CN105758905B - Method that is a kind of while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions - Google Patents
Method that is a kind of while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions Download PDFInfo
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- CN105758905B CN105758905B CN201610239124.4A CN201610239124A CN105758905B CN 105758905 B CN105758905 B CN 105758905B CN 201610239124 A CN201610239124 A CN 201610239124A CN 105758905 B CN105758905 B CN 105758905B
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- G—PHYSICS
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- 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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Abstract
The invention discloses a kind of methods for detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions simultaneously.Using by working electrode, reference electrode and the three-electrode system sensor constituted to electrode.Wherein, working electrode uses graphene/tantalum, graphene/boron-doped diamond film, graphene/titanium；Reference electrode uses saturated calomel electrode, Ag/AgCl electrode, mercury/Mercurous sulfate electrode；Platinum plate electrode is used to electrode.Pass through the mixed solution of five kinds of determinands of various concentration first, working curve is obtained by differential pulse voltammetry, then human serum is diluted, it is adjusted to concentration within the scope of working curve, the peak current obtained by differential pulse voltammetry obtains five kinds of testing concentrations in serum compared with working curve.The present invention relates to having the characteristics that quickly while to detect five kinds of testing concentrations, and high sensitivity, detection limit is low.
Description
Technical field
The present invention relates to a kind of methods by electrochemistry to detect ascorbic acid, dopamine, uric acid, tryptophan and Asia simultaneously
Five kinds of substances such as nitrate anion.
Background technique
This five kinds of substances of ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions play weight to the metabolism of human body
Act on, and by the detections of these substances, can by judge scurvy, schizophrenia and senile dementia, diabetes,
The content of contained above-mentioned substance in patients' body fluid such as hepatic insufficiency, food poisoning, and play prevention effect.
Currently, commonly using oxidimetry, spectrophotometry, uricase-peroxidase conjugate in clinical medicine
Method, fluorescence spectrophotometry, colorimetric method etc..In clinical application or to different material using different detection means or
Sample handling processes are cumbersome or are marked using the biological substance of the easy inactivation such as enzyme, this to detection speed and effect all
Bring adverse effect.In addition, since, there is interaction, only real-time synchronization measures between various determinands, it could be anti-
Answer respective substance in body fluid really horizontal.
Clinically the positive low cost of the detection means of body fluid, multinomial synchronization, highly sensitive, accurate and quick direction are developed.
And electrochemical sensor is the electrochemical response spike potential between sensor electrode and determinand for qualitative index, response peak is electric
Stream is quantitative target, can detect many kinds of substance simultaneously.
Electrochemical sensor of the present invention, it is simple process, at low cost, it can be quick by the electrochemical sensor
The content of this five kinds of substances of contained ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions in human body fluid is detected simultaneously.
Summary of the invention
It is an object of the invention to construction work electrode, reference electrode and to electrode three-electrode system, while realizing detection
This five kinds of substances of ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions realize the purpose of highly sensitive, low detection limit.
In order to achieve the above object, technical scheme is as follows:
Method that is a kind of while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions, by by work electricity
Pole, reference electrode, the electrochemical sensor constituted to three electrodes of electrode are realized, comprising the following steps:
(1) using phosphate buffer solution as bottom liquid, configure the ascorbic acid of different pH value, dopamine, uric acid, tryptophan and
The three electrodes connection electrochemical workstation of five kinds of determinand mixed solutions of nitrite anions, electrochemical sensor is inserted into the solution,
It is tested using differential pulse voltammetry, the voltage-to-current working curve with 5 electrochemical response characteristic peaks is obtained, in difference
The size that peak current is compared under pH value, finds out the pH value of maximum peak current；
(2) using phosphate buffer solution as bottom liquid, dilute serum simultaneously adjusts pH value, and the pH value of serum is made to be adjusted to step
The pH value of the maximum peak current of middle determination, ascorbic acid, dopamine, uric acid, five kinds of substances of tryptophan and nitrite anions solution
In concentration dilution to step (1) working curve concentration range；
In (3) three electrode insertion step (2) solution, tested using differential pulse voltammetry, wherein peak current size and work
Compare as curve, after being diluted in serum 5 determinands concentration, which can be obtained in serum multiplied by extension rate
The concentration of ascorbic acid, dopamine, five kinds of uric acid, tryptophan and nitrite anions substances.
