CN106544008B - A kind of chromium ion detection fluorescent probe molecule, Preparation method and use based on rhodamine 6G - Google Patents
A kind of chromium ion detection fluorescent probe molecule, Preparation method and use based on rhodamine 6G Download PDFInfo
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- CN106544008B CN106544008B CN201610868383.3A CN201610868383A CN106544008B CN 106544008 B CN106544008 B CN 106544008B CN 201610868383 A CN201610868383 A CN 201610868383A CN 106544008 B CN106544008 B CN 106544008B
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- rhodamine
- chromium ion
- probe molecule
- fluorescent probe
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- 229910001430 chromium ion Inorganic materials 0.000 title claims abstract description 102
- 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 title claims abstract description 70
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 62
- 238000001514 detection method Methods 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 52
- 239000003480 eluent Substances 0.000 claims description 21
- 239000000523 sample Substances 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 14
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 150000001875 compounds Chemical class 0.000 claims description 12
- FVAUCKIRQBBSSJ-UHFFFAOYSA-M sodium iodide Chemical compound [Na+].[I-] FVAUCKIRQBBSSJ-UHFFFAOYSA-M 0.000 claims description 12
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical class CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 11
- 238000010992 reflux Methods 0.000 claims description 11
- -1 rhodamine 6G hydrazide compound Chemical class 0.000 claims description 11
- ICSNLGPSRYBMBD-UHFFFAOYSA-N 2-aminopyridine Chemical compound NC1=CC=CC=N1 ICSNLGPSRYBMBD-UHFFFAOYSA-N 0.000 claims description 10
- 239000002904 solvent Substances 0.000 claims description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 235000019441 ethanol Nutrition 0.000 claims description 8
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 8
- 238000010898 silica gel chromatography Methods 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 239000005725 8-Hydroxyquinoline Substances 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 6
- 229960000583 acetic acid Drugs 0.000 claims description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 6
- 239000012074 organic phase Substances 0.000 claims description 6
- 229960003540 oxyquinoline Drugs 0.000 claims description 6
- VGCXGMAHQTYDJK-UHFFFAOYSA-N Chloroacetyl chloride Chemical compound ClCC(Cl)=O VGCXGMAHQTYDJK-UHFFFAOYSA-N 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- 239000008247 solid mixture Substances 0.000 claims description 5
- 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
- 241000208125 Nicotiana Species 0.000 claims description 4
- 235000002637 Nicotiana tabacum Nutrition 0.000 claims description 4
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 4
- 235000009518 sodium iodide Nutrition 0.000 claims description 4
- 238000005292 vacuum distillation Methods 0.000 claims description 4
- 238000009835 boiling Methods 0.000 claims description 3
- 239000000126 substance Substances 0.000 claims description 3
- 241000244203 Caenorhabditis elegans Species 0.000 claims description 2
- 239000002027 dichloromethane extract Substances 0.000 claims description 2
- 125000005909 ethyl alcohol group Chemical group 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- 239000003292 glue Substances 0.000 claims 1
- 229910052710 silicon Inorganic materials 0.000 claims 1
- 239000010703 silicon Substances 0.000 claims 1
- 238000005406 washing Methods 0.000 claims 1
- 239000000243 solution Substances 0.000 description 33
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 21
- 239000011651 chromium Substances 0.000 description 20
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 14
- 229910052804 chromium Inorganic materials 0.000 description 14
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 9
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 8
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical class [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 description 8
- 238000012360 testing method Methods 0.000 description 8
- 241000244206 Nematoda Species 0.000 description 6
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 238000002189 fluorescence spectrum Methods 0.000 description 6
- 150000002500 ions Chemical class 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 230000000875 corresponding effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 229910021645 metal ion Inorganic materials 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 239000010453 quartz Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 238000002211 ultraviolet spectrum Methods 0.000 description 4
- 230000000536 complexating effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-M perchlorate Inorganic materials [O-]Cl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-M 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000000825 ultraviolet detection Methods 0.000 description 2
- NBYLBWHHTUWMER-UHFFFAOYSA-N 2-Methylquinolin-8-ol Chemical compound C1=CC=C(O)C2=NC(C)=CC=C21 NBYLBWHHTUWMER-UHFFFAOYSA-N 0.000 description 1
