CN106220640A - One class mercury ion fluorescence probe and its preparation method and application - Google Patents
One class mercury ion fluorescence probe and its preparation method and application Download PDFInfo
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Abstract
The invention discloses class mercury ion fluorescence probe and preparation method thereof and use, belong to environmental analysis, the fluorescent probe technique field of bioanalysis detection.Technical scheme main points are: a class mercury ion fluorescence probe, are the base ring modified derivatives of rhodamine B, and structural formula is, ortho position that wherein substituent group F is positioned on phenyl ring, meta or para position.The invention also discloses preparation method and the application in mercury ion content in detection water environment system, medicine or biological cell system thereof of this fluorescent probe.Synthesis step of the present invention is simple, and the response time is short, it is easy to purifying and yield is higher, this compound is the Fluorescence Increasing type probe of mercury ion, mercury ion is had high specificity, can work under the conditions of physiological environment, have the prospect being applied to bioluminescence imaging.
Description
Technical field
The invention belongs to environmental analysis, the fluorescent probe technique field of bioanalysis detection, be specifically related to a class height and select
Property, mercury ion content in sample can be carried out mercury ion fluorescence probe of detection by quantitative and preparation method thereof and use.
Background technology
Hydrargyrum is a heavy metal species, is commonly called as hydrargyrum.The Hg of the dissolved2+There is higher chemism, be the master of mercury contaminants
Want existence form, when concentration is the lowest, organism is just had extremely strong physiological-toxicity.Inorganic mercury enters ring by all means
Behind border, it is methyl mercury by the role transformation of microorganism in deposit, threatens the existence of aqueous bio, cause a large amount of Fish to be poisoned
Or it is dead.The toxicity of hydrargyrum finally can be transferred in mankind's body on food chain top by the bioconcentration of food chain,
Hydrargyrum cannot be excreted by the mankind by own metabolism effect, and then causes multiple cardiovascular and cerebrovascular disease and hormonal system disease
Disease, the serious harm mankind's is healthy.Therefore, the analysis detection always people of hydrargyrum in organism and environment are paid special attention to
Problem, set up mercury ion detecting method fast and efficiently, in fields such as life sciences, medical science and environmental conservation, there is great meaning
Justice.
It has been reported that the method for multiple detection mercury ion in recent decades.Such as ultraviolet spectrophotometry, atom are inhaled
Transmit/receive the method for penetrating, hexavalent chrome bio-removal, inductively coupled plasma mass spectrometry and electrochemical process (voltammetry and polarography) etc., on
Although state several traditional detection method can complete the mercury content of different system samples is detected, but they equipment to instrument
Require the highest, expensive, and require there is substantial amounts of testing sample, and do not allow continuous detecting, self office of these methods
Sex-limited govern the instant detection in various testing samples particularly organism and trace analysis.
Fluorimetry has that selectivity is good, highly sensitive, detection limit is low and consumes the advantages such as sample size is few, at the trace of hydrargyrum
Amount detection plays effect greatly, and fluorescence molecule can be with penetration cell wall, to the metal in living cells and organism
Ion carries out the Tracing detection that develops, and this is that above-mentioned traditional analysis institute is the most replaceable.At present, fluorimetry is applied
For being fluorescent molecular probe widely.Fluorescent molecular probe is divided into solvay-type and response type two kinds, solvay-type fluorescent probe based on
The supermolecule of non-covalent bond interacts, mainly coordinate bond and hydrogen bond action；The identification process of response type fluorescent probe is different
In solvay-type, it is probe and detection object generation chemical reaction, generates the Photoactive compounds of different structure, makes fluorescence molecule
The spectral quality (mainly color) of probe changes, thus realizes optical sensing.Response type fluorescent molecular probe utilizes choosing
Select the advantage identification predetermined substance of reaction, there is the advantage such as specificity and irreversibility, and detection signal can be retained for a long time,
Receive more and more attention in recent years.Based on fluorescent parent such as rhodamine, Coumarins, cyanine dye and fluoresceins anti-
Type fluorescent molecular probe is answered to quickly grow in the context of detection of heavy metal ion.
