CN103901033A - Method for detecting lead ion concentration in sample - Google Patents
Method for detecting lead ion concentration in sample Download PDFInfo
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- CN103901033A CN103901033A CN201410164473.5A CN201410164473A CN103901033A CN 103901033 A CN103901033 A CN 103901033A CN 201410164473 A CN201410164473 A CN 201410164473A CN 103901033 A CN103901033 A CN 103901033A
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Abstract
The invention relates to a method for detecting lead ion concentration in a sample. The method comprises the following steps: (1) mixing gold nanoparticles and a DNA sequence rich in G bases, wherein the DNA sequence rich in G bases is adsorbed to the surfaces of the gold nanoparticles, and the lead ion can be specifically identified; (2) mixing a sample to be detected with the mixture, wherein a G-quadruplet is formed by the DNA sequence rich in G bases and is adsorbed from the surfaces of the gold nanoparticles under the condition that the lead ion exists; (3) aggregating the gold nanoparticles, and changing the color of the system from red to blue; and (4) determining the lead ion concentration in the sample based on the color change in the step (3). According to the method, a biosensor which is brand new, simple, rapid, high in selectivity and high in sensitivity can be provided and is used for detecting the lead ion concentration in the sample.
Description
Technical field
The invention belongs to chemical field, relate to a kind of method that detects plumbum ion concentration in sample, particularly a kind of lead ion detection method based on golden nanometer particle and G-tetrad.
Background technology
Nano material, due to its unique physical property and chemical property, makes its application in biology sensor become international research forward position and study hotspot.Golden nanometer particle is to occur the earliest, one of nano material of most study.It has important application in fields such as biomarker, sensor structure and biochip test.Conventional golden nanometer particle is prepared easy and controlled, and long-term dispersed, good stability, has good biocompatibility, and have unique optical property, makes it be able to widespread use in analyzing and testing field.
The detection method of lead ion mainly contains: dithizone spectrophotometry, atomic fluorescence spectrometry, inductively coupled plasma emission spectrography, anodic stripping voltammetry, oscilloscopic polarography, biological stain test paper method etc.But these methods not only need a large amount of pre-service, and operating personnel are had to very high technical requirement.Therefore the concentration method of, studying quick, easy detection lead ion still has important practical significance.
Therefore, need to do further improvement to the detection method of lead ion.
Summary of the invention
The object of this invention is to provide a kind of method that detects plumbum ion concentration in sample, in order to realize the simple and quick detection of lead ion in water, to overcome the unhandy problem of existing lead ion detection method.
The present invention is achieved through the following technical solutions: a kind of method that detects plumbum ion concentration in sample, it is characterized in that, and comprising:
(1) golden nanometer particle and the DNA sequence dna that is rich in G base are mixed, the DNA sequence dna that is rich in G base is adsorbed on surface and the energy specific recognition lead ion of golden nanometer particle;
(2) testing sample is mixed with said mixture, under the condition existing at lead ion, the DNA sequence dna that is rich in G base can form G-tetrad and get off from golden nanometer particle surface desorption is attached;
(3) golden nanometer particle is assembled, and system color becomes blueness by redness;
(4) variation based on color in step (3), determines the plumbum ion concentration in described sample.
The present inventor finds, G-tetrad is the DNA sequence dna that is rich in G base by a section, under specific ionic strength and pH value condition, by forming Hoogsteen base pairing between the corresponding G base between strand or in strand, thereby make 4 or the 4 sections DNA single chains that are rich in G base revolve the G-tetrad that is polymerized to one section of parallel dextrorotation.G-tetrad has been widely used in biomedical and bioassay technique field now, as is applied to the detection to nucleic acid, protein, metallic ion and organic molecule.When golden nanometer particle exists in solution with the DNA sequence dna that is rich in G base, DNA is adsorbed on golden nanometer particle and it is played to stabilization around.When existing after lead ion, the DNA that is rich in G base can form G-tetrad and get off from golden nanometer particle surface desorption is attached, causes subsequently the gathering of golden nanometer particle, and system color becomes blueness by redness.The present inventor is based on this principle, according to the variation of system color, developed the biology sensor of a kind of easy, quick, high selectivity, high sensitivity, in order to detect the concentration of lead ion in water.According to embodiments of the invention, can utilize the type of the sample that method of the present invention detects and be not particularly limited.According to a particular embodiment of the invention, can be aqueous solution, such as potable water, underground water, sewage etc.
