CN106867513A - A kind of cell membrane localization zinc ion fluorescent and its preparation method and application - Google Patents
A kind of cell membrane localization zinc ion fluorescent and its preparation method and application Download PDFInfo
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Abstract
A kind of cell membrane localization zinc ion fluorescent and its preparation method and application.With 1,8- naphthalimides for fluorescent parent, acid amides-lutidines amine is recognition group to the fluorescence probe, and long alkyl chain is seeking group, and long alkyl chains are n, and fluorescence probe general structure is as follows：Wherein 4≤n≤18, and n are integer.Probe synthesis step is simple, and yield is high, good light stability, with Zn2+With reference to rear maximum emission wavelength red shift to 512nm, about 18 times of Fluorescence Increasing.With existing Zn2+Probe is compared, and the probe is not only to Zn2+There is high selectivity, while cell membrane localization can be realized, have extremely important application value in biomedical sector.
Description
Technical field
This is the invention belongs to bioanalysis detection field, and in particular to a kind of detection zinc ion of cell membrane localization is glimmering
Light probe, its synthetic method and biomedical applications.
Background technology
Fluorescence probe has turned into the important tool in RESEARCH ON CELL-BIOLOGY, and it enables researcher in complex biological ring
Specific ion, molecule and biochemical reaction process are intuitive to see in border.Positioned by subcellular organelle, it is glimmering
Light probe can detect the target analytes in subcellular organelle, and can disclose different physics in cell specific region
And chemical property.Cell membrane is the barrier for preventing ECM to be freely accessible to cell, it is ensured that intracellular loops
The stabilization in border.In addition to barrier action is provided, the field that cell membrane or various metabolism and signal transduction occur
Institute.Therefore, the life in helping to understand cell to the real-time monitoring of target analytes and imaging on cell membrane
Change course of reaction, while also can in time reflect the growth conditions of cell, so as to further enter to relevant disease
Row early stage is diagnosed and treated.
Zinc ion has been demonstrated influence multiple cellular physiological processes such as including propagation, differentiation, apoptosis, it is considered to be
The indispensable trace element of organism.In vivo, zinc ion with protein binding except forming metal egg
White compound, also there are the free zinc ion of free state.In the different tissues of organism, free zinc from
Sub- content differs widely, and with nervous system, islet tissue, content is enriched the most in prostata tissue, and
The specific function of these tissues is inseparable with the presence of zinc ion.By to these organize in free zinc from
The detection of son helps to diagnose the early stage of A Ermozihaimo diseases, diabetes, prostate cancer.
The content of the invention
It is an object of the invention to provide a kind of cell membrane localization zinc ion fluorescent and preparation method and application,
The probe can realize cell membrane localization under complex environment and have single-minded selectivity and higher to zinc ion
Sensitivity, its synthetic method has that raw material is cheap and easy to get, synthesis step simple, is adapted to amplify the excellent of synthesis
Point.
The invention provides a kind of detection zinc ion fluorescent that can realize cell membrane localization, the fluorescence probe with
1,8- naphthalimide is fluorescent parent, and acid amides-lutidines amine is recognition group, and long alkyl chain is seeking group,
Long alkyl chains are n, and fluorescence probe general structure is as follows：
Wherein 4≤n≤18, and n are integer.
The present invention provides the preparation method of cell membrane localization zinc ion fluorescent, and the method is comprised the following steps that：
1) synthesis of 4- nitros -1,8- naphthalene anhydrides：Sodium dichromate glacial acetic acid is dissolved in 0.1-0.5g/mL ratios, then
Add the 5- nitro acenaphthenes relative to 0.1-0.5 times of quality of sodium dichromate, back flow reaction 3-8 at 120-150 DEG C
Hour.Reaction solution is mixed with the frozen water of 2-5 times of volume, filter cake is filtrated to get and is recrystallized, obtained
4- nitro -1,8- naphthalene anhydrides.
