CN106841129B - The method for measuring ion concentration of mercury in Studies on Avian Habitat water source - Google Patents
The method for measuring ion concentration of mercury in Studies on Avian Habitat water source Download PDFInfo
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- 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
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- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The present invention relates to the methods of ion concentration of mercury in measurement Studies on Avian Habitat water source.In ethanol/water (1/1, v/v) system (pH=7.4, PBS buffer solution), the detection to mercury ion is may be implemented in the method for the present invention.Method of the invention is to the response of mercury ion not by coexisting ion K+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+Interference, to Hg2+Selectivity it is fine.When ion concentration of mercury is within the scope of 0-4 μm of ol/L, the concentration and fluorescence intensity of mercury ion are at good linear relationship, regression equation are as follows: y=107305x+99254 (R2=0.9875), detection is limited to 0.07 μm of ol/L.
Description
Technical field
The present invention relates to a kind of methods of ion concentration of mercury in measurement Studies on Avian Habitat water source.
Technical background
Mercury is element a kind of volatile and with serious toxicity, and mercury element once enters water body, the mercury ion in environment
It will be converted into organic mercury (such as methyl mercury), and can be accumulated always in vivo by bacterium, through food chain enrichment,
It is finally absorbed by the mankind and other animals such as birds and generates harm.Mercury and its derivative can destroy organism molecule knot
Structure, this is because mercury ion can be combined with the group of the sulfur-bearing in protein, enzyme.Especially methyl mercury can be poisoned seriously dynamic
The nerve of object causes the behavior of animal to perceive disorder, neurotoxic and kidney, the production to the mankind and other animals such as birds
Raw very big threat.
The feather and ovum of bird are high to mercury enrichment degree, shadow of the mercury ion content to composition of bird community or population in water source
Sound is it will be apparent that in addition birds are the indicator of environment.So the content of mercury is to pass in government department's monitoring water environment
Important.Therefore, research and develop it is more highly selective, more quickly, it is more highly sensitive can be in environment such as natural waters
In the method for mercury ion content detection in real time be very important, because of that one can track in water source mercury ion content or
It changes the influence to composition of bird community or population.
Currently, common mercury element detection means is mainly traditional dithizone method colorimetric method, anodic stripping voltammetry, oxygen
Change reduction potential method, inductively coupled plasma mass spectrometry, Atomic absorption/emission spectrometry, x ray fluorescence spectrometry etc..But
In practical application, these detection methods are costly and complicated for operation.And fluorescence probe detection method is due to, selection good with convenience
The advantages that property, high sensitivity and it is extremely noticeable.In recent years, exactly just make mercury ion glimmering under above-mentioned various backgrounds
The research of light probe obtains the concern of more and more scholars.
Chinese patent application 201610463837.9 disclose a kind of mercury ion fluorescence probe based on naphthalimide and its
Preparation method and application, prepared mercury ion fluorescence probe be with 1,8- naphthalimide be fluorescent parent, pass sequentially through with just
Butylamine reaction, obtains bromo- 1, the 8- naphthalimide of intermediate N butyl -4- of fluorescent both, gained intermediate and 2- methoxybenzyl
Amine reaction introduces nitrogen-atoms and oxygen atom, enhances the coordination ability of the compound and metal ion, and that invents in detection process should
Nitrogen-atoms and oxygen atom on small molecule sensor can be soon coordinated with mercury ion, enhance fluorescence rapidly, thus real
Now to the detection of mercury ion.
Chinese patent application 201610397020.6 discloses a kind of citric acid ester type fluorescent chemicals, with dicyclohexyl carbon
Diimine not only can play activation as catalyst with activated carboxyl, be also used as tiing up aqua, absorb esterification
The water generated in the process carries out so that reacting to the direction for generating ester；Pass through simple preparation method, lower temperature and shorter
Time can be obtained by the citric acid ester type fluorescent chemicals of the higher yellow green of fluorescence intensity, fluorescence quantum yield is reachable
5.0%.