- KZMAWJRXKGLWGS-UHFFFAOYSA-N 2-chloro-n-[4-(4-methoxyphenyl)-1,3-thiazol-2-yl]-n-(3-methoxypropyl)acetamide Chemical compound S1C(N(C(=O)CCl)CCCOC)=NC(C=2C=CC(OC)=CC=2)=C1 KZMAWJRXKGLWGS-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 206010007269 Carcinogenicity Diseases 0.000 description 1
- 206010064571 Gene mutation Diseases 0.000 description 1
- 241000482313 Globodera ellingtonae Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 231100000260 carcinogenicity Toxicity 0.000 description 1
- 230000007670 carcinogenicity Effects 0.000 description 1
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- LBJNMUFDOHXDFG-UHFFFAOYSA-N copper;hydrate Chemical compound O.[Cu].[Cu] LBJNMUFDOHXDFG-UHFFFAOYSA-N 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229910052564 epsomite Inorganic materials 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000000695 excitation spectrum Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229940042795 hydrazides for tuberculosis treatment Drugs 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229910000474 mercury oxide Inorganic materials 0.000 description 1
- UKWHYYKOEPRTIC-UHFFFAOYSA-N mercury(ii) oxide Chemical compound [Hg]=O UKWHYYKOEPRTIC-UHFFFAOYSA-N 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
- C07D491/10—Spiro-condensed systems
- C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
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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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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Abstract
The invention discloses a kind of, and the chromium ion based on rhodamine 6G detects fluorescent probe molecule, which has structure as follows.The invention also discloses described, and the chromium ion based on rhodamine 6G detects the preparation method of fluorescent probe molecule and the purposes for chromium ion detection.
Description
Technical field
The invention belongs to organic functional material fields, and in particular to a kind of chromium ion detection fluorescence spy based on rhodamine 6G
Needle molecule and preparation method and its purposes detected for chromium ion.
Background technology
Trivalent chromium is primarily present in glucose tolerance factor, is played a role as active constituent.It is current studies have shown that
Chromium in general food is trivalent chromium, and trivalent chromium rises in the anabolism of carbohydrate, protein, fat, nucleic acid and amino acid
To important role.Chromium content is little in human body, adult's total amount containing chromium be 1.7~6mg, be primarily present in lung, muscle, liver, the heart,
In the organs such as kidney, brain.The shortage of chromium can cause glucose tolerance to reduce, the speed of growth and service life decline, serum cholesterol
The illnesss such as level raising.Research has confirmed that the compound of Cr VI is toxic, has the function of the mutation of carcinogenic and modificator gene,
And the effect of carcinogenicity and the modificator gene mutation of trivalent chromium is confirmed by experiments not yet.But high concentration trivalent chromium in vitro
Under the conditions of existing, trivalent chromium also can induce and generate free radicals, and have an effect with DNA, cause mankind aging and illness.
Currently, in the common test method of chromium ion, although generally having the characteristics that in higher sensitivity, detection
Of high cost, the problems such as sample preparation is complicated, test process is long, is still widely present.A kind of simple, quick, sensitive detection side of invention
Method is of great significance to the detection of chromium ion in environment and tobacco leaf and correlated product.Inspection based on fluorescence probe
Survey is one developed in recent years with high sensitivity, good, the easily operated visual inspection method of selectivity, can be compared with
Good simple, quick, the sensitive testing requirements of satisfaction.
Invention content
In this invention, we have studied Rhodamine Derivatives, preparation method and applications.Rhodamine is as typical
On/off-type fluorogen, it is contemplated that cause C-N keys to disconnect after being combined with specific metal ion (such as chromium ion), to generate face
The Selective recognition to chromium ion in solution system and detection are realized in the variation of color and fluorescence.The compound synthesis method is simple,
High income, compound small toxicity itself have efficient selectivity to the identification of chromium ion, and resolving time is short, color and fluorescence
Variation is apparent.Since there is the compound good cell-penetrating ability to be realized to chromium on the basis of solution experiments
Fluorescence imaging in the online polypide of ion.Based on the above advantages, it is believed that the Rhodamine Derivatives 1 are highly selective glimmering in chromium ion
It has a good application prospect in terms of light recognition detection.
Technical problem to be solved by the invention is to provide a kind of highly selective chromium ion detections based on rhodamine 6G
Fluorescent probe molecule and preparation method and its purposes detected for chromium ion.
