CN108395427A - Kit for detecting cysteine - Google Patents
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- CN108395427A CN108395427A CN201810155918.1A CN201810155918A CN108395427A CN 108395427 A CN108395427 A CN 108395427A CN 201810155918 A CN201810155918 A CN 201810155918A CN 108395427 A CN108395427 A CN 108395427A
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- 0 CC1(C2[C@@](*)C1)C(C)=*(C)C1=C2C(*)C(*)C(*)=*(*)*1 Chemical compound CC1(C2[C@@](*)C1)C(C)=*(C)C1=C2C(*)C(*)C(*)=*(*)*1 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/10—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing aromatic rings
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- 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"
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
- C09K2211/1037—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom with sulfur
Abstract
The present invention relates to the kits for detecting cysteine.The kit of the present invention can be acted on cysteine, and have the characteristics that high sensitivity and selectivity, can be applied in cysteine high sensitivity and highly selective identification or its can in determination sample cysteine concentration.
Description
Technical field
The present invention relates to the kits for detecting cysteine.The kit of the present invention can be acted on cysteine,
And have the characteristics that high sensitivity and selectivity, it can be applied in cysteine high sensitivity and highly selective identification, or
It can in determination sample cysteine concentration.
Background technology
There are many kinds of the types of mercaptan, but there are three types of small molecule biological thiols, respectively homocysteine, glutathione
And cysteine.And wherein cysteine is present in many small molecules and protein of biological tissue cell, is to form them
Important substance, and cysteine played in mammal regulating system adjusts a variety of physiology and pathologic process it is quite important
Effect, mainly by the reproducibility speciality of cysteine can be oxidized to oxidisability substance maintain cellular environment in
Redox equilibrium.Homocysteine plays an important role in organism, it is methionine in some reactions
Intermediate product, rear once being formed there are two types of outlets, first, methionine is re-converted into, second is that the shape under the catalysis of some substances
At cysteine.Glutathione is non-protein sulfydryl most abundant in human body, in the cell there are many function, including removing toxic substances
Free radical and peroxide adjust the function of cell growth and protein, and safeguard immune function, so, glutathione it is dense
Spend index that can be as assessment redox state and in biologic detoxication state status in organism.But cysteine is in people
It is to play one of vital amino acid in body, it can change the disulfide bond between protein molecule to weaken protein sky
Between structure, to enable protein to extend.In addition cysteine also have removing toxic substances, anti-aging, change inflammation and in advance
Anti- treatment radiation disease.A large amount of investigation show that the biological thiol beyond normal level has been demonstrated the generation with human diseases
It is related, it such as slowly grows, liver is impaired and skin damage.Cysteine deficiency can lead to many diseases, including retarded growth, liver damage
Bad, drowsiness, muscle weakness and fat loss.The homocysteine of human blood middle and high concentration is to cause nerve channel effect, bone
The risk factor of the diseases such as matter osteoporosis and inflammatory bowel disease.
Currently, the method for quantitatively detection cysteine is varied, main method includes:Titration method, ultraviolet suction
Receipts method, molecular fluorescent method, emission spectrometry method, mass spectrometry etc..Several method in contrast, utilizes molecular fluorescence
The fluorescence probe of analysis has an apparent advantage, including it is highly selective, have it is highly sensitive, can fast reaction, detection in time,
Cell imaging etc..The probe for being used for detecting cysteine at present has very much, but the fluorescence probe of Ratio-type near-infrared and can
Carry out the seldom of bio-imaging.
On the other hand, Ratiometric fluorescent probe can mitigate ambient enviroment using the variable value between two adjacent peaks
Influence to cysteine detection, so the practical value with bigger.
To sum up, it is very intentionally to establish a kind of kit for the Ratio-type near infrared fluorescent probe that can detect cysteine
Justice.
