CN109959688A - The method for detecting dopamine and uric acid simultaneously based on poly- l-Glutathione modified glassy carbon electrode - Google Patents
The method for detecting dopamine and uric acid simultaneously based on poly- l-Glutathione modified glassy carbon electrode Download PDFInfo
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Abstract
The invention discloses one kind based on poly-LThe method that glutathione modified glassy carbon electrode detects dopamine and uric acid simultaneously.The method first immerses clean glass-carbon electrodeLIn the phosphate buffer of glutathione, lead to nitrogen deoxygenation, electrochemical polymerization is carried out using cyclic voltammetry, is gatheredLGlutathione modified electrode, then will gatherLGlutathione modified electrode is placed in the phosphate buffer solution of sample to be tested, using differential pulse voltammetry, detect electrochemical response electric current, the linear relationship of the linear relationship and response current of electric current and dopamine concentration and uric acid concentration according to response, is calculated the concentration of the dopamine in sample to be tested and uric acid.The method of the present invention has many advantages, such as that high sensitivity, easy to operate, stability and reproducible, the linear detection range to dopamine and uric acid are respectively 2~300 μM and 10~500 μM, has actual application prospect in the clinical detection of dopamine and uric acid.
Description
Technical field
The invention belongs to electrochemical analysis detection technique fields, and in particular to one kind is based on poly-LGlutathione modifies glass carbon
The method that electrode detects dopamine and uric acid simultaneously.
Background technique
Dopamine (DA) is a kind of important neurotransmitter, plays the work of regulation physiology and cognitive function in human body
With.Monitoring organism dopamine live signal variation to find its human body function and further appreciate that brain function have weight
Want meaning.Uric acid (UA) is one of the final product of purine metabolism in human body, it in human body fluid concentration variation can reflect
The situation of the functions such as people's vivo immunization and metabolism.DA and UA are architectures normally co-exist in body fluid (such as blood and urine), Yi Xieji
Sick such as gout, epilepsy, Parkinson's disease can be judged by their contents in human body fluid.Therefore, dopamine and urine
The quantitative detection of acid has vital meaning in clinical analysis and diagnosis.
DA and UA is the biomolecule with electro-chemical activity, in recent years, for electrochemical method to dopamine and urine
There are many method for measuring of acid.Electrochemical detection method has easy to operate a, response quickly, high sensitivity and at low cost etc. excellent
Point, it is more attractive compared to traditional detection method.The overpotential of DA and UA is all relatively high, and oxidizing potential relatively connects
Closely, when being measured with traditional electrode to it, it can generate interfere with each other therebetween, it is selective and less reproducible, it is difficult to real
It is measured while existing DA and UA.At present about the report of the DA and UA modified electrode detected simultaneously, but mostly exist
Defect.If Xu et al. takes off the glass-carbon electrode that alloyage is prepared for the modification of platinum titanium alloy by two steps, realize to the same of DA and UA
When detect (Zhao, D., et al, 2016.A highly sensitive and stable electrochemical
sensor for simultaneous detection towards ascorbic acid,dopamine,and uric
acid based on the hierarchical nanoporous PtTi alloy.Biosens Bioelectron 82,
119-126.), but this method preparation step is cumbersome and detection limit is higher.Ma et al. is prepared for 3D graphene hydrogel/gold nano
(Zhu, Q., et al, 2017.3D graphene is measured while particle composite material modified glassy carbon electrode is used for DA and UA
hydrogel–gold nanoparticles nanocomposite modified glassy carbon electrode
for the simultaneous determination of ascorbic acid,dopamine and uric
Acid.Sens.Actuators B:Chem, 238,1316-1323.), but this method is relatively narrow to the range of linearity of DA and UA, point
It Wei not be 0.2~30 μM and 1~60 μM.Therefore, measurement side while establishing simple, quick, the sensitive dopamine of one kind and uric acid
Method is very necessary.
Summary of the invention
The purpose of the present invention is to provide a kind of simple and efficient based on poly-LGlutathione modified glassy carbon electrode is examined simultaneously
The method for surveying dopamine and uric acid.