First aspect present invention is related to a kind of chromium ion detection fluorescent probe molecule based on rhodamine 6G, chromium ion inspection
Fluorescent probe molecule is surveyed to have the following structure：
Second aspect of the present invention is related to the preparation method of the chromium ion detection fluorescent probe molecule based on rhodamine 6G,
Include the following steps：
(1) rhodamine 6G and hydrazine hydrate are heated to reflux 4~6 hours in methyl alcohol, obtained flocculent deposit after suction filtration,
For by silica gel chromatography column with the first eluant, eluent separating-purifying, obtained solid matter is rhodamine 6G hydrazide compound；
(2) rhodamine 6G hydrazide compound and 2- aldehyde radicals -8-hydroxyquinoline that step (1) obtains are dissolved into the second of boiling
In alcohol, glacial acetic acid is added, is heated to reflux under the protection of nitrogen 6~10 hours, obtained deposit is washed through solvent and filtered
Afterwards, for by silica gel chromatography column with the second eluant, eluent separating-purifying, obtained solid matter is the derivative of rhodamine 6G；
(3) by 2-aminopyridine and chloracetyl chloride, room temperature reaction 3~5 is small in the dichloromethane added with a few drop triethylamines
When, obtained reaction solution is extracted three times with water, and obtained organic phase is dry with anhydrous sodium sulfate again and filters, and is then evaporated under reduced pressure
Solid mixture is obtained, by silica gel chromatography column third eluant, eluent separating-purifying obtains the chloro- N- pyridineacetamides of compound 2-
Close object；
(4) derivative for the rhodamine 6G that potassium carbonate, sodium iodide and step (2) obtain is dissolved into acetone, is flowed back
After 0.5~2 hour, the chloro- N- pyridineacetamides compounds of 2- that step (3) obtains are added, are heated to reflux under the protection of nitrogen
12~14 hours；Reaction solution is cooled to room temperature after reaction, with saturation NH under condition of ice bath4It is 7 or so that Cl, which adjusts pH,
Then three times (3 × 20mL) with dichloromethane extraction, obtained organic phase is filtered after being dried with anhydrous sodium sulfate, is evaporated under reduced pressure to
To solid mixture, the 4th eluant, eluent separating-purifying of by silica gel chromatography column, obtained solid matter is to be based on rhodamine 6G
Chromium ion detect fluorescent probe molecule.
Preferably, the ratio of rhodamine 6G, hydrazine hydrate and methanol is 3g in the step (1):1mL:50mL, described first
Eluant, eluent is n-hexane：Dichloromethane：Methanol volume ratio is 10:4:1 mixture.
Preferably, rhodamine 6G hydrazide compound and 2- aldehyde radicals -8-hydroxyquinoline and glacial acetic acid in the step (2)
Molar ratio is 3:2:0.1, the solvent is ethyl alcohol:Ether product is than being 1:1 mixture, second eluant, eluent are dichloromethane
Alkane.
Preferably, the molar ratio of 2-aminopyridine, triethylamine and chloracetyl chloride is 1 in the step (3):1.1:1.1, institute
It is dichloromethane to state third eluant, eluent:Ethyl acetate volume ratio is 3:1 mixture.
Preferably, in the step (4) rhodamine 6G derivative 4, the chloro- N- pyridineacetamides 5 of 2-, potassium carbonate and iodine
The molar ratio for changing sodium is 1:1.2:2:0.56, the 4th eluant, eluent is dichloromethane:Ethyl acetate volume ratio is 3:1 mixing
Object.
Third aspect present invention is related to the chromium ion detection fluorescent probe molecule based on rhodamine 6G and is used for water body
Sample, tobacco sample, the purposes that in vivo chromium ion detects.
Preferably, the live body is caenorhabditis elegan.
Beneficial effects of the present invention：
1, the chromium ion detection fluorescent probe molecule synthetic method based on rhodamine 6G of the invention is simple, high income.
2, the chromium ion detection fluorescent probe molecule excitation and emission spectra based on rhodamine 6G of the invention is in visual field,
It is insensitive to solvent polarity, and chemical stability is good.
3, the chromium ion detection fluorescent probe molecule based on rhodamine 6G of the invention contains and multiple can occur with chromium ion
The amine groups of complexing can form multiple hydrogen bonds and then realize complexing recognition reaction, have good selection to chromium ion
Property, to Na+,K+,Ca2+,Mg2+Etc. common metal ions and its anion there is good anti-interference ability.
4, fluorescent emission before and after the chromium ion detection fluorescent probe molecule complexing chromium ion based on rhodamine 6G of the invention
There are about 13 times of growth, detection sensitivity is high, and resolving time is short, can be applied to environment water body example, tobacco sample, in vivo
Chromium ion detection, have wide application prospect.
