CN102735662A - High sensitivity and high selectivity fluorescence emission spectrum analysis method for zinc ions - Google Patents
High sensitivity and high selectivity fluorescence emission spectrum analysis method for zinc ions Download PDFInfo
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- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 title claims abstract description 25
- 238000002189 fluorescence spectrum Methods 0.000 title claims abstract description 8
- 230000035945 sensitivity Effects 0.000 title abstract description 7
- 238000004458 analytical method Methods 0.000 title abstract 2
- 239000000523 sample Substances 0.000 claims abstract description 61
- 239000011701 zinc Substances 0.000 claims abstract description 57
- 150000001875 compounds Chemical class 0.000 claims abstract description 30
- 239000000243 solution Substances 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 22
- 150000007857 hydrazones Chemical class 0.000 claims abstract description 15
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 230000002708 enhancing effect Effects 0.000 claims abstract description 4
- 239000012086 standard solution Substances 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 150000002500 ions Chemical class 0.000 claims description 12
- 238000011088 calibration curve Methods 0.000 claims description 11
- 238000007805 zymography Methods 0.000 claims description 10
- 125000001622 2-naphthyl group Chemical group [H]C1=C([H])C([H])=C2C([H])=C(*)C([H])=C([H])C2=C1[H] 0.000 claims description 9
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 8
- 239000012488 sample solution Substances 0.000 claims description 7
- -1 Bis (1-hydroxy-2-naphthyl)-methylene hydrazone Chemical class 0.000 claims description 6
- 239000007983 Tris buffer Substances 0.000 claims description 6
- 238000007792 addition Methods 0.000 claims description 6
- 230000005284 excitation Effects 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 4
- 238000013459 approach Methods 0.000 claims description 4
- 238000004364 calculation method Methods 0.000 claims description 4
- 238000013016 damping Methods 0.000 claims description 4
- 239000012530 fluid Substances 0.000 claims description 4
- 238000005259 measurement Methods 0.000 claims description 4
- 229910052751 metal Inorganic materials 0.000 claims description 4
- 239000002184 metal Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 229910001429 cobalt ion Inorganic materials 0.000 claims description 3
- XLJKHNWPARRRJB-UHFFFAOYSA-N cobalt(2+) Chemical compound [Co+2] XLJKHNWPARRRJB-UHFFFAOYSA-N 0.000 claims description 3
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 229910002651 NO3 Inorganic materials 0.000 claims description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 claims description 2
- 229910052783 alkali metal Inorganic materials 0.000 claims description 2
- 150000001340 alkali metals Chemical class 0.000 claims description 2
- 238000011095 buffer preparation Methods 0.000 claims description 2
- 230000008859 change Effects 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 238000013213 extrapolation Methods 0.000 claims description 2
- 229910001385 heavy metal Inorganic materials 0.000 claims description 2
- 239000004615 ingredient Substances 0.000 claims description 2
- 150000002739 metals Chemical class 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims description 2
- 229910052761 rare earth metal Inorganic materials 0.000 claims description 2
- 150000002910 rare earth metals Chemical class 0.000 claims description 2
- 229910052723 transition metal Inorganic materials 0.000 claims description 2
- 150000003624 transition metals Chemical class 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 10
- 239000007853 buffer solution Substances 0.000 abstract 1
- 239000003086 colorant Substances 0.000 abstract 1
- 150000001455 metallic ions Chemical class 0.000 description 10
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 6
- 229910052725 zinc Inorganic materials 0.000 description 6
- UFWIBTONFRDIAS-UHFFFAOYSA-N Naphthalene Chemical compound C1=CC=CC2=CC=CC=C21 UFWIBTONFRDIAS-UHFFFAOYSA-N 0.000 description 4
- WHMDKBIGKVEYHS-IYEMJOQQSA-L Zinc gluconate Chemical compound [Zn+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O WHMDKBIGKVEYHS-IYEMJOQQSA-L 0.000 description 4