2) synthesis of 4- amino -1,8- naphthalene anhydrides：By 4- nitro -1,8- naphthalene anhydrides with absolute ethyl alcohol in 0.1-0.5g/mL ratios
Dissolving, will press 1-2g/mL relative to 4- nitro -1, the stannous chloride of 3-5 times of quality of 8- naphthalene anhydrides with concentrated hydrochloric acid
After dissolving, above-mentioned 4- nitro -1 is dropwise instilled, in 8- naphthalene anhydrides and absolute ethyl alcohol, back flow reaction 8- at 80-100 DEG C
12 hours overnight.Reaction solution separates out product with the sodium carbonate liquor tune neutrality of 1-5mol/L, filters, filter
Cake is recrystallized, and obtains 4- amino -1,8- naphthalene anhydrides.
3) synthesis of 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides：4- amino -1,8- naphthalene anhydrides are pressed into 0.01- with dry THF
0.05g/mL ratios, add relative to 4- amino -1, the chloracetyl chloride of 5-10 times of quality of 8- naphthalene anhydrides, at room temperature
After stirring 3-10 hours, reaction system is become by muddiness to be clarified.Solvent is spin-dried for, column chromatography for separation is spin-dried for obtaining
4- (2- chloracetyls) amino -1,8- naphthalene anhydrides.
4) synthesis of 4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides：By 4- (2- chloracetyls) amino -1,8- naphthalenes
Acid anhydride, DPA, DIPEA and KI in mass ratio 1:0.5-0.8:1.5-2:0.2-0.5 mixes, and adds acetonitrile to press
0.01-0.05g/mL ratios dissolve 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides, the lower 80-100 DEG C of backflow of nitrogen protection
8-12 hours overnight.Reaction is spin-dried for solvent after terminating, with the recrystallizing methanol for being spin-dried for 5-10 times of quality of product,
Obtain 4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides.
5) synthesis of probe molecule：4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides, alkylamine are pressed into quality
Than 1:1-2 mixes, and adds ethanol to dissolve 4- (2- lutidines amidos acetyl) in 0.01-0.05g/mL ratios
Amino -1,8- naphthalene anhydrides are heated to 70-90 DEG C and flow back 8-12 hours overnight.After being spin-dried for solvent, column chromatography for separation
Obtain probe molecule.Above-mentioned steps 5) in, the integer between a length of 4-18 of alkylamine carbochain.
Above-mentioned steps 5) in, the eluant, eluent of the column chromatography is dichloromethane:Methyl alcohol=50：1, flow velocity 10-20
Ml/min, 20-25 DEG C of temperature.
Above-mentioned steps 3) described in column chromatography eluant, eluent be dichloromethane, flow velocity 10-20ml/min, temperature 20
-25℃。
Specific synthetic route is as follows：
The fluorescence probe that the present invention is provided can be applied to cell membrane localization detection zinc ion and its in biomedicine
Detection to zinc ion.
The present invention has following characteristics：
Probe molecule preparing raw material is easy to get, and synthetic route is simple, convenient post-treatment.Probe recognizes energy to zinc ion
Power is single-minded, fast response time.Compared with existing zinc ion probe, the probe can be realized positioning to cell membrane,
Release of the monitoring cell to zinc ion, has extremely important application value in biomedical sector.
