CN109580573B - Detection method of chromium ions - Google Patents

Detection method of chromium ions Download PDF

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CN109580573B
CN109580573B CN201910052588.8A CN201910052588A CN109580573B CN 109580573 B CN109580573 B CN 109580573B CN 201910052588 A CN201910052588 A CN 201910052588A CN 109580573 B CN109580573 B CN 109580573B
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chromium ions
concentration
reaction system
nucleic acid
fluorescence
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CN109580573A (en
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李旺
杨涛
林亲录
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Central South University of Forestry and Technology
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
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    • G01N21/6402—Atomic fluorescence; Laser induced fluorescence

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Abstract

The invention discloses a detection method of chromium ions. The detection method of the chromium ions comprises the following steps: placing a proper amount of cerium dioxide nano particles, ethylene diamine tetraacetic acid and fluorescent molecule modified nucleic acid into a container containing buffer solution, and uniformly mixing; placing the solution to be detected in the mixed solution obtained in the step S1 to prepare and form a reaction system; in a reaction system, the concentration of cerium dioxide nano particles is 10mg/L, the concentration of ethylene diamine tetraacetic acid is 0.1-1.0mM, and the concentration of nucleic acid modified by fluorescent molecules is 100 nM; the reaction system of step S2 is subjected to fluorescence detection, and the concentration of chromium ions in the reaction system is measured by the change in the intensity of the fluorescence signal. The invention also provides a detection method of the chromium ions, which is used for rapidly detecting the trivalent chromium ions and has the characteristics of strong selectivity and high sensitivity.