Rhodamine B has the superior Photophysics such as good light stability and fluorescence quantum yield height and becomes a class relatively
Good fluorescent probe parent, is of very high actual application value, and its optical property is by external condition such as illumination, temperature and substituent group
Impact relatively big, can be with open loop under hydrion or some metal cation effect, fluorescence is obviously enhanced and along with color
Change.The rhodamine B fluorescent probe kind of common reporter is a lot, is roughly divided into complexation-volution open ring type and open loop-cyclisation
Response type.Wherein response type metal-ion fluorescent probe based on rhodamine volution " off-on " mechanism is the weight of Recent study
Point and focus, due to Hg2+Having thiophilicity, it can promote thiocarbonyls generation desulphurization reaction, generates corresponding carbonyl
Compound, such as Hg2+Induction thiourea derivatives devulcanization generates guanidine.The rhodamine fluorescence molecule of design sulfur-bearing carbonyl structure
Probe can identify Hg well2+, reacted Hg2+Presented in HgS precipitates.Research mechanism shows, rhodamine is specific
Spirane structure in owing to containing atoms such as N, O and S, making Hg2+First be combined with rhodamine, and promote that volution is unfolded into first
Individual balance, generates the rhodamine derivant with hyperfluorescence character further.Utilize fluorescent material optical physics before and after reaction
Matter is different, can realize Hg2+Detection.It addition, selectivity experiment proves that this kind of response type fluorescent probe is to Hg2+Show height
Degree specificity, significant in terms of chemistry and biological detection.
Summary of the invention
Present invention solves the technical problem that and there is provided class mercury ion fluorescence probe and preparation method thereof and use, this hydrargyrum from
Sub-fluorescent probe uses the most looped ester group-aminated for rhodamine B base ring method, and synthesis has the mercury ion of specificly-response
Fluorescent probe molecule, the selectivity fluorescence for mercury ion quickly detects.
The present invention solves that above-mentioned technical problem adopts the following technical scheme that, a class mercury ion fluorescence probe, its feature exists
Then the base ring modified derivative of rhodamine B, structural formula is, wherein substituent group F
The ortho position that is positioned on phenyl ring, meta or para position.
The preparation method of mercury ion fluorescence probe of the present invention, it is characterised in that concrete steps:
Step (1), is dissolved in rhodamine B in ethanol in reaction vessel, adds hydrazine hydrate aqueous solution, in the oil bath of 85 DEG C
Back flow reaction, TLC monitoring raw material reaction adds distilled water cancellation reaction the most afterwards, is extracted with ethyl acetate reaction system, collects
Distillation of reducing pressure after organic facies obtains rhodamine B hydrazide compound；
Step (2), adds fluorine-based substituted benzene isosulfocyanate compound and rhodamine that step (1) obtains in reaction vessel
B hydrazide compound, adds isopropanol, is stirred at reflux reaction in the oil bath of 85 DEG C, and TLC monitoring raw material reaction drops the most afterwards
To room temperature, reduce pressure and solvent is distilled off and adds neutral alumina its dry method is mixed sample, use neutral alumina as fixing phase
Carry out column chromatographic isolation and purification and obtain fluorine-based replacement fluorescent probe.
Further preferably, described in step (1), the mass concentration of hydrazine hydrate aqueous solution is 80%, hydrazine hydrate and rhodamine B
Mol ratio is 20:3.
Further preferably, fluorine-based substituted benzene isosulfocyanate compound described in step (2) and rhodamine B hydrazides chemical combination
The mol ratio of thing is 1.5:1.
Mercury ion fluorescence probe of the present invention is mercury ion in detection water environment system, medicine or biological cell system
Application in content, it is characterised in that detailed process is: mercury ion fluorescence probe is dissolved in dimethyl sulphoxide solution by (1), joins
The mercury ion fluorescence probe standard solution becoming molar concentration to be 0.1 mmol/L, with N-(2-ethoxy) piperazine-N'-2-ethane sulphur
The mercury ion storing solution of the buffer preparation variable concentrations of acid, takes 30 μ L mercury ion fluorescence probe standard solution respectively with 3
The mercury ion storing solution mixing of mL variable concentrations, left at room temperature 10 min, surveys glimmering at wavelength 589 nm of mixed liquor respectively
Light emission intensity, with the concentration of mercury ion as abscissa, draws standard curve with fluorescence intensity for vertical coordinate, or matching obtains again
Standard straight-line equation；(2) the dimethyl sulphoxide solution 30 μ L of the mercury ion fluorescence probe that molar concentration is 0.1 mmol/L is added
Enter in 3 mL testing sample solutions, left at room temperature 10 min, measure detected solution fluorescence at wavelength 589 nm and send out
Penetrate intensity, then contrast with standard curve or substitute into standard straight-line Equation for Calculating, obtain the concentration of mercury ion in testing sample solution.