Brief description of the drawings
The structural formula of Fig. 1 fluorescence probes of the present invention；
Fig. 2 fluorescence probe synthetic route charts of the present invention；
Alkyl carbon chain n=12 fluorescence probes C12 nuclear magnetic spectrograms hydrogen spectrum long prepared by Fig. 3 embodiment of the present invention 1；
Alkyl carbon chain n=14 fluorescence probes C14 nuclear magnetic spectrograms hydrogen spectrum long prepared by Fig. 4 embodiment of the present invention 2；
Alkyl carbon chain n=18 fluorescence probes C18 nuclear magnetic spectrograms hydrogen spectrum long prepared by Fig. 5 embodiment of the present invention 3；
Alkyl carbon chain n=12 fluorescence probes C12 nuclear magnetic spectrograms carbon spectrum long prepared by Fig. 6 embodiment of the present invention 1；
Alkyl carbon chain n=14 fluorescence probes C14 nuclear magnetic spectrograms carbon spectrum long prepared by Fig. 7 embodiment of the present invention 2；
Alkyl carbon chain n=18 fluorescence probes C18 nuclear magnetic spectrograms carbon spectrum long prepared by Fig. 8 embodiment of the present invention 4；
The fluorescence probe of n=12 prepared by Fig. 9 embodiments 1 adds the change in fluorescence feelings after different metal ions
Condition；
The fluorescence probe of n=12 prepared by Figure 10 embodiments 1 and the Zn of various concentrations2+Fluorescence light after effect
Spectrogram；
The fluorescence probe of n=12 prepared by Figure 11 embodiments 1 cell membrane localization effect in colon cancer cell；
The fluorescence probe of n=12 prepared by Figure 12 embodiments 1 co-focusing imaging in prostatic cell RWPE-1
As a result, wherein, a, b, c are respectively the bright field figure of cell, and d is to add the cell after probe culture 30 minutes
Imaging, e is addition Zn2+Cell imaging after cultivating 10 minutes, after e after addition EDTA to cultivate 5 minutes
Cell imaging.
Specific embodiment
The following examples will be further described to the present invention, but not thereby limiting the invention.
Embodiment 1：The preparation of cell membrane localization zinc ion fluorescent, basic building-up process is as follows：
(1) synthesis of 4- nitros -1,8- naphthalene anhydrides：
In single port bottle, 11.2g (37.5mmol) sodium dichromate is dissolved in 50ml glacial acetic acid, be slow added into
The 5- nitro acenaphthenes of 3g (15mmol), back flow reaction 3 hours.Reaction solution is mixed with 200mL frozen water,
It is filtrated to get filter cake to be recrystallized with water, obtains 4- nitro -1,8- naphthalene anhydride 2.35g, yield 65%.
(2) synthesis of 4- amino -1,8- naphthalene anhydrides：
In flask add 1.2g (5mmol) 4- nitro -1,8- naphthalene anhydrides, 10mL absolute ethyl alcohols as solvent,
Then the stannous chloride of 4g (18mmol) 3.5mL concentrated hydrochloric acids are dissolved, is dropwise instilled in flask, at 90 DEG C
Back flow reaction 12 hours is overnight.Reaction solution is filtered after adjusting neutrality with the sodium carbonate liquor of 2mol/L, and filter cake is used
Water is recrystallized, and obtains 4- amino -1,8- naphthalene anhydride 780mg, yield 73%.
(3) synthesis of 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides：
4- amino -1 of 430mg (2mmol) is dissolved with the dry THF of 30mL in flask, 8- naphthalene anhydrides are added
The chloracetyl chloride of 1mL, after stirring 3 hours at room temperature, reaction system is become by muddiness to be clarified.It is spin-dried for solvent,
Column chromatography for separation at 25 DEG C, obtains yellow solid 242mg after being spin-dried for, i.e. 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides,
Eluant, eluent is dichloromethane, yield 42%.1H NMR (400MHz, CDCl3) δ 9.25 (s, 1H), 8.71 (d, J=
7.2Hz, 1H), 8.69-8.60 (m, 2H), 8.31 (d, J=8.8Hz, 1H), 7.94-7.90 (m, 1H), 4.42 (s,
2H).