Realize the technical scheme is that
Based on poly-LThe method that glutathione modified glassy carbon electrode detects dopamine and uric acid simultaneously, the specific steps are as follows:
Step 1, glass-carbon electrode is polished, and ultrasonic cleaning is dried with nitrogen, and clean glass-carbon electrode is immersedLGluathione
In the phosphate buffer of peptide, lead to nitrogen deoxygenation, electrochemical polymerization is carried out using cyclic voltammetry, using glass-carbon electrode as work electricity
Pole, platinum electrode are to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, scanning speed
For 10~500mVs-1, scanning circle number is 5~15, and obtained modified electrode washing is dried with nitrogen, is gatheredLGlutathione is repaired
Adorn electrode；
Step 2, will gatherLGlutathione modified electrode is placed in the phosphate buffer solution of sample to be tested, and logical nitrogen removes
Oxygen detects electrochemical response electric current, according to response the linear relationship I of electric current and dopamine concentration using differential pulse voltammetry
(μ A)=0.541CDALinear relationship I (μ A)=0.104C of the concentration of (μM)+1.421 and response current and uric acidUA(μM)+
1.082, the concentration of the dopamine in sample to be tested and uric acid is calculated.
Preferably, in step 1, the pH of the phosphate buffer is 6~8.
Preferably, in step 1, the scanning circle number is 12 circles.
Preferably, in step 2, the differential pulse voltammetry, current potential increment is 4mV, amplitude 50mV, pulse width
For 0.05s, pulse spacing 0.5s.
The present invention is formed on glass-carbon electrode poly- by electrochemical methodLGluathione peptide film, the polymer film is to dopamine
There is apparent catalytic action with the electrochemical redox of urea, can be realized detection while to dopamine and uric acid.
Compared with prior art, the invention has the advantages that (1) realizes in aqueous solutionLThe electrochemistry of glutathione is poly-
It closes, has the characteristics that green non-pollution；(2) it obtains compound uniform poly-LGlutathione film, and drug dosage is few, preparation
Method is simple and quick, reproducible；(3) modified electrode prepared can quickly detect dopamine and uric acid, the line of dopamine simultaneously
Property detection range be 2~300 μM, the linear detection range of uric acid is 10~500 μM.
Detailed description of the invention
Fig. 1 is 10mM'sLElectrochemistry of the 0.1M phosphate buffer solution (pH7) of glutathione on glass-carbon electrode is poly-
The CV of conjunction schemes, and sweeps fast 100mVs-1。
Fig. 2 is 100 μM of DA and 300 μM of UA in bare glassy carbon electrode (curve a) and poly-LGlutathione modified glassy carbon electrode
(the differential pulse voltammetry figure on curve b).
Fig. 3 is the differential pulse voltammetry figure that DA is individually measured when DA and UA is existed simultaneously.
Fig. 4 is the linear relationship chart of dopamine differential pulse voltammetry peak current and concentration in the presence of 100 μM of uric acid.
Fig. 5 is the differential pulse voltammetry figure that UA is individually measured when DA and UA is existed simultaneously.
Fig. 6 is the linear relationship chart of uric acid differential pulse voltammetry peak current and concentration in the presence of 50 μM of dopamines.
Fig. 7 is poly-LGlutathione containing 100 μM of dopamines pH7.4 phosphate buffer difference sweep speed (20~
500mVs-1) cyclic voltammogram.
Fig. 8 is 100 μM of dopamines peak current and linear relationship chart for sweeping speed in pH7.4 phosphate buffer.
Fig. 9 is poly-LGlutathione containing 300 μM of uric acid pH7.4 phosphate buffer difference sweep speed (20~
500mVs-1) cyclic voltammogram.
Figure 10 is 300 μM of uric acid peak current and linear relationship chart for sweeping speed in pH7.4 phosphate buffer.
Specific embodiment
Below with reference to embodiment and attached drawing, the invention will be further described.
Embodiment 1
(1) respectively with 1.0,0.3,0.05 μm of α-A12O3Glass-carbon electrode is polished, uses deionization after polishing every time
Water ultrasonic cleaning, is finally dried up with high pure nitrogen；
(2) glass-carbon electrode immersion is contained into 10mMLIn the electrolytic cell of glutathione phosphate buffer, logical nitrogen is removed
Oxygen；
(3) electrochemical polymerization is carried out using cyclic voltammetry, using three-electrode system, glass-carbon electrode is working electrode, platinum
Plate electrode is to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, and scanning speed is
100mVs-1, 12 circle of scanning, obtained modified electrode is rinsed with deionized water, and with being dried with nitrogen, is gatheredLGlutathione is repaired
Adorn electrode.
