CN101493453A - Thin layer detecting pool for detecting dopamine and method for making same - Google Patents
Thin layer detecting pool for detecting dopamine and method for making same Download PDFInfo
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- CN101493453A CN101493453A CN 200810205060 CN200810205060A CN101493453A CN 101493453 A CN101493453 A CN 101493453A CN 200810205060 CN200810205060 CN 200810205060 CN 200810205060 A CN200810205060 A CN 200810205060A CN 101493453 A CN101493453 A CN 101493453A
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Abstract
The invention discloses a thin-layer detection pool which is used for dopamine detection in the electrochemical detection technology field and a preparation method thereof. The detection pool comprises a working electrode, a reference electrode and an auxiliary electrode, wherein the working electrode, the reference electrode and the auxiliary electrode are respectively connected with a liquid chromatogram and an electrochemical system; the working electrode comprises a platinum particle layer, a polyvinyl alcohol layer and an electrode matrix, wherein the polyvinyl alcohol layer and the platinum particle layer are coated on the external part of the electrode matrix from the inside to the outside in sequence. The thin-layer detection pool which is prepared by the invention can realize the detection for the concentration of dopamine with simple steps, with the linear detection range up to 1.0 multiplied by 10<-9>-1.0 multiplied by 10<-5> mol/L and the detection resolution of 1.0 multiplied by 10<-10> mol/L.
Description
Technical field
What the present invention relates to is the device in a kind of electrochemical measuring technique field and preparation method thereof, specifically is a kind of thin layer detection cell that is used for the dopamine detection and preparation method thereof.
Background technology
Studies show that dopamine is as a kind of very important neurotransmitter in the biosome, its concentration and Parkinsonian substantial connection arranged.This shows that the detection levels of dopamine is significant to the research of Parkinson's pathology.At present, there have been some researchs to report the method for measuring dopamine, for example: fluorescent spectrometry, ultraviolet spectrophotometry, voltammetry, polarography and GC-MS(gas chromatography-mass spectrography) etc.Though said method successfully has been applied to the detection of monoamine neurotransmitter in biological and the drug sample, because monoamine neurotransmitter lacks luminophore, this has limited the application of fluorescence method and other spectral method of detection.Characteristics such as liquid chromatography-electrochemical measuring technique is because to have a required sample few, highly sensitive, and the high and sampling volume of separation efficiency is little and be widely used in the detection of biological sample.But because the content of dopamine is very low in the biosome, the electrochemical sensor of therefore developing high sensitivity (being used for the detection of trace dopamine in the body) mensuration dopamine has caused people's research interest.
Retrieval through prior art is found, people such as Jianshe Huang propose a kind of method that detects dopamine in Biosensors andBioelectronics magazine (2008 the 24th volume 4 phase 632-637 pages or leaves) " utilizing palladium and charcoal nanofiber modified electrode to detect dopamine simultaneously; uric acid and ascorbic acid ", but this technology is to dopamine, and the detection of uric acid and ascorbic acid is limited to 0.2 * 10
-6Mol/L.
Find by retrieval in addition, people such as Shurong Hou in Analytical Biochemistry magazine (2008 381 the 2nd phase 179-184 pages or leaves) " poly-3; the film modified electrode of 5-dihydroxy-benzoic acid detects dopamine in the presence of ascorbic acid " to gather 3, the film modified electrode detection dopamine of 5-dihydroxy-benzoic acid, its detection is limited to 6.0 * 10
-8Mol/L.Because dopamine content in biological samples such as cerebrospinal fluid is extremely low, therefore above-mentioned detection method is difficult to the accurate mensuration of realization to DOPAMINE CONTENT IN RABBIT in the biological cerebrospinal fluid.
Summary of the invention
The present invention is directed to the prior art above shortcomings, a kind of thin layer detection cell that is used for the dopamine detection and preparation method thereof is provided, by the detection of easy steps realization dopamine concentration in the biological sample.
The present invention is achieved by the following technical solutions, the present invention relates to be used for the thin layer detection cell that dopamine detects comprises: working electrode, contrast electrode and auxiliary electrode, wherein: working electrode, contrast electrode, auxiliary electrode are connected with liquid chromatography and electro-chemical systems respectively.
