CN112557471B - Preparation method and detection method of modified electrode capable of simultaneously detecting vitamins B6 and B9 - Google Patents
Preparation method and detection method of modified electrode capable of simultaneously detecting vitamins B6 and B9 Download PDFInfo
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- CN112557471B CN112557471B CN202011532099.1A CN202011532099A CN112557471B CN 112557471 B CN112557471 B CN 112557471B CN 202011532099 A CN202011532099 A CN 202011532099A CN 112557471 B CN112557471 B CN 112557471B
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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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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- 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
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
Abstract
The invention belongs to the technical field of vitamin detection, and in particular relates to a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9, which comprises the following steps: a glassy carbon electrode, a gold electrode or a copper electrode is selected as a matrix electrode, and the surface of the matrix electrode is polished and cleaned for later use; dripping or electroplating a modification solution on the surface of the substrate electrode prepared in the step (1), and drying by using an infrared lamp to prepare the modification electrode; wherein the modification solution consists of one or more of sulfuric acid, graphene, carbon nano tubes, vitamin B12, acetic acid, citric acid, gold nano particles, silver nitrate, sodium sulfate, sodium acetate, potassium chloride and amino black 10B, and the concentration range of each component is 0.0002-3 mol/L. The modified electrode can be used for simultaneously detecting the vitamins B6 and B9, has higher sensitivity, ensures that the peaks of the two vitamins are not interfered with each other, has good reproducibility and can greatly reduce the detection limit.
Description
Technical Field
The invention belongs to the technical field of vitamin detection, and particularly relates to a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9.
Background
With the progress of society, people are paying more attention to self health. As vitamins with important physiological functions of the organism, their rational application, therapeutic and diagnostic monitoring are also becoming more and more important. Wherein, vitamin B6 is also called pyridoxine, which comprises pyridoxine, pyridoxal and pyridoxamine, exists in the form of phosphate in vivo, and vitamin B6 is a constituent component of certain coenzymes in human bodies, and participates in various metabolic reactions, especially has close relation with amino acid metabolism. Vitamin B6 functions include antibody synthesis, gastric acid production in the digestive system, fat and protein utilization (especially for weight loss, supplementation), and sodium/potassium balance maintenance (nervous system stabilization). General deficiency of vitamin B6 may be caused by inappetence, low food utilization, weightlessness, emesis, diarrhea, etc. Severe deficiency may have acne, anemia, arthritis, cramps in children, depression, headache, hair loss, susceptibility to inflammation, learning disability, weakness, etc. However, excessive vitamin B6 can cause somnolence, and can form addiction after long-term administration.
Vitamin B9, also known as folic acid, is a water-soluble vitamin with the effect of promoting the maturation of young cells in bone marrow, and human beings such as deficiency of folic acid can cause megaloblastic anemia and leukopenia, and is particularly important for pregnant women. Folic acid can maintain growth and intestinal function, and pregnant women can frequently supplement folic acid to prevent congenital malformations such as premature birth of newborn infants and cleft palate of infants. At the same time, folic acid has anti-tumor and anticancer effects, and also has effects of promoting the development of nerve cells and brain cells of infants. Deficiency of folic acid can cause fetal nerve tube deformity, cause hyperhomocysteinemia and increase the risk of cardiovascular disease; excessive amounts can induce convulsions in the patient, affecting zinc absorption and resulting in zinc deficiency, which can lead to damage to the nervous system.
Therefore, the quantitative detection of the vitamins B6 and B9 has great clinical significance. The existing methods for detecting the vitamin B6 and the vitamin B9 are different and mainly comprise a gravimetric method, a titration method, a fluorescence analysis method, a high performance liquid chromatography method, an electrochemical method and the like. The weight method and the titration method are complex in operation, the measurement error of low-content components is large, the large error can be caused if the end point judgment is not sharp, and the vitamins B6 and B9 can not be detected simultaneously; the pretreatment process of the spectrometry and the chromatography is too tedious and time-consuming, uses more organic solvents, and requires more blood samples; the electrochemical method is sensitive and quick, has little need for samples, does not need pretreatment, and is particularly suitable for detecting vitamins in biological samples.