According to embodiments of the invention, said method can also have following additional technical feature:
Further, the described DNA sequence dna that is rich in G base has the nucleotide sequence as shown in SEQ ID NO:1.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
Further, the concentration of described golden nanometer particle is 0.075nM.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
Further, the concentration of the described DNA sequence dna that is rich in G base is 100nM.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
Further, in described testing sample, plumbum ion concentration is 1-1000 nM.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
Further, in described testing sample, plumbum ion concentration is 5-500 nM.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
Further, based on following linear equation, determine the concentration of lead ion in described sample: y=6.233 × 10
-4x+0.0727, y is the relative absorbance under different plumbum ion concentrations, the concentration that x is corresponding lead ion.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
According to embodiments of the invention, based on the change color of system, determine that the plumbum ion concentration in described sample is by the color of described system and typical curve have been compared, wherein, the standard model that described typical curve is respectively 1nM, 5nM, 10nM, 50nM, 100 nM, 200nM, 500nM, 1000 nM based on known plumbum ion concentration carries out parallel laboratory test and sets up.Thus, can further improve the efficiency and the sensitivity that utilize the inventive method to carry out plumbum ion concentration detection.
Additional aspect of the present invention and advantage in the following description part provide, and part will become obviously from the following description, or recognize by practice of the present invention.
Brief description of the drawings
Above-mentioned and/or additional aspect of the present invention and advantage accompanying drawing below combination is understood becoming the description of embodiment obviously and easily, wherein:
Fig. 1 utilizes different plumbum ion concentration standard models to detect obtained ultraviolet absorption curve, and wherein plumbum ion concentration is got respectively 1nM, 5nM, 10nM, 50nM, 100 nM, 200nM, 500nM, 1000 nM.
Fig. 2 has shown specificity analyses figure according to an embodiment of the invention.
Embodiment
According to following embodiment, the present invention may be better understood.But, those skilled in the art will readily understand, the described content of embodiment is only for the present invention is described, and should also can not limit the present invention described in claims.
The design of the corresponding G-DNA fragment of embodiment 1 is with synthetic.
Design one section of energy specific recognition lead ion, walk abreast into the DNA fragmentation of G-tetrad, DNA sequence dna is prepared by DNA synthesizer.
G-DNA：5’-GGAAGGTGTGGAAGG-3’（SEQ ID NO：1）
Synthesizing of embodiment 2 golden nanometer particles.
The golden nanometer particle of experiment use is synthetic with reference to the trisodium citrate reduction method of Frens.Concrete operation step is as follows: 1.0 mM chlorauric acid solution 50 mL of preparation are added in conical flask bottle, be placed on thermostatic electromagnetic stirrer and be heated to continue 5 minutes after boiling, add rapidly 0.6 mL, the citric acid three sodium solution of 38.8 mM, continues agitating heating 6 minutes, and the color of solution is by the faint yellow claret that is converted to, now golden nanometer particle generates, continue heating and stop after 10 minutes, continue stirring and make it cooling, 4 ° of C save backup.
The foundation of embodiment 3 lead ion typical curves.
In centrifuge tube, adding golden nanometer particle successively, with Tris-HAc(20mM, pH 7.0) damping fluid is settled to 2mL, and wherein the concentration of golden nanometer particle is 0.075nM.To the G-DNA 20 μ L that add 10 μ M in centrifuge tube, making its ultimate density is 100nM, adds NaCl successively subsequently in centrifuge tube, and making its ultimate density is 50mM, mixes, and surveys the absorbance at its 521nm wavelength place.With adding successively appropriate lead ion in backward centrifuge tube, make its ultimate density be respectively 1nM, 5nM, 10nM, 50nM, 100 nM, 200nM, 500nM, 1000 nM, in centrifuge tube, add NaCl successively, making its ultimate density is 50mM, react 30 minutes, then survey the absorbance at its 680nm place.According to the relation of the concentration of relative absorbance and lead ion, make the canonical plotting of lead ion.The range of linearity 5nM-500nM of the method, detects and is limited to 3nM.The linear equation of typical curve is y=6.233 × 10
-4x+0.0727, y is the relative absorbance under different plumbum ion concentrations, the concentration that x is corresponding lead ion, linear dependence >0.99.
Embodiment 4 specific assay.
