CN110357817A - A kind of invertibity detects pyroracemic aldehyde and glyoxal fluorescence probe and its preparation method and application - Google Patents
A kind of invertibity detects pyroracemic aldehyde and glyoxal fluorescence probe and its preparation method and application Download PDFInfo
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- CN110357817A CN110357817A CN201910713163.7A CN201910713163A CN110357817A CN 110357817 A CN110357817 A CN 110357817A CN 201910713163 A CN201910713163 A CN 201910713163A CN 110357817 A CN110357817 A CN 110357817A
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- C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
- C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
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- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The invention discloses a kind of invertibitys to detect pyroracemic aldehyde and glyoxal fluorescence probe and its preparation method and application, this method includes by 4- bromo- 1, 8- naphthalene anhydride and amine flow back obtain N-R1-4- bromo- 1 after 3.5h in ethanol, 8- naphthalimide, then, under the conditions of nitrogen protection, N-R1-4- bromo- 1, 8- naphthalimide and guanidine are stirred overnight to obtain invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds under the conditions of ethylene glycol monomethyl ether is at 100 DEG C, with 1, 8- naphthalimide is fluorogen, using forming reversible dihydroxy imidazolidine after guanidine radicals and pyroracemic aldehyde and glyoxal reaction, charge transfer effect increases in excited state molecule, to generate intense fluorescence.Therefore fluorescence probe of the invention can quick, reversible detection pyroracemic aldehyde and glyoxal.
Description
Technical field
The present invention relates to organic syntheses and technical field of analytical chemistry, more particularly to a kind of invertibity detection pyroracemic aldehyde and second
Dialdehyde fluorescence probe and its preparation method and application.
Background technique
Active carbonyl compound, such as formaldehyde, acetaldehyde, pyroracemic aldehyde, glyoxal, methacrylaldehyde etc., play in life system
Important role.Due to the high reaction activity of these compounds and DNA, protein, so these active carbonyl groups
The occurrence and development of other chronic diseases such as compound and diabetes, cancer, obesity, alzheimer's disease, aging are related.Its
Middle pyroracemic aldehyde and glyoxal can induce advanced glycation end products as active carbonyl compound.
Pyroracemic aldehyde mainly passes through and lysine, arginine, and cysteine residues reaction generates glycosylation end products, this is anti-
Answering process mainly influences different protein, such as insulin, hemoglobin and growth factor, can also damage to DNA,
So as to cause cytotoxicity.It is reported that the content of pyroracemic aldehyde is 0.4-1.0 μM in normal human's serum, and diabetic blood
It can reach 2.2-3.8 μM (AnalyticalLetters.2006,39 (10): 2205-2215.) in clear.In order to preferably
Understand function of these active carbonyl compounds under physiology and pathological conditions, need to develop selective enumeration method and monitors biological sample
The method of active carbonyl compound level in product.Most of detection pyroracemic aldehyde is that o-phenylenediamine and acetone aldehyde reaction generate and stablize
Group, such as HPLC, LC-MS, GC-MS is all based on this principle and is used to detect pyroracemic aldehyde.However, these detection methods all need
Cell is cracked, be not suitable for applying in complicated life system.And fluorescence detection is since its is simple, sensitive, has
Biocompatibility can detecte the analyte in life system, to have received widespread attention.
So far, fluorescence detection pyroracemic aldehyde is all using o-phenylenediamine as detection moiety, with pyroracemic aldehyde irreversible reaction
After generate fluorescence, all there is disadvantage in terms of selectivity and reaction speed and dynamic detection in this method.Therefore, develop one newly
Detection pyroracemic aldehyde group be necessary and also be rich in challenge work.
Summary of the invention
The present invention with 1,8- naphthalimide for fluorogen, using forming invertibity after guanidine radicals and pyroracemic aldehyde and glyoxal reaction
Dihydroxy imidazolidine, ICT effect increase, to generate intense fluorescence.Therefore fluorescence probe of the invention can be quick, reversible
The detection pyroracemic aldehyde and glyoxal of property.