5, the present invention is based on the chromium ion of rhodamine 6G detection fluorescent probe molecule cell permeability it is good, to cell itself poison
The fluoroscopic examination of chromium ion in nematode body may be implemented in Small side effects.
Description of the drawings
Fig. 1 is the synthetic route that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule；
Fig. 2 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule1H-NMR spectrum；
Fig. 3 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule13C-NMR spectrograms；
Fig. 4 is HRMS (ESI) spectrums that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule
Figure；
Fig. 5 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule in water solution system
To the ultraviolet variation spectrogram of chromium ion response；
Fig. 6 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule in water solution system
To different metal ions at 528nm UV intensity block diagram；
Fig. 7 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule in water solution system
Fluorescence spectrum variation diagram is responded to chromium ion；
Fig. 8 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule 1 in water solution system
In to different metal ions at 556nm fluorescence intensity block diagram；
Fig. 9 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule in water solution system
To various concentration chromium ion ultraviolet spectra variation diagram；
Figure 10 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule extinction at 528nm
Degree and corresponding Cr3+Ion concentration matched curve figure；
Figure 11 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule in water solution system
In to various concentration chromium ion fluorescence spectrum variation diagram；
Figure 12 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule fluorescence at 556nm
Intensity and corresponding Cr3+Ion concentration matched curve figure；
Figure 13 is right in the chromium ion detection online polypide of fluorescent probe molecule according to the present invention based on rhodamine 6G
Chromium ion fluorescence identifying figure.(wherein, a-c is light field, and d is the fluorescence picture for the probe compound that 10 μm of ol/L are only added, and e is
750 μm of ol/L Cr are first added3+The probe compound that nurture 3h adds 10 μm of ol/L feeds the fluorescence picture of 2h again, f be first plus
Enter 1500 μm of ol/L Cr3+The probe compound 1 that nurture 3h adds 10 μm of ol/L feeds the fluorescence picture of 2h again.)
Specific implementation mode
Embodiment 1：The synthesis of chromium ion detection fluorescent probe molecule based on rhodamine 6G
Specific synthetic route is shown in Fig. 1.
(1) preparation of 2- aldehyde radicals -8-hydroxyquinoline：2- methyl -8-hydroxyquinoline (2mmol) of 0.32g is taken to be dissolved into
In the dioxane of 20mL.60 DEG C are heated to, then weighs the SeO of 0.27g2It is added in above-mentioned solution, then rises to temperature
80 DEG C, reaction mixture is heated to reflux 10 hours under the protection of nitrogen.After reaction, reaction solution is cooled to room temperature,
Gained precipitation is cleaned with the dichloromethane of the dioxane of 8mL and 8mL, is filtered, and is dried in vacuo.Gained crude product passes through silica gel
Chromatographic column uses petroleum ether：Ethyl acetate=9:1(v:V) it elutes, eluent is concentrated to obtain the light yellow compound of 0.14g.Production
Rate 40.5%.1H-NMR(500MHz,CDCl3):δ10.2(s,1H),8.31-8.33(d,1H),8.15(s,1H),8.05-8.06
(d,1H),7.6-7.64(m,1H),7.42-7.44(m,1H),δ7.2-7.3(m,1H)。
(2) preparation of rhodamine 6G hydrazide compound：0.3g rhodamine 6Gs (0.63mmol) are weighed, the first of 5mL is dissolved into
In alcohol, 0.1mL hydrazine hydrates are added.Reflux 4~6 hours, obtains flocculent deposit.After gained precipitation filters, by silica gel chromatography column,