Described working electrode comprises: platinum grain sublayer, polyvinyl alcohol layer and electrode matrix, wherein: polyvinyl alcohol layer and platinum grain sublayer are coated on the electrode matrix outside successively from inside to outside, the thickness of platinum grain sublayer is 50～120nm, and the thickness of polyvinyl alcohol layer is 50～130nm.
Described electrode matrix is the electrode matrix of glass carbon material;
Described contrast electrode is the Ag-AgCl electrode;
Described auxiliary electrode is the electrode that stainless steel is made;
Described liquid chromatography adopts Luna 5 μ m C
18Reverse-phase chromatographic column;
Described electro-chemical systems adopts CHI832 type galvanochemistry instrument.
The present invention relates to be used for the preparation method of the thin layer detection cell that dopamine detects, may further comprise the steps:
The first step: the preparation of polyvinyl alcohol (PVA) modified electrode:
At first glass-carbon electrode is carried out ultrasonic cleaning in redistilled water after the polishing;
Glass-carbon electrode after cleaning is then put into phosphate buffered solution ,-0.2V～+ carry out scan round with the sweep speed of 20mV/s in the potential range of 2.0V to handle 10～20min;
Again the glass-carbon electrode after the scan round is put into solute and is mass percent and be the phosphate buffered solution of the lithium perchlorate of 0.1%～1.0% polyvinyl alcohol (PVA) and 0.01mol/L, 0.0V～+ carry out scan round once more with the sweep speed of 20mV/s in the potential range of 1.0V to handle 2～8 cycles;
To take out the back through the glass-carbon electrode after the secondary cycle scan process and clean, make the polyvinyl alcohol (PVA) modified electrode with redistilled water.
It is that the alumina powder of 0.3 μ m is polished to minute surface on chamois leather that described polishing is meant with abrasive paper for metallograph and fineness.
Described redistilled water is meant: the pure water that obtains after twice distillation;
Described phosphate buffered solution is meant: phosphate concentration is potassium dihydrogen phosphate and the disodium phosphate soln of 0.2mol/L, utilizes phosphoric acid to regulate different pH values.
Described scan round processing is meant working electrode, contrast electrode and auxiliary electrode is linked to each other with electro-chemical systems respectively, carries out cycle potentials scanning with certain sweep speed in the voltage range of setting.
Second step: platinum particles loads:
At first the polyvinyl alcohol (PVA) modified electrode is placed and contain 1.0 * 10
-3Mol/L～5.0 * 10
-3The K of mol/L
2PtCl
6Concentration be in the Klorvess Liquid of 0.1mol/L, with the sweep speed of 0.1V/s-0.4V～+ carry out scan round in the potential range of 0.3V to handle 10～20 cycles;
Take out the polyvinyl alcohol (PVA) modified electrode through overscanning then, wash several times successively in turn with absolute ethyl alcohol and redistilled water, the polyvinyl alcohol (PVA) modified electrode after promptly obtaining platinum particles and loading is as working electrode.
The 3rd step: successively contrast electrode, working electrode and auxiliary electrode are linked to each other with liquid chromatography and electro-chemical systems, wherein: contrast electrode is the Ag-AgCl electrode; Auxiliary electrode is the electrode that stainless steel is made; Liquid chromatography adopts Luna 5 μ m C
18The moving phase that reverse-phase chromatographic column, liquid chromatography use is the phosphate buffered solution of pH5.0～7.0 of containing 10% methyl alcohol and 0.2mol/L, and the flow velocity of this moving phase is 1.0mL/min, makes to be used for the thin layer detection cell that dopamine detects.
The present invention detects by following steps:
At first open liquid chromatography and electro-chemical systems, make the dopamine sample of different concentration known with phosphate buffered solution through Luna 5 μ m C
18Reverse-phase chromatographic column flows through the surface of contrast electrode, working electrode and auxiliary electrode successively;
Utilize CHI832 type electro-chemical systems to note the current signal that different samples produce then, carry out match according to the current value of the response current of known dopamine concentration under the different samples and corresponding actual measurement as coordinate in length and breadth, the thin layer detection cell is carried out sample learning;
Again brain microdialysis liquid sample 20 μ L are placed and prepare test environment, record the current value of response current, read contained dopamine concentration in the brain microdialysis liquid sample of this current value correspondence according to the linear relationship chart of the dopamine concentration-corresponding electric current that obtains through sample learning.
