州罗兰生物科技有限公司 | Fluorescent molecular probe for detecting fluoride ions in aqueous solutions as well as synthesis method and application thereof |
| CN106588912A (en) * | 2016-11-30 | 2017-04-26 | 济南大学 | Fluorescent probe capable of separately detecting cysteine/homocysteine, glutathione and sulfuretted hydrogen and preparation method and application of fluorescent probe |
| CN106950210A (en) * | 2017-03-27 | 2017-07-14 | 山西大学 | A kind of reagent for detecting glutathione and its synthetic method and application |
| CN107033111A (en) * | 2017-04-21 | 2017-08-11 | 台州学院 | A kind of synthesis and application of the near infrared fluorescent probe for detecting hydrogen sulfide |
| CN107056769A (en) * | 2017-04-07 | 2017-08-18 | 济南大学 | A kind of L cysteines fluorescence probe and preparation method thereof |
| CN107216270A (en) * | 2017-07-24 | 2017-09-29 | 南京微瑞莱电子科技有限公司 | A kind of application for detecting hydrogen sulfide high selectivity response type fluorescence probe |
| US20180074068A1 (en) * | 2016-09-14 | 2018-03-15 | Washington State University | Fluorescent probes for detecting hydrogen polysulfides |
| CN108285789A (en) * | 2018-04-20 | 2018-07-17 | 济南大学 | A kind of hydrogen peroxide fluorescence probe and its preparation method and application |
| KR101879726B1 (en) * | 2017-08-08 | 2018-07-18 | 건국대학교 산학협력단 | A novel coumarin derivative, composition for detecting cyanide ion comprising the same and method for detecting cyanide ion using the same |
| CN108358906A (en) * | 2018-03-14 | 2018-08-03 | 中南大学 | One species specificity distinguishes the fluorescence probe of different mercaptan |
| CN108727326A (en) * | 2018-07-06 | 2018-11-02 | 广西师范学院 | Identify fluorescence probe and preparation method and the application of cysteine and glutathione |
| CN109574977A (en) * | 2019-01-04 | 2019-04-05 | 福州大学 | A kind of Coumarins hypochlorous acid fluorescence probe and preparation method thereof |
-
2019
- 2019-04-19 CN CN201910318929.1A patent/CN110016008B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104418874A (en) * | 2013-08-28 | 2015-03-18 | 苏州罗兰生物科技有限公司 | Fluorescent molecular probe for detecting fluoride ions in aqueous solutions as well as synthesis method and application thereof |
| US20180074068A1 (en) * | 2016-09-14 | 2018-03-15 | Washington State University | Fluorescent probes for detecting hydrogen polysulfides |
| CN106588912A (en) * | 2016-11-30 | 2017-04-26 | 济南大学 | Fluorescent probe capable of separately detecting cysteine/homocysteine, glutathione and sulfuretted hydrogen and preparation method and application of fluorescent probe |
| CN106950210A (en) * | 2017-03-27 | 2017-07-14 | 山西大学 | A kind of reagent for detecting glutathione and its synthetic method and application |
| CN107056769A (en) * | 2017-04-07 | 2017-08-18 | 济南大学 | A kind of L cysteines fluorescence probe and preparation method thereof |
| CN107033111A (en) * | 2017-04-21 | 2017-08-11 | 台州学院 | A kind of synthesis and application of the near infrared fluorescent probe for detecting hydrogen sulfide |
| CN107216270A (en) * | 2017-07-24 | 2017-09-29 | 南京微瑞莱电子科技有限公司 | A kind of application for detecting hydrogen sulfide high selectivity response type fluorescence probe |
| KR101879726B1 (en) * | 2017-08-08 | 2018-07-18 | 건국대학교 산학협력단 | A novel coumarin derivative, composition for detecting cyanide ion comprising the same and method for detecting cyanide ion using the same |
| CN108358906A (en) * | 2018-03-14 | 2018-08-03 | 中南大学 | One species specificity distinguishes the fluorescence probe of different mercaptan |
| CN108285789A (en) * | 2018-04-20 | 2018-07-17 | 济南大学 | A kind of hydrogen peroxide fluorescence probe and its preparation method and application |
| CN108727326A (en) * | 2018-07-06 | 2018-11-02 | 广西师范学院 | Identify fluorescence probe and preparation method and the application of cysteine and glutathione |
| CN109574977A (en) * | 2019-01-04 | 2019-04-05 | 福州大学 | A kind of Coumarins hypochlorous acid fluorescence probe and preparation method thereof |
Non-Patent Citations (9)
| Title |
|---|
| H.LI，等: "A novel dual-emission fluorescent probe for the simultaneous detection of H2S and GSH", 《CHEMICAL COMMUNICATIONS 》 * |
| HYOCKMAN KWON,等: "Coumarin–malonitrile conjugate as a fluorescence turn-on probe for biothiols and its cellular expression", 《CHEMICAL COMMUNICATIONS》 * |
| LI FU，等: "The unique substitution-cyclization reaction cascade inspired highly selective H2Sn probe development", 《SENSORS AND ACTUATORS B CHEMICAL》 * |
| WENQIANG CHEN,等: "Rational development of a highly selective ratiometric fluorescent probe for hydrogen polysulfides", 《SENSORS AND ACTUATORS B: CHEMICAL》 * |
| WENQIANG CHEN,等: "Unexpected reaction patterns enable simultaneous differentiation of H2S, H2Sn and biothiols", 《CHEMICAL COMMUNICATIONS》 * |
| 傅立: "基于香豆素母体的活性硫荧光探针的设计、合成及应用", 《中国优秀硕士学位论文数据库 工程科技I辑》 * |
| 夏玉婷,等: "一种用于检测硫化氢和生物硫醇化学传感器的合成与应用", 《遵义医学院学报》 * |
| 张鹏，等: "用于检测多硫化氢和亚硝酰氢的小分子荧光探针", 《影像科学与光化学》 * |
| 陈逢灶: "生物硫醇荧光探针的构建及生物成像研究", 《中国优秀硕士学位论文数据库 工程科技I辑》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110669043A (en) * | 2019-10-12 | 2020-01-10 | 南宁师范大学 | Fluorescent probe for identifying cysteine, homocysteine, glutathione and hydrogen sulfide and preparation method thereof |
| CN111718319A (en) * | 2020-07-27 | 2020-09-29 | 山西大学 | Fluorescent probe for distinguishing and detecting mercaptan and monitoring Cys/GSH metabolism and preparation method thereof |
| CN112939918A (en) * | 2021-02-05 | 2021-06-11 | 山西大学 | Coumarin derivative CTT and synthesis method and application thereof |
| CN112939918B (en) * | 2021-02-05 | 2022-07-19 | 山西大学 | Coumarin derivative CTT and synthesis method and application thereof |
| WO2023097820A1 (en) * | 2021-12-03 | 2023-06-08 | 德州学院 | Flavonol compound, and preparation method therefor and use thereof in detection of biological mercaptan |
| CN115490664A (en) * | 2022-09-29 | 2022-12-20 | 湖南师范大学 | Synthesis and application of fluorescent probe for selectively detecting homocysteine |
| CN115490664B (en) * | 2022-09-29 | 2024-02-13 | 湖南师范大学 | Synthesis and application of fluorescent probe for selectively detecting homocysteine |
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