(4) synthesis of 4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides：
4- (2- chloracetyls) amino -1 of 150mg (0.5mmol), 8- naphthalene anhydrides, 94mg (0.45 are added in flask
Mmol) DPA, 220mg (1.5mmol) DIPEA, 40mg (0.24mmol) KI, 50mL acetonitrile as solvents,
Night is flowed through in nitrogen protection next time.After being spin-dried for solvent, 1mL recrystallizing methanols are used, obtain yellow waxy thing 175mg,
That is 4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides, yield 78%.1H NMR(400MHz,DMSO)
δ 11.62 (s, 1H), 9.08 (d, J=8.5Hz, 1H), 8.62 (d, J=7.1Hz, 1H), 8.55 (dd, J=17.9,8.4
Hz, 2H), 8.40 (d, J=4.5Hz, 2H), 8.09-8.03 (m, 1H), 7.75 (t, J=7.5Hz, 2H), 7.46 (d, J
=7.7Hz, 2H), 7.29-7.20 (m, 2H), 4.06 (s, 4H), 3.66 (s, 2H)
(5) synthesis of probe molecule：
4- (2- lutidines amidos acetyl) amino -1 of addition 90mg (0.2mmol) in flask, 8- naphthalene anhydrides,
90mg (0.48mmol) lauryl amine, 20mL ethanol makees solvent, 90 DEG C of heated overnight at reflux 12 hours.
After being spin-dried for solvent, at 25 DEG C after column chromatography for separation, it is spin-dried for obtaining yellow, waxy solid 97mg i.e. probe molecule,
Eluant, eluent is dichloromethane:Methyl alcohol=50：1, yield 80%.1H NMR(400MHz,CDCl3)δ11.73
(s, 1H), 9.08 (d, J=8.6Hz, 1H), 8.69 (d, J=7.2Hz, 1H), 8.61 (dd, J=23.5,8.2Hz, 1H),
8.45 (d, J=4.2Hz, 1H), 7.89-7.78 (m, 1H), 7.62 (td, J=7.6,1.6Hz, 1H), 7.31 (d, J=
7.6Hz, 1H), 7.15 (dd, J=7.4,5.0Hz, 1H), 4.22-4.12 (m, 1H), 4.08 (s, 2H), 3.62 (s,
1H), (t, J=6.7Hz, the 3H) of 1.77-1.69 (m, 2H), 1.62 (s, 5H), 1.25 (s, 11H), 0.87
The preparation of the cell membrane localization zinc ion fluorescent of embodiment 2, basic building-up process is as follows：
With the step 5 of tetradecylamine alternate embodiment 1) in lauryl amine, remaining condition is with embodiment 1
Reacted, yield 75%.1H NMR (400MHz, CDCl3) δ 9.08 (d, J=8.5Hz, 1H), 8.68 (d, J
=7.2Hz, 1H), 8.60 (dd, J=27.5,8.3Hz, 2H), 8.45 (d, J=4.5Hz, 2H), 7.91-7.78 (m,
1H), 7.62 (td, J=7.6,1.4Hz, 2H), 7.32 (d, J=7.7Hz, 2H), 7.15 (dd, J=6.9,5.3Hz, 2H),
4.22–4.13(m,2H),4.08(s,4H),3.62(s,2H),1.81–1.68(m,3H),1.25(s,20H),0.87
(t, J=6.7Hz, 4H) ..
The preparation of the cell membrane localization zinc ion fluorescent of embodiment 3, basic building-up process is as follows：
With the step 5 of octadecylamine alternate embodiment 1) in lauryl amine, remaining condition is with embodiment 1
Reacted, yield 73%.1H NMR (400MHz, CDCl3) δ 11.74 (s, 1H), 9.09 (d, J=8.5Hz,
1H), 8.69 (d, J=7.0Hz, 1H), 8.62 (dd, J=25.9,8.3Hz, 2H), 8.45 (d, J=4.4Hz, 2H),
7.86-7.81 (m, 1H), 7.62 (t, J=6.9Hz, 2H), 7.31 (d, J=7.7Hz, 2H), 7.18-7.12 (m,
2H), 4.20-4.14 (m, 2H), 4.08 (s, 4H), 3.62 (s, 2H), 1.73 (d, J=6.7Hz, 2H), 1.25 (s,
30H), 0.88 (t, J=6.7Hz, 3H)
Fig. 3-5 is respectively the nucleus magnetic hydrogen spectrum of n=12,14,18 probes.
Fig. 6-8 is respectively the nuclear-magnetism carbon spectrum of n=12,14,18 probes.
