CN106841129A - The method for determining ion concentration of mercury in Studies on Avian Habitat water source - Google Patents
The method for determining ion concentration of mercury in Studies on Avian Habitat water source Download PDFInfo
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- 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
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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
Method the present invention relates to determine ion concentration of mercury in Studies on Avian Habitat water source.In ethanol/water (1/1, v/v) system (pH=7.4, PBS), the inventive method can realize the detection to mercury ion.Coexisting ion K is not received in response of the method for the present invention to mercury ion+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+Interference, to Hg2+Selectivity it is fine.When ion concentration of mercury is in the range of 04 μm of ol/L, the concentration of the mercury ion linear relationship into good with fluorescence intensity, regression equation is：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 method for determining ion concentration of mercury in 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
Will be organic mercury (such as methyl mercury) by Bacterial Transformation, and can accumulate always in vivo, through food chain enrichment,
Finally absorbed by the mankind and other animals such as birds and produce 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 seriously poison dynamic
The nerve of thing, causes the behavior of animal to perceive disorder, neurotoxic and kidney, to the mankind and the product of other animals such as birds
The very big threat of life.
The feather and ovum of bird are high to mercury enrichment degree, and mercury ion content is to composition of bird community or the shadow of population in water source
Sound is it will be apparent that birds are the indicator of environment in addition.So the content of mercury is to pass in government department's monitoring water environment
Important.Therefore, research and development have more high selectivity, it is more quick, more highly sensitive can be in environment such as natural waters
In mercury ion content detection in real time method be it is very important, because of that one can follow the trail of in water source mercury ion content or
Its influence of change to composition of bird community or population.
At present, 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 complex operation.And fluorescence probe detection method due to convenience it is good, selection
It is property, the advantages of sensitivity is high and extremely noticeable.In recent years, just cause that mercury ion is glimmering exactly under above-mentioned a variety of backgrounds
The research of light probe obtains the concern of increasing scholar.
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- naphthalimides be fluorescent parent, pass sequentially through with just
Butylamine reacts, and obtains bromo- 1, the 8- naphthalimides of intermediate N butyl -4- of fluorescent both, gained intermediate and 2- methoxybenzyls
Amine reaction introduces nitrogen-atoms and oxygen atom, strengthens the coordination ability of the compound and metal ion, and that is invented in detection process should
Nitrogen-atoms and oxygen atom on small molecule sensor soon can be coordinated with mercury ion, fluorescence is strengthened rapidly, so that 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
During produce water so that react to generation ester direction carry out；By simple preparation method, relatively low temperature and shorter
Time can be obtained by the citric acid ester type fluorescent chemicals of fluorescence intensity yellow green higher, fluorescence quantum yield is reachable
5.0%.
Chinese patent application 201610112770.4 disclose it is a kind of detect mercury ion new Rhodamine fluorescent probe and
Its preparation method, belongs to fluorescence probe and preparation method thereof technical field.This application is except 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
Generation 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 is high and easily prepared to mercury ion selectively good, sensitivity, is
A kind of preferable mercury ion sensor molecules.
According to mercury ion and the type of action of probe molecule, mercury ion fluorescence probe can be divided into corrdination type and response type
With mercury ion be mutually complexed fluorescence probe by the form of non-covalent bond by two major classes, the fluorescence probe of corrdination type, but this network
Active force is when mercury ion identification is applied to, and single-minded selectivity is not high, and the probe of response type is based on change therebetween
Reaction is learned, new product can be generated, and then cause the fluorescence or ultra-violet absorption spectrum of system to change, there is very strong special
One selectivity, so such probe possesses sensitivity and good anti-Cation Interferences ability higher.
The content of the invention
It is badly in need of a kind of by high selectivity colorimetric ratio measure Hg in this area2+Method such that it is able to effective detection is special
It is the Hg that can detect (such as water source in Studies on Avian Habitat) in environment2+.The present inventor's design has synthesized a kind of deselenization
Act on recognizing the probe of mercury ion, establish a kind of method that can be used for mercury ion content detection.Therefore, the present invention is proposed
A kind of novel high selectivity colorimetric ratio test Hg2+Probe, the preparation of the probe is simple, and/or selectivity is high, and profit
With the Hg in the probe assay sample2+Concentration.
The Hg at (such as water source in Studies on Avian Habitat) in detection sample2+Method, it includes will be as follows
The step of probe of structure is contacted with sample.
Exemplarily, heretofore described environment refers to water head site, such as water source in 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, for example, dilutes 1-20 times, it is dilute to cause
Detection range of the concentration of mercury ion in probe of the present invention in sample after releasing.
