CN105891286A - Lead ion direct electrochemical detection method for probe-integrated functional nucleic acid modified electrode - Google Patents

Lead ion direct electrochemical detection method for probe-integrated functional nucleic acid modified electrode Download PDF

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CN105891286A
CN105891286A CN201610372050.1A CN201610372050A CN105891286A CN 105891286 A CN105891286 A CN 105891286A CN 201610372050 A CN201610372050 A CN 201610372050A CN 105891286 A CN105891286 A CN 105891286A
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probe
nucleic acid
functional nucleic
modified electrode
electrode
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CN105891286B (en
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洪华
张爱平
施筱迪
王海天
洪昱
王红卫
王超颖
申晓萍
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Nantong Entry-Exit Inspection And Quarantine Bureau Of People's Republic Of
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Priority to CN201811265189.1A priority Critical patent/CN109060903A/en
Priority to CN201811258669.5A priority patent/CN109060902A/en
Priority to CN201811258719.XA priority patent/CN109187695A/en
Priority to CN201610372050.1A priority patent/CN105891286B/en
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
    • G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28—Electrolytic cell components
    • G01N27/30—Electrodes, e.g. test electrodes; Half-cells
    • G01N27/333—Ion-selective electrodes or membranes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage

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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses a lead ion direct electrochemical detection method for a probe-integrated functional nucleic acid modified electrode. The probe-integrated functional nucleic acid modified electrode is immersed in a buffering solution of to-be-detected lead ions for 20-30 min, the electrode is taken out, washed and subjected to DPV determination in a 0.1 M NaClO4 electrolyte solution, and the lead ion concentration in a sample is calculated according to the change rate of DPV peak current signals scanned and recorded in the range of 0.1-0.6 V. The method is good in selectivity, sensitive, simple and quick.