Chinese patent application 201610112770.4 disclose it is a kind of detect mercury ion novel Rhodamine fluorescent probe and
Preparation method belongs to fluorescence probe and preparation method thereof technical field.This application is in addition to disclosing Rhodamine fluorescent probe
Structure also discloses the preparation method of Rhodamine fluorescent probe: by rhodamine B and hydrazine hydrate, back flow reaction generates sieve in ethanol
Red bright hydrazides；Then with phenyl glyoxal back flow reaction in ethanol, through being recrystallized to give schiff bases；Again through sodium borohydride reduction
Generate fluorescence probe.
Chinese patent application 201511031104.X discloses a kind of mercury ion fluorescence probe compound, preparation method and answers
With being related to technical field of analytical chemistry.The chemical molecular formula of the mercury ion fluorescence probe is to be by 2- (2 '-hydroxyphenyl) benzo
Imidazoles is precursor structure, is obtained by derivatization.This probe selectively good, high sensitivity and easily prepared to mercury ion is
A kind of ideal mercury ion sensor molecules.
According to the type of action of mercury ion and probe molecule, mercury ion fluorescence probe can be divided into corrdination type and response type
Two major classes, fluorescence probe is mutually complexed with mercury ion by way of non-covalent bond for the fluorescence probe of corrdination type, but this network
For active force when being applied to mercury ion identification, single-minded selectivity is not high, and the probe of response type is based on change between the two
Reaction is learned, new reaction product can be generated, and then the fluorescence of system or ultra-violet absorption spectrum is caused to change, is had very strong special
One selectivity, so such probe has higher sensitivity and good anti-Cation Interferences ability.
Summary of the invention
It is badly in need of a kind of by highly selective colorimetric ratio measurement Hg in this field2+Method, so as to effectively detect especially
It is the Hg for being able to detect in environment (such as water source in Studies on Avian Habitat)2+.The present inventor's design has synthesized a kind of deselenization
The probe to identify mercury ion is acted on, a kind of method that can be used for mercury ion content detection is established.For this purpose, the present invention proposes
A kind of novel highly selective colorimetric ratio test Hg2+Probe, preparing for the probe is simple, and/or selectivity is high, and benefit
With the Hg in the probe assay sample2+Concentration.
The present invention provides the Hg at (such as water source in Studies on Avian Habitat) in test sample2+Method comprising will be as follows
The step of probe and sample of structure contact.
Illustratively, heretofore described environment refers to water source in water head site, such as Studies on Avian Habitat.The water source
Existence form can be river, lake, irrigation canals and ditches etc..
Preferably, after probe and sample contact, its fluorescence intensity at 456nm is detected.
Optionally further, sample is diluted before probe and sample contact, such as 1-20 times of dilution, so that dilute
Detection range of the concentration of mercury ion in probe of the present invention in sample after releasing.
Preferably, in test sample when ion concentration of mercury, setting compares and detects its fluorescence intensity at 456nm.
Detailed description of the invention
Fig. 1: probe (5 μm of ol/L) is in the buffer solution of pH 7.4PBS ethanol/water (5:5) with ion concentration of mercury (0-10
μm ol/L) fluorescence emission spectrogram of compound that changes, insertion figure indicates the fluorescence intensity at probe (5 μm of ol/L) 456nm with mercury ion
The change of concentration (0-10 μm of ol/L).The concentration of mercury ion is respectively 0,0.5,1,2,2.5,3,3.5,4,4.5,5 from top to bottom,
6,7,8,9,10 μm of ol/L.
Fig. 2: the working curve that probe responds mercury ion.
Fig. 3: probe (5 μm of ol/L) is in the buffer solution of pH 7.4PBS ethanol/water (5:5) to mercury ion (2.5 μm of ol/
L) response changes over time relationship.
Fig. 4: different metal ions (3 μm of ol/L) are to probe (5 μ in the buffer solution of pH 7.4PBS ethanol/water (5:5)
Mol/L) the influence of fluorescence intensity.F: the fluorescence intensity of different metal ions is added；F0: the fluorescence intensity of metal ion is not added.
