CN103675054B - A kind of electrolytic cell for detecting dopamine and method thereof - Google Patents
A kind of electrolytic cell for detecting dopamine and method thereof Download PDFInfo
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- CN103675054B CN103675054B CN201210361734.3A CN201210361734A CN103675054B CN 103675054 B CN103675054 B CN 103675054B CN 201210361734 A CN201210361734 A CN 201210361734A CN 103675054 B CN103675054 B CN 103675054B
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Abstract
The invention discloses a kind of electrolytic cell for detecting dopamine and method thereof, electrolytic cell comprises upper part and lower part, upper part comprises rounding mesa-shaped pond body and Chi Gai with holes, lower part comprises at the bottom of cylindrical pond, in the middle of bottom at the bottom of pond, shrinkage pool is built with ferromagnetism magnet, upper part and lower part are by screens and screw fixing and locking, and the junction of upper part and lower part is provided with rubber blanket, and in electrolytic cell, pond body is teflon.Under buffer system, dopamine solution to be measured and magnetic microsphere generation coupling reaction is caused with activator, reaction product is enriched in printing carbon electrodes under magnetic field added by electrolytic cell, peak current is detected by Differential Pulse Voltammetry, the linear relationship set up by peak current and standard specimen concentration, draws testing sample concentration by set up linear relationship.The present invention utilizes magnetic microsphere can produce magnetic under the effect of electrolytic cell externally-applied magnetic field, does displacement and is enriched in the surface of working electrode to improve the sensitivity of its detection.
Description
Technical field
The present invention relates to a kind of testing equipment and detection method thereof of electrochemical field, more specifically relating to a kind of electrolytic cell for detecting dopamine and method thereof.
Background technology
Dopamine is the Catecholamines Neurotransmitters in Blood be present in mammalian central nervous system, and the change of its amount can cause some important diseases as schizophrenia and Parkinson's disease.When electrochemically measuring dopamine at present, because the overpotential of dopamine on solid electrode is high, electrode reaction is slow, and its electrochemical reaction product is easily adsorbed in electrode surface, result in electrode surface passivation, and detection sensitivity reduces.
Summary of the invention
According to an aspect of the present invention, a kind of electrolytic cell for detecting dopamine is provided, comprise the working electrode that Chi Tiji is positioned over Chi Tizhong, to electrode and contrast electrode, pond body comprises Chi Gai, at the bottom of solution pool and pond, cover strip hole, pond, solution pool is rounding mesa-shaped, be cylindrical at the bottom of pond, shrinkage pool is provided with in the middle of bottom at the bottom of pond, shrinkage pool is built with magnet, magnet lower end is provided with spring, working electrode crosses at junction at the bottom of solution pool and pond, the working portion of working electrode is towards solution pool, electrode and contrast electrode are inserted by the hole of Chi Gai, the working portion of electrode and contrast electrode is entered in solution pool.
In some embodiments, be fixed by screw at the bottom of solution pool and pond, at the bottom of solution pool and pond, junction is provided with rubber blanket.
In some embodiments, adopt teflon material at the bottom of solution pool, Chi Gai and pond, the bottom steel plate of electrolytic cell is coated.
In some embodiments, working electrode selects printing carbon electrode, and the carbon paste district of printing carbon electrode is circular, and select platinum electrode to electrode, contrast electrode selects mercury electrode.
A kind of detection method for detecting dopamine, comprise: under buffer system, by the magnetic microsphere after activation and dopamine solution generation coupling reaction, coupling reaction product is enriched in working electrode surface under magnetic field added by electrolytic cell, peak current is detected by Differential Pulse Voltammetry, set up linear relationship by peak current and standard specimen concentration, draw testing sample concentration by set up linear relationship.
In some embodiments, working electrode selects printing carbon electrode.
In some embodiments, buffer system selects Acetic acid-sodium acetate damping fluid.
In some embodiments, the activator used in reactivation process is carbodiimide.
