CN104062339A - [Ru(tatp)3]<2+> modified electrode and method for detecting glucose, uric acid and ascorbic acid in blood by adopting electrode - Google Patents
[Ru(tatp)3]<2+> modified electrode and method for detecting glucose, uric acid and ascorbic acid in blood by adopting electrode Download PDFInfo
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Abstract
The invention relates to the technical field of electrochemical detection, and discloses a [Ru(tatp)3]<2+> modified electrode and a method for detecting glucose, uric acid and ascorbic acid in blood by adopting electrode. The [Ru(tatp)3]<2+> modified electrode is obtained by dropping a [Ru(tatp)3]<2+> solution on a work electrode and then performing constant-temperature drying at a temperature of 40-80 DEG C, wherein the solution is prepared by dissolving dimethyl sulfoxide. The method for detecting the glucose, the uric acid and the ascorbic acid in the blood by adopting the [Ru(tatp)3]<2+> modified electrode comprises the steps: connecting the [Ru(tatp)3]<2+> modified electrode into an electrochemical work station, respectively enabling potentials to be 0.25-0.35 V, 0.65-0.75 V and 0.95-1.05 V by using a potential step jump method, then dropping a blood sample, measuring response currents corresponding to different potentials, and respectively comparing with respective concentration-current standard curves, and calculating actual concentrations of the glucose, the uric acid and the ascorbic acid in the blood.
Description
Technical field
The invention belongs to electrochemical measuring technique field, be specifically related to one [Ru (tatp)
3]
2+modified electrode and for detection of the method for glucose in blood, uric acid and ascorbic acid.
Background technology
Glucose, uric acid and ascorbic acid are present in blood of human body conventionally jointly, and the physiological metabolism of human body is played a very important role.In these three kinds of components, any concentration abnormality all can produce serious harm to the mankind's health.
The first, the concentration of glucose in blood is too high will lead diabetogenic generation, and diabetes are a kind of due to defect of insulin secretion or the impaired metabolic disease causing of its biological agent.It is to various functional lesion or the obstacles that all can produce in various degree organized in human body.Therefore, diabetic is carried out to effective external blood glucose value and measure and early stage examination, in active prevention and treatment diabetes, tool is of great significance; Second, the too high meeting of uric acid concentration causes human body fluid souring, affect the normal function of human body cell, ignore for a long time and may cause some serious diseases, as gout and hyperuricemia etc., these have all caused great threat to patient's health, so prevention and control uric acid are too high very important concerning the mankind's health; The 3rd, ascorbic acid concentrations is too low will cause scorbutic appearance, thereby accurate detection to ascorbic acid concentrations in blood of human body, can active prevention scurvy and carry out the various acute and chronic communicate illnesss of supplemental treatment by controlling ascorbic acid.
At present, the method that detects glucose in blood mainly contains high performance liquid chromatography, spectrophotometric method, vapor-phase chromatography and electrochemica biological sensor etc., the method that detects uric acid mainly contains high performance liquid chromatography, phosphotungstic acid reducing process, colourimetry and electrochemica biological sensor etc., and the method that detects ascorbic acid mainly contains high performance liquid chromatography, molybdenum blue colorimetric method, polarimetry and electrochemica biological sensor etc.As jointly detecting one of two kinds of main method of glucose, uric acid and ascorbic acid, high performance liquid chromatography applied range, but need expensive large-scale instrument, and be not easy to carry; Another kind method is electrochemica biological sensor detection method, and electrochemica biological sensor is divided into organized enzyme and non-enzyme sensor, organized enzyme electrochemica biological sensor exists that sensitivity is low, poor reproducibility and cost high-leveled and difficult in problems such as production in enormous quantities.By contrast, the electrochemica biological sensor of non-enzyme has the outstanding advantage that simultaneously detects glucose, uric acid and ascorbic acid, and linear detection range is wide, easy and simple to handle, stable performance and highly sensitive, in the detection of glucose, uric acid and ascorbic acid, there is good application prospect.
Summary of the invention
In order to solve the shortcoming and defect part of prior art, primary and foremost purpose of the present invention is to provide a kind of [Ru (tatp)
3]
2+modified electrode.
Another object of the present invention is to provide a kind of with above-mentioned [Ru (tatp)
3]
2+modified electrode detects the method for glucose in blood, uric acid and ascorbic acid.
