CN113237935B - Cysteine detection method - Google Patents
Cysteine detection method Download PDFInfo
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- CN113237935B CN113237935B CN202110472486.9A CN202110472486A CN113237935B CN 113237935 B CN113237935 B CN 113237935B CN 202110472486 A CN202110472486 A CN 202110472486A CN 113237935 B CN113237935 B CN 113237935B
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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
- 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
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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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
- 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
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
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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
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- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
Abstract
The invention discloses a cysteine detection method. The method comprises the following steps: s1, preparation of modifier solution: dispersing the C-N @ GC nano material in a water-ethanol solution, dropwise adding a naphthol solution with the weight percentage of 5 wt% into the solution, and performing ultrasonic degradation for 2 hours by using an ultrasonic degradation method to obtain a uniform modifier solution; s2, modification of the surface of the glassy carbon electrode: uniformly dripping the modifier solution in the S1 on the surface of the polished glassy carbon electrode, and drying at room temperature to finally obtain the glassy carbon electrode modified by C-N @ GC; s3, using the glassy carbon electrode modified by the C-N @ GC obtained in the S2 for electrochemical detection of cysteine; and modifying the surface of a glassy carbon electrode by using C-N @ GC for cysteine detection. The invention realizes qualitative and quantitative detection of cysteine by applying the nano modified electrode, and provides a new idea and a new method for discovering and constructing a novel electrochemical detection probe.
Description
Technical Field
The invention relates to the technical field of electroanalysis, in particular to a cysteine detection method.
Background
Cysteine (Cys) is a small thiol-containing molecule that, because of its chemical instability, plays a variety of roles in physiological and biological processes. In biological processes, cysteine is a precursor for the synthesis of glutathione, acetyl-coa, cystine, and the like. It can undergo reversible redox reactions under physiological conditions, which is essential for maintaining the tertiary and quaternary structure of proteins. In addition, cysteine can be catalytically metabolized as a substrate into bioactive molecules such as pyruvic acid, sulfenic acid, hydrogen sulfide, and the like, and further, can exert various functions in vivo. Cysteine deficiency in humans can lead to hair loss, slow growth, liver damage, edema, weakness, and skin damage. Elevated cysteine levels are also directly associated with several diseases, including L-cystinuria, aids and cancer. Therefore, it is essential to maintain normal levels of cysteine in humans, and plays a crucial role in maintaining human health.
At present, there are many methods for detecting cysteine, mainly including: fluorescence spectroscopy, colorimetry, chromatography, capillary electrophoresis, chemiluminescence, and the like. Although these methods achieve detection of cysteine, they have problems such as long detection time, expensive equipment and the like. The electrochemical sensor is a preferred method for detecting cysteine due to the advantages of simplicity, high sensitivity, easy operation, miniaturization and the like.
Disclosure of Invention
1. Technical problem to be solved
The invention aims to provide a synthesis method of a C-N @ GC nano material and detection application of the C-N @ GC nano material in cysteine, so as to solve the problems of long detection time, expensive required instruments and equipment and the like in the background technology.
2. Technical scheme
In order to solve the problems, the invention adopts the following technical scheme:
a cysteine detection method comprises the following steps:
s1, preparation of modifier solution: dispersing the C-N @ GC nano material in a water-ethanol solution, dropwise adding a naphthol solution with the weight percentage of 5 wt% into the solution, and performing ultrasonic degradation for 2 hours by using an ultrasonic degradation method to obtain a uniform modifier solution;
s2, modification of the surface of the glassy carbon electrode: uniformly dripping the modifier solution in the S1 on the surface of the polished glassy carbon electrode, and drying at room temperature to finally obtain the glassy carbon electrode modified by C-N @ GC;
S3, using the glassy carbon electrode modified by the C-N @ GC obtained in the S2 for electrochemical detection of cysteine;
the method for synthesizing the C-N @ GC nano material in the S1 comprises the following steps of:
(1) synthesis of ZIF-8: pouring the methanol solution of 2-methylimidazole into Zn (NO)2·6H2In methanol solution of O; secondly, stirring the mixed solution at room temperature for 24 hours to obtain milky white liquid; then washing the milky white liquid with methanol for 3 times, and centrifuging and collecting the milky white liquid by a centrifuge; finally, vacuum drying at 60 ℃ overnight to obtain ZIF-8;
(2) synthesis of ZIF-8@ ZIF-67: taking ZIF-8 and dispersing in methanol solution with ultrasonic wave, stirring for 10min to obtain ZIF-8 solution, mixing methanol solution of 2-methylimidazole with Co (NO)2·6H2Pouring the methanol solution of O into the ZIF-8 solution to obtain a mixed solution; secondly, stirring the mixed solution at room temperature for 24 hours to obtain a purple solution; then washing the purple solution with methanol for 3 times, and centrifuging and collecting the solution by a centrifuge; finally, vacuum drying at 60 ℃ overnight to obtain ZIF-8@ ZIF-67;
(3) synthesis of C-N @ GC: and (3) putting the ZIF-8@ ZIF-67 synthesized in the step (2) into a muffle furnace, heating at a heating rate of 2 ℃ per minute in an Ar atmosphere, calcining at a constant temperature of 920 ℃ for 2 hours, and cooling the finished product to room temperature to obtain a black solid C-N @ GC nano material.