In order to realize that the main object of the present invention, the present invention provide:
A kind of fluorescence probe of invertibity detection pyroracemic aldehyde and glyoxal, the fluorescence probe have following general formula I or logical
Formula II or general formula III:
Wherein, R1, R2 are alkyl；N is natural number 1,2 or 3.
Preferably, the R1, R2 are methyl independent, ethyl, propyl.
Preferably, the Formulas I is following formula I-1:
Preferably, the Formula II is following formula II -1:
Preferably, the formula III is following formula II I-1:
The method of fluorescence probe shown in above-mentioned general formula I is synthesized by following route:
This method includes obtaining the bromo- 1,8- naphthoyl of N-R1-4- after the bromo- 1,8- naphthalene anhydride of 4- and amine to be flowed back to 3.5h in ethanol
Imines, then, under the conditions of nitrogen protection, bromo- 1, the 8- naphthalimide of N-R1-4- and guanidine are in ethylene glycol monomethyl ether in 100 DEG C of conditions
Under be stirred overnight to obtain invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds.
Wherein, the molar ratio of bromo- 1, the 8- naphthalene anhydride of 4- and amine is 1:5, obtains intermediate N R1The bromo- 1,8- naphthalimide of -4-
Molar ratio with guanidine is 1:5.
The method of fluorescence probe shown in above-mentioned general formula II is synthesized by following route:
This method is included under the conditions of nitrogen protection, by N-R2The bromo- 1,8- naphthalimide of -4- and methylguanidine are in ethylene glycol monomethyl ether
It is stirred overnight to obtain invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds under the conditions of 100 DEG C, wherein N-R2-4-
The molar ratio of bromo- 1,8- naphthalimide and methylguanidine is 1:5.
The method of fluorescence probe shown in above-mentioned general formula III is synthesized by following route:
This method is included under the conditions of nitrogen protection, and compound 1 and guanidine are stirred under the conditions of ethylene glycol monomethyl ether is at 100 DEG C
Night obtains invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds, wherein the molar ratio of the compound 1 and guanidine is 1:
5.Preferably, the invention also discloses fluorescence probes prepared by the above method third in detection solution, cell or organism
Application in keto-aldehyde and glyoxal.
Invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe of the present invention, make probe in conjunction with pyroracemic aldehyde or glyoxal
Afterwards, ICT effect greatly increases, and fluorescence intensity is caused to increase.Identification reaction is as follows:
Fluorescence probe: there are characteristic fluorescence, and its photoluminescent property (excitation and transmitted wave in ultraviolet-visible-near infrared region
Length, intensity, service life, polarization etc.) it can be with the property of local environment, delicately changing as polarity, refractive index, viscosity etc. change
A kind of fluorescence molecule；Make one kind with nucleic acid (DNA or RNA), protein or other macromolecular structure noncovalent interactions
Or the small-molecule substance that several photoluminescent properties change.It can be used for studying the property and behavior of macromolecular substances.
Compared with prior art, the advantageous effects that the present invention has:
(1) invertibity, invertibity detect the fluorescence probe of pyroracemic aldehyde and glyoxal, realize in intracellular and serum quickly
The level of invertibity detection pyroracemic aldehyde and glyoxal；
(2) high sensitivity, the fluorescence probe property of can choose and pyroracemic aldehyde and glyoxal of invention quickly occur specific anti-
It answers, generates the product with hyperfluorescence, compared to common other aldehydes, amino acid and metal ion, fluorescence of the invention is visited
It is directed to pyroracemic aldehyde and glyoxal shows higher selectivity and sensitivity；
(3) reaction time is fast, and fluorescence probe of the invention and pyroracemic aldehyde and glyoxal react basic at room temperature
It can produce the product with hyperfluorescence in 15min；
(4) it is widely used, potentiality are big, which has good optical stability and specificly-response pyroracemic aldehyde and second
Dialdehyde can be used for the detection of pyroracemic aldehyde and glyoxal level in diabetic animal models serum, have potential practical application valence
Value；
(5) preparation advantage is significant, and fluorescence probe preparation method is simple, and yield is higher.