The existing electrochemical analysis technology for detecting vitamins mainly comprises the traditional linear sweep voltammetry, the conventional pulse voltammetry, the differential pulse voltammetry, the square wave voltammetry, the stripping voltammetry and the like, and the methods can be used for detecting the vitamins, but have lower sensitivity, and particularly when the vitamins B6 and B9 in a blood sample are detected simultaneously, the detection limit is higher, the peaks of the two substances are not separated, and the concentration level of the vitamins B6 and B9 in the blood sample cannot be reached.
Disclosure of Invention
The invention aims to provide a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9, which overcomes the defects of the prior art, and the modified electrode can be used for simultaneously detecting the vitamins B6 and B9, has higher sensitivity, has good reproducibility and can greatly reduce the detection limit, and the peaks of the two vitamins are not interfered with each other.
In order to solve the problems, the technical scheme adopted by the invention is as follows:
a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9 comprises the following steps:
(1) Pretreatment of a matrix electrode: a glassy carbon electrode, a gold electrode or a copper electrode is selected as a matrix electrode, and the surface of the matrix electrode is polished and cleaned for later use;
(3) Preparing a modified electrode: dripping or electroplating a modification solution on the surface of the substrate electrode prepared in the step (1), and drying by using an infrared lamp to prepare the modification electrode;
wherein the modification solution consists of one or more of sulfuric acid, graphene, carbon nano tubes, vitamin B12, acetic acid, citric acid, gold nano particles, silver nitrate, sodium sulfate, sodium acetate, potassium chloride and amino black 10B, and the concentration range of each component is 0.0002-3 mol/L.
Further, the polishing cleaning treatment of the base electrode specifically includes: firstly, polishing the substrate electrode on metallographic sand paper of different types to enable the surface of the substrate electrode to be smooth like a mirror surface; then sequentially carrying out mirror polishing on the polished substrate electrode on alumina polishing powder with the particle diameters of 1.0 mu m,0.3 mu m and 0.05 mu m; and finally, respectively placing the polished substrate electrode in ethanol and deionized water for 2 minutes by ultrasonic treatment to remove residual organic matters and adsorbed alumina powder on the surface of the substrate electrode, and drying the substrate electrode by nitrogen for later use.
Further, the method of electroplating in the step (2) is selected from one of cyclic voltammetry or potential step method.
Further, the modification solution dripped on the surface of the substrate electrode in the step (2) is 5-800 microliters.
The invention also provides a detection method of the modified electrode capable of simultaneously detecting the vitamins B6 and B9, which comprises the following steps:
taking a blood sample to be tested and reacting with vitamin B6 and B9 releasing agents to convert the vitamins B6 and B9 in a combined state in the blood sample to be tested into vitamins B6 and B9 in a free state;
the optimized modified electrode is selected as a working electrode, and the oxidation-reduction current generated on the working electrode by vitamins B6 and B9 in a blood sample to be detected is detected simultaneously by using a convolution current voltammetry; the two vitamins respectively show peaks at specific peak potential, and the peak type is very good, so that the peak current corresponding to each vitamin can be conveniently measured, and the two peaks are not interfered with each other;
the current signal values of vitamin B6 and vitamin B9 standard samples with different concentrations are respectively compared, a standard curve is manufactured with the corresponding vitamin B6 and vitamin B9 standard liquid concentrations, and the contents of vitamin B6 and vitamin B9 in the blood sample to be measured are respectively calculated through a linear equation according to the current signal values generated by the sample to be measured under the same conditions.
