CN109060902A - The functional nucleic acid modified electrode that lead ion detection method is integrated with probe in lake water water sample - Google Patents

The functional nucleic acid modified electrode that lead ion detection method is integrated with probe in lake water water sample Download PDF

Info

Publication number
CN109060902A
CN109060902A CN201811258669.5A CN201811258669A CN109060902A CN 109060902 A CN109060902 A CN 109060902A CN 201811258669 A CN201811258669 A CN 201811258669A CN 109060902 A CN109060902 A CN 109060902A
Authority
CN
China
Prior art keywords
electrode
probe
nucleic acid
buffer solution
modified electrode
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201811258669.5A
Other languages
Chinese (zh)
Inventor
王红卫
张爱平
王海天
申晓萍
王超颖
洪华
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nantong Entry-Exit Inspection And Quarantine Bureau Of People's Republic Of
Original Assignee
Nantong Entry-Exit Inspection And Quarantine Bureau Of People's Republic Of
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nantong Entry-Exit Inspection And Quarantine Bureau Of People's Republic Of filed Critical Nantong Entry-Exit Inspection And Quarantine Bureau Of People's Republic Of
Priority to CN201811258669.5A priority Critical patent/CN109060902A/en
Publication of CN109060902A publication Critical patent/CN109060902A/en
Pending legal-status Critical Current

Links

Classifications

    • 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

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses the functional nucleic acid modified electrodes that lead ion detection method in a kind of lake water water sample is integrated with probe, the detection method is that the functional nucleic acid modified electrode for integrating probe 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 functional nucleic acid modified electrode that lead ion detection method is integrated with probe in lake water water sample
The application is application number: 201610372050.1, the applying date: 2016-05-31, title " the integrated function core of probe The divisional application of sour modified electrode Direct Electrochemistry detection 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: by probe collection At functional nucleic acid modified electrode immerse added with buffer 20 ~ 30 minutes of lead ion to be measured, take out washing and 0.1 M's 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 calculates the plumbum ion concentration in sample；The buffer is pH 7.5,20 containing 140 mM NaCl and 5 mM KCl MM Tris-HCl solution；
The integrated functional nucleic acid modified electrode of the probe is prepared by following methods: gold nanoparticle modified electrode is placed in Self assembly 10 ~ 18 hours in buffer solution added with 1 μM of mercapto-modified 17E DNAzyme；The buffer be pH 7.5, The 20 mM Tris-HCl solution containing 140 mM NaCl and 5 mM KCl；It takes out electrode to be cleaned with the buffer solution, then will be electric Pole is placed in the above-mentioned buffer solution added with 1mM sulfydryls hexanol and is passivated 1-3 hours, cleans removing not with same buffer solution The mercaptoethanol of assembling；Finally electrode is transferred in the substrate 17DS solution containing 1 μM and is assembled 10 ~ 18 hours；The substrate The terminal modified ferrocene probe molecule in the 3 ' of 17DS；Electrode is washed 2-3 times with above-mentioned buffer solution again, obtains the integrated function of probe 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, to environment It 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 self assembly 12 in the buffer solution added with 1 μM of mercapto-modified 17E DNAzyme Hour；The buffer is pH 7.5, the 20 mM Tris-HCl solution containing 140 mM NaCl and 5 mM KCl；Take out electrode It 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, used Same buffer solution cleaning removes unassembled mercaptoethanol；Electrode is finally transferred to the substrate 17DS solution containing 1 μM Middle assembling 12 hours；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, 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. the functional nucleic acid modified electrode that lead ion detection method is integrated with probe in a kind of lake water water sample, it is characterized in that: described Lead ion detection method is in lake water water sample: the modified electrode prepared is immersed in a series of lead ions added with various concentration After impregnating 25 min in the buffer solution of standard water sample, with above-mentioned buffer solution cleaning electrode；With the NaClO of 0.1 M4As branch Electrolyte solution is held, DPV sweep test, test parameter are carried out are as follows: adjustment time is 50 ms, and interval time is 0.5 s, modulation Amplitude is 50 mV, and step potential is 5 mV, and scanning range is 0.1~0.6V；Using DPV peak current as ordinate, plumbum ion concentration For abscissa, standard working curve is drawn；
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；
The modified electrode is the functional nucleic acid modified electrode that probe integrates, and is prepared by following method: by gold nanoparticle Modified electrode is placed in self assembly about 12 hours in the buffer solution added with 1 μM of mercapto-modified 17E DNAzyme；The buffering Solution is pH 7.5, the 20 mM Tris-HCl solution containing 140 mM NaCl and 5 mM KCl；It is molten with the buffering to take out electrode Liquid cleaning, then electrode is placed in the above-mentioned buffer solution added with 1mM sulfydryls hexanol and is passivated, it is cleaned with same buffer solution Remove unassembled mercaptoethanol；Finally electrode is transferred in the substrate 17DS solution containing 1 μM and is assembled；The substrate The terminal modified ferrocene probe molecule in the 3 ' of 17DS；Electrode is washed with above-mentioned buffer solution again, obtains the integrated functional nucleic acid of probe Modified electrode.
CN201811258669.5A 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 Pending CN109060902A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
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

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
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
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

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN201610372050.1A Division CN105891286B (en) 2016-05-31 2016-05-31 The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method