Further, working electrode using graphene/tantalum electrode or graphene/boron-doped diamond film electrode or graphene/
Ti electrode；Reference electrode is using saturated calomel electrode or Ag/AgCl electrode or mercury/Mercurous sulfate electrode；Platinized platinum is used to electrode
Electrode.
Further, working electrode is realized using following steps: (1) pre- in the substrate of tantalum or boron-doped diamond film or titanium
One layer of nickel particle is deposited, as catalyst needed for graphene growth；(2) Dc arc plasma jet CVD chemical gaseous phase is used
Deposition technique prepares graphene, and wherein reaction pressure is 100Pa~4000Pa, and base reservoir temperature is 500 DEG C~1200 DEG C, when growth
Between be 0.5min~30min, arc voltage be 50V~70V, arc current be 80A~130A.
Further, step (1) and (2) indsole liquid pH value is 4.0~10.0, concentration is 0.1mol/L~0.2mol/L's
Phosphate buffer solution.
Further, the concentration range of ascorbic acid is 5~1500 μm of ol/L, the concentration range of dopamine in step (1)
For 0.25~75 μm of ol/L, the concentration range of uric acid is 0.4~120 μm of ol/L, and the concentration range of tryptophan is 0.25~75 μ
Mol/L, the concentration range of nitrite anions are 2~600 μm of ol/L.
Further, the potential range of differential pulse voltammetry is set as -1.0V~1.2V, current potential increment in step (1)
It is set as 0.001V~0.05V, amplitude is set as 0.01V~0.05V, and 0.005s~0.25s, pulse period is arranged in pulse width
0.01s~0.5s is set.
Further, serum diluting multiple selects 1~100 times in step (2).
The invention has the characteristics that:
(1) five kinds of objects such as Ascorbic Acid in Blood Serum, dopamine, uric acid, tryptophan and nitrite anions can be quickly detected simultaneously
The concentration of matter.
(2) the detection range of linearity is wide, high sensitivity, and detection limit is low.
(3) reproducibility, repeatability, stability of sensor are good, and the service life is long.
(4) at low cost, be conducive to be miniaturized.
Detailed description of the invention
Fig. 1 is the electron scanning micrograph of working electrode surface graphene prepared by the present invention.
Fig. 2 is to use the graphene/tantalum wire prepared on the basis of the present invention as working electrode, and saturated calomel electrode is reference
Electrode, platinized platinum are the differential pulse voltammetry curve and working curve obtained under various concentration to electrode.
Wherein, ascorbic acid concentrations are 5~1500 μm of ol/L, and dopamine concentration is 0.25~75 μm of ol/L, uric acid concentration
For 0.4~120 μm of ol/L, Tryptophan concentration is 0.25~75 μm of ol/L, and nitrite anions concentration is 2~600 μm of ol/L.In figure, a
For differential pulse voltammetry curve, b~f is followed successively by the working curve of ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions.
Specific embodiment
It elaborates below to the embodiment of the present invention, the present embodiment carries out under the premise of the technical scheme of the present invention
Implement, the detailed implementation method and specific operation process are given, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1:
(1) preparation of working electrode
1. configuration concentration is the Ni (NO of 0.6mol/L3)2Ethanol solution, and drop coating is the tantalum wire surface of 0.6mm in diameter,
Drying is placed on the sample stage of Dc arc plasma jet CVD chemical gas-phase deposition system, is adjusted sample stage height, is made it
Apart from plasma jet mouth about 1.5cm.
2. closing reaction chamber, system is vacuumized, when vacuum degree reaches 1Pa or less, is passed through the hydrogen that flow is 2L/min
The methane of gas, the argon gas of 1.5L/min and 200mL/min, wherein reaction pressure is 100Pa~4000Pa, arc voltage 50V
~70V, arc current are 80A~130A, and base reservoir temperature is 500 DEG C~1200 DEG C, after growth time is 0.5min~30min,
Graphene/tantalum wire sample is taken out, as working electrode.The present invention preferably when chamber pressure and pump pressure reach 3000Pa and 13000Pa when,
Apply the arc current of 110A, sample stage base reservoir temperature is 950 DEG C, and when growing graphene 15min, gained working electrode is managed the most
Think.The electron scanning micrograph of gained working electrode surface graphene is as shown in Fig. 1.