Use n-hexane：Dichloromethane：Methanol=10:4:1(v:v:V) it elutes.Eluent is concentrated, dry rhodamine 6G hydrazides chemical combination
Object.1H-NMR(500MHz,CDCl3):δ7.96-7.94(m,1H),7.46-7.44(m,2H),7.06-7.05(m,1H),6.39
(s,2H),6.26(s,2H),3.57(s,4H),3.23-3.20(m,4H),1.96-1.88(s,6H),1.36-1.25(m,6H)。
(3) synthesis based on rhodamine 6G derivative：Weigh 0.21g 2- aldehyde radicals -8-hydroxyquinoline (1.2mmol) and
0.307g rhodamine 6Gs hydrazide compound (0.8mmol) is dissolved into the ethyl alcohol of boiling, and 3 drop glacial acetic acid are added, reaction is mixed
Object is heated to reflux 8 hours under the protection of nitrogen.Gained yellow mercury oxide ethyl alcohol：Ether=1:1(v:V) solvent cleaning and mistake
It filters, then by silica gel chromatography column, with dichloromethane eluent, concentrate eluant obtains 0.23g orange-yellow compounds.1H-NMR
(400MHz,DMSO-d6):δ9.87(s,1H),8.69(s,1H),8.24-8.22(d,1H),7.98-7.96(d,1H),7.87-
7.85(d,1H),7.62-7.57(m,2H),7.40-7.38(d,2H),7.33-7.31(d,1H),7.09-7.05(t,2H),
6.40(s,2H),6.27(s,2H),5.10(s,2H),3.17-3.12(m,4H),1.85(s,6H),1.23-1.19(t,6H)。
(4) synthesis of the chloro- N- pyridineacetamides compounds of 2-：The round-bottomed flask for taking a 50mL adds under the conditions of 0 DEG C
The 2-aminopyridine for entering 2mmol (188mg) adds the anhydrous methylene chloride (15mL) for being mixed with triethylamine (0.3mL).Fully stir
It mixes 30 minutes.Then under condition of ice bath, chloracetyl chloride 2.2mmol (0.175mL, mixing is added dropwise dropwise into above-mentioned mixed liquor
5mL dichloromethane), rear reaction mixture is added dropwise and is reacted 3 hours in nitrogen protection and under room temperature, contact plate confirms reaction
After, two secondary response mixed liquors first are cleaned with 30mL saturated salt solutions, then washed with 20mL, and extract, repeatedly three times.
It obtains filtering after organic phase is dried with anhydrous sodium sulfate, vacuum distillation obtains solid mixture, finally uses dichloromethane：Acetic acid second
Ester=3:1(v:V) it crosses column as eluant, eluent and obtains 0.27g khaki sterlings.Yield is 78.5%.1H-NMR(500MHz,
CDCl3):δ8.87(s,1H),8.33-8.32(d,1H),8.21-8.18(d,1H),7.77-7.72(m,1H),7.09-7.13
(m,1H),4.21(s,2H)。
(5) synthesis of the chromium ion detection fluorescent probe molecule (i.e. the compound of the present invention) based on rhodamine 6G：Take one
The round-bottomed flask of a 100mL is separately added into the rhodamine 6G derivative 4 of 1mmol (0.584g), the K of 2mmol (0.276g)2CO3
With the NaI of 0.56mmol (0.084g).Be added 30mL acetone be used as solvent, fully react 1 hour after, into above-mentioned mixed liquor by
The chloro- N- pyridineacetamides of 2- (compound 5,0.2050g, mixing 5mL acetone) of 1.2mmol are added dropwise in drop, are heated to reflux stirring 12
Hour.Contact plate confirmation is cooled to room temperature after reaction, with saturation NH under condition of ice bath4It is 7 or so that Cl, which adjusts pH, is then used
Dichloromethane extracts (3 × 20mL), and obtained organic phase filters after being dried with anhydrous sodium sulfate, and vacuum distillation obtains solid mixing
Object finally uses dichloromethane：Ethyl acetate=3:1(v:V) it crosses column as eluant, eluent and obtains the orange-yellow sterlings of 0.43g, yield is
60%.1H NMRδ(DMSO-d6):δ10.72(s,1H),8.76(s,1H),8.32-8.28(m,2H),7.98-7.96(d,1H),
7.90-7.88(d,1H),7.62(d,1H),7.57(m,1H),7.50-7.48(m,3H),7.21(t,1H),7.06(t,1H),
7.05(d,1H),6.35(s,2H),6.26(s,2H),5.08-5.06(t,2H),5.03(s,2H),3.12-3.09(m,4H),
1.83(s,6H),1.20-1.16(m,6H).13C-NMR(100MHz,DMSO-d6)δ:165.19,152.34,151.84,
147.96,141.75,141.44,122.78,129.89,129.45,128.70,128.22,128.12,127.33,124.16,
123.75,118.35,106.67,97.19,66.44,38.78,17.10,15.16.HRMS(ESI):calcd for
C43H40N7O4[M+H]+=718.3136, found m/z 718.3135. wherein, DMSO-d6For deuterated dimethyl sulfoxide.It is related
Spectrogram is shown in Fig. 2~4.