- 229960000306 zinc gluconate Drugs 0.000 description 4
- 235000011478 zinc gluconate Nutrition 0.000 description 4
- 239000011670 zinc gluconate Substances 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000012046 mixed solvent Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 2
- MWPLVEDNUUSJAV-UHFFFAOYSA-N anthracene Chemical compound C1=CC=CC2=CC3=CC=CC=C3C=C21 MWPLVEDNUUSJAV-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- BBEAQIROQSPTKN-UHFFFAOYSA-N pyrene Chemical compound C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 BBEAQIROQSPTKN-UHFFFAOYSA-N 0.000 description 2
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical compound [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 2
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 1
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 1
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 1
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- 238000001636 atomic emission spectroscopy Methods 0.000 description 1
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N benzo-alpha-pyrone Natural products C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- VTJUKNSKBAOEHE-UHFFFAOYSA-N calixarene Chemical class COC(=O)COC1=C(CC=2C(=C(CC=3C(=C(C4)C=C(C=3)C(C)(C)C)OCC(=O)OC)C=C(C=2)C(C)(C)C)OCC(=O)OC)C=C(C(C)(C)C)C=C1CC1=C(OCC(=O)OC)C4=CC(C(C)(C)C)=C1 VTJUKNSKBAOEHE-UHFFFAOYSA-N 0.000 description 1
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- 238000000605 extraction Methods 0.000 description 1
- 229910001447 ferric ion Inorganic materials 0.000 description 1
- GVEPBJHOBDJJJI-UHFFFAOYSA-N fluoranthrene Natural products C1=CC(C2=CC=CC=C22)=C3C2=CC=CC3=C1 GVEPBJHOBDJJJI-UHFFFAOYSA-N 0.000 description 1
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- 238000004445 quantitative analysis Methods 0.000 description 1
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- JWVCLYRUEFBMGU-UHFFFAOYSA-N quinazoline Chemical compound N1=CN=CC2=CC=CC=C21 JWVCLYRUEFBMGU-UHFFFAOYSA-N 0.000 description 1
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention provides a fluorescence spectrum analysis method for detecting zinc ions by adopting a compound di[(1-hydroxy-2-naphthyl)- methenyl]hydrazone as a probe. According to the method, a Tris-HCl buffer solution is adopted to adjust the pH value to 8 in a DMF/H2O mixing solution with a strictly-controlled H2O content of 10%, the compound di[(1-hydroxy-2-naphthyl)- methenyl]hydrazone is adopted as a fluorescent coloring agent for detecting Zn<2+>, and the fluorescence intensity of the solution can be measured to determine the Zn<2+> content in the solution. According to the present invention, the specific media condition is designed, such that the compound is adopted as the fluorescence enhancing probe for detecting zinc ions so as to perform high sensitivity and high selectivity identification detection on zinc ions in different samples.
Description
Technical field
Highly sensitive, the high fluorescence emission Zymography of selecting of a kind of zinc ion of the present invention belongs to the analytical chemistry field.
Background technology:In the molecular recognition field; Fluorescence probe with molecular device character through with target substance selectivity bonding; Fluorescence enhancing or parameters such as quencher, fluorescence polarization, wavelength and peak shape, fluorescence lifetime before and after combining are changed; The effect of microscopic fields is showed through optical information, thereby the original position that is implemented on the molecular level detects in real time, reaches the effective identification to metallic ion, organic molecule, biomolecule etc.Utilize probe molecule to combine to form fluorescigenic complex, supermolecule or aggregation with covalency or other forms and measure promptly so-called " fluorescence probe " technology with non-fluorescence or hypofluorescence material.Because the highly sensitive and high selectivity of fluorescence analysis; Real-time in-situ detects; Equipment is simple; And abundant spectral information can be provided, important effect is being brought into play in aspects such as various ion detection, protein molecular marker, DNA and cell detection, immunoassay in fields such as analytical chemistry, biological chemistry, environmental science, medicine and pharmacology.