Description

Detection method of chromium ions
Technical Field
The invention relates to the technical field of photochemistry and biochemical sensing, in particular to a detection method of chromium ions.
Background
Along with the continuous improvement of the living standard of people and the continuous enhancement of environmental protection consciousness, the heavy metal pollution in the water body is more and more emphasized. The accumulation of heavy metal ions in organs and tissues such as liver and kidney of human body can cause injury and chronic poisoning of various organs and tissues. Therefore, the method has important significance for protecting human beings from being damaged by heavy metal pollution and finding a simple, convenient and efficient method for detecting heavy metal residues in the environment.
Chromium is one of environmental pollutants, mainly comes from industries of alloy steel smelting, electroplating, paint, tanning, medicine, textile, printing and the like, and waste water and waste gas of the industries flow through farmlands, rivers and soil, so that plants such as crops, vegetables and the like are polluted by chromium, and harm to human beings and animals is caused. Toxic chromium is generally classified into trivalent and hexavalent chromium, with trivalent chromium being mainly present in surface water and hexavalent chromium being mainly present in groundwater. The conventional method for detecting chromium ions includes methods such as chromatography, spectrophotometry, atomic spectrometry and the like. However, these methods have two major problems: firstly, the selectivity is poor, and secondly, the sensitivity is low.
In view of the above, it is an urgent need in the art to provide a simple method for detecting chromium ions with high selectivity and high sensitivity.
Disclosure of Invention
The invention aims to provide a detection method of chromium ions, which is used for rapidly detecting trivalent chromium ions and has the characteristics of strong selectivity and high sensitivity.
In order to solve the above problems, the present invention provides a method for detecting chromium ions, which has the following technical scheme:
a method for detecting chromium ions comprises the following steps:
step S1, placing a proper amount of cerium dioxide nano particles, ethylene diamine tetraacetic acid and fluorescent molecule modified nucleic acid into a container containing buffer solution, and uniformly mixing;
step S2, placing the solution to be detected in the mixed solution obtained in the step S1 to prepare and form a reaction system; in a reaction system, the concentration of cerium dioxide nano particles is 10mg/L, the concentration of ethylene diamine tetraacetic acid is 0.1-1.0mM, and the concentration of nucleic acid modified by fluorescent molecules is 100 nM;
and a step S3 of detecting fluorescence of the reaction system obtained in the step S2 and measuring the concentration of chromium ions in the reaction system based on the change in the intensity of the fluorescence signal.
Furthermore, in the reaction system, the concentration of the cerium dioxide nano particles is 10mg/L, the concentration of the ethylene diamine tetraacetic acid is 0.5mM, and the concentration of the fluorescent molecule modified nucleic acid is 100 nM.
Further, in step S2, the reaction time is 10-20min during the preparation of the reaction system.
Further, in step S2, the reaction temperature is 20-30 ℃ during the preparation of the reaction system.
Further, the measured chromium ions are trivalent chromium ions.
Further, in step S3, the excitation wavelength for fluorescence detection is 485nm and the emission wavelength is 535 nm.
Further, in step S1, the buffer used is HEPES buffer.
Compared with the prior art, the detection method of the chromium ions provided by the invention has the beneficial effects that:
the detection method of chromium ions provided by the invention utilizes the characteristic that chromium has a very small ligand exchange rate. The electronegative ethylene diamine tetraacetic acid radical can act on the surface of the cerium dioxide nanoparticle with positive electricity to prevent the nanoparticle from adsorbing electronegative nucleic acid molecules, so that the nucleic acid modified by fluorescent molecules is far away from the cerium dioxide nanoparticle to prevent the fluorescence of the nucleic acid from being quenched by the nanoparticle. When positively charged chromium ions exist in the reaction system, the chromium ions can be a bridge to adsorb nucleic acid molecules with negative electricity to the surface of the ethylene diamine tetraacetic acid-cerium dioxide nano particle composite material with negative electricity, so that the fluorescence of the composite material is quenched by the nano particles; the size of the adsorption capacity can be regulated and controlled through target chromium ions to be detected, and the change of a fluorescence signal is realized, so that the content of the chromium ions is determined.
Because the ligand exchange rate of other metals is at least 5 orders of magnitude higher than that of chromium, the other metals are quickly chelated with the EDTA, and the selectivity of the method can be ensured. Therefore, the chromium ion detection method provided by the invention has the advantages of convenience in operation, high selectivity, high sensitivity and the like.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a schematic diagram illustrating the detection principle of the detection method of chromium ions according to the present invention;
FIG. 2 is a diagram of fluorescence quenching rate analysis of different concentrations of EDTA disodium reaction system;
FIG. 3 is a fluorescence kinetic spectrum of a chromium ion reaction system with different concentrations;
FIG. 4 is a standard curve diagram of the detection method of chromium ions according to the present invention;
fig. 5 is a schematic diagram of a selective test result of the detection method of chromium ions provided by the present invention.
Detailed Description
In order to make the technical solutions in the embodiments of the present invention better understood and make the above objects, features, and advantages of the present invention more comprehensible, specific embodiments of the present invention are described below with reference to the accompanying drawings.
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual values, and between the individual values may be combined with each other to yield one or more new ranges of values, which ranges of values should be considered as specifically disclosed herein.
Example 1
The invention provides a method for detecting chromium ions, which comprises the following steps:
step S1, placing a proper amount of cerium dioxide nano particles, ethylene diamine tetraacetic acid and fluorescent molecule modified nucleic acid into a container containing buffer solution, and uniformly mixing;
wherein, the nucleic acid modified by the fluorescent molecule is sold in the market, the product is provided by the company of biological engineering (Shanghai), the number of the basic groups of the nucleic acid is 24, and the basic group sequence is as follows: 5 '-FAM-ACG CAT CTG TGA AGA GAA CCT GGG-3' (FAM is modified by fluorescent molecules);
the buffer solution is HEPES buffer solution; the container is a 96-hole enzyme label plate.
Step S2, placing the solution to be detected in the mixed solution obtained in the step S1 to prepare and form a reaction system; the total volume of the reaction system is 100 mu L, the concentration of the prepared cerium dioxide nano particles is 10mg/L, the concentration of the ethylene diamine tetraacetic acid is 0.1-1.0mM, and the concentration of the nucleic acid modified by the fluorescent molecules is 100 nM;
wherein the solution to be detected is a solution containing trivalent chromium ions; in the preparation process of the reaction system, the reaction time is 10-20min, and the reaction temperature is 20-30 ℃.
And a step S3 of detecting fluorescence of the reaction system obtained in the step S2 and measuring the concentration of chromium ions in the reaction system based on the change in the intensity of the fluorescence signal.
