CN104151325B - Fluorescent probe with rhodamine fluorophore as matrix and preparation method of fluorescent probe with rhodamine fluorophore as matrix - Google Patents
Fluorescent probe with rhodamine fluorophore as matrix and preparation method of fluorescent probe with rhodamine fluorophore as matrix Download PDFInfo
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- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 239000011159 matrix material Substances 0.000 title abstract 3
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- 229940043267 rhodamine b Drugs 0.000 claims description 13
- 229940042795 hydrazides for tuberculosis treatment Drugs 0.000 claims description 11
- VYXSBFYARXAAKO-WTKGSRSZSA-N chembl402140 Chemical compound Cl.C1=2C=C(C)C(NCC)=CC=2OC2=C\C(=N/CC)C(C)=CC2=C1C1=CC=CC=C1C(=O)OCC VYXSBFYARXAAKO-WTKGSRSZSA-N 0.000 claims description 10
- KFZMGEQAYNKOFK-UHFFFAOYSA-N 2-propanol Substances CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 238000010992 reflux Methods 0.000 claims description 8
- 238000010898 silica gel chromatography Methods 0.000 claims description 7
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 claims description 4
- LRWZZZWJMFNZIK-UHFFFAOYSA-N 2-chloro-3-methyloxirane Chemical compound CC1OC1Cl LRWZZZWJMFNZIK-UHFFFAOYSA-N 0.000 claims description 4
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims description 4
- 238000001953 recrystallisation Methods 0.000 claims description 4
- 238000002390 rotary evaporation Methods 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 abstract description 21
- 239000010949 copper Substances 0.000 abstract description 14
- 229910001431 copper ion Inorganic materials 0.000 abstract description 14
- 238000001514 detection method Methods 0.000 abstract description 13
- 230000015572 biosynthetic process Effects 0.000 abstract description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 7
- 238000000799 fluorescence microscopy Methods 0.000 abstract description 7
- 238000003786 synthesis reaction Methods 0.000 abstract description 7
- 201000010099 disease Diseases 0.000 abstract description 5
- 238000013399 early diagnosis Methods 0.000 abstract description 3
- 230000008506 pathogenesis Effects 0.000 abstract description 3
- 238000001917 fluorescence detection Methods 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 210000002700 urine Anatomy 0.000 abstract description 2
- 239000003344 environmental pollutant Substances 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 21
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 7
- 229910052802 copper Inorganic materials 0.000 description 7
- 238000005481 NMR spectroscopy Methods 0.000 description 6
- 241001597008 Nomeidae Species 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 230000003834 intracellular effect Effects 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 4
- 238000004448 titration Methods 0.000 description 4
- 238000001816 cooling Methods 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000001636 atomic emission spectroscopy Methods 0.000 description 2
- 238000001391 atomic fluorescence spectroscopy Methods 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000003501 hydroponics Substances 0.000 description 2
- 230000002452 interceptive effect Effects 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- OALHHIHQOFIMEF-UHFFFAOYSA-N 3',6'-dihydroxy-2',4',5',7'-tetraiodo-3h-spiro[2-benzofuran-1,9'-xanthene]-3-one Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 OALHHIHQOFIMEF-UHFFFAOYSA-N 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 102000012437 Copper-Transporting ATPases Human genes 0.000 description 1
- 206010058314 Dysplasia Diseases 0.000 description 1
- 208000002972 Hepatolenticular Degeneration Diseases 0.000 description 1
- 241000784728 Lycaena virgaureae Species 0.000 description 1
- 208000008948 Menkes Kinky Hair Syndrome Diseases 0.000 description 1
- 208000012583 Menkes disease Diseases 0.000 description 1
- -1 Na+ Chemical class 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 208000018839 Wilson disease Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- XJHABGPPCLHLLV-UHFFFAOYSA-N benzo[de]isoquinoline-1,3-dione Chemical compound C1=CC(C(=O)NC2=O)=C3C2=CC=CC3=C1 XJHABGPPCLHLLV-UHFFFAOYSA-N 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 239000010453 quartz Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
-
- 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
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to a fluorescent probe with a rhodamine fluorophore as a matrix and a preparation method of the fluorescent probe. The fluorescent probe has a structural formula as shown in the specification. The preparation method of the fluorescent probe is mild in reaction condition and few and simple in synthesis step; the yield of the fluorescent probe is high; and raw materials are cheap and easy to obtain. The fluorescent probe related by the invention can be applied to detection of environmental pollutants and copper ions in urine and has the advantages of strong anti-jamming capability, short response time, high selectivity and sensitivity and the like, and the detection limit can be as low as the nM grade. In addition, the fluorescent probe can be applied to fluorescence imaging of Cu<2+> in living cells and has a potential application value on the aspects of fluorescence detection of Cu<2+> in a life body as well as early diagnosis and pathogenesis research on diseases related to Cu<2+>.