Embodiment 4：The fluorescence probe of n=12 prepared by embodiment 1 is to Zn2+Selective enumeration method
Data in Fig. 9 are at HEPES (concentration 20mM, pH=7.4):CH3CN mixed solution (volumes
Than 5:5) test gained in, excitation wavelength is 365nm.Concentration and probe concentration is in 10 μM, then each experimental group
It is separately added into other metal ion (concentration are 30 μM respectively) Ag+,Au+,Ca2+,Cd2+,Co2+,Cr3+,Cu+,
Cu2+,Fe2+,Fe3+,Hg2+,K+,Mg2+,Mn2+,Na+,Ni2+,Pb2+,Pd2+,Pt2+,Zn2+.Result shows, only
Be happens is that in the presence of there is Fluorescence Increasing, and zinc ion under zinc ion and chromium ion existence condition obvious
Red shift enhancing changes, and chromium ion happens is that blue shift enhancing change, and other metal ions are without too substantially change
Change, thus above-mentioned interfering ion presence under conditions of, probe to zinc ion still have preferably selectivity and
Sensitivity.Fig. 9 abscissas are wavelength, and ordinate is fluorescence intensity；
Embodiment 3：The response condition that the fluorescence probe of n=12 prepared by embodiment 1 is determined to zinc ion
Data in Figure 10 are at HEPES (concentration 20mM, pH=7.4):CH3CN mixed solution (volumes
Than 5:5) test gained in, excitation wavelength is 365nm.Concentration and probe concentration is 10 μM, when the concentration of zinc ion
It it is 0,0.1,1,1.5,2,3,4,5,6,8,10,20,30 μM when increasing successively, its fluorescence spectrum gradually occurs bright
Aobvious red shift enhancing change.Abscissa is wavelength in Figure 10, and ordinate is fluorescence intensity.
Embodiment 4：The fluorescence probe of n=12 prepared by embodiment 1 cell membrane localization effect in colon cancer cell
Cell is colon cancer cell in Figure 11, and concentration and probe concentration is 5 μM, and zinc ion concentration is 25 μM.Altogether
Colon cancer cell is inoculated with the concentration of 20,000 cells/culture dish in focusing capsule, after 1640 medium cultures 5 days.
Culture medium is sucked, after PBS cushioning liquid washes twice, fresh 1640 culture mediums of 2mL is added, 2mM is added
The fluorescence probe solution of concentration, make its in the medium concentration be 5 μM, CO2Cultivated at 37 DEG C in incubator
After 30min, culture medium is sucked, PBS is washed 2 times, add 2mL fresh cultures, add 10mM dense
The zinc ion solution of degree, make its in the medium concentration be 25 μM.CO2In incubator 5min is cultivated at 37 DEG C
Afterwards, it is imaged under Laser Scanning Confocal Microscope 405nm laser excitations and is taken pictures.Can substantially observe that probe is gathered in cell
On film, and the increase and red shift of wavelength for causing fluorescence strong are combined with the zinc ion on cell membrane.
Embodiment 5：The fluorescence probe of n=12 prepared by embodiment 1 is focused into altogether in prostatic cell RWPE-1
As effect
Cell is prostate normal cell strain RWPE-1 in Figure 12, and concentration and probe concentration is 5 μM, zinc ion concentration
It is 25 μM.RWPE-1 cells, 1640 are inoculated with the concentration of 20,000 cells/culture dish in copolymerization Jiao's capsule
After medium culture 5 days.Culture medium is sucked, after PBS cushioning liquid washes twice, 2mL fresh 1640 is added
Culture medium, adds the fluorescence probe solution of 2mM concentration, make its in the medium concentration be 5 μM, CO2
After cultivating 30min at 37 DEG C in incubator, culture medium is sucked, PBS is washed 2 times, add 2mL fresh cultureds
Base, observation is taken pictures under Laser Scanning Confocal Microscope.The zinc ion solution of 10mM concentration is added, makes it in culture
Concentration is 25 μM in base, and observation is taken pictures under Laser Scanning Confocal Microscope.The EDTA solution of 20mM concentration is added,
Make its concentration in the medium for 50 μM, observation is taken pictures under Laser Scanning Confocal Microscope.Wherein, a, b, c distinguish
It is the bright field figure of cell, d is to add the cell imaging after probe culture 30 minutes under 405nm laser excitations,
E is addition Zn2+Cell imaging after cultivating 10 minutes under 405nm laser excitations, after e is to add EDTA
Cell imaging after cultivating 5 minutes under 405nm laser excitations；Can substantially observe that the cell for adding probe exists
Under conditions of zinc ion, fluorescent value is very low, and after zinc ion is added, fluorescence is greatly improved, afterwards
Add EDTA fluorescence to disappear, illustrate that probe is present on cell membrane.