Detailed description of the invention
Fig. 1 is the nuclear magnetic resonance spectroscopy of probe I -1；
Fig. 2 is the nuclear magnetic resonance spectroscopy of probe I I-1；
Fig. 3 is the nuclear magnetic resonance spectroscopy of probe I II-1；
Fig. 4 is the mass spectrum of probe I -1；
Fig. 5 is the mass spectrum of probe I I-1；
Fig. 6 is the mass spectrum of probe I II-1；
Fig. 7 is the change in fluorescence that probe I -1 is acted on the pyroracemic aldehyde of various concentration；
Fig. 8 is the change in fluorescence that probe I -1 is acted on the glyoxal of various concentration；
Fig. 9 is the change in fluorescence of probe I -1 and pyroracemic aldehyde effect at any time；
Figure 10 is the change in fluorescence of probe I -1 and glyoxal effect at any time；
Figure 11 is probe I -1 and the invertibity research of acetone aldehyde reaction；
Figure 12 is probe I -1 and the reversible Journal of Sex Research of glyoxal reaction；
Figure 13 is the selection Journal of Sex Research of probe I -1；
Figure 14 is the change in fluorescence that probe I I-1 is acted on the pyroracemic aldehyde of various concentration；
Figure 15 is the selection Journal of Sex Research of probe I I-1；
Figure 16 is the common location fluorescence imaging of probe I -1 Yu pyroracemic aldehyde and glyoxal.
Specific embodiment
Below by specific embodiment, the present invention will be further elaborated, it should be appreciated that following the description merely to
In illustrating the present invention, summary of the invention is not defined.
Raw materials used and equipment is that those skilled in the art are known in embodiment, and be can buy in the market or
It is easy to get or is made.
Embodiment 1
The synthesis of fluorescence probe I -1:
Bromo- 1, the 8- naphthalene anhydride (1.1g, 4mmol) of 4- is suspended in 10mL ethyl alcohol, is added dropwise n-propylamine (1.18g, 20mmol),
Heating reflux reaction 3.5h.It puts to room temperature, filtering is washed three times with ethyl alcohol, filter cake drying, obtain product, N- propyl -4- bromo- 1,
8- naphthalimide is faint yellow solid (1.02g, 80.5%).Under the protection of nitrogen, by compound N-propyl -4- bromo- 1,8-
Naphthalimide (382mg, 1.2mmol) is suspended in 5mL ethylene glycol monomethyl ether, then guanidine (355mg, 6mmol) is dissolved in 3mL ethylene glycol first
In ether, at room temperature, the ethylene glycol monomethyl ether solution of guanidine is slowly added dropwise to the suspension of bromo- 1, the 8- naphthalimide of N- propyl -4-
In.Reaction mixture is stirred overnight at 100 DEG C.Ethyl acetate is added, water is extracted.Organic layer is dry with anhydrous sodium sulfate
It is dry, it filters, be spin-dried for, obtain compound I-1 (138mg, 39%) by column chromatography (methylene chloride: methanol=10:1, volume ratio),
For Orange red solid.
The nuclear magnetic resonance spectroscopy of compound I-1 and the structural characterization of high resolution mass spectrum are as shown in Fig. 1 and Fig. 4.Wherein, nuclear-magnetism
Resonate hydrogen modal data:1HNMR(400MHz,DMSO-d6) δ 8.56 (dd, J=8.2,1.1Hz, 1H), 8.44 (dd, J=7.4,
1.1Hz, 1H), 8.32 (d, J=8.2Hz, 1H), 7.72 (dd, J=8.2,7.4Hz, 1H), 7.33 (d, J=8.2Hz, 1H),
6.70 (bs, 4H), 4.00 (m, 2H), 1.63 (h, J=7.4Hz, 2H), 0.90 (t, J=7.4Hz, 3H)；
High resolution mass spectrum (ESI-HRMS): m/z is according to molecular formula C16H17N4O2[M+H] calculated+The theoretical value at peak is
297.1346 measured value is 297.1353.