Description
Technical field
The present invention relates to a kind of fluorescent molecular probe and preparation method thereof, particularly a kind of with rhodamine fluorogen as parent
Fluorescent probe and preparation method thereof.
Background technology
Copper is requisite trace element in human body, in nervous system, gene expression, the side such as structure of protein
Face plays an important role.Lacking copper in human body, to easily cause scarce copper anemia, dysplasia of children, hypomnesis, reaction slow
Blunt, woman infertilities etc., the long-time copper that lacks also can cause arteriosclerosis, cause the generation of coronary heart disease.On the other hand, the copper of excess is taken the photograph
Entering and also can produce toxicity, this is to can result in the oxidation of other biological molecule etc. owing to copper catalysis creates active oxygenated species
Damage.Therefore, the imbalance of the homeostasis of intracellular copper will cause its growth and the disorder of metabolism, even causes serious disease
Sick and dead.The disease relevant to copper includes Menkes syndrome, Wilson disease, Alzheimer, ALS disease etc..
Relative to atomic emission spectrometry (AES), atomic absorption spectrography (AAS) (AAS), atomic fluorescence spectrometry (AFS) and
The detection methods such as electrochemical method, fluorescence detection has detection highly sensitive, real-time, to advantages such as the basic not damageds of sample,
The Common fluorescent group being currently used for fluoroscopic examination mainly has rhodamine fluorogen, naphthalimide fluorescent group, fluoresceins glimmering
Light blob etc..Wherein, rhodamine and derivant thereof are owing to having higher specific absorbance, stronger fluorescence quantum yield and photochemistry
The feature that stability etc. are good, has been widely used in the design of ion probe.
Current already present organic molecular probe mainly there is also following problem: I. synthesis aspect, probe molecule
Design synthesis there is also the deficiency that structure is complicated, step is cumbersome, productivity is relatively low.II. fluoroscopic examination aspect, organic point of part
There is the problems such as detection sensitivity is low, response time length, capacity of resisting disturbance are low in sub-probe.Therefore, the fluorescence of synthesizing new is designed
Probe molecule, it is achieved the trace detection of copper ion and intracellular Fluorescence imaging, will have important Research Significance and using value.
Summary of the invention
An object of the present invention is the deficiency overcoming existing organic probes molecule to exist, it is provided that a class is glimmering with rhodamine
Light blob is the fluorescent probe of parent.
The two of the purpose of the present invention are to provide the preparation method of this fluorescent probe.
For reaching above-mentioned purpose, the present invention uses following reaction mechanism:
；
Or
According to above-mentioned reaction mechanism, the present invention adopts the following technical scheme that
A kind of fluorescent probe with rhodamine fluorogen as parent, it is characterised in that the structural formula of this fluorescent probe:
Or。
A kind of method preparing the above-mentioned fluorescent probe with rhodamine fluorogen as parent, it is characterised in that the method
Concretely comprise the following steps:
A. salicylide is soluble in water according to the mol ratio of 2:1 ~ 5:1 with epoxychloropropane, back flow reaction 4 h, after cooling
It is filtrated to get product, then purifies by the method for recrystallization and obtain 1,3-bis-(2-carboxaldehyde radicals phenoxy group)-2-propanol, its structural formula
For:；
B. by uniform according to the mixed in molar ratio of 1:10 ~ 1:40 with hydrazine hydrate to rhodamine B or rhodamine 6G, and it is dissolved in ethanol
In, then it is heated to reflux 8 h, the product utilization silica gel column chromatography isolated rhodamine B hydrazides obtained or rhodamine 6G hydrazides,
Its structural formula is:Or；
C. by the rhodamine B of 1,3-bis-(2-carboxaldehyde radicals the phenoxy group)-2-propanol of above-mentioned steps a gained Yu step b gained
Hydrazides is uniform according to the mixed in molar ratio of 1:3 ~ 1:10, and is dissolved in ethanol, is heated to reflux 24 h, and rotary evaporation removes solvent,
Utilize silica gel column chromatography isolated rhodamine B derivative probe molecule.
