CN103254128A - Water-soluble fluorescent probe for naked eye and light enhancement detection of mercury ions and synthesis and application thereof - Google Patents

Abstract

The utility model relates to a novel naked-eye and fluorescence-enhancement type mercury ion fluorescent probe and its preparation and an application, specifically to a preparation based on 7-hydroxy-1-alkyl quinoline derivative and its high-sensitivity detection of mercury ions in a pure water solution. The fluorescent probe provided by the invention is obtained by a reaction between 7-ethylnenoxy quinoline derivative and an alkylating reagent, and a probe pure product is obtained through column chromatography isolation. The synthesis of the fluorescent probe is simple and requires mild reaction condition. The fluorescent probe is easy to purify, and has advantages of high yield, good water solubility, stable optical performance and high fluorescence quantum efficiency. An aqueous solution of probe molecules is colorless and transparent, and almost has no fluorescence; and after mercury ions are added, the solution changes from colorless to yellow and presents strong green fluorescence. The probe molecules have strong recognition capability of mercury ions and have high sensitivity. Response range is 0-3uM, and the lower limit of detection is low and reaches 0.22nM. The probe provided by the invention has important practical application value in the fields of biological and medicinal chemistry, environmental sciences and the like.

Description

A kind of water-soluble bore hole and light strengthen fluorescent probe and the synthetic and application thereof that detects mercury ion

Technical field

What the present invention relates to is chemical analysis detection technique field, is specifically related to a kind of bore hole and fluorescence and strengthens detection mercury ion fluorescent molecular probe and preparation method thereof and the application aspect the detection mercury ion.

Background technology

Mercury pollution has become a global environment problem.Mainly be to be discharged in the physical environment by a series of artificial activity to go among inorganic mercury such as Hg (0) and the Hg (II), the for example burning of coal stone wet goods fossil oil, the incineration of the smelting of gold mine and solid waste matter all can be discharged into inorganic mercury in the large natural environment.Mercury ion can be accumulated in vivo by various animals and plants in physical environment, and forms the organic mercury of severe toxicity, finally enters in the human body by food chain then.Methyl mercury is very remarkable to the harm of human body, and it can be absorbed by the body and break through the hemato encephalic barrier of human body easily, directly acts on our central nervous system then, and human body is caused huge injury.For example occurring in the minamata disease event of Japan and occurring in Irak mercury poisoning event is exactly the most serious example of mercury pollution harm.Therefore the detection to mercury is very necessary.[J]Environment?Intefnational.2001，27，285-290.Harada?M，Nakaishi?J，Yasoda?E，Pinheiro?MCN，Oikawa?T.Mercury?pollution?in?the?Tapajos?River?basin，Amazon：Mercury?level?of?head?hair?and?health?effects.[J]Science?of?The?Total?Environment.Olivero?J，Johnson?B，Arguello?E.Human?exposure?to?mercury?in?San?Jorge?river?basin，Colombia(South?America).[J]Chem.Rev.2008，108，3443-3480.Nolan?EM，Lippard?SJ.Tools?and?tactics?for?the?optical?detection?of?mercuric?ion.