Further, when the content of vitamin B6 and B9 in the blood sample is detected by a convolution current method, the potential of an initial electrode of instrument parameters is-0.4-0.2V, the potential of a termination electrode is 0.8-1.4V, the scanning speed is 30-200 mV/s, the working mode is a differential mode of 0.5 times, 1.5 times or 2.5 times, and the noise filtering is a low-pass mode; the vitamin B6 and B9 releasing agent consists of a solvent and an inert supporting electrolyte; the solvent is one or more aqueous solutions of sodium hydroxide, potassium hydroxide, citric acid, hydrochloric acid, acetic acid, phosphoric acid and boric acid, the concentration of the solvent is 0.00005-0.1 mol/L, the inert supporting electrolyte is one or more of sodium acetate, potassium chloride, sodium citrate, dipotassium hydrogen phosphate, sodium sulfate, disodium hydrogen phosphate and sodium dihydrogen phosphate, and the concentration of the inert supporting electrolyte is 0.001-5 mol/L.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention is modified on the basis of the existing glassy carbon electrode, gold electrode or copper electrode, not only can detect the vitamins B6 and B9 simultaneously, but also has higher sensitivity, and the peaks of the two vitamins are not interfered with each other, so that the invention has good reproducibility and can greatly reduce the detection limit.
2. The invention adopts the convolution current voltammetry to simultaneously measure the contents of vitamin B6 and B9 in the blood sample, has the advantages of high sensitivity, good reproducibility, more convenient and rapid operation, shorter detection time and the like, and can simultaneously and accurately detect the contents of vitamin B6 and B9 in the blood sample in the same system.
3. The method adopts the standard addition method to simultaneously determine the contents of the vitamins B6 and B9 in the sample, eliminates the interference caused by the matrix effect of the sample, reduces the detection limit of the vitamins B6 and B9, and has accurate and reliable measurement result.
Drawings
FIG. 1 is a linear equation corresponding to vitamin B6 and vitamin B9 detected using the modified electrode prepared in example 1.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
The embodiment discloses a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9, which comprises the following steps:
(1) Pretreatment of a matrix electrode: a glassy carbon electrode, a gold electrode or a copper electrode is selected as a matrix electrode, and the surface of the matrix electrode is polished and cleaned for later use;
(2) Preparing a modified electrode: dripping or electroplating a modification solution on the surface of the substrate electrode prepared in the step (1), and drying by using an infrared lamp to prepare the modification electrode; the modification solution was prepared by mixing 0.0002mol/L sulfuric acid, 0.001mol/L carbon nanotubes, and 0.005mol/L potassium chloride.
Wherein the electroplating method in the step (2) is selected from one of cyclic voltammetry or potential step method; the modification solution dripped on the surface of the substrate electrode in the step (2) is 5-800 microliters.
Example 2
The embodiment discloses a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9, which comprises the following steps:
(1) Pretreatment of a matrix electrode: a glassy carbon electrode, a gold electrode or a copper electrode is selected as a matrix electrode, and the surface of the matrix electrode is polished and cleaned for later use;
(2) Preparing a modified electrode: dripping or electroplating a modification solution on the surface of the substrate electrode prepared in the step (1), and drying by using an infrared lamp to prepare the modification electrode; the modification solution was prepared by mixing 0.5mol/L acetic acid, 0.03mol/L gold nanoparticles, and 0.2mol/L sodium acetate.
Wherein the electroplating method in the step (2) is selected from one of cyclic voltammetry or potential step method; the modification solution dripped on the surface of the substrate electrode in the step (2) is 5-800 microliters.
Example 3
The embodiment discloses a preparation method of a modified electrode capable of simultaneously detecting vitamins B6 and B9, which comprises the following steps:
(1) Pretreatment of a matrix electrode: a glassy carbon electrode, a gold electrode or a copper electrode is selected as a matrix electrode, and the surface of the matrix electrode is polished and cleaned for later use;
(2) Preparing a modified electrode: dripping or electroplating a modification solution on the surface of the substrate electrode prepared in the step (1), and drying by using an infrared lamp to prepare the modification electrode; the modification solution was prepared by mixing 1mol/L of citric acid, 3mol/L of sodium sulfate and 0.5mol/L of amino black 10B.