During fluorescence detection, the excitation wavelength and the emission wavelength used by the instrument are determined according to the selected fluorescent organic small molecules.
Fig. 1 is a schematic diagram illustrating a detection principle of the method for detecting chromium ions according to the present invention. In the process of fluorescence detection, when no chromium ions exist in a reaction system, the fluorescence signal of the solution is strong; when chromium ions exist in the reaction system, the chromium ions can cause the fluorescence of the fluorescent molecules modified on the nucleic acid molecules to be quenched by the cerium dioxide nano particles, and the fluorescence signal of the solution is low; and the fluorescence signal obtained by detection is smaller and smaller as the concentration of the chromium ions is increased.
The optimum experimental conditions for the reaction system were obtained by adjusting the concentration of disodium ethylenediaminetetraacetate in the reaction system, and examples 2-7 were obtained.
Examples 2 to 7
The used solutions to be detected are respectively a chromium ion-containing solution and a blank group; adjusting the concentration of the disodium ethylene diamine tetraacetate in the reaction system to be 0.05mM, 0.1mM, 0.3mM, 0.5mM, 0.7mM and 1.0mM respectively, wherein the concentration of chromium ions in the chromium-containing solution to be tested is 0.5 mu M; controlling the reaction time to be 20min and the reaction temperature to be 25 ℃; the remaining parameters refer to example 1;
fluorescence detection: and (3) placing the reaction systems into a microplate reader respectively to perform kinetic test on the intensity of fluorescence emission. The detection excitation wavelength of the microplate reader is 485nm, the emission wavelength is 535nm, the test time is 20min, and the temperature is normal temperature.
Please refer to fig. 2, which is a graph illustrating fluorescence quenching rate analysis of different concentrations of edta disodium reaction system. As can be seen from FIG. 2, when the concentration of disodium EDTA is 0.5mM, the corresponding quenching rate of fluorescence signal is the highest, and therefore, the detection sensitivity is higher when detecting trivalent chromium ions.
In summary, in the detection method of chromium ions provided by the present invention, the optimized reaction conditions in the reaction system are as follows:
the concentration of the cerium dioxide nano particles is 10mg/L, the concentration of the ethylene diamine tetraacetic acid is 0.5mM, and the concentration of the nucleic acid modified by the fluorescent molecules is 100 nM; the reaction time is 20min, and the reaction temperature is normal temperature.
Examples 8 to 18
Determination of standard curve chart
The volume of the reaction system is 100 mu L, the concentration of cerium dioxide nano particles is 10mg/L, the concentration of ethylene diamine tetraacetic acid is 0.5mM, the concentration of nucleic acid modified by fluorescent molecules is 100nM, and the concentrations of chromium ions in the solution to be detected are 0 mu M, 0.05 mu M, 0.1 mu M, 0.2 mu M, 0.5 mu M, 1.0 mu M, 2.0 mu M, 5.0 mu M, 10 mu M, 15 mu M and 20 mu M respectively; the reaction time is 20min, and the reaction temperature is normal temperature; the remaining parameters refer to example 1;
the reaction systems of examples 8 to 18 were subjected to fluorescence detection using a microplate reader, the excitation wavelength for detection of the microplate reader was 485nm, the emission wavelength was 535nm, and the detection temperature was room temperature.
Please refer to fig. 3 and fig. 4 in combination, wherein fig. 3 is a fluorescence kinetic spectrum of a reaction system with different concentrations of chromium ions; fig. 4 is a standard curve diagram of the detection method of chromium ions provided by the present invention. As can be seen from fig. 3 and 4, the fluorescence value becomes smaller as the concentration of chromium ions increases. As can be seen from fig. 4, in the detection method of chromium ions provided by the present invention, the linear equation is: F/F0＝0.0261+0.2633C(R20.995), the linear relationship is good, the linear detection range is 0-2 μ M, and the lower detection limit is 0.02 μ M.
Therefore, the detection method of the chromium ions provided by the invention has a lower detection limit and can meet the requirement of actual detection.
Examples 19 to 30
Selectivity test
In order to test the selectivity of the detection method of chromium ions provided by the invention, several common metal ions Ca are selected2+、Mg2+、Cd2+、Cu2+、Pb2+、Zn2+、Ni2+、Mn2+、Al3+、Fe3+、Cr3+Adding the sample into a detection environment, and performing an interference experiment; wherein Ca2+、Mg2+、Cd2+、Cu2+、Pb2+、Zn2+、Ni2+、Mn2+、Al3+、Fe3+Has a concentration of 20. mu.M, Cr3+The concentration is 5 mu M; and blank groups were added for comparative experiments.
The volume of the reaction system is 100 mu L, the concentration of the cerium dioxide nano particles is 10mg/L, the concentration of the ethylene diamine tetraacetic acid is 0.5mM, and the concentration of the nucleic acid modified by the fluorescent molecules is 100 nM; the reaction temperature is room temperature, and the reaction time is 20 min; the remaining parameters refer to example 1.
Please refer to fig. 5, which is a schematic diagram of a selective experiment result of the method for detecting chromium ions according to the present invention. As can be seen from FIG. 5, Ca2+、Mg2+、Cd2+、Cu2+、Pb2+、Zn2+、Ni2+、Mn2+、Al3+、Fe3+Plasma metal ion pair Cr3+The detection of (a) is essentially non-interfering, mainly because the ligand exchange rate of the other metal is at least 5 orders of magnitude higher than that of chromium, and therefore the other metal is rapidly chelated with ethylenediaminetetraacetic acid. Further, the detection method of the chromium ions provided by the invention has good selectivity.
The detection method of chromium ions provided by the invention utilizes the characteristic that chromium has a very small ligand exchange rate. The electronegative ethylene diamine tetraacetic acid radical can act on the surface of the cerium dioxide nanoparticle with positive electricity to prevent the nanoparticle from adsorbing electronegative nucleic acid molecules, so that the nucleic acid modified by fluorescent molecules is far away from the cerium dioxide nanoparticle to prevent the fluorescence of the nucleic acid from being quenched by the nanoparticle. When positively charged chromium ions exist in the reaction system, the chromium ions can be a bridge to adsorb nucleic acid molecules with negative electricity to the surface of the ethylene diamine tetraacetic acid-cerium dioxide nano particle composite material with negative electricity, so that the fluorescence of the composite material is quenched by the nano particles; the size of the adsorption capacity can be regulated and controlled through target chromium ions to be detected, and the change of a fluorescence signal is realized, so that the content of the chromium ions is determined.
Since the ligand exchange rate of other metals is at least 5 orders of magnitude higher than that of chromium, other metals are rapidly chelated with ethylenediaminetetraacetic acid.
Therefore, the chromium ion detection method provided by the invention has the advantages of convenience in operation, high selectivity, high sensitivity and the like.
The embodiments in the present specification are described in a progressive manner, and the same and similar parts among the embodiments are referred to each other, and each embodiment focuses on the differences from the other embodiments.
The embodiments of the present invention are described in detail above with reference to the drawings, but the present invention is not limited to the described embodiments. Various changes, modifications, substitutions and alterations to these embodiments will occur to those skilled in the art without departing from the spirit and scope of the present invention.