The above-mentioned fluorescent probe with rhodamine fluorogen as parent is answering in the trace detection of copper ion in determinand
With.
The above-mentioned fluorescent probe with rhodamine fluorogen as parent is realizing in living cells in the fluorescence imaging of copper ion
Application.
The fluorescent probe of the present invention is the trace detection of copper ion in determinand, has the feature that 1. selectivitys are good,
The detection of single copper ion can be realized, there is not coexisting ion impact；The most highly sensitive, detection limits as little as nanomole rank；3.
Response time is fast, and reaction moment completes, and can realize detecting in real time；4. fluorescence and color change naked eyes are visible.
According to the present invention, such fluorescent probe, for the determinand of copper ion detection, includes but not limited to environmental contaminants
Sample and human urine sample.
According to the present invention, such living cells selected by fluorescent probe copper ion fluorescence imaging, include but not limited to Hela
Cell.
The preparation method of the class provided by the present invention fluorescent probe with rhodamine fluorogen as parent, reaction condition temperature
With, synthesis step is few, and products collection efficiency is high, and raw material is cheap and easy to get.Fluorescent probe provided by the present invention is to copper ion response time
Short, selectivity and highly sensitive, detection limits as little as nM rank, and can with the naked eye identify the change of fluorescence and color, is one
Plant the fluorescent probe of good detection copper ion.Fluorescent probe provided by the present invention applies also for Cu in living cells2+Glimmering
Photoimaging and in life entity Cu2+Fluoroscopic examination, with Cu2+Have on the early diagnosis of relevant disease and pathogenesis
There is potential using value.
Accompanying drawing explanation
Fig. 1 is selectivity experiment uv-visible absorption spectra corresponding in embodiment 3.
Fig. 2 is the fluorescence spectrum of titration experiments corresponding in embodiment 3.
Fig. 3 is the cell fluorescence imaging of rhodamine B derivative probe molecule corresponding in embodiment 4.
Detailed description of the invention
For making the present invention easier to understand, describe in detail below in conjunction with drawings and Examples.These embodiments are only
Play illustrative effect, it is not limited to the range of application of the present invention.
Embodiment 1: the preparation of rhodamine B derivative probe molecule
(1) salicylide is soluble in water according to the mol ratio of 2:1 ~ 5:1 with epoxychloropropane, reacting by heating 4 h, after cooling
It is filtrated to get product, then purifies by the method for recrystallization and obtain 1,3-bis-(2-carboxaldehyde radicals phenoxy group)-2-propanol.
(2) rhodamine B is uniform according to the mixed in molar ratio of 1:10 ~ 1:40 with hydrazine hydrate, and be dissolved in ethanol, then add
Hot reflux 8 h, the product utilization silica gel column chromatography isolated rhodamine B hydrazides obtained.
(3) 1,3-bis-(2-carboxaldehyde radicals phenoxy group)-2-propanol above-mentioned steps (1) prepared is prepared with above-mentioned steps (2)
Rhodamine B hydrazides uniform according to the mixed in molar ratio of 1:3 ~ 1:10, and be dissolved in ethanol, be heated to reflux 24 h, rotary evaporation
Remove solvent, utilize silica gel column chromatography isolated rhodamine B derivative probe molecule, then use nuclear magnetic resonance analyser
(Bruker AVANCE 500MHz type nuclear magnetic resonance analyser, Bruker company, Switzerland) pure to prepared rhodamine B derivative
Product carry out nuclear magnetic resonance spectroscopy, and concrete analysis result is as follows:
1H NMR: δ (deuterated DMSO) 9.14(s, 2H, N=C-H), 7.90(m, 2H, ArH), 7.57(m, 6H,
ArH), 7.29(m, 2H, ArH), 6.99(t, 2H, ArH) and, 6.29-6.50(m, 12H, ArH), 4.30-4.54
(m, 5H, CH2CHCH2), 3.35(s, 1H, OH), 3.23 (q, 16H, NCH 2 CH3), 1.09(t,24H,NCH2 CH 3 ).