Preferably, in ion concentration of mercury in detecting sample, setting is compareed and detects its fluorescence intensity at 456nm.
Brief description of the drawings
Fig. 1：Probe (5 μm of ol/L) is in pH 7.4PBS ethanol/waters (5：5) with ion concentration of mercury (0-10 in cushioning liquid
μm ol/L) fluorescence emission spectrogram of compound that changes, insertion figure represents 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 is responded to mercury ion.
Fig. 3：Probe (5 μm of ol/L) is in pH 7.4PBS ethanol/waters (5：5) to mercury ion (2.5 μm of ol/ in cushioning liquid
L) response changes over time relation.
Fig. 4：In pH 7.4PBS ethanol/waters (5：5) different metal ions (3 μm of ol/L) are to probe (5 μ in cushioning liquid
Mol/L) the influence of fluorescence intensity.F：Add the fluorescence intensity of different metal ions；F0：It is not added with the fluorescence intensity of metal ion.
Fig. 5：Different metal ions coexist the influence to fluorescence probe intensity.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 reaction 24h, is spin-dried for solvent, plus
Enter 25mL toluene water knockout drum back scrubbing 4h, remove solid, organic phase is spin-dried for, use column chromatography purification target product (dichloro
Methane：Petroleum ether=1：1) 1.01g red powder fluorescence probes, yield 52%, are obtained.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, find 427.0.
Embodiment 2：The preparation of reaction reagent
Probe storing solution and probe test liquid：Synthesized probe in the accurate embodiment 1 for weighing 4.26mg, first with a small amount of
Dichloromethane is dissolved, and then moves it into 10mL volumetric flasks, and scale is settled to ethanol, is mixed, and now obtaining concentration is
The probe storing solution liquid of 1mmol/L.And take the storing solution of 1mL and be added in the ethanol of 100mL, first plus some deionized waters, then
The PBS (pH=7.4) of 5mL is added, is continued plus deionized water is settled to 200mL, obtain 5 μm of probe tests of ol/L
Liquid.
Mercury ion storing solution：Accurately weigh 32.46mg mercuric nitrates (Hg (NO3)2), add deionized water to be dissolved, treat sample
After product are completely dissolved, all move into 10mL volumetric flasks, be settled to scale, mix, the concentration of this solution is 10mmol/L.Take
The above-mentioned solution of 1mL is diluted to 1mmol/L.
Embodiment 3：The measure of fluorescence probe spectrum
Take 15 colorimetric cylinders of 10mL respectively, be then numbered, be separately added into 5 μm of probe test liquid 5mL of ol/L, 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.Solution 4mL is in colorimetric in taking above-mentioned 15 colorimetric cylinders respectively
Ware, carries out fluorescence spectrometry at 456nm.
The change of various concentrations 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, its fluorescence intensity is also increasing, when the concentration of mercury ion is more 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) as 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 of mercury ion is into preferable with fluorescence intensity
Linear relationship, regression equation is：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：Fluorescence intensity level of 20 blank solutions of measure at 456nm, then calculates σ, and by work
Make curve and understand that slope K=107305 of working curve show that the detection for calculating the method is limited in substituting into the formula in Fig. 2
0.07μmol/L。
Above test result indicate that, synthesized probe can be responded in aqueous with mercury ion in embodiment 1, can be with
The mercury ion in water is detected with fluorescence signal.
Embodiment 4：The determination in reaction time
Take out a colorimetric cylinder of 10mL, add 1mmol/L probe storing solution 5mL, be subsequently adding the mercury of 1mmol/L from
The μ L of sub- storing solution 12.5, shake up.Above-mentioned solution 4mL is taken in cuvette, detects the fluorescence intensity at 456nm in 30min at any time
Between change.The situation that record fluorescence intensity level is varied over, so that it is determined that the Best Times of reaction.
Fluorescence intensity level is determined under the conditions of the different reaction time containing mercury solution with same concentration, is plotted in different
The change curve of the fluorescence intensity level under the conditions of the reaction time, as a result as shown in Figure 3.From figure 3, it can be seen that mercury ion and reality
Apply fluorescence intensity in synthesized probe reaction 1min in example 1 to sharply increase and reach stabilization, illustrate that the reaction is transient response.
Embodiment 5：Ion interference is tested
Consider in the practical application of mercury ion assay method, may be disturbed by some other metal ions,
Accordingly, it would be desirable 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 the probe test liquid 5mL of 5 μm of ol/L and then respectively different
Ion：K+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+、Hg2+The μ L of (concentration is 1mmol/L) solution 15, shake up.Point
Solution 4mL carries out fluorescence spectrometry in cuvette at 456nm in not taking above-mentioned 10 colorimetric cylinders.