The organic fluorescence probe serves as main representative with luciferin, rhodamine, acridine, Coumarins, has the fluorescence quantum yield height, and excitation wavelength is long, and the scope of application is wide.Wherein, can detect Hg
2+, Cu
2+, Pb
2+, Cr
3+, Fe
3+Isoionic fluorescence strengthens or the existing a large amount of reports of quencher type rhodamine class probe.Aromatics often is used as the fluorescence signal generation group of organic fluorescence probe.With naphthalene, anthracene, pyrene, quinoline, cyanines class, pyrroles's methine class etc. serves as that the main condensed-nuclei aromatics of representing has strong and stable fluorescence; Stokes shift is big; In the fluorescence probe field, as the model fluorophore, constructed fluorescence probe is the sensor of one type of function admirable.For example, be the isostructural fluorescence probe of calixarenes, tripod of fluorophor with the naphthalene, show Zn
2+, Hg
2+, Cu
2+Isoionic identification, quinazoline are as Co
2+The research of ion fluorescence quencher type probe also has report.
Zinc is one of human body trace elements necessary, is distributed in widely in the cell and body fluid of human body.In transition metal ion, its content is only second to ferric ion, and is closely related with health.Need set up trace Zn in fields such as plant, food medicine are clinical
2+Detection method.
Highly sensitive, high selectivity detection and Identification research in fields such as living things system, physiology course, environment and food securities has application fields to trace metal ion.Discovery is different from traditional organic fluorescent dye molecule, makes that its sample preparation is simple, assay method is quick, testing cost is cheap, and it is significant to detect the fluorescence probe of multiple metallic ion, superior performance simultaneously respectively.The trace zinc detection method has methods such as atomic absorption spectrography (AAS), atomic emission spectrometry, inductively coupled plasma, mass spectroscopy, UV-VIS spectrophotometry, fluorescence method.Fluorescent spectrometry does not need expensive instrument and equipment owing to simple to operate, has more using value.But because most developer poorly water-soluble needs just can be used for detecting through complicated pretreatment such as extraction, separation, crucial problem is that the sensitivity and the selectivity that detect can not satisfy increasingly high demand.Fluorescence signal has unrivaled superiority in sensitivity, use relevant report that fluorescent method detects zinc ion content also seldom at present, seeks new for Zn
2+There is the fluorescence probe of sufficiently high selectivity and sensitivity to have challenge and using value.Inventor of the present invention finds compound two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone in research process, at DMF/H
2In the mixed solvent of O (V/V, 9/1), on fluorospectrophotometer, be excitation wavelength with 438nm, 516nm is the fluorescent emission wavelength, and Zn is being arranged
2+When existing, the fluorescence of probe compound is strengthened, at Zn
2+In the finite concentration scope, Zn
2+Concentration is directly proportional with fluorescent emission intensity, and common coexistent metallic ion is interference measurement not, has high sensitivity and high selectivity, for the highly sensitive, high of a kind of zinc ion of the present invention selects the fluorescence emission Zymography to lay a good foundation.
Summary of the invention: the objective of the invention is to use the fluorescence spectrophotometry method, detection by quantitative sample zinc ion content is created a kind of new for Zn
2+Concentration has the fluorescence emission spectrum quantitative analysis method of sufficiently high selectivity and sensitivity.