The detection by quantitative of the mercury ion of traditional water sample kind generally adopts atomic absorption, means such as cold steam atomic fluorescence spectroscopy and vapor-phase chromatography.These methods often need sample is compared complicated pretreatment, need expensive precision instrument simultaneously.Comparatively speaking, the small molecules fluorescent probe is when detecting mercury ion, and sample preparation is succinct, and it is low to detect lower limit, and a large amount of mercury ion fluorescent probe reports has been arranged at present.Wherein, most mercury ion fluorescent probes all are to utilize that the change to fluorescence realizes one of mercury ion quantitative measurement behind mercury ion and some the heteroatomic complex coordinations.This often have reasonable sensitivity based on the mercury ion probe with the heteroatoms coordination, reaches the reaction times fast; But be subjected to silver easily, gold, lead, cadmium, the interference of metal ions such as copper, selectivity is often relatively poor.Therefore in recent years, the mercury ion probe based on mechanism of catalytic reaction had developed gradually.Mainly contain two kinds of mechanisms of catalytic reaction at present: one, utilize the sulphur property of having a liking for of mercury ion, allow mercury ion catalysis thiocarbonyl group be hydrolyzed into carbonyl, thereby realize the change of fluorophore molecular fluorescence, though the mercury ion fluorescent probe based on this mechanism increases on selectivity, but still be subjected to the interference of metal ions such as silver easily, and this hydrolysis reaction is at room temperature slow, needs the long reaction times.Its two, be to utilize mercury ion katalysis in solution that vinyloxy group is hydrolyzed into hydroxyl, thereby change the fluorescence of fluorophore that often have sensitivity preferably and good selectivity based on such mercury ion fluorescent probe, the reaction times is moderate.Yet, the solubleness of mercury ion fluorescent probe in water of the type of report is not fine at present, when the mercury ion in the aqueous solution is carried out quantitative measurment, often need to add some organic solvents (as DMSO, CH3CN, DMF, THF etc.) as solubility promoter, also there is not to measure in 100% the aqueous solution the inside this type fluorescent probe report of mercury ion at present.Therefore, a kind of novel mercury ion fluorescent probe that has highly sensitive, highly selective and have a good aqueous solubility of exploitation has very important meaning.[J]Org.Lett.2007.9.2313-2316.Yuan?M，LiY，Li?J，Li?C，Liu?X，Lv?J，Xu?J，Liu?H，Wang?S，Zhu?D.A?colorimetric?and?fluorometric?dual-modal?assay?for?mercury?ion?by?a?molecule.[J]Org.Lett.2009.11.2740-2743.Suresh?M，Mishra?S，Mishra?SK，Suresh?E，Mandal?AK，Shrivastav?A，Das?A.Resonance?energy?transfer?approach?and?a?new?ratiometric?Probe?for?Hg ²⁺?in?aqueous?media?and?living?organism.[J]Org.Lett.2011.13.1422-1425.Kumar?M，Kumar?N，Bhalla?V，?Singh?H，Sharma?PR，Kaur?T.Naphthalimide?appended?rhodamine?derivative：through?bond?energy?transfer?for?sensing?of?Hg ²⁺?ions.[J]J.Am.Chem.Soc.2005.127.16760-16761.Yang?YK，Yook?KJ，Tae?J.A?rhodamine-based?fluorescent?and?colorimetric?chemodosimeter?for?the?rapid?detection?of?Hg ²⁺ions?in?aqueous?media.[J]Angew.Chem.Int.Ed.2008.47.8025-8029.A?ratiometric?fluorescent?probe?based?on?FRET?for?imaging?Hg ²⁺ions?in?living?cells.[J]Chem.Comm.2012.48.8371-8373.Jiang?J，Liu?W，?Cheng?J，Yang?L，Jiang?H，Bai?D，Liu?W.A?sensitive?colorimetric?and?ratiometric?fluorescent?probe?for?mercury?species?in?aqueous?solution?and?living?cells.[J]Org.Lett.2011.13.3422-3425.Santra?M，Roy?B，Ahn?KH.A“Reactive”ratiometric?fluorescent?probe?for?mercury?species.

Summary of the invention

For above-mentioned situation, one of purpose of the present invention provide a kind of synthetic easy, reaction conditions is gentle, the synthetic method of lower-cost mercury ion fluorescent probe; Two of purpose provides that a kind of selectivity is good, highly sensitive, good water solubility and the mercury ion fluorescent probe.

The open hole detection that the present invention uses and fluorescence strengthen detection mercury ion fluorescent molecular probe, adopt 7-quinolinol derivative and alkylating reagent synthetic, and the vinyloxy group part is as recognition group.