Description

The functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe is integrated
Technical field
The present invention relates to a kind of functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe is integrated.
Background technology
Lead ion is common a kind of harmful heavy metal ions, and its monitoring/detection suffers from highly important meaning to human health and environmental monitoring.In Guo Qujishinian, people have been developed the multiple methods such as Atomic absorption/emission spectrum, fluorescence spectrum, mass spectrum, photonic crystal such as and are monitored lead ion/detect.In recent years, Lu Yi research group of the U.S. obtains the functional nucleic acid 17E DNAzyme sensitive to lead ion by in-vitro screening technology, and they are using 17E DNAzyme as lead ion recognition component, with golden nanometer particle as sensing element, develop colorimetric sensor detection lead ion (Liu, J.; Lu, Y., Nature Protocols 2006, 1, 246-252.).Wang group based on 17E DNAzyme with and double-stranded DNA fluorescence molecule chelating dyes Picogreen developed label-free fluorescent optical sensor and detected lead ion (Zhang, L.; Han, B.; Li, T.; Wang, E., Chemical Communications 2011, 47, 3099–3101.).Subsequently, Yin group based on 17E DNAzyme developed electrochemical luminous sensor detection lead ion (Gao, A.; Tang, C.; He, X.; Yin, X., Analyst, 2013, 138, 263–268).The extensive concern of domestic and international expert is received hence with the functional nucleic acid 17E DNAzyme detection lead ion that lead ion is sensitive.
Owing to fluoroscopic examination, ultraviolet detection and electrochemical luminescence detection are required for the detecting instrument that comparison is huge, valuable, therefore these methods are not suitable for the occasion of some simpler and cruder yard field works in one's power.Electrochemical detection method due to have highly sensitive, simple to operate quickly, low cost, energy consumption are low, be prone to the advantages such as instrumentation, thus by the favor day by day of researcher.
Summary of the invention
It is an object of the invention to provide the functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that a kind of selectivity probe good, sensitive, easy, quick is integrated.
The technical solution of the present invention is:
A kind of functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe is integrated, is characterized in that: integrated for probe functional nucleic acid modified electrode immerses the buffer solution 20 ~ 30 minutes added with lead ion to be measured, takes out washing the NaClO at 0.1 M4Electrolyte solution carries out DPV mensuration, the plumbum ion concentration calculating in sample according to the rate of change of the DPV peak current signal scanning record in the range of 0.1～0.6V；Described buffer solution is pH 7.5, containing 140 mM NaCl and 20 mM of 5 mM KCl Tris-HCl solution；
The integrated functional nucleic acid modified electrode of described probe is prepared by following methods: be placed in by golden nanometer particle modified electrode added with self assembly 10 ~ 18 hours in the cushioning liquid of the 17E DNAzyme of 1 μM of sulfydryl modification；Described buffer solution is pH 7.5, containing 140 mM NaCl and 20 mM of 5 mM KCl Tris-HCl solution；Take out electrode to clean with described cushioning liquid, then electrode is placed in the above-mentioned cushioning liquid added with 1mM sulfydryl hexanol passivation 1-3 hour, clean with same cushioning liquid and remove unassembled mercaptoethanol；Finally transfer to electrode the substrate 17DS solution containing 1 μM assembles 10 ~ 18 hours；3 ' the terminal modified ferrocene probe molecules of described substrate 17DS；Wash electrode 2-3 time with above-mentioned cushioning liquid again, obtain the functional nucleic acid modified electrode that probe is integrated.
The integrated functional nucleic acid modified electrode of probe immerses the buffer solution 25 minutes added with lead ion to be measured.
Golden nanometer particle modified electrode is placed in added with self assembly 12 hours in the cushioning liquid of the 17E DNAzyme of 1 μM of sulfydryl modification.
Finally transfer to electrode the substrate 17DS solution containing 1 μM assembles 12 hours.
The invention have the benefit that
1. The present invention uses the identification molecule that functional nucleic acid 17E DNAzyme detects as lead ion, the best, to environment non-secondary pollution.
2. Employing electrochemical method detects, have highly sensitive, instrument is simple, simple to operate quickly, the advantage such as low cost and low energy consumption.
3. This method uses the method that probe and substrate chain 17DS integrate, and is integrated into electrode surface, and without adding extra electrochemical probe reagent in electrolyte solution during detection, the degree of integration of electrode is high, functional by force, detection convenience improves.
Accompanying drawing explanation
The invention will be further described with embodiment below in conjunction with the accompanying drawings.
Fig. 1 is the DPV curvilinear motion figure in the present invention to the modified electrode measured during lead at different concentrations ion detection.
Fig. 2 is the standard working curve figure that lead ion is detected by the modified electrode of the present invention.
Detailed description of the invention
(1) The preparation of the functional nucleic acid modified electrode that probe is integrated
Golden nanometer particle modified electrode is placed in added with self assembly 12 hours in the cushioning liquid of the 17E DNAzyme of 1 μM of sulfydryl modification；Described buffer solution is pH 7.5, containing 140 mM NaCl and 20 mM of 5 mM KCl Tris-HCl solution；Take out electrode to clean with described cushioning liquid, then electrode is placed in the above-mentioned cushioning liquid added with 1mM sulfydryl hexanol passivation 2 hours, clean with same cushioning liquid and remove unassembled mercaptoethanol；Finally transfer to electrode the substrate 17DS solution containing 1 μM assembles 12 hours；3 ' the terminal modified ferrocene probe molecules of described substrate 17DS；Wash electrode 3 times with above-mentioned cushioning liquid again, obtain the functional nucleic acid modified electrode that probe is integrated.
(2) detection of lead ion in lake water water sample
The modified electrode prepared is immersed in a series of added with the cushioning liquid of the lead ion standard water sample of variable concentrations soaks 25 min after, with above-mentioned cushioning liquid cleaning electrode 2 times；NaClO with 0.1 M4As supporting electrolyte solution, carrying out DPV sweep test, test parameter is: the adjustment time is 50 ms, and interval time is 0.5 s, and modulated amplitude is 50 mV, and step electromotive force is 5 mV, and sweep limits is 0.1～0.6V.With DPV peak current as ordinate, plumbum ion concentration is abscissa, draws standard working curve, as shown in Figure 2.
Using lake water as actual sample, lake water with 0.22 μm membrane filtration, then after being centrifuged with 15000 turns/min, takes supernatant and is measured after activated carbon decolorizing.By 50 The lake water water sample that μ L handles well joins in above-mentioned cushioning liquid, according to said determination step operation, measures its DPV peak current, according to DPV peak current rate of change (I0-I)/I0, the linear equation of standard curve fit can calculate plumbum ion concentration in water sample is 327 pM.