Highly sensitive, the high fluorescence emission spectral analysis method of selecting of a kind of zinc ion of the present invention; It is characterized in that with compound two [(1-hydroxyl; The 2-naphthyl)-and methine] hydrazone is probe; Adopt the fluorescence emission spectrum standard measure to measure the concentration of zinc ion, the chemical structural formula of compound two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone does
Molecular formula C
22H
16N
2O
2, English name is Bis (1-hydroxy-2-naphthyl)-methylene hydrazone.Analytical approach adopts calibration curve method: a peek 10.0ml volumetric flask, adding concentration in each volumetric flask successively is 0.10 mmolL
-1Probe compound solution 1ml, Tris-HCl damping fluid 1ml, pH ~ 8.The Zn that in each volumetric flask, adds variable concentrations then
2+Standard solution adds sample liquid in one of them volumetric flask, uses DMF and H at last
2The mixed liquor constant volume of O shakes up to 10.0ml, and 25 ℃ of constant temperature are placed half an hour, on fluorospectrophotometer, are excitation wavelength with 438nm, and 516nm is that emission wavelength is measured fluorescence intensity, draws fluorescence intensity to standard Zn
2+The calibration curve of concentration, the fluorescent emission intensity of solution utilizes calibration curve to try to achieve Zn in the sample solution per sample
2+Concentration, calculation sample content, Zn in the standard solution
2+Concentration is 8.00 * 10
-6~ 2.00 * 10
-4MolL
-1In the scope, Zn
2+Make the fluorescence probe intensity enhancing linear; Other coexisting ion comprises that alkaline metal, earth alkali metal, transition metal, heavy metal, rare earth metal and cobalt ions are at concentration and Zn
2+When concentration was identical, the influence of the fluorescence intensity that zinc ion is detected was lower than 5%.The DMF chemical name is N, dinethylformamide, and molecular formula is HCON (CH
3)
2, be common organic solvent.DMF and H
2The volume ratio of the mixed solution of O is 9:1, and the probe compound solution of indication is the DMF solution of two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone; Indication Tris is a trishydroxymethylaminomethane.The present invention is equally applicable to standard addition method: get 6 10.0ml volumetric flasks, add sample solution 1ml in each volumetric flask successively, concentration is 0.10 mmolL
-1Probe compound solution 1ml, Tris-HCl damping fluid 1ml, pH ~ 8.In first volumetric flask, do not add Zn then
2+Standard solution adds the Zn of same concentrations different volumes successively in all the other each volumetric flasks
2+Standard solution is used DMF and H at last
2The mixed liquor constant volume of O shakes up to 10.0ml, and 25 ℃ of constant temperature are placed half an hour, on fluorospectrophotometer, are excitation wavelength with 438nm, and 516nm is an emission wavelength sequentially determining fluorescence intensity, draws fluorescence intensity to adding Zn
2+The volume calibration curve of standard solution is zero and horizontal ordinate intersection according to straight-line extrapolation to ordinate, can try to achieve Zn in the sample solution
2+Concentration, calculation sample content.
Above-mentioned is that the compound method of all ingredients is:
(1) compound method of probe compound solution: take by weighing two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone of 3.4 mg, with the DMF dissolving, be mixed with 100mL solution, concentration is 0.100 mmolL
-1
(2) Zn
2+Standard solution: take by weighing the pure ZnCl of top grade
227.2 mg with the DMF dissolving, and is mixed with 100mL solution, Zn
2+Concentration is 2.00 * 10
-3MolL
-1Arrive suitable concentration with the DMF stepwise dilution as required;
(3) Tris-HCl buffer preparation: take by weighing 0.242 g Tris and dissolve with DMF, being mixed with 500 mL concentration is 0.004 molL
-1Solution, pipette certain volume Tris and mix mutually with an amount of HCl, regulate pH value to 8.
(4) preparation of other coexisting ion solution: get the nitrate or the hydrochloride of the pure various metals of top grade, with DMF dissolving, and to be mixed with concentration be 2.00 * 10
-3MolL
-1DMF solution.
The above-mentioned analytical approach of the present invention is when getting volume change, and various solution additions will be done corresponding variation in proportion.
When the present invention measures zinc ion, coexisting ion Na
+, K
+, Rb
+, Mg
2+, Ca
2+, Sr
2+, Ba
2+, Pb
2+, Bi
2+, Mn
2+, Ni
2+, Cu
2+, Cd
2+, Hg
2+, Al
3+, Fe
3+, Sc
3+, Ti
3+, Cr
3+And Co
2+, at concentration and Zn
2+In the time of quite, to Zn
2+The relative deviation of the fluorescence intensity influence that detects is all in 5%.Equal interference measurements not.