The present invention's technical scheme that adopts of dealing with problems is that a kind of open hole detection and fluorescence strengthen the fluorescent molecular probe that detects mercury ion, have following general structure:

Wherein:: R ₁, R ₂Be H atom or methyl, R ₃A kind of for H atom or chlorine atom, R ₄For-CH ₃,-CH ₂CH ₃,-CH ₂CH ₂CH ₃Perhaps-CH ₂CH ₂CH ₂SO ₃H's is a kind of.

R in formula I ₁, R ₂, R ₃Be the H atom, R ₄For methyl is, the synthetic route of the representational compound of the present invention is shown in following formula:

(1) 7-hydroxyquinoline, salt of wormwood and glycol dibromide are dissolved in the acetone together, reflux is 48 hours then.With thin-layer chromatography monitoring reaction process, treat that raw material reaction finishes, cool to room temperature, distillation desolventizing.Use dichloromethane extraction again, organic phase is used saturated ammonium chloride solution respectively, saturated aqueous common salt and water washing.Separate organic phase, add anhydrous sodium sulfate drying, the distillation desolventizing, column chromatographic isolation and purification gets pure 7-(2-bromine oxethyl)-quinoline.

(2) 7-(2-bromine oxethyl)-quinoline is dissolved in the dimethyl sulfoxide (DMSO), adds potassium tert.-butoxide, stirring at room 2 hours is with thin-layer chromatography monitoring reaction process.Reaction is poured into whole system in the water after finishing, and uses ethyl acetate extraction, isolates organic layer, uses saturated ammonium chloride solution respectively, saturated aqueous common salt, water washing.Anhydrous sodium sulfate drying, the distillation desolventizing obtains pure 7-vinyloxy group quinoline.(3) 7-vinyloxy group quinoline and methyl iodide are dissolved in the acetonitrile.Reflux 6 hours, the thin-layer chromatography monitoring reaction.After treating that raw material point disappears, stop heating, be cooled to room temperature.The distillation desolventizing, resistates uses column chromatography and obtains pure probe compound.

In the embodiment of this specification sheets, understand the synthetic and detection method of this probe in more detail.Fluorescent molecular probe using method of the present invention does not have particular restriction, can not need extra organic solvent with in the direct dissolved aqueous solution of probe molecule usually, tests under the room temperature.When adding mercury ion, because the hydrolysis of mercury ion catalyzed ethylene oxygen base becomes hydroxyl, thereby improved power supply performance, make fluorescence strengthen, follow the change of this solution colour simultaneously.Obvious variation has taken place in ultraviolet and fluorescence spectrum.

It is as follows that open hole detection of the present invention and fluorescence enhancing detect the concrete feature that contains the mercaptoamino-acid fluorescent molecular probe:

This molecular fluorescence probe solvability in the aqueous solution is better, and absorbing wavelength is 349nm, does not almost observe fluorescent signal; Along with the adding of mercury ion, probe molecule descends gradually in the maximum absorption at 349nm place, and a new absorption peak that strengthens gradually appears in the 404nm place simultaneously, and the absorption spectrum blue shift is about 55nm, and the color of solution is by the colourless yellow that become; In fluorescence spectrum, maximum emission peak appears in the 507nm place, and along with the increase of ion concentration of mercury, its fluorescence intensity also increases thereupon, and maximum the enhancing is 127 times.Under the irradiation of ultraviolet lamp, probe presents stronger green fluorescence.

Probe molecule raw material of the present invention is easy to get, and synthetic yield is higher, and optical property is stable, and sensitivity is higher, and strong to the mercury ion recognition capability, response speed is very fast, and responding range is 0-3 μ M, and detectability is low to be 0.22nM.Therefore the type probe has actual using value in fields such as biological chemistry, environmental sciences.

Description of drawings

Fig. 1 be fluorescent probe of the present invention in 100% aqueous solution, the changing conditions that ultra-violet absorption spectrum increases with ion concentration of mercury, X-coordinate is wavelength, ordinate zou is absorbancy.