Invention content
A kind of high sensitivity is badly in need of in this field and highly selective Ratio-type measures the kit of cysteine, so as to have
Effect detection especially can detect cysteine in the sample.For this purpose, the present invention proposes a kind of novel detection cysteine
Kit, can highly sensitive and highly selective Ratio-type identify cysteine.The kit of the present invention is related to using probe
Carry out high sensitivity to cysteine and highly selective measurement.
Specifically, the present invention provides a kind of kit of detection cysteine, it includes the spies of identification cysteine
Needle, structure are as follows:
Wherein:R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R13For independently selected from by hydrogen atom, straight chain or
The group of branched alkyl, straight or branched alkoxyl, sulfonic group, ester group and hydroxyl composition;And R therein1, R2, R3, R4, R5, R6,
R7, R8, R9, R10, R11, R12And R13It can be identical or different.
Preferably, heretofore described linear or branched alkyl group, straight or branched alkoxyl are the alkyl or alcoxyl of C1-10
Base.
Preferably, heretofore described linear or branched alkyl group, straight or branched alkoxyl are the alkyl or alcoxyl of C1-5
Base.
Preferably, heretofore described linear or branched alkyl group, straight or branched alkoxyl are the alkyl or alcoxyl of C1-3
Base.
Preferably, heretofore described linear or branched alkyl group be methyl, ethyl, propyl, n-pentyl, 2- methyl butyls,
Isobutyl group or 4- methylheptyls.
Preferably, heretofore described straight or branched alkoxyl is methoxyl group, ethyoxyl or propoxyl group.
Preferably, R in the present invention1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R3For independently selected from hydrogen original
Son, methyl, ethyl, propyl, methoxyl group, ethyoxyl or propoxyl group.
Preferably, the Ratio-type probe in kit of the present invention is:
Ratiometric fluorescent probe in kit of the present invention can be acted on cysteine, lead to the change of fluorescence spectrum
Change, the Ratio-type of cysteine is quantitatively detected to realize.
Specifically, cysteine Ratiometric fluorescent probe in kit of the present invention respectively with Cys, Gly, Pro, Leu,
Val, Thr and Ala, which are acted on, cannot lead to substantially changeing for fluorescence spectrum, to realize the selectivity to cysteine
Identification, and then can be optionally used for excluding the interference of the quantitative determination of the presence of these amino acid to cysteine.
The stability of cysteine colorimetric probe in kit of the present invention is good, being capable of long-term preservation use.
Further, the cysteine probe in kit of the present invention being capable of highly sensitive and highly selective Ratio-type measurement half
Cystine, and further organic synthesis modification need not be done, be conducive to commercialized popularization and application.
Description of the drawings
Fig. 1:Probe has good response for the cysteine of various concentration.Probe (5 μM) and cysteine (0,
0.1,0.2,0.3 0.4,0.5,0.6,0.7,0.8,0.9,1,1.5,2,2.5,3,3.5,4,4.5,5,5.5,6,6.5,7,
7.5,8 μM) it is put into sepectrophotofluorometer and is detected after reaction 10min.
Fig. 2:Have at a concentration of 0 to 8 μM linear well, detection is limited to 1 μM.Probe (5 μM) and cysteine (0,
0.1,0.2,0.3 0.4,0.5,0.6,0.7,0.8,0.9,1,1.5,2,2.5,3,3.5,4,4.5,5,5.5,6,6.5,7,
7.5,8 μM) it is put into sepectrophotofluorometer and is detected after reaction 10min.
Fig. 3:When other amino acid (Cys, Gly, Pro, Leu, Val, Thr, Ala) are 100 μM a concentration of, probe (5 μM)
To their fluorescence response value, and the response of probe after cysteine is inside added again.It is second to detect solution system
Alcohol:Water=5:5 (v/v) contain PBS=5mM, pH=7.4, and test temperature is 25 DEG C.
Specific implementation mode
The present invention provides the kits that a kind of highly sensitive and highly selective Ratio-type measures cysteine.