Embodiment 2：Selectivity of the chromium ion detection fluorescent probe molecule based on rhodamine 6G to chromium ion ultraviolet detection
Using CH3CN (acetonitrile)：Tris-HCl buffer solutions (trishydroxymethylaminomethane) (0.01mol/L, pH=7.4)=
9:1(v:V) solution controls experiment condition.
By the chromium ion detection fluorescent probe molecule CH based on rhodamine 6G3CN:Tris-HCl=9:1(v:V) molten
In agent dissolving and constant volume to the volumetric flask of 100mL, it is configured to the solution of a concentration of 10 μm of ol/L of fluorescent probe molecule.
Sample bottle is divided into 12 groups, every group of each sample bottle be separately added into a concentration of 10 μm of ol/L of 5mL based on rhodamine 6G
Chromium ion detection fluorescent probe molecule CH3CN：Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution, the
One bottle of solution is as blank group, the other 11 groups K for being separately added into 75 a concentration of 0.1mol/L of μ L+,Na+,Co2+,Ni2+,Zn2+,Pb2 +,Cd2+,Li+,Fe3+,Hg2+And Cr3+Perchlorate aqueous solution.Stand 3 minutes after, by each test job liquid be transferred to 1cm ×
In the standard quartz cuvette of 1cm, its ultraviolet spectra is measured.
Ultraviolet selective enumeration method such as Fig. 5 institute of the chromium ion detection fluorescent probe molecule based on rhodamine 6G to chromium ion
Show.The result shows that only there is chromium ion significantly at 528nm in the chromium ion detection fluorescent probe molecule based on rhodamine 6G
Ultraviolet absorption peak.This result shows that it is according to the present invention based on rhodamine 6G chromium ion detection fluorescent probe molecule to chromium from
Sublist reveals the ultraviolet selectivity of height.
We select the UV absorption intensity value of each ion at 528nm to make associated bars, as shown in fig. 6, from
Can visually see in Fig. 6 probe ultraviolet selectivity it is very good.
Embodiment 3：Selectivity of the chromium ion detection fluorescent probe molecule based on rhodamine 6G to chromium ion fluoroscopic examination
Using CH3CN (acetonitrile)：Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution control experiment item
Part.
By the chromium ion detection fluorescent probe molecule CH based on rhodamine 6G3CN:Tris-HCl=9:1(v:V) molten
In agent dissolving and constant volume to the volumetric flask of 100mL, it is configured to the solution of a concentration of 10 μm of ol/L of fluorescent probe molecule.
Sample bottle is divided into 12 groups, every group of each sample bottle be separately added into a concentration of 10 μm of ol/L of 5mL based on rhodamine 6G
Chromium ion detection fluorescent probe molecule CH3CN:Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution, the
One bottle of solution is as blank group, the other 11 groups K for being separately added into 75 a concentration of 0.1mol/L of μ L again respectively+,Na+,Co2+,Ni2+,
Cu2+,Zn2+,Pb2+,Cd2+,Li+,Fe3+,Hg2+And Cr3+Perchlorate aqueous solution.After standing 3 minutes, by each test job liquid
It is transferred in the standard quartz cuvette of 1cm × 1cm, measures its fluorescence spectrum.Excitation wavelength is 495nm, and launch wavelength is
556nm.Chromium ion detection fluorescent probe molecule based on rhodamine 6G detects chromium ion fluorescence selectivity as shown in Figure 7.It can
See that the chromium ion based on rhodamine 6G detects fluorescent probe molecule at 556nm only to Cr3+There is apparent Enhancement of Fluorescence (about
13 times of enhancings), show that the chromium ion detection fluorescent probe molecule according to the present invention based on rhodamine 6G shows chromium ion
Go out the fluorescence selectivity of height.
We select the fluorescence intensity level of each ion at 556nm to make dependent cylindrical figure (Fig. 8), and Fig. 8 can be intuitive
Find out that the fluorescence selectivity of probe is very good in ground.
Embodiment 4：Quantitative ultraviolet detection of the chromium ion detection fluorescent probe molecule based on rhodamine 6G to chromium ion
Using CH3CN (acetonitrile)：Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution control experiment item
Part.
By the chromium ion detection fluorescent probe molecule CH based on rhodamine 6G3CN：Tris-HCl (0.01mol/L, pH=
7.4)=9:1(v:V) solvent dissolving simultaneously in constant volume to the volumetric flask of 100mL, is configured to a concentration of 10 μ of fluorescent probe molecule
The solution of mol/L.