Fig. 2 be fluorescent probe of the present invention in 100% aqueous solution, adding behind the mercury ion of 1.25 times of equivalents at wavelength is 507nm place fluorescence intensity variation relation in time.X-coordinate is the time, the ordinate zou fluorescence intensity.

Fig. 3 be fluorescent probe of the present invention in 100% aqueous solution, the changing conditions that fluorescence emission spectrum increases with ion concentration of mercury, X-coordinate is wavelength, ordinate zou is fluorescence intensity.

Fig. 4 be fluorescent probe of the present invention in 100% the aqueous solution, the linear relationship of fluorescence intensity ratio and ion concentration of mercury before and after the reaction.X-coordinate is the concentration of mercury ion, and ordinate zou is F/F ₀

Fig. 5 is in the aqueous solution of fluorescent probe 100% of the present invention, and fluorescence spectrum is with the different metal ion that adds same concentration: K ⁺, Na ⁺, Ca ²⁺, Cd ²⁺, Fe ³⁺, Ag ⁺, Co ²⁺, Pb ²⁺, Cu ²⁺, Zn ²⁺, Cr ³⁺, Mg ²⁺, Pd ²⁺, Al ³⁺, Mn ²⁺, Au ³⁺After changing conditions.X-coordinate is wavelength, and ordinate zou is fluorescence intensity.

Fig. 6 is that fluorescent probe of the present invention is in 100% aqueous solution, at other competing ions K ⁺, Na ⁺, Ca ²⁺, Cd ²⁺, Fe ³⁺, Ag ⁺, Co ²⁺, Pb ²⁺, Cu ²⁺, Zn ²⁺, Cr ³⁺, Mg ²⁺, Pd ²⁺, Al ³⁺, Mn ²⁺, Au ³⁺Exist the fluorescence intensity that adds the mercury ion front and back down to change.X-coordinate is the concentration of different metal ion, and ordinate zou is fluorescence intensity.

Fig. 7 be fluorescent probe of the present invention in 100% aqueous solution, fluorescence intensity is with adding isoconcentration different anions tester: I ^-, Br ^-, F ^-, SO ₃ ^2-, NO ₂ ^-, ClO ₄ ^-, AcO ^-After the fluorescence spectrum changing conditions, X-coordinate is wavelength, ordinate zou is fluorescence intensity.

Fig. 8 be fluorescent probe of the present invention in 100% the aqueous solution, fluorescence intensity is with different testers: I ^-, Br ^-, F ^-, SO ₃ ^2-, NO ₂ ^-, ClO ₄ ^-, AcO ^-Add the changing conditions that the back increases, X-coordinate is coexistence tester, the fluorescence intensity at ordinate zou 507nm place.

Fig. 9 be fluorescent probe of the present invention in 100% aqueous solution, add different positively charged ion testers: K ⁺, Na ⁺, Ca ²⁺, Cd ²⁺, Fe ³⁺, Ag ⁺, Co ²⁺, Pb ²⁺, Cu ²⁺, Zn ²⁺, Cr ³⁺, Mg ²⁺, Pd ²⁺, Al ³⁺, Mn ²⁺, Au ³⁺After the ultra-violet absorption spectrum changing conditions.Ordinate zou is absorbancy, and X-coordinate is wavelength.

Probe fluorescence intensity under different pH conditions that Figure 10 synthesizes for the present invention reaches the variation of fluorescence intensity behind the mercury ion that adds 3 times of amounts under the different pH.Wherein ordinate zou is fluorescence intensity, and X-coordinate is pH.

The proton nmr spectra of the 7-that Figure 11 synthesizes for the present invention (2-bromine oxethyl)-quinoline.

The proton nmr spectra of the 7-vinyloxy group quinoline that Figure 12 synthesizes for the present invention.