Cysteine Ratio-type probe used in kit of the present invention has following structure general formula:
Wherein:R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R13For independently selected from by hydrogen atom, straight chain or
The group of branched alkyl, straight or branched alkoxyl, sulfonic group, ester group and hydroxyl composition;And R therein1, R2, R3, R4, R5, R6,
R7, R8, R9, R10, R11, R12And R13It can be identical or different.
Preferably, heretofore described linear or branched alkyl group, straight or branched alkoxyl are the alkyl or alcoxyl of C1-10
Base.
Preferably, heretofore described linear or branched alkyl group, straight or branched alkoxyl are the alkyl or alcoxyl of C1-5
Base.
Preferably, heretofore described linear or branched alkyl group, straight or branched alkoxyl are the alkyl or alcoxyl of C1-3
Base.
Preferably, heretofore described linear or branched alkyl group be methyl, ethyl, propyl, n-pentyl, 2- methyl butyls,
Isobutyl group or 4- methylheptyls.
Preferably, heretofore described straight or branched alkoxyl is methoxyl group, ethyoxyl or propoxyl group.
Preferably, R in the present invention1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R13For independently selected from hydrogen original
Son, methyl, ethyl, propyl, methoxyl group, ethyoxyl or propoxyl group.
Preferably, R in the present invention1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R13Selected from hydrogen, methyl, second
Base or propyl;Most preferably, R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R13It is H.
Kit of the present invention can highly sensitive and highly selective Ratio-type measure cysteine and/or in other amino acid
In the presence of can accurately to cysteine carry out quantitative analysis.
The kit of the present invention has the following advantages that:
1, there is very good selectivity to cysteine, and for other amino acid then without apparent response;
2, the probe response time is 20mi n, therefore can be as the means of quick detection cysteine;And/or
3, the detection of the kit is limited to 1 μM, the cysteine for 1 to 8 μM of the detection that can be quantified, and sensitivity is relatively high.
There are one the blue shifts of 170nm after probe in the kit is reacted with cysteine, this is in detection cysteine
It is fewer in the probe of Ratio-type near-infrared class.
It below will be by the way that the present invention be described in more detail by following embodiment.Following embodiment is merely illustrative,
It should be understood that the present invention is not limited by following embodiments.
It is prepared by 1 probe of embodiment
2,6- diformyl phenol synthesizes and purification
It weighs in the balance and phenol 940.5mg and hexamethylenetetramine 2.80g is taken to be put into the glass reaction bottle of 100mL, be added
The trifluoroacetic acid of 30mL is put into the small magneton of a wash clean, is heated to reflux later using 84-1 magnetic agitation control-temperature electric heating sets,
Recording reacting time tracks process in reaction process using tlc silica gel plate, about needs reaction 6h, after the reaction was complete, turns off
Reaction.Synthetic reaction is shown in Formulas I:
Formulas I:The synthesis of 2,6- diformyl phenol
The beaker for preparing a 100mL, pours the distilled water of 50mL or so into wherein.Stop in reaction, it, will be anti-after cooling
Solution is answered to be added dropwise in preprepared beaker,
It is stirred evenly with dropper, the liquid in beaker is poured into separatory funnel, then add 20mL's or so wherein
Dichloromethane, fastening plug rock with strength it is several under, separatory funnel is placed on brandreth stands a few minutes later, make liquid point
Layer.Since the density ratio water of dichloromethane is big, so dichloromethane solution is below, our product is organic matter, generally not
It is dissolved in water, to be completely dissolved in dichloromethane during extraction.The piston for opening separatory funnel makes dichloromethane be flowed from lower part
Go out.Then the dichloromethane extraction of 20mL or so is added in remaining liquid in separatory funnel again, repeatedly repeatedly 3 to 5
It is secondary, make product completely into dichloromethane.