In centrifuge tube, adding golden nanometer particle successively, with Tris-HAc(20mM, pH 7.0) damping fluid is settled to 2mL, and wherein the concentration of golden nanometer particle is 0.075nM.To the G-DNA 20 μ L that add 10 μ M in centrifuge tube, making its ultimate density is 100nM, adds NaCl successively in centrifuge tube, and making its ultimate density is 50mM, adds NaCl successively subsequently in centrifuge tube, and making its ultimate density is 50mM, surveys its absorbance.With adding successively appropriate lead ion, mercury ion, cadmium ion, copper ion, nickel ion in backward centrifuge tube, making its ultimate density is 500nM, reacts 30 minutes, then surveys its absorbance.Check that result shows, the method has reasonable specificity.
The detection of embodiment 5 actual samples.
To the lead ion that adds respectively 5nM, 10nM, 20nM and 50 nM in tap water sample, adopt said method to determine the plumbum ion concentration in sample, result is as table 1:
The mensuration of table 1 lead ion water sample
Result demonstration, the interpolation recovery that the lead ion detection method based on golden nanometer particle and G-tetrad is measured the lead ion in actual sample is between 94.50%-103.3%, and standard deviation is 4.67%, can meet the detection demand to lead ion in actual life completely.
Sequence table
<110> Changshu Institute of Technology
Mono-kind of <120> detects the method for plumbum ion concentration in sample
<130> xb14042301
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 15
<212> DNA
<213> Artificial
<220>
<223> is rich in the DNA sequence dna of G base
<400> 1
ggaaggtgtg gaagg 15
Claims (7)
1. a method that detects plumbum ion concentration in sample, is characterized in that, comprising:
(1) golden nanometer particle and the DNA sequence dna that is rich in G base are mixed, the DNA sequence dna that is rich in G base is adsorbed on surface and the energy specific recognition lead ion of golden nanometer particle;
(2) testing sample is mixed with said mixture, under the condition existing at lead ion, the DNA sequence dna that is rich in G base can form G-tetrad and get off from golden nanometer particle surface desorption is attached;
(3) golden nanometer particle is assembled, and system color becomes blueness by redness;
(4) variation based on color in step (3), determines the plumbum ion concentration in described sample.
2. method according to claim 1, is characterized in that: the described DNA sequence dna that is rich in G base has the nucleotide sequence as shown in SEQ ID NO:1.
3. method according to claim 1, is characterized in that: the concentration of described golden nanometer particle is 0.075nM.
4. method according to claim 1, is characterized in that: the concentration of the described DNA sequence dna that is rich in G base is 100nM.
5. method according to claim 1, is characterized in that: in described testing sample, plumbum ion concentration is 1-1000 nM.
6. method according to claim 5, is characterized in that: in described testing sample, plumbum ion concentration is 5-500 nM.
7. method according to claim 1, is characterized in that: based on following linear equation, determine the concentration of lead ion in described sample:
y=6.233×10
-4x+0.0727，
Y is the relative absorbance under different plumbum ion concentrations, the concentration that x is corresponding lead ion.
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Cited By (4)
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| CN108251446A (en) * | 2018-01-12 | 2018-07-06 | 天津大学 | A kind of construction method of lead ion responsive type whole-cell biological sensor |
| CN108801997A (en) * | 2018-06-11 | 2018-11-13 | 岭南师范学院 | A kind of no quencher fluorescence probe CP-DNA and preparation method thereof, application and detection micro Pb2+Method |
| CN111638209A (en) * | 2019-03-01 | 2020-09-08 | 中国科学院上海应用物理研究所 | Lead ion visual detection method based on nano golden flower and DNA ribozyme and application thereof |
| CN113252758A (en) * | 2021-04-08 | 2021-08-13 | 陕西省石油化工研究设计院 | Method for non-marking electrochemical detection of lead ions |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108251446A (en) * | 2018-01-12 | 2018-07-06 | 天津大学 | A kind of construction method of lead ion responsive type whole-cell biological sensor |
| CN108801997A (en) * | 2018-06-11 | 2018-11-13 | 岭南师范学院 | A kind of no quencher fluorescence probe CP-DNA and preparation method thereof, application and detection micro Pb2+Method |
| CN111638209A (en) * | 2019-03-01 | 2020-09-08 | 中国科学院上海应用物理研究所 | Lead ion visual detection method based on nano golden flower and DNA ribozyme and application thereof |
| CN113252758A (en) * | 2021-04-08 | 2021-08-13 | 陕西省石油化工研究设计院 | Method for non-marking electrochemical detection of lead ions |
| CN113252758B (en) * | 2021-04-08 | 2023-09-05 | 陕西省石油化工研究设计院 | Method for detecting lead ions through unlabeled electrochemistry |
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Application publication date: 20140702 |