The proton nmr spectra of the molecular fluorescence probe that Figure 13 synthesizes for the present invention.

Embodiment

The preparation of embodiment 1:7-(2-bromine oxethyl)-quinoline

With the 7-hydroxyquinoline (145mg, 1mmol), salt of wormwood (276mg, 2mmol) and glycol dibromide (752mg 4mmol) joins in the 10mL acetone, behind the reflux 48h, is cooled to room temperature, the distillation desolventizing.With methylene dichloride (2x50mL) extracting twice, organic phase is used saturated ammonium chloride solution, saturated aqueous common salt and water washing respectively successively.Tell organic phase, add anhydrous sodium sulfate drying, the distillation desolventizing, column chromatography for separation (5: 1 sherwood oil of volume ratio: ethyl acetate is eluent) gets pure white solid 122mg (yield: 51%), be 7-(2-bromine oxethyl)-quinoline.

¹HNMR(300MHz，CDCl ₃)：δ _ppm＝8.84(1H，d，J＝6Hz)，8.09(1H，d，J＝6Hz)，7.72(1H，d，J＝9Hz)，7.40(1H，s)，7.29-7.30(1H，m)，7.24(1H，d，J＝6Hz)，4.47(2H，t，J＝6Hz)，3.75(2H，t，J＝9Hz，J＝6Hz).

The preparation of embodiment 2:7-vinyloxy group quinoline

(62mg 0.25mmol) is dissolved in the 2mL dimethyl sulfoxide (DMSO), and (56mg, 0.5mmol) stirring at room is 2 hours, with thin-layer chromatography monitoring reaction process to add potassium tert.-butoxide with 7-(2-bromine oxethyl)-quinoline.Reaction is poured into reaction solution in the 50mL water after finishing, and with ethyl acetate (2x75mL) extracting twice, isolates organic layer, uses saturated ammonium chloride solution, saturated aqueous common salt water washing respectively.Separate organic phase, anhydrous sodium sulfate drying.The underpressure distillation desolventizing gets water white oily liquids 41mg (yield: 95%), be pure 7-vinyloxy group quinoline.

HNMR(300MHz，CDCl ₃)：δ _ppm＝8.87(1H，d，J＝6Hz)，8.11(1H，d，J＝9Hz)，7.77(1H，d，J＝9Hz)，7.59(1H，s)，7.32-7.30(1H，m)，7.27(1H，d，J＝6Hz)，6.85(1H，t，J＝9Hz，J＝6Hz)，4.93(1H，d，J＝12Hz)，4.62(1H，d，6Hz).

Embodiment 3: the preparation of molecular fluorescence probe

With 7-vinyloxy group quinoline (35mg, 0.2mmol) and methyl iodide (284mg 2mmol) is dissolved in the 3mL acetonitrile.After the reflux 12 hours, be cooled to room temperature.The distillation desolventizing, resistates uses column chromatography (volume ratio is 15: 1 methylene dichloride: methyl alcohol is eluent) and gets pale solid 25mg (yield: 40%), be probe molecule.

HNMR(300MHz，CDCl ₃)：δ _ppm＝9.39(1H，d，J＝6Hz)，9.18(1H，d，J＝6Hz)，8.47(1H，d，J＝9Hz)，8.03(1H，t，J＝6Hz)，7.93(1H，s)，7.82(1H，d，J＝9Hz)，7.46-7.39(1H，m)，5.12(1H，d，J＝12Hz)，4.88(1H，d，J＝6Hz)，4.57(3H，s).