The used reagent of the inventive method is AR, and used water is redistilled water.
The used instrument of the present invention is a fluorospectrophotometer, and model is a Cary Eclipse fluorospectrophotometer, and U.S. VARIAN company produces.
The inventive method has big Stokes displacement, high selectivity, detect thresold low, do not need to separate, the response time is fast, near advantages such as neutral test conditions.Can be used for Environmental Water monitoring (contaminant water), Pharmaceutical Analysis (zinc supplemental treatment medicament), Food Inspection fields such as (containing the zinc beverage) detection Zn
2+Content.
Utilize compound two [(1-hydroxyl 2-naphthyl)-methine] hydrazone molecule at optimum solvent (DMF/H
2O, 9/1) presents hypofluorescence in, can form the complex of confirming coordination ratio with the metallic zinc ion under given conditions.Gordian technique in the patent of the present invention is the different DMF/H of control
2O mixed solvent ratio and certain pH value just can make compound two [(1-hydroxyl 2-naphthyl)-methine] hydrazone become and detect Zn
2+The fluorescence enhancement mode probe of ion.Operation and control method are easy, and unique properties is the practical application of this organic molecule as fluorescence probe.
Description of drawings:
Fig. 1 compound two [(1-hydroxyl 2-naphthyl)-methine] hydrazone (10 μ M, DMF/H
2O, 10% H
2O, Tris-HCl, pH ~ 8) detect the fluorescence spectrum of identification zinc ion (200 μ M), other metallic ion (200 μ M): Na
+, K
+, Rb
+, Mg
2+, Ca
2+, Sr
2+, Ba
2+, Pb
2+, Bi
2+, Mn
2+, Ni
2+, Cu
2+, Cd
2+, Hg
2+, Al
3+, Fe
3+, Sc
3+, Ti
3+, Cr
3+And Co
2+Interference measurement not in error range can be known the control pH value of solution about 8, DMF/H from figure
2O mixed solvent volume ratio is 1:9, the fluorescence intensity of probe compound very a little less than, with this understanding, add Zn
2+Ion can make the very hyperfluorescence of probe compound solution in the emission of 516nm wavelength, and other experiment metallic ion comprises Co
2+Ion all can not make its fluorescence significantly strengthen.
Fig. 2 compound two [(1-hydroxyl 2-naphthyl)-methine] hydrazone is probe in detecting Zn
2+The time coexistent metallic ion fluorescence intensity.White: probe; Or probe+metallic ion; Black: probe+Zn
2++ metallic ion.From figure, can know the DMF/H of probe compound at pH 8
2O (V/V, 1:9) in the medium, fluorescent emission very weak (left 1 white bars among the figure), add behind other experiment coexisting ion to fluorescence probe still very a little less than, to its intensity effect little (among the figure from a left side 3 to 20, the white bars that metallic ion is corresponding), add Zn
2+After make fluorescence probe significantly strengthen (left 2 white bars among the figure), the influence (black bar among the figure) in error range that fluorescence probe is strengthened to zinc ion after in probe and the solution that has cobalt ions to exist, adding other metallic ion.
Fig. 3 detects Zn
2+Calibration curve.Ordinate is represented fluorescence intensity, and horizontal ordinate is represented the Zn that adds
2+Concentration of standard solution.
Detect Zn in Fig. 4 lake water sample
2+Fluorescence spectrum.The Zn that in sample liquid, adds different amounts by standard addition method successively
2+The spectrogram that standard solution records.
Zn in Fig. 5 zinc gluconate sample
2+The fluorescence spectrum of content detection.Press the spectrogram of twice sample of calibration curve replicate determination.
The practical implementation method:
Embodiment 1: pollute Zn in the lake water
2+Content detection.