The object of the invention is achieved through the following technical solutions:
A kind of [Ru (tatp)
3]
2+modified electrode, prepares by following preparation method:
With dmso solution [Ru (tatp)
3] Cl
2preparation [Ru (tatp)
3]
2+solution, then by above-mentioned [Ru (tatp)
3]
2+solution is added drop-wise on working electrode, and this working electrode is put into baking oven freeze-day with constant temperature under 40～80 DEG C of conditions, obtains [Ru (tatp)
3]
2+modified electrode.
[Ru (tatp) of the present invention
3] Cl
2prepare by the preparation method described in document (2012 discipline formula ripple master thesis of South China Normal University " DNA and BSA promote assembling and the photoelectric properties research of ruthenium (II) complex on ITO and nano TiO 2 particles " p30).
The preferred graphite electrode of described working electrode or glass-carbon electrode, working electrode is preferably dimensioned to be 5cm × 1cm.
Described [Ru (tatp)
3]
2[Ru (tatp) in solution
3]
2+concentration be preferably 0.5mmol/L, [Ru (tatp)
3]
2+the dripping quantity of solution is preferably 20 μ L.
A kind of with above-mentioned [Ru (tatp)
3]
2+modified electrode detects the method for glucose in blood, uric acid and ascorbic acid, comprises the following steps:
(1) by above-mentioned [Ru (tatp)
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system, then in electrolytic cell, add trishydroxymethylaminomethane buffer solution (pH=7.2), adopt cyclic voltammetry to carry out the scan round activation of 15 circles at the potential region of 0～1.2V;
(2) be 0.25～0.35V by potential step method constant potential, in the time that tending towards stability, electric current drips blood sample to be measured in electrolytic cell, obtain the response current of glucose in blood, this response current and concentration of glucose-response current typical curve are compared, calculate the concentration of glucose in blood sample;
(3) be 0.65～0.75V by potential step method constant potential, in the time that tending towards stability, electric current drips blood sample to be measured in electrolytic cell, obtain the response current of uric acid in blood, this response current and uric acid concentration-response current typical curve are compared, calculate the concentration of uric acid in blood sample;
(4) be 0.95～1.05V by potential step method constant potential, in the time that tending towards stability, electric current drips blood sample to be measured in electrolytic cell, obtain the response current of ascorbic acid concentrations in blood, this response current and ascorbic acid concentrations-response current typical curve are compared, calculate the concentration of ascorbic acid in blood sample.
The preferred 0.3V of constant potential described in step (2); The preferred 0.7V of constant potential described in step (3); The preferred 1.0V of constant potential described in step (3); The amount that drips blood sample to be measured described in step (2), (3) and (4) is preferably 10 μ L.
Have the following advantages and beneficial effect by preparation method of the present invention and the product tool that obtains:
(1) the present invention is based on [Ru (tatp)
3]
2+modified electrode, not only avoided the defect that organized enzyme electrode may inactivation, and be conducive to reusing of non-enzyme electrode, improved and improved sensitivity and reappearance that glucose, uric acid and ascorbic acid detect;
(2) the simple portable of electrode that prepared by the present invention, easy to use;
(3) detection method provided by the present invention integrates glucose, uric acid and ascorbic acid three's detection, and detection efficiency is high;
(4) detection method of glucose in blood provided by the present invention, uric acid and ascorbic acid is simple to operate, detects linear interval wide, and required blood volume is few, and measuring accuracy is high, and antijamming capability is strong, has good application prospect;
(5) select [Ru (tatp)
3]
2+as the decorative layer of electrode, it is not only strong in the absorption affinity of graphite electrode surface, and there is good redox active, there is a pair of good redox peak at about 0.48V in it, this current potential is in electrostatic double layer district, have energy consumption low, subsidiary reaction is few and disturb few feature, is conducive to catalytic oxidation and Electrochemical Detection to biomolecule.
