CN106290539B - The method for detecting tyrosine concentration in solution - Google Patents
The method for detecting tyrosine concentration in solution Download PDFInfo
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- CN106290539B CN106290539B CN201610619910.7A CN201610619910A CN106290539B CN 106290539 B CN106290539 B CN 106290539B CN 201610619910 A CN201610619910 A CN 201610619910A CN 106290539 B CN106290539 B CN 106290539B
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a kind of methods of tyrosine concentration in detection solution, the tyrosine in solution to be measured is detected by differential pulse voltammetry using three-electrode system, the concentration of tyrosine in solution to be measured is obtained according to the linear equation of tyrosine, the working electrode in three-electrode system is glass-carbon electrode.Method of the invention can quickly detect tyrosine concentration, and high sensitivity, accuracy height.The present invention constructs relevant sensing interface as three-electrode system sensor and is used to detect, and is quantified using potential difference, and the sensitivity and accuracy of detection tyrosine are improved, minimum to can detecte out 3 × 10‑5The tyrosine of mol/L.Method of the invention greatly reduces testing cost, simple to operate.
Description
Technical field
The present invention relates to heterogeneous amino acid detection techniques fields.It is more particularly related to which a kind of utilize difference
The method that pulse voltammetry detects tyrosine concentration in solution.
Technical background
Tyrosine is the important semi-dispensable amino acid of human body, the amino acid metabolisms such as tyrosinemia, alkaptonuria disease, liver
Kidney diaseases, nervous system degeneration disease, malignant tumour etc. can lead to tyrosine metabolic disorder, so as to cause junket ammonia in body fluid
The variation of acid and its metabolite content.Therefore, research establishes easy, quick, sensitive analysis method and is applied to body
The analysis of tyrosine content in liquid has highly important clinical meaning for the auxiliary diagnosis of certain diseases, treatment and monitoring.
Summary of the invention
It is an object of the invention to solve at least the above problems or defect, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide a kind of methods of tyrosine concentration in detection solution, can be quickly and fixed
The content of tyrosine in amount detection solution establishes a kind of simple, quickly and sensitivity and the high tyrosine detection side of accuracy
Method.
In order to realize these purposes and other advantages according to the present invention, tyrosine concentration in a kind of detection solution is provided
Method, wherein include: the linear equation of preparatory building tyrosine, using by working electrode, reference electrode and auxiliary electrode structure
At three-electrode system the tyrosine in solution to be measured is detected to obtain relevant parameter by differential pulse voltammetry, by phase
The linear equation for answering parameter to substitute into the tyrosine obtains the concentration of tyrosine in solution to be measured；
Wherein, the working electrode glass-carbon electrode.
Preferably, the linear equation of tyrosine is constructed in advance in the method for tyrosine concentration in the detection solution
The following steps are included:
Step 1: compound concentration is 0~1.2 × 10-4The standard solution of at least 5 parts tyrosine of mol/L, including one
The standard solution that part tyrosine concentration is 0mol/L；
Step 2: being carried out using differential pulse voltammetry to resulting every part of standard solution using the three-electrode system
Detection, obtains the differential pulse voltammetry curve of every part of standard solution, records the difference of every part of standard solution respectively in the process
Current potential in Pulse Voltammetry curve at appearance；
Step 3: with the corresponding potential value of differential pulse voltammetry curve of resulting every part of standard solution and the tyrosine
Concentration is the difference of the potential value of the standard solution of 0mol/L as ordinate, is drawn using the concentration of every part of standard solution as abscissa
Standard curve processed and the linear equation that the tyrosine is calculated.
Preferably, using the three-electrode system, difference is utilized in the method for tyrosine concentration in the detection solution
Divide pulse voltammetry to detect the solution to be measured containing unknown concentration tyrosine, obtains the differential pulse voltammetry of solution to be measured
Curve, the standard solution for being 0mol/L by the corresponding potential value of differential pulse voltammetry curve of solution to be measured and tyrosine concentration
The difference of potential value substitutes into linear equation obtained in step 3, and the concentration of tyrosine in solution to be measured is calculated.
Preferably, in the detection solution in the method for tyrosine concentration, the standard of tyrosine in the step 1
Solution allocation method are as follows: the standard solution of the tyrosine of concentration 0mol/L is prepared using trishydroxymethylaminomethane buffer solution,
And mix the tyrosine solution that trishydroxymethylaminomethane buffer solution and concentration are 0.01mol/L, it is prepared into respectively dense
Degree is 3 × 10-5mol/L、6×10-5mol/L、9×10-5mol/L、1.2×10-4The standard solution of four tyrosine of mol/L,
And all containing concentration in every portion solution is 1 × 10-4Mol/L tryptophan；Wherein the trishydroxymethylaminomethane buffering is molten
The pH of liquid is 6.5~7.5, concentration 0.01mol/L.