Weigh 0.9169g Cr (ClO)3·6H2O is configured into a concentration of 0.1mol/L's with 20mL deionized water dissolvings
Cr3+Aqueous solution.
Sample bottle is divided into 12 groups, every group of each sample bottle be separately added into a concentration of 10 μm of ol/L of 5mL based on rhodamine 6G
Chromium ion detection fluorescent probe molecule CH3CN：Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution, then
It is separately added into the Cr of 0 a concentration of 0.1mol/L of the μ of μ L~75 L3+Aqueous solution, make in test system chromium ion concentration be 0 μm of ol/L~
1500μmol/L.After standing 3 minutes, each test job liquid is transferred in the standard quartz cuvette of 1cm × 1cm, it is measured
Ultraviolet spectra.
Fig. 9 is that the chromium ion according to the present invention based on rhodamine 6G detects fluorescent probe molecule in water solution system
The ultraviolet spectrogram changed with chromium ion concentration.Absorbance at ultraviolet spectra 528nm is fitted with corresponding chromium ion concentration,
It is to obtain a matched curve (Figure 10) within the scope of 0 μm of ol/L~1800 μm ol/L in chromium ion concentration, shows involved in the present invention
Chromium ion based on rhodamine 6G detect fluorescent probe molecule and can quantify detection chromium ion concentration in water solution system.
Embodiment 5：Chromium ion detection fluorescent probe molecule based on rhodamine 6G detects the quantitative fluorescence of chromium ion
Using CH3CN (acetonitrile)：Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution control experiment item
Part.
Chromium ion based on rhodamine 6G is detected into fluorescent probe molecule, uses CH3CN：Tris-HCl(0.01mol/L,pH
=7.4)=9:1(v:V) solvent dissolving simultaneously in constant volume to the volumetric flask of 100mL, is configured to a concentration of 10 μ of fluorescent probe molecule
The solution of mol/L.
Weigh 0.9169g Cr (ClO)3·6H2O is configured into a concentration of 0.1mol/L's with 20mL deionized water dissolvings
Cr3+Aqueous solution.
Sample bottle is divided into 12 groups, every group of each sample bottle be separately added into a concentration of 10 μm of ol/L of 5mL based on rhodamine 6G
Chromium ion detection fluorescent probe molecule CH3CN：Tris-HCl (0.01mol/L, pH=7.4)=9:1(v:V) solution, then
It is separately added into the Cr of 0 a concentration of 0.1mol/L of the μ of μ L~75 L3+Aqueous solution, make in test system chromium ion concentration be 0 μm of ol/L~
1500μmol/L.After standing 3 minutes, each test job liquid is transferred in the standard quartz cuvette of 1cm × 1cm, it is measured
Fluorescence spectrum.Fluorometric investigation grating gap size is 5 × 5nm.Figure 11 be it is according to the present invention based on the chromium of rhodamine 6G from
The fluorescence spectra that son detection fluorescent probe molecule changes in water solution system with chromium ion concentration.By fluorescence spectrum 556nm
The fluorescence intensity at place is fitted with corresponding chromium ion concentration, is within the scope of 0 μm of ol/L~1800 μm ol/L in chromium ion concentration
A matched curve (Figure 12) is obtained, shows that the chromium ion detection fluorescent probe molecule according to the present invention based on rhodamine 6G exists
Detection chromium ion concentration can be quantified in water solution system.
Embodiment 6：To the fluorescence of chromium ion in the chromium ion detection online polypide of fluorescent probe molecule based on rhodamine 6G
Identification
Nematode in culture dish M9 buffer solutions (are contained into 15.12 grams of Na in every liter of M9 buffer solution2HPO4·12H2O；3 grams
KH2PO4；5 grams of NaCl；0.25 gram of MgSO4·7H2O it) comes out, divides three to be assembled into centrifuge tube.Again into this three groups of centrifuge tubes
1mL M9 solution is added, 0 μ L of chromium ion solution, 7.5 μ of a concentration of 0.1mol/L are then separately added into this three groups of centrifuge tubes
L, 15 μ L are incubated 3 hours at ambient temperature.Nematode is washed 3 times with M9 buffer solutions, and 3 points are centrifuged under 3000r/min speed
Clock, the backward centrifuge tube equipped with nematode in the chromium ion detection fluorescence based on rhodamine 6G of a concentration of 1mmol/L be added visit
10 μ L of needle molecule, and be incubated 2 hours at 20 DEG C.Nematode is washed 3 times with M9 solution again, and 3 points are centrifuged under 3000r/min speed
It is transferred to after clock on glass slide and carries out fluorescence imaging experiments.The result is shown in Figure 13.It can be seen that the chromium based on rhodamine 6G in Figure 13 c
Ion detection fluorescent probe molecule itself basic unstressed configuration transmitting in nematode, when chromium ion concentration reaches 1500 μm of ol/L,
Launch yellow-green fluorescence (Figure 13 f) in nematode body.The experiment shows the chromium ion according to the present invention based on rhodamine 6G
Fluorescence identifying can be carried out in online polypide to chromium ion by detecting fluorescent probe molecule.Experiment instrument is OlympusBX51
Fluorescence microscope.