Brief description of the drawings
Fig. 1 is [Ru (tatp) in embodiment 1
3]
2+the cyclic voltammetry scan figure of modified electrode under different potentials rate of change;
Fig. 2 is [Ru (tatp) in the embodiment 1 obtaining according to Fig. 1 result
3]
2+the graph of relation of the oxidation peak current-sweep velocity of modified electrode;
Fig. 3 is [Ru (tatp) in embodiment 1
3]
2+the differentiated pulse voltammogram that modified electrode obtains in the mixed solution of glucose, uric acid and ascorbic acid;
Fig. 4 is [Ru (tatp) in embodiment 1
3]
2+the response current of glucose solution and the relation of time that modified electrode is measured;
Fig. 5 is the concentration of glucose-response current typical curve obtaining according to Fig. 4 result;
Fig. 6 is [Ru (tatp) in embodiment 1
3]
2+the response current of uric acid solution and the relation of time that modified electrode is measured;
Fig. 7 is the uric acid concentration-response current typical curve obtaining according to Fig. 6 result;
Fig. 8 is [Ru (tatp) in embodiment 1
3]
2+the response current of ascorbic acid solution and the relation of time that modified electrode is measured;
Fig. 9 is the ascorbic acid concentrations-response current typical curve obtaining according to Fig. 8 result.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1
One [the Ru (tatp) of the present embodiment
3]
2+modified electrode, prepares by following preparation method:
With dmso solution [Ru (tatp)
3] Cl
2[the Ru (tatp) of preparation 0.5mmol/L
3]
2+solution, then by above-mentioned 20 μ L [Ru (tatp)
3]
2+solution is added drop-wise on graphite electrode, and this graphite electrode is put into baking oven freeze-day with constant temperature under 40～80 DEG C of conditions, obtains [Ru (tatp)
3]
2+modified electrode.
[the Ru (tatp) of the present embodiment
3] Cl
2by document, (" DNA and BSA promote ruthenium (II) complex at ITO and nano-TiO to 2012 discipline formula ripple master thesis of South China Normal University
2assembling on particle and photoelectric properties research " p30) described in preparation method prepare, its preparation process is: by 0.065g (0.25mmol) RuCl
33H
2o, 0.116g (0.5mmol) tatp (1,4,8,9-tetra-nitrogen terphenyls), 1.5mL37% concentrated hydrochloric acid and 1.5mL pure water mix and add reactor, be warmed up to 200 DEG C of reactions four hours, cooling after completion of the reaction, product cold water washing, 60 DEG C of vacuum drying 24h obtain [Ru (tatp)
2] Cl
2.Then by [Ru (tatp)
2] Cl
2add reactor intensification 200 DEG C to react four hour than 1:1 and concentrated hydrochloric acid with the mixed solvent (volume ratio 1:1) of water by amount of substance with tatp part, cooling washing vacuum drying obtains black crystals shape product [Ru (tatp)
3] Cl
2;
[the Ru (tatp) of the present embodiment
3]
2+the electrochemical Characterization of modified electrode, operation steps is as follows:
By [the Ru (tatp) of the present embodiment
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system; Then in the groove of electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10mmol/L; Scan by cyclic voltammetry, respectively with 0.6,0.5,0.4,0.3,0.2,0.1,0.05 and the scan speed measurement electric current of 0.02V/s (being numbered 1 to 8 in corresponding diagram 1) situation about changing with the change of current potential, the relation that obtains electric current corresponding to different scanning speed and current potential, result is as shown in Figure 1; Utilize origin mapping software to measure respectively oxidation peak current corresponding under different scanning speed, and utilize the matching of origin mapping software to obtain the relation of oxidation peak current-sweep velocity, result as shown in Figure 2, the relational expression that is obtained oxidation peak current-sweep velocity by Fig. 2 is I (μ A)=3.30+57.80v (V/s), degree of correlation R=0.993.
Can be drawn the decorative layer [Ru (tatp) of detecting electrode prepared by the present embodiment by Fig. 1 and Fig. 2
3]
2+stably be fixed on electrode surface, be subject to electrochemical process surface and control, functional.
Adopt [the Ru (tatp) of the present embodiment
3]
2+the selection of modified electrode constant potential during for detection of glucose in blood, uric acid and ascorbic acid, concrete operation step is as follows:
By [the Ru (tatp) of the present embodiment
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system; Then in the groove of electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10L-1; In the mixed solution of 100mmol/L glucose, 10mmol/L uric acid and ascorbic acid, utilize Differential Pulse Voltammetry to measure the situation that electric current changes with potential change, obtain differentiated pulse voltammogram as shown in Figure 3, can be obtained by Fig. 3, the oxidation peak current potential that glucose, uric acid and ascorbic acid are corresponding is respectively near 0.3V, 0.7V and 1.0V.Therefore adopt [Ru (tatp) of the present invention
3]
2+modified electrode detect glucose in blood, uric acid and ascorbic acid constant potential be respectively 0.25～0.35V, 0.65～0.75V and 0.95～1.05V, best constant potential is chosen as respectively 0.3V, 0.7V and 1.0V.