Embodiment 2
The synthesis of fluorescence probe II-1:
Under the protection of nitrogen, bromo- 1, the 8- naphthalimide (100mg, 0.314mmol) of compound N-propyl -4- is suspended
It is dissolved in 1mL ethylene glycol monomethyl ether in 1mL ethylene glycol monomethyl ether, then by methylguanidine (115mg, 1.57mmol), at room temperature, slowly by first
The ethylene glycol monomethyl ether solution of guanidine is added dropwise in the suspension of the bromo- 1,8- naphthalimide of N- propyl -4-.By reaction mixture 100
It is stirred overnight at DEG C.Ethyl acetate is added, water is extracted.Organic layer is dried, filtered with anhydrous sodium sulfate, is spin-dried for, by column
Chromatography (methylene chloride: methanol=10:1, volume ratio) obtains compound II-1 (27mg, 28%), is Orange red solid.
The structural characterization of the nuclear magnetic resonance spectroscopy of compound II-1 and high resolution mass spectrum is as shown in figures 2 and 5:1HNMR
(400MHz,DMSO-d6) δ 8.63 (dd, J=8.2,1.2Hz, 1H), 8.40 (dd, J=7.4,1.2Hz, 1H), 8.24 (d, J=
8.3Hz, 1H), 7.62 (dd, J=8.2,7.4Hz, 1H), 7.08 (d, J=8.3Hz, 1H), 6.41 (bs, 1H), 6.07 (bs,
2H), 3.99 (m, 2H), 2.82 (s, 3H), 1.62 (h, J=7.4Hz, 2H), 0.90 (t, J=7.4Hz, 3H)；
High resolution mass spectrum (ESI-HRMS): m/z is according to molecular formula C17H19N4O2[M+H] calculated+The theoretical value at peak is
311.1503 measured value is 311.1510.
Embodiment 3
The synthesis of fluorescence probe III-1:
Under the protection of nitrogen, compound a (947mg, 2mmol) is suspended in 15mL ethylene glycol monomethyl ether, then by guanidine
(591mg, 10mmol) is dissolved in 3mL ethylene glycol monomethyl ether, at room temperature, the ethylene glycol monomethyl ether solution of guanidine is slowly added dropwise to a's
In suspension.Reaction mixture is stirred overnight at 100 DEG C.Ethyl acetate is added, water is extracted.The anhydrous sulphur of organic layer
Sour sodium is dried, filtered, is spin-dried for, and obtains compound III-1 by column chromatography (methylene chloride: methanol: triethylamine=200:20:1)
(160mg, 18%) is Orange red solid.
Compound III-1 structural characterization is as shown in Figure 3 and Figure 6:1HNMR(400MHz,DMSO-d6) δ 8.61 (dd, J=
8.3,1.2Hz, 1H), 8.36 (dd, J=7.3,1.2Hz, 1H), 8.22 (d, J=8.3Hz, 1H), 7.72 (bs, 1H), 7.64-
7.59 (m, 3H), 7.26 (d, J=8.1Hz, 2H), 7.12 (d, J=8.3Hz, 1H), 6.17 (bs, 4H), 4.09 (t, J=
6.7Hz,2H),3.03(bs,2H),2.28(s,3H)；
High resolution mass spectrum (ESI-HRMS): m/z is according to molecular formula C22H22N5O4[M+H] that S is calculated+The theoretical value at peak is
452.1387 measured value is 452.1393.
Embodiment 4
The fluorescence spectrum of fluorescence probe I-1 and various concentration pyroracemic aldehyde and glyoxal reaction changes
The fluorescence probe I-1 prepared in Example 1 is dissolved in dimethyl sulfoxide (DMSO), and it is the glimmering of 1mM that concentration, which is made,
Light probe mother liquor；The pyroracemic aldehyde solution that mass fraction is 40% is added in distilled water, it is 10mM's and 100mM that concentration, which is made,
Pyroracemic aldehyde mother liquor；8.8M glyoxal solution is added in distilled water, the glyoxal mother liquor for being 10M and 100M at concentration.According to
Fluorescence probe and the good required PBS aqueous solution (10mM, pH=7.4) of pyroracemic aldehyde/glyoxal concentration calculation be added to 1cm ×
In 1cm quartz colorimetric utensil (volume 3.5mL), takes 6 μ L fluorescence probe mother liquors to be added in PBS aqueous solution, add various concentration
Pyroracemic aldehyde/pyroracemic aldehyde mother liquor (0-500 μM), being configured to concentration and probe concentration is 2 μM of the total 3mL of test solution.After reaction 30 minutes,
Change (excitation wavelength with Fluorescence Spectrometer test fluorescence probe I-1 and various concentration pyroracemic aldehyde/glyoxal reaction fluorescence spectrum
For 425nm).Fluorescence spectrum variation is as shown in Figure 7 and Figure 8.It can be seen that as pyroracemic aldehyde/glyoxal concentration gradually increases, probe
Photoluminescence peak of the solution at 564nm gradually increases.