The present invention is by after dopamine and magnetic microsphere coupling, detect with the enrichment of printing carbon electrode under electrolytic cell externally-applied magnetic field condition, the carbon paste district of printing carbon electrode is circular, make the rich region of coupled product larger, and magnetic microsphere has larger specific surface, thus can carry high density functional group and can be highly dispersed in the characteristic in solution, under additional magnetic fields, soft magnetism polymer microsphere can produce magnetic, does displacement and is enriched in the surface of electrode to improve the sensitivity of its detection.
Accompanying drawing explanation
Fig. 1 is the structural representation of a kind of electrolytic cell for detecting dopamine of the present invention;
Fig. 2 is the differentiated pulse voltammogram of an embodiment of a kind of electrolytic cell for detecting dopamine of the present invention and detection method thereof;
Fig. 3 is the time m-map of current of coupling reaction of the present invention after 5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes and 50 minutes;
Fig. 4 is that dopamine concentration of the present invention is respectively 1 × 10
-7mol/L, 2.5 × 10
-7mol/L, 5 × 10
-7mol/L, 7.5 × 10
-7mol/L, 10 × 10
-7concentration-the map of current be depicted as during mol/L.
Embodiment
Embodiment one
As shown in Figure 1, a kind of electrolytic cell for detecting dopamine, comprise pond body 1 and be positioned over Chi Tizhong working electrode 2, to electrode 3 and contrast electrode 4.Pond body 1 to comprise at the bottom of pond lid 11, solution pool 12 and pond 13.Pond lid 11 band circular hole, solution pool 12 is rounding mesa-shaped, and at the bottom of pond, 13 is cylindrical.At the bottom of pond bottom 13 in the middle of be provided with shrinkage pool, shrinkage pool is built with magnet 131, and magnet 131 lower end is provided with spring 132.Working electrode 2 crosses at 13 junctions at the bottom of solution pool 12 and pond, and the working portion of working electrode 2 is towards solution pool 12.Electrode 3 and contrast electrode 4 are inserted by the circular hole of pond lid 11, the working portion of electrode 3 and contrast electrode 4 is entered in solution pool.At the bottom of solution pool 12 and pond, 13 are fixed by screw, and at the bottom of solution pool 12 and pond, 13 junctions are provided with rubber blanket 5.In electrolytic cell, at the bottom of solution pool 12, pond lid 11 and pond, the material of 13 is teflon.The bottom steel plate 6 of electrolytic cell is coated.Working electrode 2 selects printing carbon electrode, and the carbon paste district of printing carbon electrode is circular, and contrast electrode 4 selects mercury electrode, selects platinum electrode to electrode 3.
First be the ethanol water ultrasonic cleaning 30 minutes of 90% by solution pool mass concentration, then after ultrapure water cleaning at 80 DEG C dry for standby.Working electrode is faced up insert at the bottom of solution pool 12 and pond 13 junction, and adjust working electrode 2 position working portion 13 to be exposed completely at the bottom of pond, lock screw; Finally, dopamine solution to be measured is poured in solution pool 12, put into electrode 3 and contrast electrode 4 by the circular hole of pond lid 11, make to enter in solution pool 12 perform region of electrode 3 and contrast electrode 4.Connect galvanochemistry platform to test.
The activation of carboxyl magnetic microsphere: measure 50uL carboxyl magnetic microsphere emulsion (solid content 5 ~ 20g/L, surface carboxyl groups amount >=0.25mmol/g, mean grain size 60nm) in test tube, add 5mL NaAc_HAc buffer solution (PH=4.6) dilution shake up after, add 1-(3-the dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride activation that 0.05mL concentration is 100mg/L, magnetic-adsorption activation products are cleaned with ultrapure water after 30 minutes, abandoning supernatant, 2mL hac buffer (pH=3.6) is used to wash 3 times more respectively, finally in test tube, add 5mL hac buffer (pH=3.6).