Fig. 5: the influence to fluorescence probe intensity coexists in different metal ions.1: blank；2: mercury ion；3: mercury ion+nickel
Ion；4: mercury ion+magnesium ion；5: mercury ion+sodium ion；6: mercury ion+chromium ion；7: mercury ion+zinc ion；8: mercury ion
+ lead ion；9: mercury ion+copper ion；10: mercury ion+potassium ion；11: mercury ion+cadmium ion.
Embodiment 1: the synthesis of fluorescence probe
The synthetic route and method of mercury ion fluorescence probe of the invention are as follows:
Specifically, umbelliferone (0.49g, 3mmol) is dissolved in THF (2mL), it is slow under conditions of nitrogen protection
It is slow that selenium powder (0.19g, 24mmol) THF (5mL) solution is added dropwise, 10min is stirred at room temperature, addition is dissolved in triethylamine (2mL) and THF
The diphenyl phosphine chloride (0.53g, 24mmol) of (20mL) mixed solution, stirred under nitrogen atmosphere react for 24 hours, are spin-dried for solvent, add
Enter 25mL toluene water segregator back scrubbing 4h, remove solid, organic phase is spin-dried for, uses column chromatography purification target product (dichloro
Methane: petroleum ether=1:1), obtain 1.01g red powder fluorescence probe, yield 52%.1H NMR(400MHz,D2O)δ
7.77-7.68 (m, 6H), 7.62 (dd, J=10.9,4.0Hz, 1H), 7.55-7.43 (m, 8H) [M+H]+C21H16O3PSe
427.0 discovery 427.0.
Embodiment 2: the preparation of reaction reagent
Probe stock solution and probe test fluid: accurately weighing probe synthesized in the embodiment 1 of 4.26mg, first with a small amount of
Methylene chloride dissolution, then moves it into 10mL volumetric flask, is settled to scale with ethyl alcohol, mixes, obtaining concentration at this time is
The probe stock solution liquid of 1mmol/L.And the stock solution of 1mL is taken to be added in the ethyl alcohol of 100mL, first plus some deionized waters, then
The PBS buffer solution (pH=7.4) of 5mL is added, continues plus deionized water is settled to 200mL, obtains the probe test of 5 μm of ol/L
Liquid.
Mercury ion stock solution: 32.46mg mercuric nitrate (Hg (NO is accurately weighed3)2), deionized water is added and is dissolved, to sample
It after product are completely dissolved, all move into 10mL volumetric flask, is settled to scale, mix, the concentration of this solution is 10mmol/L.It takes
The above-mentioned solution of 1mL is diluted to 1mmol/L.
Embodiment 3: the measurement of fluorescence probe spectrum
The colorimetric cylinder for taking 15 10mL respectively, is then numbered, be separately added into 5 μm of ol/L probe test fluid 5mL, so
The mercury ion for being separately added into 1mmol/L afterwards is followed successively by 0 μ L, 2.5 μ L, 5 μ L, 10 μ L, 12.5 μ L, 15 μ L, 17.5 μ L, 20 μ L,
22.5 μ L, 25 μ L, 30 μ L, 35 μ L, 40 μ L, 45 μ L, 50 μ L shake up.Take in above-mentioned 15 colorimetric cylinders solution 4mL in colorimetric respectively
Ware carries out fluorescence spectrometry at 456nm.
The variation of various concentration mercury ion fluorescent intensity is added in embodiment 1 in synthesized probe, as shown in Figure 1.With
The increase of ion concentration of mercury, fluorescence intensity also increasing, when the concentration of mercury ion is greater than 4 μm of ol/L, institute in embodiment 1
The fluorescence intensity of the probe of synthesis is basicly stable, and it can be seen that ion concentration of mercury is in 0~4 μm of ol/L, ion concentration of mercury
Certain linear relationship is presented with fluorescence intensity level.
With ion concentration of mercury (0~4 μm of ol/L) for abscissa, the fluorescence intensity level at 456nm is ordinate, draws mark
Directrix curve, as a result as shown in Figure 2.
As seen from Figure 2, when ion concentration of mercury is in 0~4 μm of ol/L, the concentration and fluorescence intensity of mercury ion are at preferable
Linear relationship, regression equation are as follows: y=107305x+99254 (R2=0.9875), detection is limited to 0.07 μm of ol/L.Therefore, originally
The designed mercury ion probe of invention can determine the content of mercury ion to be measured using fluorescence spectrum.