Embodiment 5
The fluorogram that fluorescence probe I-1 and pyroracemic aldehyde and glyoxal reaction change over time changes
According to the good required PBS aqueous solution (10mM, pH=7.4) of fluorescence probe and pyroracemic aldehyde/glyoxal concentration calculation
It is added to and the fluorescence probe mother liquor in 6 μ L embodiments 1 is taken to be added to PBS water in 1cm × 1cm quartz colorimetric utensil (volume 3.5mL)
In solution, 60 μ L (10mM) pyroracemic aldehyde/glyoxal mother liquors are added, being configured to concentration and probe concentration is 2 μM, and pyroracemic aldehyde/glyoxal is dense
The total 3mL of test solution that degree is 200 μM.Make excitation wavelength with 425nm, tests its fluorogram variation changed over time.Such as
Shown in Fig. 9 and 10, as the time increases, photoluminescence peak is gradually increased at 564nm.
Embodiment 6
The reversible Journal of Sex Research of fluorescence probe I -1 and pyroracemic aldehyde and glyoxal.
According to the good required PBS aqueous solution (10mM, pH=7.4) of fluorescence probe and pyroracemic aldehyde/glyoxal concentration calculation
Be added in 1cm × 1cm quartz colorimetric utensil (volume 3.5mL), take aminoguanidine (AG) 110mg be configured into aqueous solution 10mM and
The mother liquor of 100mM.It takes the fluorescence probe mother liquor in 6 μ L embodiments 1 to be added in PBS aqueous solution, adds 60 μ L (10mM) third
Keto-aldehyde/pyroracemic aldehyde mother liquor, being configured to concentration and probe concentration is 2 μM, adds the survey of various concentration aminoguanidinium salts hydrochlorate (150 μM of -1mM)
Try the total 3mL of solution.Wherein using aminoguanidine as inhibitor.Make excitation wavelength with 425nm, tests it with different glimmering of inhibitor concentration
Spectrogram variation, such as Figure 11 and 12, it is possible to find glimmering at 564nm no matter mother liquor is pyroracemic aldehyde or glyoxal after inhibitor is added
Luminous intensity reduces really, and the amplitude reduced with the increase of inhibitor concentration increases.
Embodiment 7
The selection Journal of Sex Research of -1 pair of disturbance analyte of probe I
According to the good required PBS aqueous solution (10mM, pH=7.4) of the concentration calculation of fluorescence probe and disturbance analyte
It is added to and the fluorescence probe mother liquor in 6 μ L embodiments 2 is taken to be added to PBS water in 1cm × 1cm quartz colorimetric utensil (volume 3.5mL)
In solution, add 200 μM of different analytes: formaldehyde, salicylide, benzaldehyde, triphosgene, nitric oxide donors NOC-18,
Pyroracemic aldehyde, glyoxal.Being configured to concentration and probe concentration is 2 μM, the total 3mL of test solution that disturbance analyte concentration is 200 μM, together
Shi Liuyi only add the skip test sample of probe.After reaction 30 minutes, tested with Fluorescence Spectrometer at the 564nm of different samples
Fluorescent emission intensity (excitation wavelength 425nm).As shown in figure 13, relative to skip test solution, pyroracemic aldehyde and glyoxal
Obvious rising has occurred in the fluorescence of test solution, and the fluorescence enhancement of other analytes is few.Experimental result illustrates fluorescence probe
I-1 has good selectivity pyroracemic aldehyde and glyoxal.