Publications (1)

Publication Number Publication Date
CN109060902A true CN109060902A (en) 2018-12-21

Family

ID=56709740

Family Applications (4)

Application Number Title Priority Date Filing Date
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
CN201811258669.5A Pending 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
CN201610372050.1A Expired - Fee Related CN105891286B (en) 2016-05-31 2016-05-31 The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method
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

Family Applications Before (1)

Application Number Title Priority Date Filing Date
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

Family Applications After (2)

Application Number Title Priority Date Filing Date
CN201610372050.1A Expired - Fee Related CN105891286B (en) 2016-05-31 2016-05-31 The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method
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

Country Status (1)

Country Link
CN (4) CN109060903A (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101363795A (en) * 2008-08-11 2009-02-11 中国科学院长春应用化学研究所 Unmarked colorimetric determination metallic lead ion method based on aurum nanometer probe and nuclease
CN102375020A (en) * 2010-08-19 2012-03-14 华东师范大学 Method for detecting mercury ions through gold electrode modified by polythymidine DNA
CN102031284B (en) * 2010-11-04 2015-07-29 中国科学院上海微系统与信息技术研究所 A kind of lead ion detection chip based on picodna enzyme, making and using method
CN103123333B (en) * 2012-12-31 2016-03-02 北京师范大学 Based on the method for three electrode sensors and Differential Pulse Voltammetry Fast Measurement lead
CN104569085B (en) * 2013-10-28 2019-04-02 南京大学 The mercury metal ion electrochemical sensor of highly sensitive high selection
CN104020200B (en) * 2014-06-18 2016-05-18 青岛科技大学 A kind of electrochemical method of measuring melamine
CN104020204B (en) * 2014-06-19 2016-08-24 湖南大学 A kind of for electrochemical sensor detecting lead and its preparation method and application
CN104914151B (en) * 2015-05-08 2017-10-13 首都师范大学 It is a kind of to be used for the forming method and its electrochemical sensor of ampicillin and sulfadimethoxine electrochemical sensor self assembly passivation layer
CN105223250B (en) * 2015-09-23 2018-01-02 南京邮电大学 Double stem ring DNA electrochemical sensors of polyA overseas Chinese federations and its preparation and application
CN105445350A (en) * 2015-11-13 2016-03-30 南京理工大学 Electrochemical DNA (Deoxyribose Nucleic Acid) biosensor based on peptide nucleic acid and preparation method of electrochemical DNA biosensor
CN105606675B (en) * 2015-12-30 2018-09-18 湖南大学 Aptamer sensor and its preparation method and application for detecting lead

Also Published As

Publication number Publication date
CN105891286A (en) 2016-08-24
CN105891286B (en) 2018-12-18
CN109187695A (en) 2019-01-11
CN109060903A (en) 2018-12-21

Similar Documents

Publication Publication Date Title
CN102706940B (en) Electrochemical sensor capable of detecting trace mercury in water body, and preparation method and application thereof
Herdan et al. Field evaluation of an electrochemical probe for in situ screening of heavy metals in groundwater
Ordeig et al. Trace detection of mercury (II) using gold ultra‐microelectrode arrays
CN104569085B (en) The mercury metal ion electrochemical sensor of highly sensitive high selection
US20100089773A1 (en) Total organic compound (toc) analyzer
CN101402858A (en) Process for producing low-toxicity fluorescent carbon point
CN102375020A (en) Method for detecting mercury ions through gold electrode modified by polythymidine DNA
CN101563603A (en) Improved online water analysis
CN101563602A (en) Improved water analysis
CN105891286B (en) The integrated functional nucleic acid modified electrode Direct Electrochemistry of probe detects lead ion method
Vasjari et al. Amino acid determination using screen-printed electrochemical sensors
Barek et al. Voltammetric Determination of N, N‐Dimethyl‐4‐amine‐carboxyazobenzene at a Silver Solid Amalgam Electrode
Wu et al. Polythymine Oligonucleotide‐Modified Gold Electrode for Voltammetric Determination of Mercury (II) in Aqueous Solution
CN101382514A (en) Method for detecting oxidative metal ion in water
CN109923415A (en) System and method for determining material concentration in solution
Abdelsalam et al. Detection of Hydroxide Ions in Aqueous Solutions by Steady‐State Voltammetry
De Souza et al. Characterization and Use of Copper Solid Amalgam Electrode for Electroanalytical Determination of Triazines‐Based Herbicides
Zhang et al. Electrochemical stripping analysis of cadmium on tantalum electrode
CN205003110U (en) To hg2+, Pb2+ has electrochemistry DNA biosensing detection device of specificity detection
CN109709171A (en) A kind of optical electro-chemistry sensor and preparation method thereof for trace copper ion detection
CN111122552A (en) Method for monitoring cell to secrete copper ions by single-particle electrochemiluminescence imaging technology
CN110186972B (en) Reproducible acetamiprid electrochemical sensor and preparation method and application thereof
Li Electrogenerated chemiluminescence detection of mercury (II) ions based on DNA probe labeled with ruthenium complex
Hicks et al. Initial Trade Study for In-line Silver Sensor for Spacecraft Potable Water Systems
Navelkar et al. ZnO nanoparticles modified sensor for the electroanalysis of thiosalicylic acid

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20181221