Getting 10uL concentration is that 5mmol/L dopamine hydrochloric acid solution NaAc_HAc buffer solution (pH=3.6) constant volume in the volumetric flask of 50mL dilutes for after 1ppm.Carboxyl magnetic microsphere solution after being activated by 5mL and 5mL concentration are that the dopamine solution coupling reaction of 1ppm is after 50 minutes, magnetic microsphere is carried out Magneto separate, spend deionized water 3 times, then the magnetic microsphere after coupling is transferred in electrolytic cell, the NaAc_HAc buffer solution being 0.05mol/L with 10mL concentration (pH=3.6) is electrolytic solution, magnetic microsphere quick adsorption under the externally-applied magnetic field of the magnet 131 of electrolytic cell after coupling, to surface of printing electrode, is put into mercury electrode and platinum electrode subsequently and is carried out Differential Pulse Voltammetry and detect.Location parameter: initial potential is-0.1V, final current potential 0.7V, sweep velocity is 4mV/s, and pulse height is 50mV, and the sampling time is 20 seconds, and pulse width is 60 milliseconds, and the recurrence interval is 2000 milliseconds, and rest time is 2 seconds.
Conventional differential pulse voltammetry is tested: getting 10uL concentration is that 5mmol/L dopamine hydrochloric acid solution 0.05mol/L NaAc_HAc buffer solution (pH=3.6) constant volume dilution in the volumetric flask of 50mL, for after 1ppm, is poured in 100mL small beaker.Printing carbon electrode, mercury electrode and platinum electrode are put into solution and carries out Differential Pulse Voltammetry.Location parameter: nitrogen continues to pass into 5 minutes and carrys out deoxygenation, and initial potential is-0.1V, final current potential 0.7V, sweep velocity is 4mV/s, and pulse height is 50mV, and the sampling time is 20 seconds, pulse width is 60 milliseconds, and the recurrence interval is 2000 milliseconds, and rest time is 2 seconds.
As shown in Figure 2, peak current=1.158 × 10 of dopamine are directly measured
-6a, peak current=1.069 × 10 of magnetic field measuring dopamine after coupling
-5a.Under externally-applied magnetic field, coupled product is enriched in electrode surface, makes oxidation peak current be exaggerated 9.2 times, and spike potential is negative to be moved, and improves the sensitivity of electrode reaction.
Embodiment two
Getting 10uL concentration is that 5mmol/L dopamine hydrochloric acid solution hac buffer (pH=3.6) constant volume in the volumetric flask of 50mL dilutes for after 1ppm.Magnetic microsphere solution after being activated by 5mL and 5mL concentration are that the dopamine solution coupling reaction of 1ppm is after 5 minutes, magnetic microsphere is carried out Magneto separate, spend deionized water 3 times, then microballoon is transferred in electrolytic cell of the present invention, with acetic acid sodium acetate buffer (pH=3.6) 10ml of 0.05mol/L for electrolytic solution, magnetic microsphere quick adsorption under the externally-applied magnetic field of electrolytic cell after coupling, to surface of printing electrode, is put into mercury electrode and platinum electrode subsequently and is carried out Differential Pulse Voltammetry and detect.Location parameter: initial potential is-0.1V, final current potential 0.7V, sweep velocity is 4mV/s, and pulse height is 50mV, and the sampling time is 20 seconds, and pulse width is 60 milliseconds, and the recurrence interval is 2000 milliseconds, and rest time is 2 seconds.When each parameter constant, the time changing coupling reaction is respectively 10 minutes, 20 minutes, 30 minutes, 40 minutes and reacts for 50 minutes.
As shown in Figure 3, Fig. 3 gives the time m-map of current of coupling reaction after 5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes and 50 minutes, and as seen from Figure 3, coupling reaction 30 minutes, oxidation peak current reaches stable.
To get 10uL concentration be 5mmol/L dopamine hydrochloric acid solution NaAc_HAc buffer solution (pH=3.6) is 0.5 × 10 in dilution
-7after the concentration of mol/L.Be 0.5 × 10 with concentration successively by the 5mL magnetic microsphere solution after activation
-7the dopamine solution coupling reaction of mol/L is after 30 minutes, magnetic microsphere is carried out Magneto separate, spend deionized water 3 times, then microballoon is transferred in electrolytic cell, with acetic acid-sodium acetate buffer (pH=3.6) 10mL of 0.05mol/L for electrolytic solution, magnetic microsphere quick adsorption under the externally-applied magnetic field of electrolytic cell after coupling, to surface of printing electrode, is put into mercury electrode and platinum electrode subsequently and is carried out Differential Pulse Voltammetry and detect.Location parameter: initial potential is-0.1V, final current potential 0.7V, sweep velocity is 4mV/s, and pulse height is 50mV, and the sampling time is 20 seconds, and pulse width is 60 milliseconds, and the recurrence interval is 2000 milliseconds, and rest time is 2 seconds.