The calculating of detection limit: then fluorescence intensity level of 20 blank solutions of measurement at 456nm calculates σ, and by work
Make to obtain in the formula in slope K=107305 substitution Fig. 2 of working curve known to curve, the detection for calculating this method is limited to
0.07μmol/L。
The experimental results showed that, synthesized probe can respond in aqueous solution with mercury ion in embodiment 1 above, can be with
The mercury ion in water is detected with fluorescence signal.
Embodiment 4: the determination in reaction time
The colorimetric cylinder for taking out a 10mL, be added 1mmol/L probe stock solution 5mL, then be added 1mmol/L mercury from
Sub- 12.5 μ L of stock solution, shakes up.It takes above-mentioned solution 4mL in cuvette, detects the fluorescence intensity at 456nm in 30min at any time
Between variation.The case where record fluorescence intensity level varies over, so that it is determined that the Best Times of reaction.
Fluorescence intensity level is measured under the conditions of the different reaction time containing mercury solution with same concentration, is plotted in different
The change curve of fluorescence intensity level under the conditions of reaction time, as a result as shown in Figure 3.From figure 3, it can be seen that mercury ion and reality
It applies fluorescence intensity in probe reaction 1min synthesized in example 1 to sharply increase and reach stable, illustrates that the reaction is transient response.
Embodiment 5: ion interference test
Consider in the practical application of mercury ion measuring method, may be subjected to the interference of some other metal ions,
Therefore, it is necessary to inquire into influence of the different types of common metal ion to mercury ion measurement result.The ion of test includes: K+、Na+、Mg2+、Zn2+、Ni2+、Cd2+、Cr3+、Pb2+、Cu2+。
Take 10 10mL colorimetric cylinders to be numbered, be separately added into 5 μm of ol/L probe test fluid 5mL, then it is respectively different
Ion: K+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+、Hg2+15 μ L of (concentration 1mmol/L) solution, shakes up.Point
Solution 4mL in above-mentioned 10 colorimetric cylinders is not taken to carry out fluorescence spectrometry at 456nm in cuvette.
Probe synthesized in embodiment 1 is studied to the selectivity of mercury ion, different test ions are to fluorescence probe intensity
The influence of value.The ion of test includes: K+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+, as a result such as Fig. 4.From Fig. 4
As can be seen that mercury ion is added, fluorescence intensity level has significant change at 456nm, and is added due to the exclusive reaction of mercury and selenium
Nickel ion, magnesium ion, sodium ion, zinc ion, lead ion, potassium ion, cadmium ion and chromium ion are to spy synthesized in embodiment 1
The fluorescence intensity of needle is substantially without influence, and in contrast, copper ion has weaker response, this may be due to copper and mercury belong to it is same
Main group, the reaction of mercury ion can occur for copper ion, but compare with mercury ion is added, and influencing very little can ignore.Experiment shows
Synthesized probe can be reacted with mercury ion in specific manner in embodiment 1, to mercury ion selectivity with higher.
Continue to investigate ion and influence to fluorescence intensity coexists.It takes 9 10mL colorimetric cylinders to be numbered, is separately added into
Synthesized probe test fluid 5mL, the 1mmol/L mercury ion of 15 μ L, then respectively different ion in the embodiment 1 of 1mmol/L:
K+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+, 15 μ L of (concentration 1mmol/L) solution, shake up.Above-mentioned 9 are taken respectively
Solution 4mL carries out fluorescence spectrometry in cuvette at 456nm in a colorimetric cylinder.As a result as shown in Figure 5.As shown in Figure 5,
The addition of other metal ions will not interfere mercury to the characteristic reaction of selenium, and be individually added into the fluorescence intensity difference of mercury ion not
Greatly, illustrate that the above-mentioned coexistent metallic ion inquired into is smaller to the interference of detection mercury ion herein, it is known that the probe to mercury from
Son has preferable selectivity.