Claims (4)

1. probe is an integrated functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method, is characterized in that: functional nucleic acid modified electrode integrated for probe immerses the buffer solution 20 ~ 30 minutes added with lead ion to be measured, takes out washing the NaClO at 0.1 M4Electrolyte solution carries out DPV mensuration, the plumbum ion concentration calculating in sample according to the rate of change of the DPV peak current signal scanning record in the range of 0.1～0.6V；Described buffer solution is pH 7.5,20 mM Tris-HCl solution containing 140 mM NaCl and 5 mM KCl；
The integrated functional nucleic acid modified electrode of described probe is prepared by following methods: be placed in by golden nanometer particle modified electrode added with self assembly 10 ~ 18 hours in the cushioning liquid of the 17E DNAzyme of 1 μM of sulfydryl modification；Described buffer solution is pH 7.5, containing 140 mM NaCl and 5 The 20 mM Tris-HCl solution of mM KCl；Take out electrode to clean with described cushioning liquid, then electrode is placed in the above-mentioned cushioning liquid added with 1mM sulfydryl hexanol passivation 1-3 hour, clean with same cushioning liquid and remove unassembled mercaptoethanol；Finally transfer to electrode the substrate 17DS solution containing 1 μM assembles 10 ~ 18 hours；3 ' the terminal modified ferrocene probe molecules of described substrate 17DS；Wash electrode 2-3 time with above-mentioned cushioning liquid again, obtain the functional nucleic acid modified electrode that probe is integrated.
The functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe the most according to claim 1 is integrated, is characterized in that: the integrated functional nucleic acid modified electrode of probe immerses the buffer solution 25 minutes added with lead ion to be measured.
The functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe the most according to claim 1 and 2 is integrated, is characterized in that: be placed in by golden nanometer particle modified electrode added with self assembly 12 hours in the cushioning liquid of the 17E DNAzyme of 1 μM of sulfydryl modification.
The functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe the most according to claim 1 and 2 is integrated, is characterized in that: finally transfer to electrode assemble 12 hours in the substrate 17DS solution containing 1 μM.
CN201610372050.1A 2016-05-31 2016-05-31 The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method Expired - Fee Related CN105891286B (en)

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CN201811265189.1A CN109060903A (en) 2016-05-31 2016-05-31 The preparation method of lead ion detection method probe integrated functionality nucleic acid modified electrode in lake water water sample
CN201811258669.5A CN109060902A (en) 2016-05-31 2016-05-31 The functional nucleic acid modified electrode that lead ion detection method is integrated with probe in lake water water sample
CN201811258719.XA CN109187695A (en) 2016-05-31 2016-05-31 Application of the probe integrated functionality nucleic acid modified electrode in lake water water sample in lead ion detection
CN201610372050.1A CN105891286B (en) 2016-05-31 2016-05-31 The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method

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CN201811258719.XA Pending CN109187695A (en) 2016-05-31 2016-05-31 Application of the probe integrated functionality nucleic acid modified electrode in lake water water sample in lead ion detection
CN201811265189.1A Pending CN109060903A (en) 2016-05-31 2016-05-31 The preparation method of lead ion detection method probe integrated functionality nucleic acid modified electrode in lake water water sample
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CN107843637A (en) * 2017-10-12 2018-03-27 辽宁师范大学 For detecting the block polymer decorated electrode and preparation method of lead ion

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CN106442345A (en) * 2016-09-08 2017-02-22 常熟理工学院 Method of detecting lead ion based on CD spectrum detection technology
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CN107843637A (en) * 2017-10-12 2018-03-27 辽宁师范大学 For detecting the block polymer decorated electrode and preparation method of lead ion

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