Preferably, in the detection solution in the method for tyrosine concentration, the step 2 specifically includes following step
It is rapid:
1) three-electrode system is respectively implanted in 5 standard solution, stirs 3min at room temperature at 20~30 DEG C,
Static 3min；
2) differential pulse map is scanned, setting scanning initial potential is -0.2V, and termination current potential is 1.0V, and current potential increment is
0.004V, Sample Width 0.0167s, amplitude 0.05V, pulse width 0.05V, sensitivity 10-5A/V, waiting time 2s；
3) potential value for measuring and recording five standard solution respectively establishes the difference arteries and veins of five standard solution
Rush volt-ampere curve.
Preferably, in the detection solution in the method for tyrosine concentration, reference electrode is Ag/AgCl electrode, auxiliary
Helping electrode is platinum electrode.
Preferably, in the detection solution in the method for tyrosine concentration, the solution to be measured includes that basis is molten
Liquid, the base soln are concentration 1 × 10-3Mol/L or concentration 1 × 10-4Mol/L or concentration 1 × 10-5The tryptophan of mol/L is molten
Liquid, the three-electrode system detect solution to be measured using tryptophan as electrochemical signals.
The invention has the following advantages:
Firstly, working electrode is made using glass-carbon electrode in the method for the invention, relevant sensing interface is constructed as three
Electrode system sensor is qualitative using current potential progress tyrosine configuration progress for detecting, molten to tyrosine using potential difference
Liquid concentration is quantified, and the sensitivity and accuracy of detection tyrosine are improved, minimum to can detecte out 3 × 10-5The junket of mol/L
Propylhomoserin.
Secondly, a kind of method of method of the invention as Electrochemical Detection tyrosine, greatly reduces testing cost, behaviour
Make simple and convenient；
Finally, a step of the invention quickly detects tyrosine, after the completion of the preparation of three-electrode system sensor, it is only necessary to which several minutes just
A step may be implemented and detect tyrosine.
Detailed description of the invention
Fig. 1 is the differential pulse voltammetry curve graph of the tyrosine standard solution of various concentration in the embodiment of the present invention 1；
Fig. 2 is the canonical plotting of tyrosine in the embodiment of the present invention 1.
Specific embodiment
Elaborate below with reference to embodiment to the present invention, with enable those of ordinary skill in the art refering to this specification after
It can implement accordingly.
It should be noted that experimental method described in following embodiments is unless otherwise specified conventional method, institute
Reagent and material are stated, unless otherwise specified, is commercially obtained.
The term definition involved in the present invention arrived:
Unless otherwise defined, otherwise all technologies used herein and scientific term all have with it is of the art
Those of ordinary skill usually understands identical meaning.Although the usable and described herein in practice or test of the invention
Similar or equivalent any method, apparatus and material, but preferred method, device and material will now be described.
" working electrode " is also known as Electrode, refers to that studied reaction occurs on this electrode.In general, to work
The basic demand of electrode is: working electrode can be solid, be also possible to liquid, and miscellaneous solid material that can be conductive is equal
It can serve as electrode.It is affected due to the reaction that the electrochemical reaction studied will not be occurred by electrode itself, and can be
It is measured in biggish potential areas；Electrode must not react with solvent or electrolyte component；Electrode area should not be too
Greatly, electrode surface preferably should be uniform smooth, and can carry out surface cleaning etc. by simple method.Using solid electrode
When, in order to guarantee the reproducibility of experiment, it has to be noted that suitable electrode pre-treatment step is established, to guarantee redox, surface
Pattern and reproducible state there is no adsorbing contaminant.
A kind of method of tyrosine concentration in detection solution, using three-electrode system by differential pulse voltammetry to be measured
Tyrosine in solution is detected, and obtains the concentration of tyrosine in solution to be measured, three electrodes according to the linear equation of tyrosine
Working electrode in system is glass-carbon electrode.Three-electrode system is made of working electrode, reference electrode and auxiliary electrode.
The present invention is to be detected using Differential Pulse Voltammetry to tyrosine, and Differential Pulse Voltammetry is in linear scan wave
In shape, it is superimposed the fixed continuous impulse of an amplitude constant and pulsewidth, in scanning process, base current potential is scanned from initial potential to termination
Current potential, before potential pulse starts and at the end of carry out current sample, by the difference of the two sample rate currents to current potential map,
As DPV curve is mainly used for electrochemical analysis, background current caused by reducing because of oxidation of impurities reduction reaction, has more preferable
Detection sensitivity and lower detectable limit, relative to other methods, cost is relatively low for the Differential Pulse Voltammetry, operation letter
It is single.