Claims (8)
1. a kind of cell membrane localization zinc ion fluorescent, it is characterised in that：With following general structure (I)：
Wherein 4≤n≤18, and n are integer.
2. cell membrane localization zinc ion fluorescent according to claim 1, it is characterised in that：With 1,8- naphthoyls
Imines is fluorescent parent, and acid amides-lutidines amine is recognition group, and long alkyl chain is seeking group, alkyl
Chain length is n.
3. a kind of preparation method of the cell membrane localization zinc ion fluorescent described in claim 1 or 2, its feature
It is：Synthesis step is as follows：
1) synthesis of 4- nitros -1,8- naphthalene anhydrides：Sodium dichromate glacial acetic acid is dissolved in 0.1-0.5g/mL ratios, then
Add the 5- nitro acenaphthenes relative to 0.1-0.5 times of quality of sodium dichromate, back flow reaction 3-8 at 120-150 DEG C
Hour, reaction solution is mixed with the frozen water of 2-5 times of volume, it is filtrated to get filter cake and is recrystallized, obtain
4- nitro -1,8- naphthalene anhydrides；
2) synthesis of 4- amino -1,8- naphthalene anhydrides：By 4- nitro -1,8- naphthalene anhydrides with absolute ethyl alcohol in 0.1-0.5g/mL ratios
Dissolving, will press 1-2g/mL relative to 4- nitro -1, the stannous chloride of 3-5 times of quality of 8- naphthalene anhydrides with concentrated hydrochloric acid
After dissolving, above-mentioned 4- nitro -1 is dropwise instilled, in 8- naphthalene anhydrides and absolute ethyl alcohol, back flow reaction 8- at 80-100 DEG C
12 hours overnight, and reaction solution separates out product with the sodium carbonate liquor tune neutrality of 1-5mol/L, filters, filter
Cake is recrystallized, and obtains 4- amino -1,8- naphthalene anhydrides；
3) synthesis of 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides：4- amino -1,8- naphthalene anhydrides are pressed into 0.01- with dry THF
0.05g/mL ratios dissolve, and add relative to 4- amino -1, the chloracetyl chloride of 5-10 times of quality of 8- naphthalene anhydrides,
After stirring 3-10 hours at room temperature, reaction system is become by muddiness to be clarified, and is spin-dried for solvent, column chromatography for separation, rotation
It is dry to obtain 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides；
4) synthesis of 4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides：By 4- (2- chloracetyls) amino -1,8- naphthalenes
Acid anhydride, DPA, DIPEA and KI in mass ratio 1:0.5-0.8:1.5-2:0.2-0.5 mixes, and adds acetonitrile to press
0.01-0.05g/mL ratios dissolve 4- (2- chloracetyls) amino -1,8- naphthalene anhydrides, the lower 80-100 DEG C of backflow of nitrogen protection
8-12 hours, reaction was spin-dried for solvent after terminating, and with the recrystallizing methanol for being spin-dried for 5-10 times of quality of product, obtains
4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides；
5) synthesis of probe molecule：4- (2- lutidines amidos acetyl) amino -1,8- naphthalene anhydrides, alkylamine are pressed into quality
Than 1:1-2 mixes, and adds ethanol to dissolve 4- (2- lutidines amidos acetyl) in 0.01-0.05g/mL ratios
Amino -1,8- naphthalene anhydrides are heated to 70-90 DEG C and flow back 8-12 hours, and after being spin-dried for solvent, column chromatography for separation is obtained
Probe molecule.