After having extracted, we are extracted obtained two by the glass reaction bottle of one 25mL of weighing with this flask
Chloromethanes liquid is spin-dried for Rotary Evaporators.Then it takes the substance being spin-dried for a little in the centrifuge tube of 4mL, adds a small amount of two
Chloromethanes makes it dissolve, and a little solution is pipetted with capillary glass tube, the contact plate on tlc silica gel plate, later by thin-layer chromatography
Silica gel plate is put into the glass jar containing about 1mL dichloromethane, about waits for 3mi n quietly, until dichloromethane liquid soon will be thin
Tlc silica gel plate is taken out with tweezers when analysis silica gel plate all soaks layer by layer, with the 365nm in portable uv analyzer
Fluorescent lamp excitation in the fluorescent lamp excitation of 254nm, without other points, illustrate extraction rotation it is observed that a phosphor dot
The solid obtained after dry is purer, can directly cast single step reaction.It weighs again after equal flasks cooling, obtains the weight of product
For 976.5mg, yield 65.1%.
The synthesis and purification of 2- formoxyl -6- benzothiazole phenol
Product 2,6- diformyl phenol obtained above is weighed into 450.0mg and near amino thiophenols weigh 375.6mg
It is put into the glass reaction bottle of 100mL, adds 25mL ethanol solutions, so that two kinds of drugs is completely dissolved, be put into wash clean
Small magneton, glass reaction bottle is placed on the strong magnetic stirring apparatus of 84-B is stirred at room temperature later.It is added during stirring
37% hydrochloric acid solution of 9mmol is eventually adding 30% hydrogenperoxide steam generator of 18mmol.Continue to be stirred at room temperature
30min or so.Synthetic reaction is shown in Formula II.
Formula II:The synthesis of 2- formoxyl -6- benzothiazole phenol
Reaction is completed, and solid matter is sucked out using dropper, then filters, is rinsed with absolute ethyl alcohol.
A little upper layer product is pipetted in centrifuge tube, is dissolved in a small amount of dichloromethane, is pipetted a little with capillary glass tube
Solution, the contact plate on tlc silica gel plate put the silica gel plate put into glass wide-mouth containing about 1mL dichloromethane later
In bottle, about wait for 3min quietly, until dichloromethane liquid soon all soaks tlc silica gel plate with tweezers by thin-layer chromatography
Silica gel plate takes out, and is excited with the fluorescent lamp of the 365nm in portable uv analyzer, it is observed that a phosphor dot,
When the fluorescent lamp excitation of 254nm, there are one the stain of very little, illustrate that the solid that suction filtration obtains is purer, since this is not final
Product, purity reach 80% or more and may be used for reacting in next step, so we do not purify further, is directly used in down
Single step reaction.
A period of time is filtered, until after solid parches completely, is pipetted from funnel solid using spoon small to 25mL
It in glass reaction bottle, weighs to small glass reaction bottle, weighs again after being put into product in advance, to obtain the weight of product
For 521.6mg, yield 68.2%.
The preparation and purification of N- methyl -2- methylquinoline salt
In the glass reaction bottle for weighing the 50mL for taking 300mg 2- methylquinolines to be put into wash clean in the balance, the iodine of 20mL is poured into
Methane is put into magneton, and or so 4 hours are heated to reflux with 84-1 magnetic agitation control-temperature electric heating sets.Synthetic reaction is shown in formula III.
Formula III:The synthesis of N- methyl -2- methylquinoline salt
Flask is removed after having reacted, after cooling, has solid precipitation, is filtered with Suction filtration device, obtain solid,
About 285mg after weighing, yield 98%.
The synthesis and purification of 2- (N- methyl -2- vinyl) quinoline -6- benzothiazole phenol
Weigh product 2- formoxyl -6- benzothiazole phenol 255mg and product N- methyl -2- methylquinoline salt 285mg
It is put into the glass reaction bottle of the 50mL of wash clean, pours the absolute ethyl alcohol of 20mL or so into, be as possible completely dissolved 2 kinds of drugs,
It puts small magneton into later, is heated to reflux with 84-1 magnetic agitation control-temperature electric heating sets, react 12 hours.
Formula IV:The synthesis of 2- (N- methyl -2- vinyl) quinoline -6- benzothiazole phenol
After reaction, reaction dissolvent absolute ethyl alcohol is removed by revolving, and the dichloromethane that 15mL is added makes it dissolve.