The method of tyrosine concentration in the detection solution, comprising the following steps:
Step 1: prepare the working electrode and build three-electrode system, compound concentration 0~1.2 × 10-4Mol/L's is more
The standard solution of part tyrosine；
Step 2: using the three-electrode system, using differential pulse voltammetry to the every part of standard prepared in step 1
Solution is detected, and the differential pulse voltammetry curve of every part of standard solution is obtained, and it is molten to record every part of standard respectively in the process
The potential value of current strength in the differential pulse voltammetry curve of liquid；
Step 3: with the corresponding potential value of differential pulse voltammetry curve and junket of every part of standard solution obtained in step 2
The difference of the potential value of standard solution when propylhomoserin concentration is 0mol/L is cross with the concentration of every part of standard solution as ordinate
Coordinate draws standard curve and calculates linear equation；
Step 4: using the three-electrode system, using differential pulse voltammetry to containing unknown concentration tyrosine to
It surveys solution to be detected, the differential pulse voltammetry curve of solution to be measured is obtained, by the differential pulse voltammetry curve pair of solution to be measured
The potential value and tyrosine concentration answered are that the difference of the potential value of the standard solution of 0mol/L substitutes into obtained in step 3 linearly
In equation, the concentration of tyrosine in solution to be measured is calculated.
Standard solution is also after being mixed with certain density tryptophan solution for basic solution+buffer+tyrosine solution
It obtains.
Either standard solution or solution to be measured are all the concentration that tyrosine is measured using tryptophan as electrochemical signals
, i.e., all contain tryptophan base soln in standard solution and solution to be measured.
Embodiment 1:
A kind of method of tyrosine concentration in detection solution, comprising the following steps:
Step 1: preparing the working electrode and building three-electrode system, compound concentration is 0~1.2 × 10-4Mol/L's
The standard solution of more parts of tyrosine；
Wherein, in the step 1 tyrosine standard solution configuration method are as follows: be added into Tris-HCl buffer solution
Concentration is the tyrosine standard solution of 0.01mol/L, and preparing tyrosine concentration respectively is 0mol/L, 3 × 10-5mol/L、6×10-5mol/L、9×10-5mol/L、1.2×10-4Five standard solution of mol/L, and all contain 1 × 10 in every a solution- 4Mol/L tryptophan；Wherein the pH of the Tris-HCl buffer solution is 7.0, concentration 0.01mol/L；
Step 2: using the three-electrode system, using differential pulse voltammetry to the every part of standard prepared in step 1
Solution is detected, and the differential pulse voltammetry curve of every part of standard solution is obtained, and it is molten to record every part of standard respectively in the process
Potential value in the differential pulse voltammetry curve of liquid；
Wherein, the step 2 specifically includes the following steps:
The three-electrode system is respectively implanted in five standard solution by step 2.1, and 3min is stirred at room temperature,
Static 3min；
Step 2.2, scanning differential pulse map, setting scanning initial potential are -0.2V, and termination current potential is 1.0V, current potential
Increment is 0.004V, Sample Width 0.0167s, amplitude 0.05V, pulse width 0.05V, sensitivity 10-5A/V, waiting time
For 2s；
Step 2.3, measure and record respectively five standard solution current strength peak value, establish it is described five mark
The differential pulse voltammetry curve of quasi- solution, the differential pulse voltammetry curve of five standard solution shown in Fig. 1；
Step 3: corresponding with the differential pulse voltammetry curve of tetra- standard solution of solution b, c, d, e obtained in step 2
Potential value and solution a standard solution potential value difference as ordinate, with the concentration of tyrosine in solution b, c, d, e
Standard curve is drawn for abscissa and calculates linear equation；For example, with the corresponding potential value of differential pulse voltammetry curve of solution b
Difference with the potential value of the standard solution of solution a is that ordinate can be true using the concentration of tyrosine in solution b as abscissa
A point on calibration directrix curve changes solution b into solution c, d, e by the above method, determines standard by solution c, d, e
The other three point on curve, draws standard curve as shown in Figure 2；
Obtain linear equation by Fig. 2 standard curve: Y=-4+0.26667X, Y is current value (E-E in formula0), E is every
The corresponding potential value of differential pulse voltammetry curve of part standard solution；E0Standard solution when for tyrosine concentration being 0mol/L
Potential value, unit mV；X is tyrosine concentration in standard solution, and unit is μm ol/L, coefficient R2It is 0.9999.Work
Electrode is limited to 3.0 × 10 to the detection of tyrosine-5mol/L；
Step 4: using the three-electrode system, using differential pulse voltammetry to containing unknown concentration tyrosine to
It surveys solution to be detected, obtains the differential pulse voltammetry curve of solution to be measured, calculate the concentration of tyrosine in liquid to be detected: will be to
The potential value for the standard solution that the corresponding potential value of differential pulse voltammetry curve and tyrosine concentration for surveying solution are 0mol/L
Difference substitutes into linear equation obtained in step 3 as Y value, calculates X value and obtains the concentration of tyrosine in solution to be measured.
