CN105891286B - The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method - Google Patents

The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method Download PDF

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CN105891286B
CN105891286B CN201610372050.1A CN201610372050A CN105891286B CN 105891286 B CN105891286 B CN 105891286B CN 201610372050 A CN201610372050 A CN 201610372050A CN 105891286 B CN105891286 B CN 105891286B
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buffer solution
electrode
nucleic acid
modified electrode
functional nucleic
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CN105891286A (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 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 the functional nucleic acid modified electrode Direct Electrochemistries that a kind of probe integrates to detect lead ion method, and the functional nucleic acid modified electrode that probe is integrated immerses buffer 20 ~ 30 minutes added with lead ion to be measured, takes out washing and in the NaClO of 0.1M4DPV measurement is carried out in electrolyte solution, and the plumbum ion concentration in sample is calculated according to the change rate for the DPV peak current signal for scanning record within the scope of 0.1~0.6V.The present invention is selectively good, sensitive, easy, quick.

Description

The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method
Technical field
The present invention relates to the functional nucleic acid modified electrode Direct Electrochemistries that a kind of probe integrates to detect lead ion method.
Background technique
Lead ion is a kind of common harmful heavy metal ions, and monitoring/detection has human health and environmental monitoring Highly important meaning.In the past few decades, people are had been developed such as Atomic absorption/emission spectrum, fluorescence spectrum, matter A variety of methods such as spectrum, photonic crystal are monitored/detect to lead ion.In recent years, Lu Yi research group in the U.S. by sieving in vitro Selecting technology obtains the functional nucleic acid 17E DNAzyme sensitive to lead ion, they are identified using 17E DNAzyme as lead ion Element uses gold nanoparticle as sensing element, develops colorimetric sensor detection lead ion (Liu, J.; Lu, Y., Nature Protocols 2006, 1, 246-252.).Wang group is based on 17E DNAzyme and and double-stranded DNA fluorescence Molecule chelating dyes Picogreen has developed label-free fluorescent optical sensor detection lead ion (Zhang, L.; Han, B.; Li, T.; Wang, E., Chemical Communications 2011, 47, 3099–3101.).Then, Yin group Electrochemical luminescence sensor detection lead ion (Gao, A. are developed based on 17E DNAzyme; Tang, C.; He, X.; Yin, X., Analyst, 2013, 138, 263–268).Therefore the functional nucleic acid 17E DNAzyme of lead ion sensitivity is utilized Detection lead ion receives the extensive concern of domestic and international expert.
Since the detection of fluorescence detection, ultraviolet detection and electrochemical luminescence requires huger, valuable detecting instrument, Therefore these methods are not suitable for the occasion of some simpler and cruder yards field work in one's power.Electrochemical detection method due to Have many advantages, such as high sensitivity, it is easy to operate it is quick, at low cost, low energy consumption, is easy to instrumentation, thus day by researcher Benefit favor.
Summary of the invention
The purpose of the present invention is to provide a kind of selectivity, and the integrated functional nucleic acid of good, sensitive, easy, quick probe is repaired It adorns electrode Direct Electrochemistry and detects lead ion method.
The technical solution of the invention is as follows:
A kind of functional nucleic acid modified electrode Direct Electrochemistry detection lead ion method that probe is integrated, it is characterized in that: will visit The integrated functional nucleic acid modified electrode of needle immerses buffer 20 ~ 30 minutes added with lead ion to be measured, takes out washing and in 0.1 M NaClO4DPV measurement is carried out in electrolyte solution, according to the DPV peak current signal for scanning record within the scope of 0.1~0.6V Change rate calculate the plumbum ion concentration in sample；The buffer is pH 7.5, containing 140 mM NaCl and 5 mM KCl 20 mM Tris-HCl solution；
The integrated functional nucleic acid modified electrode of the probe is prepared by following methods: by gold nanoparticle modified electrode It is placed in self assembly 10 ~ 18 hours in the buffer solution added with 1 μM of mercapto-modified 17E DNAzyme；The buffer is pH 7.5, containing the 20 mM Tris-HCl solution of 140 mM NaCl and 5 mM KCl；Electrode is taken out to be cleaned with the buffer solution, then Electrode is placed in the above-mentioned buffer solution added with 1mM sulfydryls hexanol and is passivated 1-3 hours, is cleaned and is removed with same buffer solution Remove unassembled mercaptoethanol；Finally electrode is transferred in the substrate 17DS solution containing 1 μM and is assembled 10 ~ 18 hours；It is described The terminal modified ferrocene probe molecule in the 3 ' of substrate 17DS；Electrode is washed 2-3 times with above-mentioned buffer solution again, obtains what probe integrated Functional nucleic acid modified electrode.
The integrated functional nucleic acid modified electrode of probe immerses buffer 25 minutes added with lead ion to be measured.
Gold nanoparticle modified electrode is placed in the buffer solution added with 1 μM of mercapto-modified 17E DNAzyme from group It fills 12 hours.
Finally electrode is transferred in the substrate 17DS solution containing 1 μM and is assembled 12 hours.
The invention has the benefit that
1. the identification molecule that the present invention is detected using functional nucleic acid 17E DNAzyme as lead ion, selectivity is good, right Environment is without secondary pollution.
2. detected using electrochemical method, have high sensitivity, instrument it is simple, it is easy to operate quickly, low cost and low energy The advantages that consumption.
3. this method is integrated into electrode surface, is not necessarily in detection process using the method for probe and substrate chain 17DS integration Additional electrochemical probe reagent is added in electrolyte solution, the degree of integration of electrode is high, functional, and detection convenience mentions It is high.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples.
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 modified electrode of the invention detects lead ion.
Specific embodiment
(1) preparation of the integrated functional nucleic acid modified electrode of probe
Gold nanoparticle modified electrode is placed in the buffer solution added with 1 μM of mercapto-modified 17E DNAzyme from group It fills 12 hours；The buffer is pH 7.5, the 20 mM Tris-HCl solution containing 140 mM NaCl and 5 mM KCl；It takes out Electrode is cleaned with the buffer solution, then that electrode is placed in the above-mentioned buffer solution added with 1mM sulfydryls hexanol passivation 2 is small When, it is cleaned with same buffer solution and removes unassembled mercaptoethanol；Electrode is finally transferred to the substrate containing 1 μM It is assembled 12 hours in 17DS solution；The terminal modified ferrocene probe molecule in the 3 ' of the substrate 17DS；It is washed again with above-mentioned buffer solution It washs electrode 3 times, obtains the integrated functional nucleic acid modified electrode of probe.
(2) in lake water water sample lead ion detection
The modified electrode prepared is immersed in a series of buffer solution of lead ion standard water samples added with various concentration After 25 min of middle immersion, with above-mentioned buffer solution cleaning electrode 2 times；With the NaClO of 0.1 M4As supporting electrolyte solution, into Row DPV sweep test, test parameter are as follows: adjustment time is 50 ms, and interval time is 0.5 s, and modulated amplitude is 50 mV, step Potential is 5 mV, and scanning range is 0.1~0.6V.Using DPV peak current as ordinate, plumbum ion concentration is abscissa, draws mark Quasi- working curve, as shown in Figure 2.
Using lake water as actual sample, lake water after active carbon decoloring with 0.22 μm of membrane filtration, then with 15000 Turn/min centrifugation after, take supernatant to be measured.The 50 μ L lake water water sample handled well is added in above-mentioned buffer solution, is pressed According to said determination step operation, its DPV peak current is measured, according to DPV peak current change rate (I0-I)/I0, intended by standard curve It is 327 pM that the linear equation of conjunction, which can calculate plumbum ion concentration in water sample,.