Then solution is added dropwise in chromatographic column, the weight for obtaining sterling is 108.9mg, yield 20.8%.
1The analysis of H-NMR results
Hydrogen modal data is as follows:1H-NMR(400MHz,DMSO-d6)δ(*10-6):4.23 (s, 3H), 7.16 (t, J=
7.6Hz, 1H), 7.25-7.27 (m, 4H), 7.52 (t, J=7.0Hz, 1H), 7.59 (t, J=7.0Hz, 1H), 7.64 (d, J=
16.9Hz, 1H), 7.90 (d, J=8.0Hz, 1H), 7.99 (d, J=8.0Hz, 1H), 8.10 (d, J=8.0Hz, 1H), 8.15
(d, J=16.9Hz, 1H), 8.20 (d, J=8.0Hz, 1H), 8.22 (d, J=7.6Hz, 1H), 8.83 (d, J=7.6Hz, 1H),
13.32(s,1H).Data above meets the structural formula feature of 2- (N- methyl -2- vinyl) quinoline -6- benzothiazole phenol.
13The analysis of C-NMR results
Carbon modal data is as follows:13C-NMR(100MHz,DMSO-d6)δ(*10-6):46.28,114.26,116.27,
117.19,118.64,119.16,121.32,121.76,123.18,123.53,123.97,124.38,125.47,126.29,
127.62,130.36,131.28,131.38,134.25,144.47,150.18,152.47,155.29,167.36.The above number
According to the structural formula feature for meeting 2- (N- methyl -2- vinyl) quinoline -6- benzothiazole phenol.
2 solution of embodiment is prepared
The final products for weighing 5.22mg are configured to the probe mother liquor of 1mM.The cysteine for weighing 12.1mg is configured to
Cysteine (Cys) mother liquor of 10mM.
50 μ L liquid dilutings are pipetted from probe mother liquor to 10mL, are made into 5 μM of probe in detecting liquid.By 10mL probe in detecting
Liquid is divided into two, and is added the cysteine mother liquor of 25 μ L in portion wherein, a concentration of 50 μM.
It is above-mentioned for prepare the system of solution to be ethyl alcohol:Water=5:5 (v/v) contain PBS=5mM, pH=7.4.
3 response time of embodiment identifies probe the influence of cysteine
5 μM of probe in detecting liquid is made into as described in embodiment 2, rapidly joining cysteine mother liquor later makes its concentration
Be 10 μM, rock it is several under, it is quickly put into sepectrophotofluorometer after keeping the dispensed materials in solution uniform, be arranged fluorescence
Thus spectrophotometer obtains the response spectrum of different time in every 30 seconds run-downs, recording light spectrogram simultaneously preserves, when detection
Room temperature is 25 DEG C.Detection solution system remains as ethyl alcohol:Water=5:5 (v/v) contain PBS=5mM, pH=7.4.As a result such as Fig. 1 institutes
Show.Find out that pure probe solution peak value at 700nm is relatively high from fluorescence spectra, peak is there is no at 530nm, is added
After cysteine, spectrogram is that the peak value at 700nm slowly reduces, and the peak value at 530nm slowly increases.
Quantitative effect of 4 probe of embodiment to cysteine
The same probe in detecting liquid for preparing 5 μM, and probe in detecting liquid is added in colorimetric cylinder and is settled to 10mL.Pipette half Guang
Propylhomoserin solution sequentially adds in colorimetric cylinder, makes the concentration of cysteine be finally:0,0.1,0.2,0.3 0.4,0.5,0.6,
0.7,0.8,0.9,1,1.5,2,2.5,3,3.5,4,4.5,5,5.5,6,6.5,7,7.5,8 μM and it is made to be evenly distributed, detected
Solution system is ethyl alcohol:Water=5:5 (v/v) contain PBS=5mM, pH=7.4, are existed with sepectrophotofluorometer after placing 10min
Excitation wavelength is to be detected under 430nm and keep spectrogram.Room temperature is 25 DEG C when detection.The results are shown in Figure 2.Probe pair
There is good response in the cysteine of various concentration.Have at a concentration of 0 to 8 μM linear well, detection is limited to 1 μM.