Embodiment 2: the preparation of rhodamine 6G derivant probe molecule
(1) salicylide is soluble in water according to the mol ratio of 2:1 ~ 5:1 with epoxychloropropane, reacting by heating 4 h, after cooling
It is filtrated to get product, then purifies by the method for recrystallization and obtain 1,3-bis-(2-carboxaldehyde radicals phenoxy group)-2-propanol.
(2) rhodamine 6G is uniform according to the mixed in molar ratio of 1:10 ~ 1:40 with hydrazine hydrate, and be dissolved in ethanol, then
It is heated to reflux 8 h, after cold filtration, solid deionized water cyclic washing is obtained rhodamine 6G hydrazides.
(3) 1,3-bis-(2-carboxaldehyde radicals phenoxy group)-2-propanol above-mentioned steps (1) prepared is prepared with above-mentioned steps (2)
Rhodamine 6G hydrazides uniform according to the mixed in molar ratio of 1:3 ~ 1:10, and be dissolved in ethanol, be heated to reflux 24 h, rotary evaporation
Remove solvent, utilize silica gel column chromatography isolated rhodamine 6G derivant probe molecule, then use nuclear magnetic resonance analyser
(Bruker AVANCE 500MHz type nuclear magnetic resonance analyser, Bruker company, Switzerland) carries out nuclear magnetic resonance spectroscopy, concrete analysis
Result is as follows:
1H NMR: δ (deuterated DMSO): 9.20(s, 2H, N=C-H), 7.94(m, 2H, ArH), 7.60(m, 6H,
ArH), 7.30(m, 2H, ArH), 6.97(t, 2H, ArH) and, 6.27-6.51(m, 12H, ArH), 4.33-4.56
(m, 5H, CH2CHCH2), 3.32(s, 1H, OH), 3.20 (q, 8H, NHCH 2 CH3), 1,87(s,12H,ArCH3), 1.12
(t,12H,NHCH2 CH 3 ).
Embodiment 3: as fluorescent probe molecule, copper ion is detected using rhodamine B derivant
(1) weigh the rhodamine B derivative probe molecule of preparation in embodiment 1, be made into 1 × 10-5 The ethanol solution of M.
(2) concentrated solution (0.1M) of each metal ion species is prepared, including interfering ion such as Na+, K+, Mg2+, Ca2+, Ba2+,
Zn2+, Li+, Mn2+, Co2+, Hg2+Deng, and ion Cu to be detected2+。
(3), in selectivity experiment, take each metal ion species of certain equivalent respectively and join the rhodamine B of 2.0 mL and derive
In thing solution, then carry out absorption spectrum and fluorescence spectrum test.Result is shown in accompanying drawing 1.
(4), in titration experiments, it is placed in 1.0 cm × 1.0 cm quartz ratios with pipette, extract 2.0 mL rhodamine B derivative
In color ware, in solution, add Cu successively with microsyringe2+Solution, often adds and once carries out a spectrum test.Result is shown in
Accompanying drawing 2.
(5), in competitive assay, first certain interfering ion is joined in 2.0 mL rhodamine B derivative solution, add
Cu2+, then carry out spectrum test.
Embodiment 4: rhodamine B derivant is applied to cell imaging
(1) matched group, joins 1 × 10-5 The RPMI. 1640 hydroponics liquid of the rhodamine B derivative probe molecule of M；
(2) Hela cell is cultivated 0.5 hour in above-mentioned culture fluid；
(3) experimental group, first, with the Cu of 10 μMs2+Hydroponics Hela cell 0.5 hour, then, then with 10 μMs
Probe culture fluid is cultivated 0.5 hour；
(4) rinse cell 3 times with PBS, the culture fluid not absorbed by cell is washed away；
(5) cell after cultivating carries out imaging, with the laser excitation of 543 nm, observation of cell on Laser Scanning Confocal Microscope
Fluorescence imaging, result such as Fig. 3.