Claims (1)

1. a kind of integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method, it is characterized in that:
(1) preparation of the integrated functional nucleic acid modified electrode of probe
Gold nanoparticle modified electrode is placed in self assembly 12 in the buffer solution added with 1 μM of mercapto-modified 17E DNAzyme Hour；The buffer solution is pH 7.5, the 20 mM Tris-HCl solution containing 140 mM NaCl and 5 mM KCl；Take out electricity Pole is cleaned with the buffer solution, then electrode is placed in the above-mentioned buffer solution added with 1mM sulfydryls hexanol and is passivated 2 hours, It is cleaned with same buffer solution and removes unassembled mercaptoethanol；Finally electrode is transferred to molten containing 1 μM of substrate 17DS It is assembled 12 hours in liquid；The terminal modified ferrocene probe molecule in the 3 ' of the substrate 17DS；Electrode 3 is washed with above-mentioned buffer solution again It is secondary, obtain the integrated functional nucleic acid modified electrode of probe；
(2) in lake water water sample lead ion detection
The modified electrode prepared is immersed in a series of buffer solutions added with the lead ion standard water sample of various concentration and is soaked After steeping 25 min, with above-mentioned buffer solution cleaning electrode 2 times；With the NaClO of 0.1 M4As supporting electrolyte solution, carry out DPV sweep test, test parameter are as follows: adjustment time is 50 ms, and interval time is 0.5 s, and modulated amplitude is 50 mV, step electricity Gesture is 5 mV, and scanning range is 0.1~0.6V；Using DPV peak current as ordinate, plumbum ion concentration is abscissa, draws standard Working curve；
Using lake water as sample, lake water after active carbon decoloring with 0.22 μm of membrane filtration, then with 15000 turns/min from After the heart, supernatant is taken to be measured；The 50 μ L lake water water sample handled well is added in above-mentioned buffer solution, according to above-mentioned survey Determine step operation, measure its DPV peak current, according to DPV peak current change rate (I0-I)/I0, by the linear of standard curve fit Equation can calculate plumbum ion concentration in water sample.
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
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CN106442345B (en) * 2016-09-08 2019-03-15 常熟理工学院 A kind of lead ion detection method based on CD spectrum detection technique
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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CN104020200A (en) * 2014-06-18 2014-09-03 青岛科技大学 Electrochemical method for measuring melamine
CN104914151A (en) * 2015-05-08 2015-09-16 首都师范大学 Forming method of ampicillin and sulfadimethoxine electrochemical sensor self-assembled passivation layer, and electrochemical sensor thereof
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