(4) will gatherLGlutathione modified glassy carbon electrode and bare glassy carbon electrode be respectively placed in containing 100 μM of dopamines and
In the phosphate buffer of the pH value 7.4 of 300 μM of uric acid, leads to nitrogen 15 minutes, use differential pulse voltammetry, detecting electrode pair
The electrochemical response of dopamine and uric acid.As shown in Fig. 2, comparing bare glassy carbon electrode, which examines dopamine and uric acid
The sensitivity of survey significantly improves.
Embodiment 2
Gather in the presence of uric acidLDetection of the glutathione modified glassy carbon electrode to dopamine.
(1) respectively with 1.0,0.3,0.05 μm of α-A12O3Glass-carbon electrode is polished, uses deionization after polishing every time
Water ultrasonic cleaning, is finally dried up with high pure nitrogen；
(2) glass-carbon electrode immersion is contained into 10mMLIn the electrolytic cell of glutathione phosphate buffer, logical nitrogen is removed
Oxygen；
(3) electrochemical polymerization is carried out using cyclic voltammetry, using three-electrode system, glass-carbon electrode is working electrode, platinum
Plate electrode is to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, and scanning speed is
100mVs-1, 12 circle of scanning, obtained modified electrode is rinsed with deionized water, and with being dried with nitrogen, is gatheredLGlutathione is repaired
Adorn electrode.
(4) will gatherLGlutathione modified glassy carbon electrode be placed in containing various concentration (2 μM, 5 μM, 10 μM, 20 μM, 40 μ
M, 60 μM, 80 μM, 100 μM, 120 μM, 150 μM, 200 μM, 250 μM, 300 μM) pH value 7.4 of dopamine and 100 μM of uric acid
In phosphate buffer, leads to nitrogen 15 minutes, use differential pulse voltammetry, electrochemistry of the detecting electrode to dopamine and uric acid
Response.Fig. 3 is the differential pulse voltammetry figure that DA is individually measured when DA and UA is existed simultaneously.UA concentration is 100 μM, and the concentration of DA is certainly
It is lower and on be followed successively by 2 μM, 5 μM, 10 μM, 20 μM, 40 μM, 60 μM, 80 μM, 100 μM, 120 μM, 150 μM, 200 μM, 250 μM,
300μM.Fig. 4 is the linear relationship chart of dopamine differential pulse voltammetry peak current and concentration in the presence of 100 μM of uric acid.Such as Fig. 3
With shown in Fig. 4, in the presence of 100 μM of uric acid, dopamine peak current and concentration are linear, linear relation be I (μ A)=
0.541CDA(μM)+1.421。
Embodiment 3
Gather in the presence of dopamineLDetection of the glutathione modified glassy carbon electrode to uric acid.
(1) respectively with 1.0,0.3,0.05 μm of α-A12O3Glass-carbon electrode is polished, uses deionization after polishing every time
Water ultrasonic cleaning, is finally dried up with high pure nitrogen；
(2) glass-carbon electrode immersion is contained into 10mMLIn the electrolytic cell of glutathione phosphate buffer, logical nitrogen is removed
Oxygen；
(3) electrochemical polymerization is carried out using cyclic voltammetry, using three-electrode system, glass-carbon electrode is working electrode, platinum
Plate electrode is to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, and scanning speed is
100mVs-1, 12 circle of scanning, obtained modified electrode is rinsed with deionized water, and with being dried with nitrogen, is gatheredLGlutathione is repaired
Adorn electrode.
(4) will gatherLGlutathione modified glassy carbon electrode is placed in containing 50 μM of dopamines of a certain concentration and various concentration
The pH value 7.4 of (10 μM, 20 μM, 40 μM, 60 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 400 μM, 500 μM) uric acid
Phosphate buffer in, lead to nitrogen 15 minutes, use differential pulse voltammetry, electrification of the detecting electrode to dopamine and uric acid
Learn response.Fig. 5 is the differential pulse voltammetry figure that UA is individually measured when DA and UA is existed simultaneously.DA concentration is 50 μM, the concentration of UA
It is followed successively by 10 μM, 20 μM, 40 μM, 60 μM, 100 μM, 150 μM, 200 μM, 250 μM, 300 μM, 400 μM, 500 μM from bottom to top.
Fig. 6 is the linear relationship chart of uric acid differential pulse voltammetry peak current and concentration in the presence of 50 μM of dopamines.Such as Fig. 5 and Fig. 6
Shown, in the presence of 50 μM of dopamines, uric acid peak current and concentration are linear, and linear relation is I (μ A)=0.104CUA
(μM)+1.082。
Embodiment 4
The peak current of dopamine and the measurement for sweeping fast relationship.