Fig. 1 is the selectivity titration experiments carrying out ultraviolet-visible spectrum in embodiment 3 with rhodamine B derivative, from figure
Can clearly be seen that in the present invention, the rhodamine B derivative probe molecule of synthesis is for Cu2+There is good selectivity.
Fig. 2 is the fluorescence titration spectrum of rhodamine B derivative in embodiment 3, as can be seen from the figure along with Cu2+Continuous
Increasing, the fluorescence intensity of rhodamine B derivative solution is the most linearly increasing.
Fig. 3 is the result that in embodiment 4, rhodamine B derivant is applied to cell imaging.It can be seen that by this
The rhodamine B derivative probe molecule of invention synthesis can be successfully applied to cell level to detect intracellular Cu2+.When with
After rhodamine B derivative probe molecule cultivates the cell cultivated in advance with copper ion, permissible by confocal microscope
Observe that cell sends obvious fluorescence, it will thus be seen that the probe molecule of present invention synthesis can successfully detect intracellular
Copper ion, this for study from now in life entity with Cu2+The early diagnosis of relevant disease and pathogenesis have important meaning
Justice.
It is only several specific embodiments of the present invention as mentioned above, is not limited to the present invention.All essences in the present invention
Within god and principle, the fluorescent probe with rhodamine fluorogen as parent obtained by same or similarity method is used to be used for
Intracellular copper ion fluorescence imaging, all in scope.
Claims (1)
1. a preparation method for the fluorescent probe with rhodamine fluorogen as parent, the structural formula of this fluorescent probe:
Or；
It is characterized in that concretely comprising the following steps of the method:
A. salicylide is soluble in water according to the mol ratio of 2:1 ~ 5:1 with epoxychloropropane, back flow reaction 4 h, cooled and filtered
Obtain product, then purify by the method for recrystallization and obtain 1,3-bis-(2-carboxaldehyde radicals phenoxy group)-2-propanol, its structural formula is:；
B. by uniform according to the mixed in molar ratio of 1:10 ~ 1:40 with hydrazine hydrate to rhodamine B or rhodamine 6G, and it is dissolved in ethanol,
Then 8 h it are heated to reflux, the product utilization silica gel column chromatography isolated rhodamine B hydrazides obtained or rhodamine 6G hydrazides, its
Structural formula is:Or；
C. by the rhodamine B hydrazides of 1,3-bis-(2-carboxaldehyde radicals the phenoxy group)-2-propanol of above-mentioned steps a gained Yu step b gained
Or rhodamine 6G hydrazides is uniform according to the mixed in molar ratio of 1:3 ~ 1:10, and it is dissolved in ethanol, is heated to reflux 24 h, rotary evaporation
Remove solvent, utilize the fluorescent probe of silica gel column chromatography isolated formula (a) and formula (b).
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| CN105754587B (en) * | 2016-03-30 | 2018-01-05 | 泰山医学院 | Imidazopyridine rhodamine hydrazides Cu-like ion ratio fluorescent probe and its application |
| CN107417694B (en) * | 2017-05-24 | 2019-04-26 | 延安大学 | A kind of colorimetric and the double response type bismuth ion detection probes of fluorescence and preparation method thereof |
| CN108440548B (en) * | 2018-03-31 | 2021-04-06 | 浙江工业大学 | Rhodamine 6G fluorescent probe containing hydrazide group, and preparation and application thereof |
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Effective date of registration: 20221212 Address after: 200444 Room 1376, Block B, Floor 5, Building 1, No. 668, Shangda Road, Baoshan District, Shanghai Patentee after: Shanghai Hupan Color Technology Co.,Ltd. Address before: 200444 No. 99, upper road, Shanghai, Baoshan District Patentee before: Shanghai University |
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Denomination of invention: Fluorescence probe based on Rhodamine fluorophore and its preparation method Granted publication date: 20170111 Pledgee: Bank of Communications Co.,Ltd. Shanghai Baoshan Branch Pledgor: Shanghai Hupan Color Technology Co.,Ltd. Registration number: Y2024310000010 |