[the Ru (tatp) of the present embodiment
3]
2+modified electrode is for the drafting of concentration of glucose-response current typical curve, and concrete operation step is as follows:
By [the Ru (tatp) of the present embodiment
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system; Then in the groove of electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10mmol/L; With potential step method constant potential be 0.3V, in the time that electric current tends towards stability every 10s to the glucose solution that drips 10 μ L10mmol/L in electrolytic cell, obtain the response current of glucose solution and the relation of time, result as shown in Figure 4; Utilize the meet with a response relation of electric current and concentration of glucose of origin mapping software matching according to Fig. 4, result as shown in Figure 5, obtaining concentration of glucose-response current typical curve formula by Fig. 5 is: I (μ A)=113.17+2.46C (mmol/L), degree of correlation R=0.995.
[the Ru (tatp) of the present embodiment
3]
2+modified electrode is for the drafting of uric acid concentration-response current typical curve, and concrete operation step is as follows:
By [the Ru (tatp) of the present embodiment
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system; Then in the groove of electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10mmol/L; With potential step method constant potential be 0.7V, in the time that electric current tends towards stability every 10s to the uric acid solution that drips 10 μ L1mmol/L in electrolytic cell, obtain the response current of uric acid solution and the relation of time, result as shown in Figure 6; Utilize the meet with a response relation of electric current and concentration of glucose of origin mapping software matching according to Fig. 6, result as shown in Figure 7, obtaining uric acid concentration-response current typical curve formula by Fig. 7 is: I (μ A)=449.04+112.10C (mmol/L), degree of correlation R=0.987.
[the Ru (tatp) of the present embodiment
3]
2+modified electrode is for the drafting of ascorbic acid concentrations-response current typical curve, and concrete operation step is as follows:
By [the Ru (tatp) of the present embodiment
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system; Then in the groove of electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10mmol/L; With potential step method constant potential be 1.0V, in the time that electric current tends towards stability every 10s to the ascorbic acid solution that drips 10 μ L1mmol/L in electrolytic cell, obtain the response current of ascorbic acid solution and the relation of time, result as shown in Figure 8; Utilize the meet with a response relation of electric current and ascorbic acid concentrations of origin mapping software matching according to Fig. 8, result as shown in Figure 9, obtaining ascorbic acid concentrations-response current typical curve formula by Fig. 9 is: I (μ A)=640.34+136.13C (mmol/L), degree of correlation R=0.997.
Embodiment 2
[the Ru (tatp) of embodiment 1
3]
2+modified electrode is for detection of the method for glucose, uric acid and ascorbic acid in blood of human body sample, and concrete steps are as follows:
(1) by [Ru (tatp) of embodiment 1
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system, then in the groove of electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10mmol/L, adopt cyclic voltammetry to carry out the scan round activation of 15 circles at the potential region of 0～1.2V;
(2) be 0.3V by potential step method constant potential, in the time that tending towards stability, electric current drips 10 μ L blood sample to be measured in electrolytic cell, obtain the response current of glucose in blood, this response current and concentration of glucose-response current typical curve are compared, and the concentration that calculates glucose in blood sample is 4.83 ± 0.052mmol/L;
(3) be 0.7V by potential step method constant potential, in the time that tending towards stability, electric current drips 10 μ L blood sample to be measured in electrolytic cell, obtain the response current of uric acid in blood, this response current and uric acid concentration-response current typical curve are compared, and the concentration that calculates uric acid in blood sample is 0.255 ± 0.004mmol/L;
(4) be 1.0V by potential step method constant potential, in the time that tending towards stability, electric current drips 10 μ L blood sample to be measured in electrolytic cell, obtain the response current of ascorbic acid concentrations in blood, this response current and ascorbic acid concentrations-response current typical curve are compared, and the concentration that calculates ascorbic acid in blood sample is 0.00038 ± 0.000002mmol/L.
The checking of the method detectability of the present embodiment:
To the glucose, the uric acid of 0.5mmol/L and the ascorbic acid of 0.001mmol/L that add the 0.5mmol/L of normal concentration in the blood of human body sample of the present embodiment, obtain the blood sample after mark-on, again detect the concentration of glucose, uric acid and ascorbic acid in the blood sample after mark-on by the method for the present embodiment, the recovery of calculating respectively the glucose, uric acid and the ascorbic acid that add by the difference of concentration before and after mark-on, result is as shown in table 1.
Table 1 recovery of standard addition the result
Can be found out by table 1 result: method of the present invention is the concentration of human body glucose in blood, uric acid and ascorbic acid accurately, and have highly sensitive, workable advantage.