(2) pH value is determined
1. the phosphate buffer solution that configuration pH value is 4.0~10.0, concentration is 0.1mol/L, as bottom liquid, configuring pH value is
4.0, the mixing of 5.0,6.0,7.0,8.0,9.0,10.0 ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions is molten
Liquid, wherein the concentration of this five kinds of substances is successively are as follows: 500 μm of ol/L, 25 μm of ol/L, 40 μm of ol/L, 25 μm of ol/L, 200 μm of ol/L.
2. graphene/tantalum wire, as working electrode, saturated calomel electrode is reference electrode, platinized platinum is to electrode, connection electricity
Chem workstation, and these three electrodes are inserted into ascorbic acid, dopamine, uric acid, tryptophan and the nitrite anions of different pH value
Mixed solution in, using in electrochemical workstation differential pulse voltammetry test current -voltage curve.Differential pulse volt
In the parameter of peace method, potential range is set as -1.0V~1.2V, and current potential increment is set as 0.001V~0.05V, and amplitude is set as
0.005s~0.25s is arranged in 0.01V~0.05V, pulse width, and 0.01s~0.5s is arranged in the pulse period.Preferred electricity in the present invention
Position increment is set as 0.001V, and amplitude is set as 0.01V, and pulse width is set as 0.005s, and 0.01s is arranged in the pulse period.
3. pH value is determined by comparing the size of the peak current obtained under different pH value, it is optimal to the electrode of this example
PH value is 7.0.
(3) working curve is established
1. using phosphate buffer solution as bottom liquid, the ascorbic acid of various concentration that configuration pH value is 7.0, dopamine, uric acid,
The mixed solution of tryptophan and nitrite anions.Wherein, ascorbic acid concentrations are 5~1500 μm of ol/L, dopamine concentration 0.25
~75 μm of ol/L, uric acid concentration are 0.4~120 μm of ol/L, and Tryptophan concentration is 0.25~75 μm of ol/L, and nitrite anions concentration is
2~600 μm of ol/L.
2. still using used three electrodes are adopted in upper step, using the differential pulse voltammetry with upper step identical parameters
Method tests current -voltage curve, referring to attached drawing 2.Since the electrochemical response characteristic of every kind of determinand is different, in above-mentioned steps
In the differential pulse voltammetry voltage-current curve of middle test, it may appear that 5 are directed to the response characteristic peak of five kinds of determinands,
Middle spike potential is used for qualitative index, and peak current is used for quantitative target, draws the working curve of each determinand, i.e. concentration is horizontal seat
Mark, peak current are the curve of ordinate.The peak of ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions is obtained from curve
Current potential is followed successively by -0.115V, 0.115V, 0.25V, 0.58V, 0.715V or so, these spike potentials are as qualitative index.
3. peak current passes through what is obtained under various concentration as vertical index using the concentration of each determinand as abscissa
The peak current size of each determinand draws working curve, the detection model as determinand.
In this example, detect ascorbic acid, dopamine, uric acid, five kinds of substances of tryptophan and nitrite anions it is minimum
Concentration can achieve 5 μm of ol/L, 0.25 μm of ol/L, 0.4 μm of ol/L, 0.25 μm of ol/L, 2 μm of ol/L.
(3) rate of recovery in serum is analyzed
Using phosphate buffer solution as bottom liquid, dilute serum, and adjusting pH value is 7.0.Due to resisting in the serum to different people
Bad hematic acid, dopamine, uric acid, tryptophan are different with the concentration of nitrite anions.In this example, the extension rate of serum uses 10
Again, 20 times, 50 times, the rate of recovery of serum can achieve 95%~105% range.