When each parameter constant, change dopamine concentration and be respectively 1 × 10
-7mol/L, 2.5 × 10
-7mol/L, 5 × 10
-7mol/L, 7.5 × 10
-7mol/L and 10 × 10
-7mol/L reacts.
Fig. 4 is that dopamine concentration is respectively 1 × 10
-7mol/L, 2.5 × 10
-7mol/L, 5 × 10
-7mol/L, 7.5 × 10
-7mol/L, 10 × 10
-7concentration-the map of current be depicted as during mol/L, dopamine concentration is 0.5 × 10
-7mol/L ~ 10 × 10
-7have good linear relationship with coupled product oxidation peak current within the scope of mol/L, obtaining linear equation by Fig. 4 is: lp=0.992 × C
dopamine+ 0.313, coefficient R
2be 0.989, lowest detection is limited to 0.5 × 10
-7mol/L.The concentration of dopamine solution to be measured can be drawn by the numerical value of the peak current of magnetic field measuring dopamine after coupling by the linear relationship of Fig. 4.
Claims (6)
1. one kind for detecting the electrolytic cell of dopamine, wherein, comprise the working electrode that Chi Tiji is positioned over Chi Tizhong, to electrode and contrast electrode, described pond body comprises Chi Gai, at the bottom of solution pool and pond, cover strip hole, described pond, described solution pool is rounding mesa-shaped, be cylindrical at the bottom of described pond, shrinkage pool is provided with in the middle of bottom at the bottom of described pond, described shrinkage pool is built with magnet, described magnet lower end is provided with spring, described working electrode crosses at junction at the bottom of described solution pool and pond, the working portion of described working electrode is towards solution pool, described electrode and contrast electrode to be inserted by the hole of Chi Gai, the described working portion to electrode and contrast electrode enters in solution pool, described working electrode selects printing carbon electrode, the carbon paste district of described printing carbon electrode is circular, described platinum electrode is selected to electrode, described contrast electrode selects mercury electrode.
2. a kind of electrolytic cell for detecting dopamine according to claim 1, wherein, is fixed by screw at the bottom of described solution pool and pond, and at the bottom of described solution pool and pond, junction is provided with rubber blanket.
3. a kind of electrolytic cell for detecting dopamine according to claim 2, wherein, adopt teflon material at the bottom of described solution pool, Chi Gai and pond, the bottom steel plate of described electrolytic cell is coated.
4. the method for the detection dopamine of the electrolytic cell of application according to any one in claim 1-3, comprise: under buffer system, by the magnetic microsphere after activation and dopamine solution generation coupling reaction, coupling reaction product is enriched in working electrode surface under magnetic field added by electrolytic cell, peak current is detected by Differential Pulse Voltammetry, set up linear relationship by peak current and standard specimen concentration, draw testing sample concentration by set up linear relationship.
5. a kind of method detecting dopamine according to claim 4, wherein, described buffer system selects Acetic acid-sodium acetate damping fluid.
6. a kind of method detecting dopamine according to claim 5, wherein, the activator used in described reactivation process is carbodiimide.
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| WO2009137713A2 (en) * | 2008-05-07 | 2009-11-12 | The Regents Of The University Of California | Particle-based electrostatic sensing and detection |
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| CN1588071A (en) * | 2004-08-16 | 2005-03-02 | 上海市胸科医院 | Enzyme labelling immunological electrochemical detecting method |
| WO2009137713A2 (en) * | 2008-05-07 | 2009-11-12 | The Regents Of The University Of California | Particle-based electrostatic sensing and detection |
| CN101576523A (en) * | 2009-06-11 | 2009-11-11 | 上海交通大学 | Method for detecting tumour cells by adopting microelectrode array impedance biosensor chip |
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