The present invention compared with prior art has the advantages that the base ring of the rhodamine B that the present invention provides is modified and spreads out
Biology, synthesis step is simple, and the response time is short, it is easy to purifying and yield is higher, this compound is the Fluorescence Increasing type of mercury ion
Probe, has high specificity to mercury ion, can work under the conditions of physiological environment, has before being applied to bioluminescence imaging
Scape.
Accompanying drawing explanation
Fig. 1 is that pH is to fluorescent probe RFS and RFS+Hg2+The influence curve of fluorescence spectrum；
Fig. 2 is that the coordination time is to fluorescent probe RFS and RFS+Hg2+The influence curve of fluorescence spectrum；
Fig. 3 is the fluorescent probe RFS Selective recognition curve to different metal ion；
Fig. 4 is the fluorescent probe RFSP Selective recognition curve to different metal ion；
Fig. 5 is the fluorescent probe RFSM Selective recognition curve to different metal ion；
What Fig. 6 was coexisting ion on fluorescent probe RFS detection mercury ion affects figure；
What Fig. 7 was coexisting ion on fluorescent probe RFSP detection mercury ion affects figure；
What Fig. 8 was coexisting ion on fluorescent probe RFSM detection mercury ion affects figure；
Fig. 9 is the fluorescent probe RFS fluorometric investigation curve to mercury ion；
Figure 10 is the fluorescent probe RFSP fluorometric investigation curve to mercury ion；
Figure 11 is the fluorescent probe RFSM fluorometric investigation curve to mercury ion；
Figure 12 is the fluorescent probe RFS working curve to mercury ion complexation；
Figure 13 is the fluorescent probe RFSP working curve to mercury ion complexation；
Figure 14 is the fluorescent probe RFSM working curve to mercury ion complexation；
Figure 15 is the fluorescence imaging experiments that mercury ion is carried out in cyton by fluorescent probe RFS, wherein the rubbing of fluorescent probe RFS
Your concentration is 5 μMs, Hg2+Molar concentration be 10 μMs.
Detailed description of the invention
By the following examples the foregoing of the present invention is described in further details, but this should be interpreted as this
The scope inventing above-mentioned theme is only limitted to below example, and all technology realized based on foregoing of the present invention belong to this
Bright scope.
Embodiment 1
The synthesis of rhodamine B hydrazide compound
Take in the round-bottomed flask that rhodamine B 1.443 g (3 mmol) moves into 100 mL, dissolve with 50 mL ethanol, then with moving
Liquid rifle pipettes hydrazine hydrate aqueous solution 3 mL (80wt%, 20 mmol), and now reaction system is burgundy color, in the oil bath of 85 DEG C
Carrying out back flow reaction, thin layer chromatography detection method (TLC) monitoring extent of reaction, developing solvent is petroleum ether and the acetic acid of volume ratio 1:1
Ethyl ester mixed solution, puts plate and finds that on lamellae, raw material point disappears substantially after reacting 6 h, Rf value sieve occurs in about 0.5 position
Red bright hydrazide product point, slight fever afterproduct point is rose, and now reaction system is khaki by burgundy complexion changed, adds 100 mL
Distilled water cancellation is reacted, and is extracted with ethyl acetate system, and after collecting organic facies, rhodamine hydrazide compound is distilled to obtain in decompression
1.2537 g, yield 91%.