1. lake water sample pre-treatments: get and pollute lake water sample 2.0 L, boiled 10 minutes, cooling is left standstill and was placed 12 hours, filters, and detects water sample pH value.Simmer down to 10ml cools off subsequent use.
2. detect: the lake water sample that pipettes the treated mistake of 1mL is in the 10mL volumetric flask, and (10 μ M, DMF), 1 mL Tris-HCl adds 0,1,2 mL Zn successively to add 1 mL probe compound
2+(10 μ M) is with moisture 10% DMF/H
2O mixed liquor constant volume, 25 ℃ of constant temperature are placed half an hour.Replicate determination 3 times.
3. the spectrogram of measuring is seen accompanying drawing 4, calculates the result according to standard addition method and sees table 1.
Detect Zn in the table 1 lake water sample
2+
Embodiment 2: zinc-content determination in the commercially available zinc gluconate.
Content to zinc in certain brand buccal zinc gluconate tablets detects:
1, pre-treatment: get a slice zinc gluconate 32mg (in zinc 0.32mg) and grind to form powdery, be dissolved in the 100mL volumetric flask.Place ultrasound wave concussion 30 minutes, leave standstill again after 2 hours and get supernatant liquor.
2, testing process: (10 μ M DMF), add 1 mL Tris-HCl (pH=8), pipette above-mentioned institute and join sample 1mL in the 10mL volumetric flask, with moisture 10% DMF/H to add 1 mL probe compound
2O mixed liquor constant volume.25 ℃ of constant temperature are placed half an hour, measure its fluorescence intensity at the 514nm place.Replicate determination 2 times.
3, the spectrogram of measuring is seen accompanying drawing 5, calculates following result according to experiment gained calibration curve, sees table 3.
Claims (7)
1. highly sensitive, the height of a zinc ion are selected the fluorescence emission Zymography; It is characterized in that with compound two [(1-hydroxyl; The 2-naphthyl)-and methine] hydrazone is probe; Adopt the fluorescence emission spectrum standard measure to measure zinc ion concentration, the chemical structural formula of compound two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone does
Molecular formula C
22H
16N
2O
2, English name is Bis (1-hydroxy-2-naphthyl)-methylene hydrazone, analytical approach adopts the standard calibration curve method: a peek 10.0ml volumetric flask, adding concentration in each volumetric flask successively is 0.10 mmolL
-1Probe compound solution 1ml, Tris-HCl damping fluid 1ml, pH ~ 8 add the Zn of variable concentrations then in each volumetric flask
2+Standard solution adds sample solution in one of them volumetric flask, use DMF and H at last
2The mixed liquor constant volume of O shakes up to 10.0ml, and 25 ℃ of constant temperature are placed half an hour, on fluorospectrophotometer, are excitation wavelength with 438nm, and 516nm is an emission wavelength sequentially determining fluorescence intensity, draws fluorescence intensity to standard Zn
2+The calibration curve of concentration, the fluorescent emission intensity of solution utilizes calibration curve to try to achieve Zn in the sample solution per sample
2+Concentration, calculation sample content, Zn in the standard solution
2+Concentration is 8.00 * 10
-6~ 2.00 * 10
-4MolL
-1In the scope, Zn
2+Make the fluorescence probe intensity enhancing in linear relation; Other coexisting ion comprises that alkaline metal, earth alkali metal, transition metal, heavy metal, rare earth metal and cobalt ions are at concentration and Zn
2+When concentration is identical, to Zn
2+All in 5%, the DMF chemical name is N to the relative deviation of the fluorescence intensity influence that detects, dinethylformamide, and molecular formula is HCON (CH
3)
2, be the organic solvent of routine, DMF and H
2The volume ratio of O mixed liquor is 9:1, and the probe compound solution of indication is the DMF solution of two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone; Indication Tris is a trishydroxymethylaminomethane.