Wherein the electroplating method in the step (2) is selected from one of cyclic voltammetry or potential step method; the modification solution dripped on the surface of the substrate electrode in the step (2) is 5-800 microliters.
Detection method
1. Simultaneous measurement of vitamin B6 and B9 contents in blood samples by single standard addition method
The specific implementation process comprises the following steps:
1. pretreatment of vitamin B6 and B9 modified electrode: the commercial glass carbon electrode, gold electrode or copper electrode with the diameter of 2mm is selected as a vitamin B6 and B9 sensor probe, and the electrodes are polished on metallographic sand paper with different types to ensure that the surfaces of the electrodes are smooth like mirror surfaces. The polished electrode was then mirror polished on alumina polishing powders having particle diameters of 1.0 μm,0.3 μm and 0.05 μm in this order. Finally, respectively placing the polished electrodes in ethanol and deionized water for 2 minutes by ultrasonic treatment to remove organic matters remained on the surfaces of the electrodes and adsorbed alumina powder, and drying the electrodes by nitrogen for later use.
2. Preparation of vitamin B6 and B9 modified electrode: the treated electrode was used as a working electrode, a platinum wire electrode was used as an auxiliary electrode, and a silver/silver chloride electrode was used as a reference electrode, and in the modification solution prepared in example 1, a potential step method was used to modify the vitamin B6, B9 sensor probes. Instrument parameter setting: the potential of the initial electrode is-0.6-0.4V (relative to the reference electrode), the potential of the final electrode is 0.8-2V (relative to the reference electrode), the scanning speed is 20-200 mV/s, the cyclic scanning is carried out for 6-15 circles under the stirring condition, and then deionized water is used for cleaning, and nitrogen is used for drying for standby.
3. Preparation of vitamin B6 and B9 releasing agent: and (3) taking a proper amount (100-1000 mL) of the single solvent or the mixed solvent in a volumetric flask, weighing the single or composite supporting electrolyte according to a proportion, adding the single or composite supporting electrolyte into the volumetric flask, covering a bottle stopper, slightly shaking, adding the same solvent into the scale mark of the volumetric flask after the supporting electrolyte is dissolved, and ensuring that the concentration of the supporting electrolyte in the solution is 0.001-5 mol/L.
4. Release of vitamins B6, B9 in blood samples: quantitatively transferring a blood sample (10-20 mu L) into a 5.0mL test tube by using a micropipette, quantitatively adding 0.1-2.0 mL of vitamin B6 and B9 specific release agent, covering a test tube plug, slightly shaking, standing for 5 minutes, and changing the vitamins B6 and B9 combined in the blood sample into free micromolecules after releasing, so that the measurement can be carried out on the machine.
5. Simultaneous determination of vitamins B6, B9 in blood samples:
firstly, a test tube containing a sample solution to be detected is fixed on a workbench of a vitamin detector, the preferable vitamin B6 and B9 modified electrodes are used as working electrodes, a platinum wire electrode is used as an auxiliary electrode, and a silver/silver chloride electrode is used as a reference electrode and is inserted into the sample solution to be detected together. The vitamin detector operating mode can be selectively set to a differential mode of 0.5 times, 1.5 times or 2.5 times, and the noise is filtered to a low-pass mode. The test parameters are as follows: the instrument parameter initial electrode potential is-0.4-0.2V, the ending electrode potential is 0.8-1.4V, the scanning speed is 30-200 mV/s, and the convolution current obtained by respectively recording and detecting the vitamin B6 and the vitamin B9 in the sample solution is recorded as h.
And then quantitatively transferring the standard solution Vs mL of the vitamins B6 and B9 by using a micropipette, adding the standard solution Vs mL into a sample solution, uniformly stirring, detecting under the same condition, and respectively obtaining convolution currents of the vitamins B6 and B9 and marking the convolution currents as H.