Claims (5)
1. a kind of method for detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions simultaneously, it is characterised in that: pass through
By working electrode, reference electrode, the electrochemical sensor that three electrodes of electrode are constituted is realized, comprising the following steps:
(1) using phosphate buffer solution as bottom liquid, ascorbic acid, dopamine, uric acid, tryptophan and the nitrous of different pH value are configured
The three electrodes connection electrochemical workstation of five kinds of determinand mixed solutions of acid group, electrochemical sensor is inserted into the solution, is used
Differential pulse voltammetry test, obtains the voltage-to-current working curve with 5 electrochemical response characteristic peaks, in different pH value
The size of lower comparison peak current, finds out the pH value of maximum peak current；
(2) using phosphate buffer solution as bottom liquid, dilute serum simultaneously adjusts pH value, and the pH value of serum is made to be adjusted in step true
The pH value of fixed maximum peak current, the solution concentration of ascorbic acid, dopamine, five kinds of uric acid, tryptophan and nitrite anions substances
It is diluted in step (1) working curve concentration range；
It in (3) three electrode insertion step (2) solution, is tested using differential pulse voltammetry, wherein peak current size and work are bent
Line compares, after being diluted in serum 5 determinands concentration, which can be obtained anti-bad in serum multiplied by extension rate
The concentration of hematic acid, dopamine, five kinds of uric acid, tryptophan and nitrite anions substances；
The working electrode is using graphene/tantalum electrode or graphene/boron-doped diamond film electrode or graphene/Ti electrode；Ginseng
Than electrode using saturated calomel electrode or Ag/AgCl electrode or mercury/Mercurous sulfate electrode；Platinum plate electrode is used to electrode；
The working electrode is realized using following steps: (1) one layer of pre-deposition in the substrate of tantalum or boron-doped diamond film or titanium
Nickel particle, as catalyst needed for graphene growth；(2) Dc arc plasma jet CVD chemical vapour deposition technique is used
Graphene is prepared, wherein reaction pressure is 100Pa~4000Pa, and base reservoir temperature is 500 DEG C~1200 DEG C, and growth time is
0.5min~30min, arc voltage are 50V~70V, and arc current is 80A~130A.
2. method that is according to claim 1 while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions,
It is characterized by: step (1) and the phosphoric acid that (2) indsole liquid pH value is 4.0~10.0, concentration is 0.1mol/L~0.2mol/L are slow
Rush solution.
3. method that is according to claim 1 while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions,
It is characterized by: the concentration range of ascorbic acid is 5~1500 μm of ol/L in step (1), the concentration range of dopamine is 0.25
~75 μm of ol/L, the concentration range of uric acid are 0.4~120 μm of ol/L, and the concentration range of tryptophan is 0.25~75 μm of ol/L, sub-
The concentration range of nitrate anion is 2~600 μm of ol/L.
4. method that is according to claim 1 while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions,
It is characterized by: the potential range of differential pulse voltammetry is set as -1.0V~1.2V in step (1), current potential increment is set as
0.001V~0.05V, amplitude are set as 0.01V~0.05V, and 0.005s~0.25s, pulse period setting is arranged in pulse width
0.01s~0.5s.
5. method that is according to claim 1 while detecting ascorbic acid, dopamine, uric acid, tryptophan and nitrite anions,
It is characterized by: serum diluting multiple selects 1~100 times in step (2).
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| CN106290500B (en) * | 2016-08-24 | 2018-11-16 | 川北医学院附属医院 | Utilize graphene-molybdenum disulfide-perfluorinated sulfonic resin detection uric acid method |
| CN107119263A (en) * | 2017-05-04 | 2017-09-01 | 天津理工大学 | A kind of preparation method of vertical graphene/boron-doped diamond sensing electrode |
| US10670580B2 (en) | 2017-06-05 | 2020-06-02 | Rutgers, The State University Of New Jersey | Quantification of inflammatory molecules in exhaled breath condensate using differential pulse voltammetry on reduced graphene oxide sensor |
| CN111517305B (en) * | 2020-04-08 | 2021-08-27 | 北京科技大学 | Preparation method of graphene/diamond composite structure with high carrier concentration |
| CN111474223A (en) * | 2020-04-22 | 2020-07-31 | 中国药科大学 | Electrochemical sensor for simultaneously detecting uric acid, ascorbic acid and dopamine, and preparation method and application thereof |
| CN113252749B (en) * | 2021-05-13 | 2023-03-21 | 天津理工大学 | Electrochemical test method for detecting ice crystal current and impedance |
| CN113804741A (en) * | 2021-09-23 | 2021-12-17 | 生态环境部南京环境科学研究所 | Electrochemical method for rapid evaluation of circulating water scale inhibitor |
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