(1) respectively with 1.0,0.3,0.05 μm of α-A12O3Glass-carbon electrode is polished, uses deionization after polishing every time
Water ultrasonic cleaning, is finally dried up with high pure nitrogen；
(2) glass-carbon electrode immersion is contained into 10mMLIn the electrolytic cell of glutathione phosphate buffer, logical nitrogen is removed
Oxygen；
(3) electrochemical polymerization is carried out using cyclic voltammetry, using three-electrode system, glass-carbon electrode is working electrode, platinum
Plate electrode is to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, and scanning speed is
100mVs-1, 12 circle of scanning, obtained modified electrode is rinsed with deionized water, and with being dried with nitrogen, is gatheredLGlutathione is repaired
Adorn electrode.
(4) will gatherLGlutathione modified glassy carbon electrode is placed in the phosphate-buffered containing 100 μM of dopamine pH value 7.4
In liquid, lead to nitrogen 15 minutes, using cyclic voltammetry, sweeping speed is 20,50,80,100,120,150,200,250,300,350,
400,450,500mVs-1, electrochemical response of the detecting electrode to dopamine.As shown in Figure 7 and Figure 8, dopamine peak current with sweep
Rapid-result linear relationship.
Embodiment 5
The peak current of uric acid and the measurement for sweeping fast relationship.
(1) respectively with 1.0,0.3,0.05 μm of α-A12O3Glass-carbon electrode is polished, uses deionization after polishing every time
Water ultrasonic cleaning, is finally dried up with high pure nitrogen；
(2) glass-carbon electrode immersion is contained into 10mMLIn the electrolytic cell of glutathione phosphate buffer, logical nitrogen is removed
Oxygen；
(3) electrochemical polymerization is carried out using cyclic voltammetry, using three-electrode system, glass-carbon electrode is working electrode, platinum
Plate electrode is to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, and scanning speed is
100mVs-1, 12 circle of scanning, obtained modified electrode is rinsed with deionized water, and with being dried with nitrogen, is gatheredLGlutathione is repaired
Adorn electrode.
(4) will gatherLGlutathione modified glassy carbon electrode is placed in the phosphate buffer containing 300 μM of uric acid pH value 7.4
In, lead to nitrogen 15 minutes, using cyclic voltammetry, sweeping speed is 20,50,80,100,120,150,200,250,300,350,
400,450,500mVs-1, electrochemical response of the detecting electrode to uric acid.As shown in Figure 9 and Figure 10, uric acid peak current and short-term training is swept
Linear relationship.
Claims (4)
1. based on poly-LThe method that glutathione modified glassy carbon electrode detects dopamine and uric acid simultaneously, which is characterized in that specific step
It is rapid as follows:
Step 1, glass-carbon electrode is polished, and ultrasonic cleaning is dried with nitrogen, and clean glass-carbon electrode is immersedLGlutathione
In phosphate buffer, lead to nitrogen deoxygenation, electrochemical polymerization is carried out using cyclic voltammetry, using glass-carbon electrode as working electrode,
Platinum electrode is to electrode, and saturated calomel electrode is reference electrode, and low potential is -1.5V, high potential 2.0V, scanning speed 10
~500mVs-1, scanning circle number is 5~15, and obtained modified electrode washing is dried with nitrogen, is gatheredLGlutathione modification electricity
Pole；
Step 2, will gatherLGlutathione modified electrode is placed in the phosphate buffer solution of sample to be tested, is led to nitrogen deoxygenation, is adopted
With differential pulse voltammetry, detect electrochemical response electric current, according to response the linear relationship I (μ A) of electric current and dopamine concentration=
0.541CDALinear relationship I (μ A)=0.104C of the concentration of (μM)+1.421 and response current and uric acidUA(μM)+1.082,
The concentration of the dopamine in sample to be tested and uric acid is calculated.
2. the method according to claim 1, wherein in step 1, the pH of the phosphate buffer is 6~
8。
3. the method according to claim 1, wherein the scanning circle number is 12 circles in step 1.
4. the method according to claim 1, wherein the differential pulse voltammetry, current potential increases in step 2
Amount is 4mV, amplitude 50mV, pulse width 0.05s, pulse spacing 0.5s.
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