Claims (7)

1. A method for detecting chromium ions is characterized by comprising the following steps:
step S1, placing a proper amount of cerium dioxide nano particles, ethylene diamine tetraacetic acid and fluorescent molecule modified nucleic acid into a container containing buffer solution, and uniformly mixing;
wherein, the nucleotide sequence modified by the fluorescent molecule is as follows: 5 '-FAM-ACG CAT CTG TGA AGA GAA CCT GGG-3';
step S2, placing the solution to be detected in the mixed solution obtained in the step S1 to prepare and form a reaction system; in a reaction system, the concentration of cerium dioxide nano particles is 10mg/L, the concentration of ethylene diamine tetraacetic acid is 0.1-1.0mM, and the concentration of nucleic acid modified by fluorescent molecules is 100 nM;
and a step S3 of detecting fluorescence of the reaction system obtained in the step S2 and measuring the concentration of chromium ions in the reaction system based on the change in the intensity of the fluorescence signal.
2. The method for detecting chromium ions according to claim 1, wherein in the reaction system, the concentration of the cerium dioxide nanoparticles is 10mg/L, the concentration of the disodium ethylenediamine tetraacetic acid is 0.5mM, and the concentration of the fluorescent molecule-modified nucleic acid is 100 nM.
3. The method for detecting chromium ions according to claim 1, wherein in step S2, the reaction time is 10-20min during the preparation of the reaction system.
4. The method for detecting chromium ions according to claim 3, wherein in step S2, the reaction temperature is 20-30 ℃ during the preparation of the reaction system.
5. The method of claim 1, wherein the detected chromium ions are trivalent chromium ions.
6. The method of claim 1, wherein in step S3, the excitation wavelength for fluorescence detection is 485nm and the emission wavelength is 535 nm.
7. The method according to any one of claims 1 to 6, wherein the buffer used in step S1 is HEPES buffer.
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Citations (3)

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Publication number Priority date Publication date Assignee Title
CN102507454A (en) * 2011-10-31 2012-06-20 中国科学院宁波材料技术与工程研究所 Method for detecting trivalent chromium ions
CN103149183A (en) * 2013-01-19 2013-06-12 桂林理工大学 Method for swiftly detecting Leersia hexandra middlechro (VI) ion concentration
CN108779455A (en) * 2016-03-17 2018-11-09 东丽株式会社 The recovery method of nucleic acid

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Publication number Priority date Publication date Assignee Title
EP2984060A4 (en) * 2013-11-01 2016-10-26 Thomas T Yamashita Compositions for controlling plant parasitic nematodes and methods for using same

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
CN102507454A (en) * 2011-10-31 2012-06-20 中国科学院宁波材料技术与工程研究所 Method for detecting trivalent chromium ions
CN103149183A (en) * 2013-01-19 2013-06-12 桂林理工大学 Method for swiftly detecting Leersia hexandra middlechro (VI) ion concentration
CN108779455A (en) * 2016-03-17 2018-11-09 东丽株式会社 The recovery method of nucleic acid

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Hydrogenated CuO\Au@CeO2 Z-scheme catalyst for photocatalytic oxidation of amines to imines;Yingying Liu等;《Catalysis Science & Technology》;20181231;第5535-5543页 *
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