Embodiment 2
The synthesis of adjacent fluorine-based rhodamine probe RFS
Take 2-fluorobenzene isothiocyanate 0.1238 g (0.75 mmol) and rhodamine hydrazide compound 0.2292 g (0.5 mmol)
And be transferred in the round-bottomed flask of 25 mL, it is subsequently adding the isopropanol that 5 mL are dried, magnetic agitation backflow in the oil bath of 85 DEG C
Reaction, carries out TLC monitoring reaction to it after reacting 5 min, developing solvent be volume ratio be that the petroleum ether of 1:1 mixes with ethyl acetate
Solution, after 10 h, raw material point disappears, stopped reaction, and after system is down to room temperature, decompression is distilled off solvent and adds neutral alumina
Its dry method is mixed sample by aluminum, uses neutral alumina to carry out column chromatography (petroleum ether: ethyl acetate=10:2, v/v) point mutually as fixing
From purification, product is the adjacent fluorine-based rhodamine probe RFS(yields 82% of white puff sprills).
Embodiment 3
Between the synthesis of fluorine-based rhodamine probe RFSM
Take 3-fluorobenzene isothiocyanate 0.1238 g (0.75 mmol) and rhodamine hydrazide compound 0.2292 g (0.5 mmol)
And be transferred in the round-bottomed flask of 25 mL, it is subsequently adding the isopropanol that 5 mL are dried, magnetic agitation backflow in the oil bath of 85 DEG C
Reaction, carries out TLC monitoring reaction to it after reacting 5 min, developing solvent be volume ratio be that the petroleum ether of 1:1 mixes with ethyl acetate
Solution, after 10 h, raw material point disappears, stopped reaction, and after system is down to room temperature, decompression is distilled off solvent and adds neutral alumina
Its dry method is mixed sample by aluminum, uses neutral alumina to carry out column chromatography (petroleum ether: ethyl acetate=10:2, v/v) point mutually as fixing
From purification, product is fluorine-based rhodamine probe RFSM(yield 74% between white puff sprills).
Embodiment 4
Synthesis to fluorine-based rhodamine probe RFSP
Take 4-fluorobenzene isothiocyanate 0.1238 g (0.75 mmol) and rhodamine hydrazide compound 0.2292 g (0.5 mmol)
And be transferred in the round-bottomed flask of 25 mL, it is subsequently adding the isopropanol that 5 mL are dried, magnetic agitation backflow in the oil bath of 85 DEG C
Reaction, carries out TLC monitoring reaction to it after reacting 5 min, developing solvent be volume ratio be that the petroleum ether of 1:1 mixes with ethyl acetate
Solution, after 10 h, raw material point disappears, stopped reaction, and after system is down to room temperature, decompression is distilled off solvent and adds neutral alumina
Its dry method is mixed sample by aluminum, uses neutral alumina to carry out column chromatography (petroleum ether: ethyl acetate=10:2, v/v) point mutually as fixing
From purification, product is that white puff sprills are to fluorine-based rhodamine probe RFSP(yield 80%).
Embodiment 5
1, fluorescence spectrometry condition
Exciting slit width 5.0 nm, launch slit width 3.0 nm, excitation wavelength lambda ex is 566 nm or 568 nm, during response
Between 1 s, wave-length coverage 500-700 nm, sensitivity is medium, sample cell select 1 cm × 4, cm × 1 cm four-way quartz
Cuvette.
Accurately weigh 0.0122 g adjacent fluorine-based rhodamine probe RFS, a fluorine-based rhodamine probe RFSM and to fluorine-based Luo Dan
Bright probe RFSP, is dissolved in 10 mL volumetric flasks and constant volume with dimethyl sulfoxide solution (DMSO) respectively, and molar concentration is 2 ×
10-3 Mol/L, takes 0.5 mL mother solution and uses DMSO constant volume, the now molar concentration of fluorescent probe in another 10 mL volumetric flask
It is 1 × 10-4 Mol/L, then by HEPES buffer preparation 1 × 10-3 The various metals saline solution of mol/L, is diluted to equally
1×10-4 mol/L。
Wherein the test condition of fluorescent probe RFS and fluorescent probe RFSP is Ex=5 nm, Em=3 nm, fluorescent probe RFSM
Test condition be Ex=3 nm, Em=3 nm, the difference of test condition shows that the fluorine-based substituted fluorescent probe RFSM of meta is relative
Fluorescence intensity is more weak.