Above-described embodiment is preferably embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under Spirit Essence of the present invention and principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (5)
1. one kind [Ru (tatp)
3]
2+modified electrode, is characterized in that preparing by following preparation method:
With dmso solution [Ru (tatp)
3] Cl
2preparation [Ru (tatp)
3]
2+solution, then by above-mentioned [Ru (tatp)
3]
2+solution is added drop-wise on working electrode, and this working electrode is put into baking oven freeze-day with constant temperature under 40～80 DEG C of conditions, obtains [Ru (tatp)
3]
2+modified electrode.
2. one according to claim 1 [Ru (tatp)
3]
2+modified electrode, is characterized in that: described working electrode is graphite electrode or glass-carbon electrode; Working electrode is of a size of 5cm × 1cm.
3. one according to claim 1 [Ru (tatp)
3]
2+modified electrode, is characterized in that: described [Ru (tatp)
3]
2+[Ru (tatp) in solution
3]
2+concentration be 0.5mmol/L, be added drop-wise to [the Ru (tatp) on working electrode
3]
2+the amount of solution is 20 μ L.
4. one kind is utilized the [Ru (tatp) described in any one in claim 1～3
3]
2+modified electrode detects the method for glucose in blood, uric acid and ascorbic acid, it is characterized in that comprising following detecting step:
(1) by dried [Ru (tatp)
3]
2+modified electrode, to electrode and contrast electrode respectively the electrode corresponding with electrochemical workstation be connected, build the electrolytic cell of a three-electrode system, then in electrolytic cell, add the trishydroxymethylaminomethane buffer solution of 10mmol/L, adopt cyclic voltammetry to carry out the scan round activation of 15 circles at the potential region of 0～1.2V;
(2) be 0.25～0.35V by potential step method constant potential, in the time that tending towards stability, electric current drips blood sample to be measured in electrolytic cell, obtain the response current of glucose in blood, this response current and concentration of glucose-response current typical curve are compared, calculate the concentration of glucose in blood sample;
(3) be 0.65～0.75V by potential step method constant potential, in the time that tending towards stability, electric current drips blood sample to be measured in electrolytic cell, obtain the response current of uric acid in blood, this response current and uric acid concentration-response current typical curve are compared, calculate the concentration of uric acid in blood sample;
(4) be 0.95～1.05V by potential step method constant potential, in the time that tending towards stability, electric current drips blood sample to be measured in electrolytic cell, obtain the response current of ascorbic acid concentrations in blood, this response current and ascorbic acid concentrations-response current typical curve are compared, calculate the concentration of ascorbic acid in blood sample.
5. [Ru (tatp) according to claim 4
3]
2+modified electrode detects the method for glucose in blood, uric acid and ascorbic acid, it is characterized in that: described in step (2), current potential is 0.3V; Described in step (3), current potential is 0.7V; Described in step (4), current potential is 1.0V; The amount that drips blood sample to be measured described in step (2), (3) and (4) is 10 μ L.
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|---|---|---|---|
| CN201410326338.6A CN104062339B (en) | 2014-07-09 | 2014-07-09 | [Ru (tatp) 3] 2+ modified electrode and the method being used for detecting glucose in blood, uric acid and ascorbic acid thereof |
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|---|---|---|---|
| CN201410326338.6A CN104062339B (en) | 2014-07-09 | 2014-07-09 | [Ru (tatp) 3] 2+ modified electrode and the method being used for detecting glucose in blood, uric acid and ascorbic acid thereof |
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Citations (2)
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|---|---|---|---|---|
| CN101377473A (en) * | 2007-08-31 | 2009-03-04 | 中国科学院过程工程研究所 | Fast quantifying electrolysis method |
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2014
- 2014-07-09 CN CN201410326338.6A patent/CN104062339B/en not_active Expired - Fee Related
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101377473A (en) * | 2007-08-31 | 2009-03-04 | 中国科学院过程工程研究所 | Fast quantifying electrolysis method |
| CN102621208A (en) * | 2012-03-21 | 2012-08-01 | 南京邮电大学 | Preparation method and application of three-dimensional graphene electrode for electrochemical biosensor |
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| Title |
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| GUI-LIAN GAN等: "Preparation of a DNA-bound [Ru(bpy)2(mbpibH2)]2+ film and its two-mode luminescence tuning by copper(II) ions and EDTA", 《SPECTROCHIMICA ACTA PART A: MOLECULAR AND BIOMOLECULAR SPECTROSCOPY》 * |
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