The fluorescence spectrum of 8 probe I I-1 various concentration pyroracemic aldehyde of embodiment changes
The fluorescence probe II-1 prepared in Example 2 is dissolved in DMSO, and it is female that the fluorescence probe II that concentration is 1mM is made
Liquid；According to the good required PBS aqueous solution (10mM, pH=7.4) of the concentration calculation of fluorescence probe and pyroracemic aldehyde be added to 1cm ×
In 1cm quartz colorimetric utensil (volume 3.5mL), takes 6 μ L fluorescence probe mother liquors to be added in PBS aqueous solution, add various concentration
Pyroracemic aldehyde mother liquor (0-1200 μM), being configured to concentration and probe concentration is 2 μM of the total 3mL of test solution.After reaction 30 minutes, fluorescence is used
Spectrometer tests fluorescence probe II-1 and the fluorescence spectrum of various concentration acetone aldehyde reaction changes (excitation wavelength 391nm).It is glimmering
Light spectrum change is as shown in figure 14.It can be seen that gradually increasing with acetone aldehyde concentration, photoluminescence peak of the probe solution at 564nm
It gradually increases.
Embodiment 8
Selection Journal of Sex Research of the probe I I-1 to disturbance analyte
According to the good required PBS aqueous solution (10mM, pH=7.4) of the concentration calculation of fluorescence probe and disturbance analyte
It is added to and the fluorescence probe mother liquor in 6 μ L embodiments 2 is taken to be added to PBS water in 1cm × 1cm quartz colorimetric utensil (volume 3.5mL)
In solution, 400 μM of different analytes are added: for formaldehyde, pyroracemic aldehyde, acetaldehyde, glyoxal, o-phthalaldehyde, glyoxalic acid, benzene
Formaldehyde, glutathione, glucose, cysteine, hydrogen peroxide, potassium ion, calcium ion, sodium ion, copper ion, zinc ion, aluminium
Ion, being configured to concentration and probe concentration is 2 μM, and disturbance analyte concentration is 400 μM of the total 3mL of test solution, while staying one
Only add the skip test sample of probe.Reaction after sixty minutes, tests fluorescent emission at the 564nm of different samples with Fluorescence Spectrometer
Intensity (excitation wavelength 391nm).As shown in figure 15, relative to skip test solution, the test solution of pyroracemic aldehyde and formaldehyde
Obvious rising has occurred in fluorescence, and the fluorescence enhancement of other analytes is few.Experimental result illustrates fluorescence probe II-1 for third
Keto-aldehyde and glyoxal have good selectivity.
Embodiment 9
The common location fluorescence imaging of pyroracemic aldehyde and glyoxal in probe I -1 and HeLa cell
By HeLa cell inoculation in containing 10% fetal calf serum and 1% glutamine DMEM culture medium (containing phenol red,
Gibco/Invitrogen in T-25 Tissue Culture Flask), 48 hours at 37 DEG C.On the day before experiment, cell is transferred to 3
So that cell adherence in Nunc35mm glass bottom Tissue Culture Dish (ThermoScientific).By probe I -1 in PBS buffer solution
Middle dilution, and be added in cell culture medium with 5 μM of final concentration.After being incubated for 1 hour in cell incubator at 37 DEG C, remove
Culture medium containing probe I -1.After being washed twice with PBS buffer solution, by the cell culture containing 20 μM of pyroracemic aldehydes or glyoxal
Base is added in pyroracemic aldehyde and glyoxal culture dish.To the same amount of fresh culture of addition in control (only probe culture dish).?
After being incubated for 30 minutes in cell incubator at 37 DEG C, by cell equipped with 40 × object lens and gain of photomultiplier is
800 (excite and launch wavelength are λex/em=405/530-590nm) LeicaTCSSP8 on be imaged.In first time image-forming step
Afterwards, 5mM acetylcysteine is added in the cell culture medium of pyroracemic aldehyde and glyoxal culture dish.Then by cell at 37 DEG C
It is incubated in cell incubator other 30 minutes.At the end of incubation, equipped with 40 × object lens and gain of photomultiplier
For 800, (excitation and launch wavelength are λex/em=405/530-590nm) LeicaTCSSP8 on capture cell image again.Such as
Shown in Figure 16, first group does not have fluorescence, and second group and the 4th group has obvious fluorescence, third group and the 5th group of ratio second group and the 5th
Group fluorescence weakens.Experimental result illustrate fluorescence probe I -1 can with the invertibity pyroracemic aldehyde intracellular to HeLa and glyoxal into
Row fluorescence imaging.