2, the screening of fluorometric investigation condition
(1) pH is to fluorescent probe RFS and RFS+Hg2+The impact of fluorescence spectrum
Under normal circumstances, the lactam nucleus of Rhodamine Derivatives is open loop in acid medium, has the strongest fluorescent emission, for
Fluorescent probe RFS is preferably applied in the complex system such as organism or environment, prepares different buffer systems such as
HAc-NaAc, HEPES (N-(2-ethoxy) piperazine-N'-2-ethane sulfonic acid), NaHCO3-Na2CO3Deng the pH value that adjustment is different,
Determine different pH value to fluorescent probe RFS and RFS+Hg2+Shadow in its maximum emission wavelength (589 nm) place's fluorescence intensity
Ringing, as it is shown in figure 1, be in the range of about 7 at pH, the fluorescence intensity of fluorescent probe RFS is almost unchanged, mainly with lactams
Stability Analysis of Structures exists.It is in the range of 6-8 at pH, RFS+Hg2+Fluorescence intensity almost unchanged.Therefore, it is the scope of 6-8 at pH
In, fluorescent probe RFS is to Hg2+Detection do not affected by pH, be conducive to its Hg in living cells and organism2+Detection.Institute
Using the HEPES buffer system (pH=7.20) of experimental selection physiological condition as the buffer solution measuring medium.
(2) coordination time and stability
Test result indicate that, the fluorescence intensity of Fig. 2 blank system is slowly increased in time, but after adding mercury ion, its fluorescence
Intensity strengthens rapidly, i.e. tends towards stability after 3 min, and during 5-10 min, fluorescence intensity is basically unchanged.Therefore select to mix at reagent
It is measured after acting on 10 min.
3, the impact of the different Selective recognition on mercury ion in fluoro substituents position on PITC
(1) fluorescent probe Selective recognition to different metal ion
In order to measure three kinds of fluorescent probes recognition performance to different metal ion, select transition metal (Fe respectively3+、Cr3+、
Co2+、Mn2+、Ni2+、Fe2+、Zn2+、Cu2+、Hg2+、Cd2+、Ag+), alkali metal (K+、Na+) and alkaline-earth metal (Ba2+、Ca2+、Mg2 +) and Hg2+For identifying object.As in Figure 3-5, except the Na that intracellular molar concentration is bigger+、Mg2+、Ca2+Molar concentration be 20
Outside μM, the concentration of remaining ion and fluorescent probe is 1 μM.Result shows, be separately added into blank fluorescence probe solution and
K+、Na+、Ag+、Mg2+、Ca2+、Ba2+、Cd2+、Co2+、Cr3+、Cu2+、Mn2+、Ni2+、Zn2+、Fe2+、Fe3+During ion, system is
Launch at wavelength 589 nm greatly colourless almost without fluorescence, solution, illustrate that now fluorescent probe is mainly presented in volution；
And adding Hg2+After, system fluorescence at maximum emission wavelength 589 nm is remarkably reinforced, and solution colour is become pink from colourless
Color, illustrates Hg2+In induced fluorescence probe electronics occur transfer thus cause spirane structure to be opened.Test result indicate that, three kinds glimmering
Light probe RFS, RFSM and RFSP are all to Hg2+There is higher single-minded identification ability.
(2) coexisting ion impact on three kinds of fluorescent probe detection mercury ions
Other metal ion and Hg are investigated further2+To Hg when coexisting2+Measure interference, make three kinds of fluorescent probes mole
Concentration is 10 μMs, except Ca2+、Mg2+、Na+Molar concentration be 0.50 mM(50 equiv) outward, other metal ion mole dense
Degree is 0.1 mM(10 equiv), as shown in figs 6-8, black histogram represent fluorescent probe molecule and different competitions from
Fluorescence intensity under son respectively Coexistence Situation, grey histogram represents and is separately added into 1 equiv Hg2+Fluorescence in rear each system
Intensity.From chart data it can be seen that at coexisting ion (especially Ca2+、Mg2+、Na+Concentration be Hg2+50 times of concentration) deposit
Under the conditions, three kinds of fluorescent probes RFS, RFSM and RFSP all show stronger capacity of resisting disturbance to other metal ion.
Therefore, fluorescent probe RFS, RFSM and RFSP is to Hg2+There is higher selective Fluorescence Increasing type probe.