4. according to the preparation method of cell membrane localization zinc ion fluorescent described in claim 1, it is characterised in that：
Step 4) in, the recrystallization system is methyl alcohol.
5. according to the preparation method of cell membrane localization zinc ion fluorescent described in claim 3, it is characterised in that：
Step 5) in, the eluant, eluent of the column chromatography is dichloromethane:Methyl alcohol=50：1, flow velocity 10-20ml/min,
20-25 DEG C of temperature.
6. according to the preparation method of cell membrane localization zinc ion fluorescent described in claim 3, it is characterised in that：
Step 5) described in a length of 4-18 of alkylamine carbochain integer.
7. according to the preparation method of cell membrane localization zinc ion fluorescent described in claim 3, it is characterised in that：
Step 3) described in column chromatography eluant, eluent be dichloromethane, flow velocity 10-20ml/min, 20-25 DEG C of temperature.
8. the zinc ion in cell membrane of the cell membrane localization zinc ion fluorescent described in a kind of claim 1 or 2
Detection or the detection in medical science to zinc ion in application.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110101876A (en) * | 2019-05-06 | 2019-08-09 | 上海师范大学 | Purposes of the novel optoacoustic probe in preparation medicine targeting photoacoustic imaging reagent or drug |
| CN110963995A (en) * | 2019-12-19 | 2020-04-07 | 山东师范大学 | Double-color fluorescent probe and synthetic method and application thereof |
| CN112209871A (en) * | 2020-10-29 | 2021-01-12 | 西北师范大学 | Zinc ion fluorescent probe based on tetraphenylethylene and preparation method and application thereof |
| CN116199669A (en) * | 2022-12-23 | 2023-06-02 | 石河子大学 | Nuclear membrane permeability zinc ion fluorescent probe and application thereof |
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| CN102603725A (en) * | 2012-03-02 | 2012-07-25 | 苏州大学 | Near-infrared fluorescence probe used for detecting zinc ions in water phase, preparation method thereof, and application thereof |
| CN103980886A (en) * | 2014-05-28 | 2014-08-13 | 国家纳米科学中心 | Fluorescent molecular probe, and preparation method and application thereof |
| CN104194773A (en) * | 2014-08-26 | 2014-12-10 | 武汉大学 | Cell-membrane target nitric oxide fluorescent probe as well as preparation method and application thereof |
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| CN102603725A (en) * | 2012-03-02 | 2012-07-25 | 苏州大学 | Near-infrared fluorescence probe used for detecting zinc ions in water phase, preparation method thereof, and application thereof |
| CN103980886A (en) * | 2014-05-28 | 2014-08-13 | 国家纳米科学中心 | Fluorescent molecular probe, and preparation method and application thereof |
| CN104194773A (en) * | 2014-08-26 | 2014-12-10 | 武汉大学 | Cell-membrane target nitric oxide fluorescent probe as well as preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110101876A (en) * | 2019-05-06 | 2019-08-09 | 上海师范大学 | Purposes of the novel optoacoustic probe in preparation medicine targeting photoacoustic imaging reagent or drug |
| CN110963995A (en) * | 2019-12-19 | 2020-04-07 | 山东师范大学 | Double-color fluorescent probe and synthetic method and application thereof |
| CN112209871A (en) * | 2020-10-29 | 2021-01-12 | 西北师范大学 | Zinc ion fluorescent probe based on tetraphenylethylene and preparation method and application thereof |
| CN116199669A (en) * | 2022-12-23 | 2023-06-02 | 石河子大学 | Nuclear membrane permeability zinc ion fluorescent probe and application thereof |
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Application publication date: 20170620 |