2. highly sensitive, the height of a kind of zinc ion according to claim 1 are selected the fluorescence emission Zymography; It is characterized in that analytical approach is equally applicable to standard addition method: get 6 10.0ml volumetric flasks, add sample solution 1ml in each volumetric flask successively, concentration is 0.10 mmolL
-1Probe compound solution 1ml, Tris-HCl damping fluid 1ml, pH
8, in first volumetric flask, do not add Zn then
2+Standard solution adds the Zn of same concentrations different volumes successively in all the other each volumetric flasks
2+Standard solution is used DMF and H at last
2The mixed liquor constant volume of O shakes up to 10.0ml, and 25 ℃ of constant temperature are placed half an hour, on fluorospectrophotometer, are excitation wavelength with 438nm, and 516nm is an emission wavelength sequentially determining fluorescence intensity, draws fluorescence intensity to adding Zn
2+The volume calibration curve of standard solution is zero and horizontal ordinate intersection according to straight-line extrapolation to ordinate, can try to achieve Zn in the sample solution
2+Concentration, calculation sample content.
3. highly sensitive, the height of a kind of zinc ion according to claim 1 and 2 are selected the fluorescence emission Zymography, it is characterized in that the compound method of all ingredients is:
(1) compound method of probe compound solution: take by weighing two [(1-hydroxyl, 2-naphthyl)-methine] hydrazone of 3.4 mg, with the DMF dissolving, be mixed with 100mL solution, concentration is 0.100 mmolL
-1
(2) Zn
2+Standard solution: take by weighing the pure ZnCl of top grade
227.2 mg with the DMF dissolving, and is mixed with 100mL solution, Zn
2+Concentration is 2.00 * 10
-3MolL
-1Arrive suitable concentration with the DMF stepwise dilution as required;
(3) Tris-HCl buffer preparation: take by weighing 0.242 g Tris and dissolve with DMF, being mixed with 500 mL concentration is 0.004 molL
-1Solution, pipette certain volume Tris and mix mutually with an amount of HCl, regulate pH value to 8;
(4) preparation of other coexisting ion solution: get the nitrate or the hydrochloride of the pure various metals of top grade, with DMF dissolving, and to be mixed with concentration be 2.00 * 10
-3MolL
-1DMF solution.
4. highly sensitive, the high fluorescence emission Zymography of selecting of a kind of zinc ion according to claim 1 and 2 is characterized in that when the volumetric flask volume change, various solution additions will be done corresponding variation in proportion.
5. highly sensitive, the height of a kind of zinc ion according to claim 1 and 2 are selected the fluorescence emission Zymography, and other coexisting ion comprises when it is characterized in that measuring zinc ion: Na
+, K
+, Rb
+, Mg
2+, Ca
2+, Sr
2+, Ba
2+, Pb
2+, Bi
2+, Mn
2+, Ni
2+, Cu
2+, Cd
2+, Hg
2+, Al
3+, Fe
3+, Sc
3+, Ti
3+, Cr
3+And Co
2+, at concentration and Zn
2+In the time of quite, equal interference measurements not.
6. highly sensitive, the high fluorescence emission Zymography of selecting of a kind of zinc ion according to claim 1 and 2 is characterized in that used reagent is AR, and institute's water is a redistilled water.
7. highly sensitive, the height of a kind of zinc ion according to claim 1 and 2 are selected the fluorescence emission Zymography, and it is characterized in that analyzing used instrument is fluorospectrophotometer.
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| CN103822905A (en) * | 2014-01-17 | 2014-05-28 | 西北师范大学 | Application of 1-naphthyl-3, 4, 5-tri(hexadecyloxyl) benzoyl hydrazone as sensor in fluorescent identification of silver ions |
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| CN106770124B (en) * | 2017-01-05 | 2019-06-11 | 昆明理工大学 | It is a kind of for detecting the fluorescent optical sensor and preparation method of zinc ion |
| CN106883846A (en) * | 2017-04-14 | 2017-06-23 | 周口师范学院 | A kind of definitely water miscible zinc ion fluorescent, preparation method and applications |
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