The mensuration of (3) three kinds of fluorescent probe detection limits
Be standard variance by formula LOD=3 σ/K(σ, n=6) the detection limit that calculates correspondence is followed successively by 0.6x10-7 mol/L、
2.7x10-7 Mol/L and 1.2x10-7Mol/L, it follows that fluorescent probe is to mercury ion when on phenyl ring, substituting group position is meta
The sensitivity identified is poor, and when benzene ring substituents is ortho para substituent group, fluorescent probe is to the sensitivity of mercury ion identification relatively
Good, wherein ortho-substituent sensitivity outline is higher than para-position.
4, fluorescent probe and Hg2+The mechanism of action inquire into
By fluorescent probe RFS (0.2 mmol) and HgCl under room temperature2(0.2 mmol) stirs 20 min in methanol, has black not
Molten solid separates out, and is spin-dried for solvent and obtains compound R FS and Hg after sucking filtration2+Reaction afterproduct RFR, 1H NMR (400 MHz,
CDCl3) δ 9.55 (s, 1H), 8.26 (d, J = 7.8 Hz, 1H), 7.84 (t, J = 7.2 Hz, 1H),
7.70 (t, J = 7.7 Hz, 1H), 7.63 (t, J = 7.5 Hz, 1H), 7.25 (d, J = 7.4 Hz, 1H),
7.08 (d, J = 9.4 Hz, 2H), 6.97 (d, J = 7.8 Hz, 1H), 6.93 (d, J = 5.5 Hz, 1H),
6.89 (d, J = 7.3 Hz, 1H), 6.77 (d, J = 9.4 Hz, 2H), 6.68 (s, 2H), 3.53 (q, J
= 6.9 Hz, 8H), 1.24 (t, J = 6.9 Hz, 12H)。
5, in living cells, fluorescent probe RFS detects Hg2+
In order to study the ability of probe RFS detection Hg 2+ in cancerous cell, first by Hela cell phosphate buffered solution
(DPBS) the HEPES buffer solution of 1% DMSO is contained after rinse with the RFS(of 5 μMs) cultivate 30 minutes at 37 DEG C, then use
PBS buffer solution cleans 3 times, the most again by Hela cell respectively with the HgCl of 10 μMs237 DEG C of HEPES buffer solution at
Cultivate 30 minutes, again clean 3 times with PBS buffer solution.Now observe with Olympus FV 1000 laser scanning confocal microscopy
Confocal fluorescent imaging.
Embodiment above describes the ultimate principle of the present invention, principal character and advantage, the technical staff of the industry should
Understanding, the present invention is not restricted to the described embodiments, and the simply explanation present invention's described in above-described embodiment and description is former
Reason, under the scope without departing from the principle of the invention, the present invention also has various changes and modifications, and these changes and improvements each fall within
In the scope of protection of the invention.
Claims (6)
1. a class mercury ion fluorescence probe, it is characterised in that being the base ring modified derivative of rhodamine B, structural formula is, ortho position that wherein substituent group F is positioned on phenyl ring, meta or para position.
2. the preparation method of the mercury ion fluorescence probe described in a claim 1, it is characterised in that concrete steps:
Step (1), is dissolved in rhodamine B in ethanol in reaction vessel, adds hydrazine hydrate aqueous solution, in the oil bath of 85 DEG C
Back flow reaction, TLC monitoring raw material reaction adds distilled water cancellation reaction the most afterwards, is extracted with ethyl acetate reaction system, collects
Distillation of reducing pressure after organic facies obtains rhodamine B hydrazide compound；
Step (2), adds fluorine-based substituted benzene isosulfocyanate compound and rhodamine that step (1) obtains in reaction vessel
B hydrazide compound, adds isopropanol, is stirred at reflux reaction, is down to after TLC monitoring raw material reaction is complete in the oil bath of 85 DEG C
Room temperature, reduces pressure and solvent is distilled off and adds neutral alumina its dry method is mixed sample, use neutral alumina to enter mutually as fixing
Row column chromatographic isolation and purification obtains fluorine-based replacement fluorescent probe.
The preparation method of mercury ion fluorescence probe the most according to claim 2, it is characterised in that: water described in step (1)
The mass concentration closing hydrazine aqueous solution is 80%, and hydrazine hydrate is 20:3 with the mol ratio of rhodamine B.