It can be seen that the probe synthesized by the present invention can detect cysteine by comparing the method for the fluorescence intensity at two neighboring peak
And there is good sensitivity.
Anti-interference when 5 probe of embodiment identifies cysteine
Prepare 7 colorimetric cylinders probe solution constant volumes be added to arrive 10mL, only 10 μM of cysteine of addition in first is remaining
Cys, Gly, Pro, Leu, Val, Thr, Ala are sequentially added, a concentration of 100 μM, then adds 10 μM of cysteine.It is same quiet
After setting 10min, since the colorimetric cylinder of first, left side, it is sequentially placed into sepectrophotofluorometer and detects, and keep data.Inspection
Survey solution system is ethyl alcohol:Water=5:5 (v/v) contain PBS=5mM, pH=7.4, and room temperature is 25 DEG C when detection.As a result such as Fig. 3 institutes
Show.
It can be seen from the figure that other thiol solutions being added in probe solution after reaction a period of time, in solution
The fluorescence intensity of probe is not responded to without too big variation, explanation, but after cysteine inside is added again, probe is just
It will appear relatively high response.Therefore, probe has good selectivity, anti-interference to cysteine.
It can be seen that the probe used in kit of the present invention be one have to cysteine it is highly selective and good
The probe of anti-interference.
Although with above embodiments describe the present invention, it should be appreciated that before the spirit without departing substantially from the present invention
It puts, the present invention further can be modified and be changed, and these modifications and variation all belong to the scope of protection of the present invention it
It is interior.
Claims (10)
1. for detecting the kit of cysteine, it includes the compound that it has following structure,
Wherein:R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R3For independently selected from by hydrogen atom, linear chain or branched chain
The group of alkyl, straight or branched alkoxyl, sulfonic group, ester group and hydroxyl composition;And R therein1, R2, R3, R4, R5, R6, R7, R8,
R9, R10, R11, R12And R13It can be identical or different.
2. kit according to claim 1, wherein the linear or branched alkyl group, straight or branched alkoxyl are C1-
10 alkyl or alkoxy.
3. kit according to claim 1, wherein the linear or branched alkyl group, straight or branched alkoxyl are C1-5
Alkyl or alkoxy.
4. kit according to claim 1, wherein the linear or branched alkyl group, straight or branched alkoxyl are C1-3
Alkyl or alkoxy.
5. kit according to claim 1, wherein the linear or branched alkyl group is methyl, ethyl, propyl, positive penta
Base, 2- methyl butyls, isobutyl group or 4- methylheptyls.
6. kit according to claim 1, wherein the straight or branched alkoxyl is methoxyl group, ethyoxyl or the third oxygen
Base.
7. kit according to claim 1, wherein:R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12And R13For
Independently selected from hydrogen atom, methyl, ethyl, propyl, methoxyl group, ethyoxyl or propoxyl group.
8. kit according to claim 1, for the compound such as lower structure:
9. kit according to claim 1 also includes specification and diluent, and the diluent is ethyl alcohol:Water
=5:The solution of 5 (v/v).
10. kit according to claim 9 also includes buffer solution.
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
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CN110885312A (en) * | 2019-12-17 | 2020-03-17 | 济南大学 | Golgi-targeted cysteine fluorescent probe, and preparation method and application thereof |
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2018
- 2018-02-23 CN CN201810155918.1A patent/CN108395427A/en active Pending
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Title |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110885312A (en) * | 2019-12-17 | 2020-03-17 | 济南大学 | Golgi-targeted cysteine fluorescent probe, and preparation method and application thereof |
CN110885312B (en) * | 2019-12-17 | 2022-04-12 | 济南大学 | Golgi-targeted cysteine fluorescent probe, and preparation method and application thereof |
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