Wherein, reference electrode is Ag/AgCl electrode, and auxiliary electrode is platinum electrode.
Although the embodiments of the present invention have been disclosed as above, but its is not only in the description and the implementation listed
With it can be fully applied to various fields suitable for the present invention, for those skilled in the art, can be easily
Realize other modification, therefore without departing from the general concept defined in the claims and the equivalent scope, the present invention is simultaneously unlimited
In specific details.
Claims (4)
1. a kind of method of tyrosine concentration in detection solution, wherein include: the linear equation of preparatory building tyrosine, use
The three-electrode system being made of working electrode, reference electrode and auxiliary electrode by differential pulse voltammetry in solution to be measured into
Row detection obtains relevant parameter, and the linear equation that relevant parameter substitutes into the tyrosine is obtained the dense of tyrosine in solution to be measured
Degree；
Wherein, the working electrode is glass-carbon electrode；
In advance building tyrosine linear equation the following steps are included:
Step 1: the standard solution of five tyrosine of configuration: being using trishydroxymethylaminomethane buffer solution preparation concentration
Trishydroxymethylaminomethane buffer solution and concentration are the junket ammonia for 0.01mol/L by the standard solution of the tyrosine of 0mol/L
Acid solution mixing, then being prepared into concentration respectively is 3 × 10-5mol/L、6×10-5mol/L、9×10-5mol/L、1.2×10- 4The standard solution of four tyrosine of mol/L, and all containing concentration in five standard solution is 1 × 10-4Mol/L tryptophan；
Wherein the pH of the trishydroxymethylaminomethane buffer solution is 6.5~7.5, concentration 0.01mol/L；
Step 2: resulting every part of standard solution is detected using differential pulse voltammetry using the three-electrode system,
The differential pulse voltammetry curve of every part of standard solution is obtained, records the differential pulse volt of every part of standard solution respectively in the process
Pacify the current potential in curve at appearance；
Step 3: with the corresponding potential value of differential pulse voltammetry curve of resulting every part of standard solution and the tyrosine concentration
For the standard solution of 0mol/L potential value difference as ordinate, drawn and marked as abscissa using the concentration of every part of standard solution
Directrix curve and the linear equation that the tyrosine is calculated；
The solution to be measured includes base soln, and the base soln is concentration 1 × 10-3Mol/L or concentration 1 × 10-4mol/L
Or concentration 1 × 10-5The tryptophan solution of mol/L, the three-electrode system is using tryptophan as electrochemical signals to solution to be measured
It is detected.
2. detecting the method for tyrosine concentration in solution as described in claim 1, wherein use the three-electrode system, benefit
The solution to be measured containing unknown concentration tyrosine is detected with differential pulse voltammetry, obtains the differential pulse of solution to be measured
Volt-ampere curve, the corresponding potential value of differential pulse voltammetry curve of solution to be measured is molten for the standard of 0mol/L with tyrosine concentration
The difference of the potential value of liquid substitutes into linear equation obtained in step 3, and the concentration of tyrosine in solution to be measured is calculated.
3. detecting the method for tyrosine concentration in solution as described in claim 1, wherein the step 2 specifically includes following
Step:
1) three-electrode system is respectively implanted in five standard solution, stirs 3min at room temperature at 20~30 DEG C, it is quiet
Only 3min；
2) differential pulse map is scanned, setting scanning initial potential is -0.2V, and termination current potential is 1.0V, and current potential increment is
0.004V, Sample Width 0.0167s, amplitude 0.05V, pulse width 0.05V, sensitivity 10-5A/V, waiting time 2s；
3) potential value for measuring and recording five standard solution respectively establishes the differential pulse volt of five standard solution
Pacify curve.
4. detecting the method for tyrosine concentration in solution as described in claim 1, wherein reference electrode is Ag/AgCl electrode,
Auxiliary electrode is platinum electrode.
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| CN104597090A (en) * | 2014-12-10 | 2015-05-06 | 中国科学院烟台海岸带研究所 | Enzyme-free potentiometric glucose sensor and detection method thereof |
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