The method of the invention can realize the detection of dopamine concentration with easy steps, detects the range of linearity and can be reached for 1.0 * 10
-9～1.0 * 10
-5Mol/L, detection resolution is 1.0 * 10
-10Mol/L.
Description of drawings
Fig. 1 is the linear relationship chart of response current and dopamine concentration among the embodiment 1;
Fig. 2 is the linear relationship chart of response current and dopamine concentration among the embodiment 2;
Fig. 3 is the linear relationship chart of response current and dopamine concentration among the embodiment 3.
Embodiment
Below in conjunction with accompanying drawing embodiments of the invention are elaborated: present embodiment is being to implement under the prerequisite with the technical solution of the present invention, provided detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
The glass-carbon electrode that pre-service is good is put into the phosphate buffer solution of pH=7.0, with the sweep speed of 20mV/s-0.2V～+ carry out scan round in the potential range of 2.0V to handle 10min.
Then electrode is put into the phosphate buffered solution of the pH=7.0 of the lithium perchlorate that contains 0.7% polyvinyl alcohol (PVA) and 0.01mol/L, with the sweep speed of 20mV/s 0.0V～+ carry out scan round in the potential range of 1.0V to handle 6 circles;
The polyvinyl alcohol (PVA) modified electrode placed contain 1.0 * 10
-3The K of mol/L
2PtCl
6In the KCl solution of 0.1mol/L, with the sweep speed of 0.1V/s-0.4V～+ carry out scan round in the potential range of 0.3V to handle 12 circles, obtain the polyvinyl alcohol (PVA) modified electrode after platinum particles loads, wherein: polyvinyl alcohol layer and platinum grain sublayer are coated on the glass-carbon electrode outside successively from inside to outside, the thickness of platinum grain sublayer is 50～70nm, and the thickness of polyvinyl alcohol layer is 70～90nm;
With the polyvinyl alcohol (PVA) modified electrode after the platinum particles loading is working electrode, the Ag-AgCl electrode is as contrast electrode, stainless steel is an auxiliary electrode, successively contrast electrode, working electrode and auxiliary electrode is linked to each other with liquid chromatography and electro-chemical systems, by Luna 5 μ m C
18Reverse-phase chromatographic column separates, and moving phase wherein is to contain in the phosphate buffered solution of pH6.0 of 10% methyl alcohol and 0.2mol/L, and flow velocity is 1.0mL/min;
Then under loading+0.6V current potential, according to the known dopamine concentration under the different samples and utilize the current value of the response current of the corresponding actual measurement of CHI832 type electro-chemical systems to carry out match, the thin layer detection cell is carried out sample learning as coordinate in length and breadth;
As shown in Figure 1, utilizing the linear relationship chart of dopamine concentration-corresponding electric current, is 3.8 * 10 with the dopamine concentration of the response current value correspondence that obtains in the rat brain spinal fluid
-8Mol/L.