Above embodiments, only preferred embodiments of the invention, be not to limit the scope of the present invention, therefore it is all according to the present invention
The equivalent change or modification that structure, feature and principle described in claim is done should be included in the invention patent Shen
It please be in range.
Claims (15)
1. a kind of fluorescence probe of invertibity detection pyroracemic aldehyde and glyoxal, which is characterized in that the fluorescence probe has as follows
General formula I or general formula II or general formula III:
Wherein, R1, R2 are alkyl；N is natural number 1,2 or 3.
2. the fluorescence probe of invertibity detection pyroracemic aldehyde and glyoxal as described in claim 1, which is characterized in that the R1,
R2 is methyl, ethyl or propyl independent.
3. invertibity as claimed in claim 2 detection pyroracemic aldehyde and glyoxal fluorescence probe, which is characterized in that the Formulas I is
Following formula I-1:
4. invertibity as claimed in claim 2 detection pyroracemic aldehyde and glyoxal fluorescence probe, which is characterized in that the Formula II is
Following formula II -1:
5. invertibity detection pyroracemic aldehyde as described in claim 1 and glyoxal fluorescence probe, which is characterized in that the formula III
For following formula II I-1:
6. a kind of method for preparing fluorescence probe shown in claim 1 formula of I, which is characterized in that this method is by following road
Line synthesis:
7. a kind of method for preparing fluorescence probe shown in claim 1 formula of II, which is characterized in that this method is by following road
Line synthesis:
8. a kind of method for preparing fluorescence probe shown in claim 1 formula of III, which is characterized in that this method is by following
Route synthesis:
9. the method for preparation invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe as claimed in claim 6, which is characterized in that
This method includes obtaining bromo- 1, the 8- naphthalimide of N-R1-4- after bromo- 1, the 8- naphthalene anhydride of 4- and amine to be flowed back to 3.5h in ethanol, so
Afterwards, under the conditions of nitrogen protection, bromo- 1, the 8- naphthalimide of N-R1-4- and guanidine are stirred under the conditions of ethylene glycol monomethyl ether is at 100 DEG C
Night obtains invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds.
10. the method for preparation invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe, feature exist as claimed in claim 9
In the molar ratio of bromo- 1, the 8- naphthalene anhydride of the 4- and amine is 1:5, obtains bromo- 1, the 8- naphthalimide of intermediate N R1-4- and guanidine
Molar ratio is 1:5.
11. the method for preparation invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe, feature exist as claimed in claim 7
In this method is included under the conditions of nitrogen protection, by bromo- 1, the 8- naphthalimide of N-R2-4- and methylguanidine in ethylene glycol monomethyl ether 100
It is stirred overnight to obtain invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds under the conditions of DEG C.
12. the method for preparation invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe, feature exist as claimed in claim 11
In the molar ratio of bromo- 1, the 8- naphthalimide of N-R2-4- and methylguanidine is 1:5.
13. the method for preparation invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe, feature exist as claimed in claim 12
In this method is included under the conditions of nitrogen protection, and compound (1) and guanidine are stirred overnight under the conditions of ethylene glycol monomethyl ether is at 100 DEG C
Obtain invertibity detection pyroracemic aldehyde and glyoxal fluorescent probe compounds.
14. the method for preparation invertibity detection pyroracemic aldehyde and glyoxal fluorescence probe, feature exist as claimed in claim 13
In the molar ratio of the compound (1) and guanidine is 1:5.
15. a kind of fluorescence probe as described in claim 1-14 Arbitrary Term pyroracemic aldehyde in detection solution, cell or organism
With the application in glyoxal.
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