Further, in the step (1) of synthesizing ZIF-8, 2-methylimidazole reacts with Zn (NO)2·6H2The mass ratio of O is 6.16: 5.95.
further, in the step (2) of synthesizing ZIF-8@ ZIF-67, 2-methylimidazole, Co (NO)2·6H2The mass ratio of O to ZIF-8 is 6.16: 5.82: 0.5.
further, before the modifying agent solution is prepared in S1, the method further includes the steps of: the C-N @ GC was transferred to a mortar, ground well to a powder, and the ground C-N @ GC was placed in a water-ethanol solution.
3. Advantageous effects
Compared with the defects and shortcomings of the prior art, the invention has the following beneficial effects: the invention aims to develop a method for electrochemically detecting cysteine, which has the advantages of simple synthesis, high sensitivity and quick detection. According to the invention, ZIF-8@ ZIF-67 is taken as a precursor, and the product C-N @ GC can be obtained through carbonization, so that the product is stable, the preparation method is simple and convenient, the cost is lower, and the industrial production is easy to realize. The C-N @ GC prepared by the invention contains metal nano particles, so that the conductivity of the material is increased, and the detection capability of the C-N @ GC is improved due to the synergistic effect between C-N and GC in the C-N @ GC. The invention can realize qualitative and quantitative detection of cysteine, provides a new idea and a new method for discovering and constructing a novel electrochemical detection probe, and promotes the development and application of an electrochemical method in cysteine detection.
Drawings
FIG. 1 shows the detection result of cysteine by a glassy carbon electrode modified by C-N @ GC;
FIG. 2 shows the detection result of a glassy carbon electrode modified by C-N @ GC and containing no cysteine solution.
Detailed Description
The present invention will be described in further detail with reference to the drawings and examples.
Example 1
A synthetic method of a C-N @ GC nanometer material comprises the following steps:
(1) synthesis of ZIF-8: a methanol solution of 2-methylimidazole (6.16g,150mL) was poured into Zn (NO)2·6H2In methanol solution of O (5.95g, 150 mL); secondly, stirring the mixed solution at room temperature for 24 hours to obtain milky white liquid; then washing the milky white liquid with methanol for 3 times, and centrifugally collecting the milky white liquid by a centrifugal machine; finally, vacuum drying at 60 ℃ overnight to obtain ZIF-8; and 2-methylimidazole with Zn (NO)2·6H2The mass ratio of O is 6.16: 5.95;
(2) synthesis of ZIF-8@ ZIF-67: the ZIF-8(0.5g) synthesized in step (1) was dispersed in a methanol solution (100mL) by ultrasonic wave, and after stirring for 10min, a methanol solution of 2-methylimidazole (6.16g,100mL) and Co (NO)2·6H2A methanol solution of O (5.82g,100mL) was poured into the above solution; secondly, stirring the mixed solution at room temperature for 24 hours to obtain a purple solution; then washing the purple solution with methanol for 3 times, and centrifuging and collecting the solution by a centrifuge; finally, vacuum drying at 60 ℃ overnight to obtain ZIF-8@ ZIF-67; and 2-methylimidazole, Co (NO) 2·6H2The mass ratio of O to ZIF-8 is 6.16: 5.82: 0.5;
(3) synthesis of C-N @ GC: and (3) putting a certain amount of ZIF-8@ ZIF-67 synthesized in the step (2) into a muffle furnace, heating at a heating rate of 2 ℃ per minute in Ar atmosphere, calcining at a constant temperature of 920 ℃ for 2 hours, and cooling the finished product to room temperature to obtain a black solid C-N @ GC nano material.
Example 2
The application of the C-N @ GC nano material in cysteine detection comprises the following steps:
s1, preparation of modifier solution: taking 5 mg of the C-N @ GC synthesized in the example 1, transferring the C-N @ GC into a mortar, fully grinding the C-N @ GC into powder, putting the ground C-N @ GC into 1 ml of water-ethanol solution (volume ratio is 3:1), uniformly dispersing the ground C-N @ GC into 1 ml of water-ethanol solution (volume ratio is 3:1), dropwise adding 20 mu L of naphthol solution (5 wt%), and performing ultrasonic degradation for 2 hours by using an ultrasonic degradation method to obtain a uniform modifier solution;
s2, modification of the surface of the glassy carbon electrode: uniformly dripping 3 mu L of modifier solution in S1 on the surface of the polished glassy carbon electrode, and drying at room temperature to finally obtain the glassy carbon electrode modified by C-N @ GC;
s3, the glassy carbon electrode modified by the C-N @ GC obtained in the S2 is used for electrochemical detection of cysteine.