Embodiment 2
The glass-carbon electrode that pre-service is good is put into the phosphate buffered solution of pH=7.0, with the sweep speed of 20mV/s-0.2V～+ carry out scan round in the potential range of 2.0V to handle 15min;
Then electrode is put into the phosphate buffered solution of the pH=7.0 of the lithium perchlorate that contains 0.5% polyvinyl alcohol (PVA) and 0.01mol/L, with the sweep speed of 20mV/s 0.0V～+ carry out scan round in the potential range of 1.0V to handle 4 circles;
The polyvinyl alcohol (PVA) modified electrode placed contain 2.5 * 10
-3The K of mol/L
2PtCl
6In the KCl solution of 0.1mol/L, with the sweep speed of 0.1V/s-0.4V～+ carry out scan round in the potential range of 0.3V to handle 16 circles, obtain the polyvinyl alcohol (PVA) modified electrode after platinum particles loads, wherein: polyvinyl alcohol layer and platinum grain sublayer are coated on the glass-carbon electrode outside successively from inside to outside, the thickness of platinum grain sublayer is 80～100nm, and the thickness of polyvinyl alcohol layer is 50～70nm;
With the polyvinyl alcohol (PVA) modified electrode after the platinum particles loading is working electrode, the Ag-AgCl electrode is as contrast electrode, stainless steel is an auxiliary electrode, successively contrast electrode, working electrode and auxiliary electrode is linked to each other with liquid chromatography and electro-chemical systems, by Luna 5 μ m C
18Reverse-phase chromatographic column separates, and moving phase wherein is to contain in the phosphate buffered solution of pH5.0 of 10% methyl alcohol and 0.2mol/L, and flow velocity is 1.0mL/min;
Then under loading+0.6V current potential, carry out match according to the current value of known dopamine concentration under the different samples and the corresponding response current that utilizes the actual measurement of CHI832 type electro-chemical systems as coordinate in length and breadth, the thin layer detection cell is carried out sample learning;
As shown in Figure 2, utilize the linear relationship chart of dopamine concentration-corresponding electric current, the dopamine concentration that records the response current value correspondence that obtains in the rat brain spinal fluid is 4.1 * 10
-8Mol/L.
The glass-carbon electrode that pre-service is good is put into the phosphate buffered solution of pH=7.0, with the sweep speed of 20mV/s-0.2V～+ carry out scan round in the potential range of 2.0V to handle 20min;
Then electrode is put into the phosphate buffered solution of the pH=7.0 of the lithium perchlorate that contains 1.0% polyvinyl alcohol (PVA) and 0.01mol/L, with the sweep speed of 20mV/s 0.0V～+ carry out scan round in the potential range of 1.0V to handle 8 circles;
The polyvinyl alcohol (PVA) modified electrode placed contain 4.5 * 10
-3The K of mol/L
2PtCl
6In the KCl solution of 0.1mol/L, with the sweep speed of 0.1V/s-0.4V～+ carry out scan round in the potential range of 0.3V to handle 18 circles, obtain the polyvinyl alcohol (PVA) modified electrode after platinum particles loads, wherein: polyvinyl alcohol layer and platinum grain sublayer are coated on the glass-carbon electrode outside successively from inside to outside, the thickness of platinum grain sublayer is 100～120nm, and the thickness of polyvinyl alcohol layer is 110～130nm;
With the polyvinyl alcohol (PVA) modified electrode after the platinum particles loading is working electrode, the Ag-AgCl electrode is as contrast electrode, stainless steel is an auxiliary electrode, successively contrast electrode, working electrode and auxiliary electrode is linked to each other with liquid chromatography and electro-chemical systems, by Luna 5 μ m C
18Reverse-phase chromatographic column separates, and moving phase wherein is to contain in the phosphate buffered solution of pH7.0 of 10% methyl alcohol and 0.2mol/L, and flow velocity is 1.0mL/min;
Then under loading+0.6V current potential, carry out match according to the current value of known dopamine concentration under the different samples and the corresponding response current that utilizes the actual measurement of CHI832 type electro-chemical systems as coordinate in length and breadth, the thin layer detection cell is carried out sample learning;
As shown in Figure 3, utilize the linear relationship chart of dopamine concentration-corresponding electric current, the crust amine concentration that records the response current value correspondence that obtains in the rat brain spinal fluid is 4.3 * 10
-8Mol/L.
Claims (10)
1, a kind of thin layer detection cell that is used for the dopamine detection, comprise: working electrode, contrast electrode and auxiliary electrode, wherein: working electrode, contrast electrode, auxiliary electrode are connected with liquid chromatography and electro-chemical systems respectively, it is characterized in that, described working electrode comprises: platinum grain sublayer, polyvinyl alcohol layer and electrode matrix, wherein: polyvinyl alcohol layer and platinum grain sublayer are coated on the electrode matrix outside successively from inside to outside.
2, the thin layer detection cell that is used for the dopamine detection according to claim 1 is characterized in that the thickness of the platinum grain sublayer in the described working electrode is 50～100nm, and the thickness of polyvinyl alcohol layer is 50～110nm.