, to the selectivity of mercury ion, different test ions are to fluorescence probe intensity for synthesized probe in research embodiment 1
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 due to mercury and the exclusive reaction of selenium, adding mercury ion, fluorescence intensity level has significant change at 456nm, and adds
Nickel ion, magnesium ion, sodium ion, zinc ion, lead ion, potassium ion, cadmium ion and chromium ion are to spy synthesized in embodiment 1
Substantially without influence, comparatively, copper ion has weaker response to the fluorescence intensity of pin, and this is probably because copper and mercury belong to same
Main group, copper ion can occur the reaction of mercury ion, but be compared with addition mercury ion, and its influence very little can be ignored.Experiment shows,
Synthesized probe can be reacted with mercury ion in specific manner in embodiment 1, have selectivity higher to mercury ion.
Continuation investigation ion coexists the influence to fluorescence intensity.Take 9 10mL colorimetric cylinders to be numbered, be separately added into
Probe test liquid 5mL, the 1mmol/L mercury ions and then respectively different ion of 15 μ L synthesized in the embodiment 1 of 1mmol/L：
K+、Na+、Mg2+、Zn2+、Ni2+、Cr3+、Cd2+、Pb2+、Cu2+, the μ L of (concentration is 1mmol/L) solution 15, shake up.Above-mentioned 9 are taken respectively
Solution 4mL carries out fluorescence spectrometry in cuvette at 456nm in individual colorimetric cylinder.Result is as shown in Figure 5.As shown in Figure 5,
The addition of other metal ions without interference with mercury to the characteristic reaction of selenium, with the fluorescence intensity difference for being individually added into mercury ion not
Greatly, illustrate that the interference of the above-mentioned coexistent metallic ion inquired into detecting mercury ion herein is smaller, it is known that the probe to mercury from
Son has preferably selectivity.
In a word, the strong compatibility based on selenium Yu mercury, successful design has simultaneously synthesized a cumarin for new Fluorescence Increasing
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 solution) in, probe can realize the detection to mercury ion.Coexisting ion K is not received in response of the probe to mercury ion+、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 of the mercury ion linear relationship into good with fluorescence intensity, regression equation is：Y=107305x+99254 (R2=
0.9875), detection is limited to 0.07 μm of ol/L.
Although with above embodiments describing the present invention, it should be appreciated that before without departing substantially from spirit of the invention
Put, the present invention can further be modified and changed, and these modification and variation belong to protection scope of the present invention it
It is interior.
Claims (10)
1. Hg in sample is detected2+Method, it includes the step of probe of following structure is contacted with sample：
2. it is according to claim 1 to detect Hg in sample2+Method, wherein the sample be Studies on Avian Habitat in water source.
3. it is according to claim 2 detection sample in Hg2+Method, the existence form at the water source can be river,
Lake, irrigation canals and ditches etc..
4. it is according to claim 1 to detect Hg in sample2+Method, wherein probe and sample contact after, detect its
Fluorescence intensity at 456nm.
5. it is according to claim 4 to detect Hg in sample2+Method, wherein probe and sample contact before sample is carried out
Dilution.
6. it is according to claim 5 to detect Hg in sample2+Method, wherein sample is diluted 1-20 times, to cause dilution
Detection range of the concentration of mercury ion in probe of the present invention in sample afterwards.
7. it is according to claim 6 to detect Hg in sample2+Method, wherein sample is diluted, to cause the sample after dilution
The concentration of mercury ion is in 0~4 μm of scope of ol/L in product.
8. it is according to claim 5 to detect Hg in sample2+Method, wherein detect sample in ion concentration of mercury when, if
Put and compare and detect its fluorescence intensity at 456nm.
9. the method according to any one of claim 1-8, wherein detecting the ethanol/water (1 in pH=7.4：1, v/v) in
Carry out.
10. method according to claim 9, wherein pH is maintained or adjusted by PBS.
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| US20120309107A1 (en) * | 2009-12-14 | 2012-12-06 | Eric Todd Kool | Direct Sensing of Molecular Species by Polyfluors on a DNA Backbone |
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| US20150053868A1 (en) * | 2012-03-30 | 2015-02-26 | Indian Institute Of Technology Madras | Visual detection of mercury ions |
| 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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| 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 |
| US20120309107A1 (en) * | 2009-12-14 | 2012-12-06 | Eric Todd Kool | Direct Sensing of Molecular Species by Polyfluors on a DNA Backbone |
| US20150053868A1 (en) * | 2012-03-30 | 2015-02-26 | Indian Institute Of Technology Madras | Visual detection of mercury ions |
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