The preparation method of mercury ion fluorescence probe the most according to claim 2, it is characterised in that: fluorine described in step (2)
Base substituted benzene isosulfocyanate compound is 1.5:1 with the mol ratio of rhodamine B hydrazide compound.
5. the mercury ion fluorescence probe described in claim 1 in detection water environment system, medicine or biological cell system hydrargyrum from
Application in sub-content.
Mercury ion fluorescence probe the most according to claim 5 is in detection water environment system, medicine or biological cell system
Application in mercury ion content, it is characterised in that detailed process is: mercury ion fluorescence probe is dissolved in dimethyl sulphoxide solution by (1)
In, it is made into the mercury ion fluorescence probe standard solution that molar concentration is 0.1 mmol/L, with N-(2-ethoxy) piperazine-N'-2-
The mercury ion storing solution of the buffer preparation variable concentrations of ethane sulfonic acid, takes 30 μ L mercury ion fluorescence probe standard solution and divides
Do not mix with the mercury ion storing solution of 3 mL variable concentrations, left at room temperature 10 min, survey mixed liquor respectively at wavelength 589 nm
The fluorescent emission intensity at place, with the concentration of mercury ion as abscissa, draws standard curve with fluorescence intensity for vertical coordinate, or intends
Conjunction obtains standard straight-line equation；(2) by the dimethyl sulphoxide solution of mercury ion fluorescence probe that molar concentration is 0.1 mmol/L
30 μ L join in 3 mL testing sample solutions, left at room temperature 10 min, measure detected solution at wavelength 589 nm
Fluorescent emission intensity, then contrast with standard curve or substitute into standard straight-line Equation for Calculating, obtain mercury ion in testing sample solution
Concentration.
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| CN107033158A (en) * | 2016-12-29 | 2017-08-11 | 济南大学 | A kind of colorimetric fluorescence probe of hypersensitive analysis mercury ion and preparation method thereof |
| CN108218880A (en) * | 2016-12-22 | 2018-06-29 | 王可 | A kind of novel mercury ion optical probe and preparation method and application |
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| CN108892681A (en) * | 2018-09-12 | 2018-11-27 | 泰山医学院 | A kind of imidazo [1,2-a] pyridines mercury ion Ratiometric fluorescent probe and its application |
| CN109320520A (en) * | 2018-09-28 | 2019-02-12 | 南京林业大学 | A kind of rhodamine B Hg based on pyridine -3- sulfonic acid chloride2+The preparation and application of fluorescent optical sensor |
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| CN109705111A (en) * | 2019-01-20 | 2019-05-03 | 湖南科技大学 | A kind of mercury ion detecting probe and its preparation method and application |
| CN110229165A (en) * | 2019-05-31 | 2019-09-13 | 苏州科技大学 | Up-conversion fluorescence probe Rhodamine Derivatives and its application |
| CN113295656A (en) * | 2020-02-22 | 2021-08-24 | 青岛科技大学 | Intracellular As3+、Pb2+And Hg2+Simultaneous fluorescence imaging method |
| CN113603702A (en) * | 2021-08-11 | 2021-11-05 | 中国矿业大学 | Colorimetric/fluorescent probe for detecting mercury ions and preparation method and application thereof |
| CN113603702B (en) * | 2021-08-11 | 2022-09-06 | 中国矿业大学 | Colorimetric/fluorescent probe for detecting mercury ions and preparation method and application thereof |
| CN114213385A (en) * | 2021-11-30 | 2022-03-22 | 浙江工业大学 | Fluorescein type ionic liquid, synthetic method thereof and application thereof in mercury ion or methyl mercury ion detection |
| CN115197230A (en) * | 2022-07-05 | 2022-10-18 | 天津科技大学 | For Hg 2+ And Pb 2+ Paper-based array sensor for visual rapid screening |
| CN115197230B (en) * | 2022-07-05 | 2024-05-31 | 天津科技大学 | Used for Hg2+And Pb2+Visual quick screening paper-based array sensor |
| CN116751210A (en) * | 2023-06-10 | 2023-09-15 | 郑州大学 | Water-soluble probe for detecting mercury ions and preparation method and application thereof |
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