Claims (7)
1. a kind of chromium ion based on rhodamine 6G detects fluorescent probe molecule, which is characterized in that have the following structure：
2. the preparation method of the chromium ion detection fluorescent probe molecule according to claim 1 based on rhodamine 6G, special
Sign is, includes the following steps：
(1) rhodamine 6G and hydrazine hydrate are heated to reflux 4~6 hours in methyl alcohol, obtained flocculent deposit is after suction filtration, through silicon
For glue chromatographic column with the first eluant, eluent separating-purifying, obtained solid matter is rhodamine 6G hydrazide compound；
(2) rhodamine 6G hydrazide compound and 2- aldehyde radicals -8-hydroxyquinoline that step (1) obtains are dissolved into the ethyl alcohol of boiling
In, be added glacial acetic acid, be heated to reflux under the protection of nitrogen 6~10 hours, obtained deposit through solvent washing filtering after,
For by silica gel chromatography column with the second eluant, eluent separating-purifying, obtained solid matter is the derivative of rhodamine 6G；
(3) 2-aminopyridine and chloracetyl chloride are reacted at room temperature 3~5 hours in the dichloromethane added with a few drop triethylamines, is obtained
The reaction solution arrived water extracts three times, and obtained organic phase is dry with anhydrous sodium sulfate again and filters, and then vacuum distillation obtains
Solid mixture, by silica gel chromatography column third eluant, eluent separating-purifying obtain the chloro- N- pyridineacetamides chemical combination of compound 2-
Object；
(4) derivative for the rhodamine 6G that potassium carbonate, sodium iodide and step (2) obtain is dissolved into acetone, reflux 0.5~
After 2 hours, the chloro- N- pyridineacetamides compounds of 2- that step (3) obtains are added, 12~14 are heated to reflux under the protection of nitrogen
Hour；Reaction solution is cooled to room temperature after reaction, with saturation NH under condition of ice bath4It is 7 or so that Cl, which adjusts pH, is then used
Dichloromethane extracts, and obtained organic phase filters after being dried with anhydrous sodium sulfate, and vacuum distillation obtains solid mixture, through silica gel
The 4th eluant, eluent separating-purifying of chromatographic column, obtained solid matter are that the chromium ion based on rhodamine 6G detects fluorescence probe
Molecule.
3. preparation method according to claim 2, which is characterized in that rhodamine 6G, hydrazine hydrate and first in the step (1)
The ratio of alcohol is 3g:1mL:50mL, first eluant, eluent are n-hexane：Dichloromethane：Methanol volume ratio is 10:4:1 it is mixed
Close object.
4. preparation method according to claim 2, which is characterized in that rhodamine 6G hydrazide compound in the step (2)
It is 3 with the molar ratio of 2- aldehyde radicals -8-hydroxyquinoline and glacial acetic acid:2:0.1, the solvent is ethyl alcohol:Ether volume ratio is 1:1
Mixture, second eluant, eluent be dichloromethane.
5. preparation method according to claim 2, which is characterized in that 2-aminopyridine in the step (3), triethylamine and
The molar ratio of chloracetyl chloride is 1:1.1:1.1, the third eluant, eluent is dichloromethane:Ethyl acetate volume ratio is 3:1 it is mixed
Close object.
6. preparation method according to claim 2, which is characterized in that the derivative of rhodamine 6G, 2- in the step (4)
The molar ratio of chloro- N- pyridineacetamides, potassium carbonate and sodium iodide is 1:1.2:2:0.56, the 4th eluant, eluent is dichloromethane
Alkane:Ethyl acetate volume ratio is 3:1 mixture.
7. a kind of chromium ion detection fluorescent probe molecule according to claim 1 based on rhodamine 6G is used for water body sample
Product, tobacco sample, in caenorhabditis elegan body chromium ion detection purposes.
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