6. Calculation of vitamin B6, B9 concentration in blood samples:
the concentrations of vitamin B2 and vitamin C in the blood sample are calculated as follows (1):
c in the formula A For blood samplesThe concentrations of the vitamins B6 and B9 are H, the convolution current values obtained by detecting the vitamins B6 and B9 in the sample solution, H, the convolution current values obtained by detecting after adding the standard solution, vx, vs, cs, k and the dilution multiple of the sample, wherein Vx is the volume of the diluent, cs is the volume of the added standard solution. The following requirements should be satisfied for the volumes and concentrations of the added vitamin B6, B9 standard solutions:
Vs<100Vx，cs>100cA
2. multi-standard addition method for measuring vitamin B6 and B9 content in blood sample
The specific implementation process comprises the following steps:
1. pretreatment of vitamin B6 and B9 modified electrode: as in example 1.
2. Preparation of vitamin B6 and B9 modified electrode: as in example 1.
3. Preparation of vitamin B6 and B9 releasing agent: as in example 1.
4. Release of vitamins B6, B9 in blood samples: as in example 1.
5. Determination of vitamins B6, B9 in blood samples:
firstly, a test tube containing a sample solution to be detected is fixed on a workbench of a vitamin detector, modified vitamin B6 and B9 sensors are used as working electrodes, a platinum wire electrode is used as an auxiliary electrode, and a silver/silver chloride electrode is used as a reference electrode and is inserted into the sample solution to be detected together. The vitamin detector operating mode can be selectively set to a differential mode of 0.5 times, 1.5 times or 2.5 times, and the noise is filtered to a low-pass mode. The test parameters are as follows: the test parameters are as follows: the instrument parameter initial electrode potential is-0.4-0.2V, the ending electrode potential is 0.8-1.4V, the scanning speed is 30-200 mV/s, and the convolution current for detecting the vitamin B6 and the vitamin B9 in the sample solution is recorded as h0.
And then quantitatively transferring the standard solutions V1mL, V2mL, V3mL, … and VnmL of the vitamins B6 and B9 by using a micropipette in sequence, respectively adding the standard solutions into the sample solutions, sequentially detecting under the same conditions, and respectively marking the obtained convolution currents as h1, h2, h3, … and hn, wherein the concentrations of the vitamins B6 and B9 which are correspondingly added are c1, c2, c3, … and cn (the convolution currents can be obtained by utilizing the volume conversion of the standard solutions which are added each time). FIG. 1 shows the linear equations corresponding to vitamin B6 and vitamin B9 detected using the modified electrode prepared in example 1.
6. Calculation of vitamin B6, B9 concentration in blood samples:
a least squares linear fit (concentration c0=0 of vitamins B6, B9 of the sample solution without standard addition) was performed using the convolution current and concentration, respectively, resulting in the following linear relationship:
h＝kc+B (2)
thus, the concentration of vitamin B6 and B9 in the blood sample can be obtained
c A ＝-B (3)
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (3)
1. A detection method of a modified electrode capable of simultaneously detecting vitamins B6 and B9 is characterized by comprising the following steps: the method comprises the following steps:
(1) Preparation of modified electrode
1) Pretreatment of a matrix electrode: a glassy carbon electrode, a gold electrode or a copper electrode is selected as a matrix electrode, and the surface of the matrix electrode is polished and cleaned for later use;
2) Preparing a modified electrode: the electroplating modification solution of the substrate electrode prepared in the step 1) is dried by an infrared lamp to prepare the modification electrode; using the treated electrode as a working electrode, a platinum wire electrode as an auxiliary electrode and a silver/silver chloride electrode as a reference electrode, and modifying the vitamin B6 and B9 sensor probes in the prepared modification solution by adopting a potential step method;
instrument parameter setting: the potential of the initial electrode is-0.6-0.4V, the potential of the final electrode is 0.8-2V, the scanning speed is 20-200 mV/s, the cyclic scanning is performed for 6-15 circles under the stirring condition, and then deionized water is used for cleaning, and nitrogen is used for drying for standby;