In short, the strong compatibility based on selenium and mercury, successful design and the cumarin for having synthesized a novel fluorescence enhancement
Class probe establishes a kind of method for detecting mercury ion content.In ethanol/water (1/1, v/v) system (pH=7.4, PBS
Buffer) in, the detection to mercury ion may be implemented in probe.Probe is to the response of mercury ion not by coexisting ion K+、Na+、Mg2 +、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+Interference, to Hg2+Selectivity it is fine.When ion concentration of mercury is in 0-4 μm of ol/L model
In enclosing, the concentration and fluorescence intensity of mercury ion are at good linear relationship, regression equation are as follows: y=107305x+99254 (R2=
0.9875), detection is limited to 0.07 μm of ol/L.
Although with above embodiments describe the present invention, it should be appreciated that before without departing substantially from spirit of the invention
It puts, the present invention further can be modified and be changed, and these modifications and variation all belong to the scope of protection of the present invention it
It is interior.
Claims (10)
1. Hg in test sample2+Method comprising the step of probe of such as flowering structure and sample are contacted:
2. Hg in test sample according to claim 12+Method, wherein the sample be Studies on Avian Habitat in water source.
3. the Hg in test sample according to claim 22+Method, the existence form at the water source is river, lake
Or irrigation canals and ditches.
4. Hg in test sample according to claim 12+Method, wherein probe and sample contact after, detect its
Fluorescence intensity at 456nm.
5. Hg in test sample according to claim 42+Method, wherein sample carried out before probe and sample contact
Dilution.
6. Hg in test sample according to claim 52+Method, wherein sample is diluted 1-20 times so that dilution
Detection range of the concentration of mercury ion in probe of the present invention in sample afterwards.
7. Hg in test sample according to claim 62+Method, wherein sample is diluted so that dilution after sample
Range of the concentration of mercury ion in 0~4 μm of ol/L in product.
8. Hg in test sample according to claim 52+Method, wherein in test sample when ion concentration of mercury, if
It sets control and detects its fluorescence intensity at 456nm.
9. method according to claim 1 to 8, wherein detecting in the ethanol/water (1:1, v/v) of pH=7.4
It carries out.
10. according to the method described in claim 9, wherein pH is maintained or is adjusted by PBS buffer solution.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5952236A (en) * | 1995-10-26 | 1999-09-14 | Thompson; Richard B. | Enzyme-based fluorescence biosensor for chemical analysis |
| CN102095723A (en) * | 2009-12-10 | 2011-06-15 | 中国科学院生态环境研究中心 | Preparation of carbohydrate fluorescence chemical sensor and quick detection of mercury ion in water by using carbohydrate fluorescence chemical sensor |
| CN104004028A (en) * | 2014-06-06 | 2014-08-27 | 南京工业大学 | Multichannel molecular probe for detecting Hg<2+> and preparation method thereof |
| CN105669689A (en) * | 2015-07-03 | 2016-06-15 | 济南大学 | Preparation and application of mercury ion fluorescent probe compound based on rhodamine B |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9416155B2 (en) * | 2009-12-14 | 2016-08-16 | The Board Of Trustees Of The Leland Stanford Junior University | Direct sensing of molecular species by polyfluors on a DNA backbone |
| US9733184B2 (en) * | 2012-03-30 | 2017-08-15 | Indian Institute Of Technology Madras | Visual detection of mercury ions |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5952236A (en) * | 1995-10-26 | 1999-09-14 | Thompson; Richard B. | Enzyme-based fluorescence biosensor for chemical analysis |
| CN102095723A (en) * | 2009-12-10 | 2011-06-15 | 中国科学院生态环境研究中心 | Preparation of carbohydrate fluorescence chemical sensor and quick detection of mercury ion in water by using carbohydrate fluorescence chemical sensor |
| CN104004028A (en) * | 2014-06-06 | 2014-08-27 | 南京工业大学 | Multichannel molecular probe for detecting Hg<2+> and preparation method thereof |
| CN105669689A (en) * | 2015-07-03 | 2016-06-15 | 济南大学 | Preparation and application of mercury ion fluorescent probe compound based on rhodamine B |
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