The C-N @ GC modified glassy carbon electrode has the specific application process in the electrochemical detection of cysteine as follows: CV tests (scan rate 50 mVs) were performed with a C-N @ GC modified glassy carbon electrode (GCE, 3 mM diameter), Pt sheet and Ag/AgCl electrode (saturated KCl) as the Working Electrode (WE), counter electrode (RE) and reference electrode (CE) for electrochemical detection, respectively, in a solution containing cysteine (5 mM PBS (pH 7.3))-1) (ii) a As shown in FIG. 1, a distinct oxidation peak can be observed on the CV curve in the presence of cysteine, indicating that the C-N @ GC modified glassy carbon electrode can detect cysteine;
comparative example
The glassy carbon electrode modified by C-N @ GC is applied to the electrochemical detection of other amino acids, and the specific application process is as follows:CV tests (scan rate of 50 mVs) were performed with a glassy carbon electrode (GCE, 3 mM diameter), Pt sheet and Ag/AgCl electrode (saturated KCl) modified with C-N @ GC as a Working Electrode (WE), counter electrode (RE) and reference electrode (CE) for electrochemical detection, respectively, in a (2mM PBS (pH 7.3)) solution containing glycine, alanine, valine, leucine, isoleucine, phenylalanine, tryptophan, tyrosine, aspartic acid, histidine, asparagine, glutamic acid, lysine, glutamine, arginine, serine, threonine, proline and methionine -1) (ii) a As shown in fig. 2, no oxidation absorption peak was observed in these CV curves, indicating that the 19 natural amino acids described above have no electrochemical response on the C-N @ GC modified glassy carbon electrode;
the invention can realize qualitative and quantitative detection of cysteine, provides a new idea and a new method for discovering and constructing a novel electrochemical detection probe, and promotes the development and application of an electrochemical method in cysteine detection.
It should be understood by those skilled in the art that the above embodiments are only for illustrating the present invention and are not to be used as a limitation of the present invention, and that changes and modifications to the above embodiments are within the scope of the claims of the present invention as long as they are within the spirit and scope of the present invention.
Claims (4)
1. A method for detecting cysteine, comprising the steps of:
s1, preparation of modifier solution: dispersing the C-N @ GC nano material in a water-ethanol solution, dropwise adding a naphthol solution with the weight percentage of 5 wt% into the solution, and performing ultrasonic degradation for 2 hours by using an ultrasonic degradation method to obtain a uniform modifier solution;
s2, modification of the surface of the glassy carbon electrode: uniformly dripping the modifier solution in the S1 on the surface of the polished glassy carbon electrode, and drying at room temperature to finally obtain the glassy carbon electrode modified by C-N @ GC;
S3, using the glassy carbon electrode modified by the C-N @ GC obtained in the S2 for electrochemical detection of cysteine;
the method for synthesizing the C-N @ GC nano material in the S1 comprises the following steps of:
(1) synthesis of ZIF-8: the methanol solution of 2-methylimidazole is poured into Zn (NO)2·6H2In methanol solution of O; secondly, stirring the mixed solution at room temperature for 24 hours to obtain milky white liquid; then washing the milky white liquid with methanol for 3 times, and centrifugally collecting the milky white liquid by a centrifugal machine; finally, vacuum drying at 60 ℃ overnight to obtain ZIF-8;
(2) synthesis of ZIF-8@ ZIF-67: taking ZIF-8 and dispersing in methanol solution with ultrasonic wave, stirring for 10min to obtain ZIF-8 solution, mixing methanol solution of 2-methylimidazole with Co (NO)2·6H2Pouring the methanol solution of O into the ZIF-8 solution to obtain a mixed solution; secondly, stirring the mixed solution at room temperature for 24 hours to obtain a purple solution; then washing the purple solution with methanol for 3 times, and centrifuging and collecting the solution by a centrifuge; finally, vacuum drying at 60 ℃ overnight to obtain ZIF-8@ ZIF-67;
(3) synthesis of C-N @ GC: and (3) putting the ZIF-8@ ZIF-67 synthesized in the step (2) into a muffle furnace, heating at a heating rate of 2 ℃ per minute in an Ar atmosphere, calcining at a constant temperature of 920 ℃ for 2 hours, and cooling the finished product to room temperature to obtain a black solid C-N @ GC nano material.
2. The method for detecting cysteine according to claim 1, wherein in the step (1) of synthesizing ZIF-8, 2-methylimidazole reacts with Zn (NO)2·6H2The mass ratio of O is 6.16: 5.95.
3. the method for detecting cysteine according to claim 1, wherein in the step (2) of synthesizing ZIF-8@ ZIF-67, 2-methylimidazole, Co (NO)2·6H2The mass ratio of O to ZIF-8 is 6.16: 5.82: 0.5.
4. the method for detecting cysteine according to claim 1, further comprising the step of, before the preparation of the modifier solution in S1: the C-N @ GC was transferred to a mortar, ground well to a powder, and the ground C-N @ GC was put into a water-ethanol solution.
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| CN109244427A (en) * | 2018-10-31 | 2019-01-18 | 北京科技大学 | Carbon coating zinc sulphide loads preparation method of the graphene as kalium ion battery cathode |
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