the modification solution consists of one or more of sulfuric acid, graphene, carbon nanotubes, vitamin B12, acetic acid, citric acid, gold nanoparticles, silver nitrate, sodium sulfate, sodium acetate, potassium chloride and amino black 10B, wherein the concentration range of each component is 0.0002-3 mol/L;
(2) Detection by modified electrode
The vitamin B6, B9 releasing agent is configured to: the vitamin B6 and B9 releasing agent consists of a solvent and an inert supporting electrolyte; the solvent is one or more aqueous solutions of sodium hydroxide, potassium hydroxide, citric acid, hydrochloric acid, acetic acid, phosphoric acid and boric acid, and the concentration of the solvent is 0.00005-0.1 mol/L; the inert supporting electrolyte is one or more of sodium acetate, potassium chloride, sodium citrate, dipotassium hydrogen phosphate, sodium sulfate, disodium hydrogen phosphate and sodium dihydrogen phosphate; the concentration of the inert supporting electrolyte is 0.001-5 mol/L; weighing 100-1000 mL of the single solvent or the mixed solvent in a volumetric flask, proportionally weighing the single or composite supporting electrolyte, adding the single or composite supporting electrolyte into the volumetric flask, covering a bottle stopper, slightly shaking, adding the same solvent into the scale mark of the volumetric flask after the supporting electrolyte is dissolved, and keeping the concentration of the supporting electrolyte in the solution at 0.001-5 mol/L;
quantitatively transferring 10-20 mu L of blood sample into a 5.0mL test tube by using a micropipette, quantitatively adding 0.1-2.0 mL of vitamin B6 and B9 specific release agent, covering a test tube plug, slightly shaking uniformly, standing for 5 minutes, and changing the vitamins B6 and B9 combined in the blood sample into small molecules in a free state after releasing, so that the small molecules can be measured on a machine;
simultaneously detecting oxidation-reduction current generated by vitamins B6 and B9 on a working electrode in a blood sample to be detected by using a convolution current voltammetry; the two vitamins respectively show peaks at specific peak potential, and the peak type is very good, so that the peak current corresponding to each vitamin can be conveniently measured, and the two peaks are not interfered with each other;
by respectively comparing the current signal values of the vitamin B6 and B9 standard samples with different concentrations and preparing standard curves with the corresponding vitamin B6 and B9 standard liquid concentrations, according to the current signal values generated by the sample to be tested under the same conditions,
calculating the contents of vitamin B6 and vitamin B9 in the blood sample to be measured through a linear equation respectively;
when the vitamin B6 and B9 contents in the blood sample are detected by a convolution current method, the instrument parameter initial electrode potential is-0.4-0.2V, the instrument parameter final electrode potential is 0.8-1.4V, the scanning speed is 30-200 mV/s, the working mode is a differential mode of 0.5 times, 1.5 times or 2.5 times, and the noise filtering is a low-pass mode.
2. The method for detecting a modified electrode capable of simultaneously detecting vitamins B6 and B9 according to claim 1, wherein: the polishing and cleaning treatment of the substrate electrode specifically comprises the following steps: firstly, polishing the substrate electrode on metallographic sand paper of different types to enable the surface of the substrate electrode to be smooth like a mirror surface; then sequentially carrying out mirror polishing on the polished substrate electrode on alumina polishing powder with the particle diameters of 1.0 mu m,0.3 mu m and 0.05 mu m; and finally, respectively placing the polished substrate electrode in ethanol and deionized water for 2 minutes by ultrasonic treatment to remove residual organic matters and adsorbed alumina powder on the surface of the substrate electrode, and drying the substrate electrode by nitrogen for later use.
3. The method for detecting a modified electrode capable of simultaneously detecting vitamins B6 and B9 according to claim 1, wherein: and (2) dripping 5-800 microlitres of the modification solution on the surface of the substrate electrode in the step (2).
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