Embodiment 4: bore hole and fluorescence strengthen the application that detection contains the mercury ion fluorescent probe

This fluorescent probe is soluble in water, add corresponding anion buffer, system is made into 1 * 10 ^-6The solution of M is tested its ultra-violet absorption spectrum and fluorescence spectrum and is changed.Fig. 1-Fig. 9 shows that probe has very high selectivity to containing mercury ion, and along with its UV spectrum of increase and the fluorescence spectrum generation considerable change of ion concentration of mercury, solution colour generation noticeable change is applicable to open hole detection; Simultaneously, add mercury ion after, fluorescence intensity strengthens 127 times, and probe is not subjected to some other metal ion, and the influence of some common negatively charged ion, as K ⁺, Na ⁺, Ca ²⁺, Cd ²⁺, Fe ³⁺, Ag ⁺, Co ²⁺, Pb ²⁺, Cu ²⁺, Zn ²⁺, Cf ³⁺, Mg ²⁺, Pd ²⁺, Al ³⁺, Mn ²⁺, Au ³⁺I ^-, Br ^-, F ^-, SO ₃ ^2-, NO ₂ ^-, ClO ₄ ^-, AcO ^-Under the condition that above-mentioned interfering ion exists, probe still has good selectivity and sensitivity to containing mercury ion.

Claims (6)

1. the present invention relates to a kind of bore hole and fluorescence and strengthen to detect the preparation method of mercury ion fluorescent molecular probe and this fluorescent molecular probe in the application that detects mercury ion, it is characterized in that having following chemical molecular general structure:

The fluorescent probe of described detection mercury ion is to be fluorophore with 7-hydroxyl-1-alkyl quinoline, and vinyloxy group is the compound of the recognition group of mercury ion.Wherein: R ₁Be H atom, R ₂For the H atom or-CH ₃Group, R ₃Be H atom or Cl atom, R ₄For-CH ₃,-CH ₂CH ₃,-CH ₂CH ₂CH ₃Perhaps-CH ₂CH ₂CH ₂SO ₃A kind of among the H.

2. the synthetic method of a fluorescent molecular probe according to claim 1 is characterized in that, described preparation method is as follows:

1) 7-quinolinol derivative, salt of wormwood and glycol dibromide are added in the acetone reflux.After reaction finishes, cool to room temperature, distillation desolventizing.Use dichloromethane extraction, use saturated ammonium chloride solution, saturated aqueous common salt and water washing respectively.Separate organic phase and use anhydrous sodium sulfate drying, the distillation desolventizing, column chromatographic isolation and purification obtains pure 7-(2-bromine oxethyl)-quinoline.

2) 7-(2-the bromine oxethyl)-quinoline with gained in the step 1) is dissolved in the dimethyl sulfoxide (DMSO), adds potassium tert.-butoxide, and stirring at room is after 2 hours, reaction solution is poured in the water, ethyl acetate extraction is isolated organic layer, uses saturated ammonium chloride solution, saturated aqueous common salt, water washing respectively.The separated and collected organic phase is also used anhydrous sodium sulfate drying, and the distillation desolventizing obtains pure 7-vinyloxy group quinoline.

3) with step 2) in 7-vinyloxy group quinoline and the alkylating reagent of gained be dissolved in the acetonitrile.After the reflux 6 hours, be cooled to room temperature.The distillation desolventizing obtains solid, uses column chromatography purifying and obtains fluorescent probe compounds.

3. method according to claim 2 is characterized in that: adopt the 7-quinolinol derivative described reactions steps 1).

4. method according to claim 2 is characterized in that described reactions steps 3) in the alkyl base reagent that adopts can be C for carbon chain lengths ₁-C ₃The straight chain alkane iodide, sulfuric acid diformazan methyl esters, ethyl sulfate, p-methyl benzenesulfonic acid methyl esters, p-methyl benzenesulfonic acid ethyl ester and 1,3-N-morpholinopropanesulfonic acid lactone.

5. method according to claim 2 is characterized in that: the solvent that adopts described reactions steps 3) can be organic solvents such as acetonitrile, methylene dichloride, chloroform.

6. the application of the described fluorescent molecular probe of claim 1 is characterized in that: selected mercury ion is mercury chloride, Mercury bisulfate, Mercury pernitrate inorganic salt.

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