3, the thin layer detection cell that is used for the dopamine detection according to claim 2 is characterized in that described electrode matrix is the electrode matrix of glass carbon material.
4, the thin layer detection cell that is used for the dopamine detection according to claim 1 is characterized in that, described liquid chromatography adopts Luna 5 μ m C
18Reverse-phase chromatographic column.
5, the preparation method who is used for the thin layer detection cell of dopamine detection according to claim 1 is characterized in that, may further comprise the steps:
The first step: the preparation of polyvinyl alcohol (PVA) modified electrode: at first glass-carbon electrode is carried out ultrasonic cleaning in redistilled water after the polishing; Glass-carbon electrode after cleaning is then put into phosphate solution and is carried out the scan round processing; Again the glass-carbon electrode after the scan round is put into the phosphate solution that solute is polyvinyl alcohol (PVA) and lithium perchlorate, carried out scan round once more and handle; To take out the back through the glass-carbon electrode after the secondary cycle scan process and clean, make the polyvinyl alcohol (PVA) modified electrode with redistilled water;
Second step: platinum particles loads: at first the polyvinyl alcohol (PVA) modified electrode is placed to contain K
2PtCl
6Klorvess Liquid in, carry out scan round and handle the back and take out polyvinyl alcohol (PVA) modified electrode through overscanning, wash several times successively in turn with absolute ethyl alcohol and redistilled water, obtain polyvinyl alcohol (PVA) modified electrode after the platinum particles loading as working electrode;
The 3rd step: successively contrast electrode, working electrode and auxiliary electrode are linked to each other with liquid chromatography and electro-chemical systems, wherein: contrast electrode is the Ag-AgCl electrode; Auxiliary electrode is the electrode that stainless steel is made, and makes to be used for the thin layer detection cell that dopamine detects.
6, the preparation method who is used for the thin layer detection cell of dopamine detection according to claim 5, it is characterized in that, handle to be meant working electrode, contrast electrode and auxiliary electrode are linked to each other with electro-chemical systems respectively in the scan round described in the first step, in the voltage range of setting, carry out cycle potentials with certain sweep speed and scan.
7, the preparation method who is used for the thin layer detection cell of dopamine detection according to claim 5, it is characterized in that, the mass percent concentration of polyvinyl alcohol (PVA) is 0.1%～1.0% in the phosphate solution described in the first step, and the concentration of lithium perchlorate is 0.01mol/L.
8, according to claim 5 or the 7 described preparation methods that are used for the thin layer detection cell of dopamine detection, it is characterized in that be meant in the phosphate buffered solution described in the first step: phosphate concentration is potassium dihydrogen phosphate and the disodium phosphate soln of 0.2mol/L.
9, the preparation method who is used for the thin layer detection cell that dopamine detects according to claim 5 is characterized in that, contains K described in second step
2PtCl
6Klorvess Liquid, K wherein
2PtCl
6Concentration be 1.0 * 10
-3Mol/L～5.0 * 10
-3Mol/L.
10, the preparation method who is used for the thin layer detection cell of dopamine detection according to claim 5, it is characterized in that, the moving phase that liquid chromatography described in the 3rd step is used is the phosphate buffered solution of pH5.0～7.0 of containing 10% methyl alcohol and 0.2mol/L, and the flow velocity of this moving phase is 1.0mL/min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105973876A (en) * | 2016-05-03 | 2016-09-28 | 国家纳米科学中心 | Electrochemiluminescence sensor for detecting dopamine, and production method and application thereof |
| CN114279968A (en) * | 2021-12-28 | 2022-04-05 | 中国科学院长春应用化学研究所 | Electrochemistry-double polarization interference photoelectric detection cell |
-
2008
- 2008-12-30 CN CN 200810205060 patent/CN101493453A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105973876A (en) * | 2016-05-03 | 2016-09-28 | 国家纳米科学中心 | Electrochemiluminescence sensor for detecting dopamine, and production method and application thereof |
| CN114279968A (en) * | 2021-12-28 | 2022-04-05 | 中国科学院长春应用化学研究所 | Electrochemistry-double polarization interference photoelectric detection cell |
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Application publication date: 20090729 |