Electrochemical detection method for detecting vitamin B2

Abstract

The invention discloses an electrochemical detection method for vitamin B2 detection, and belongs to the technical field of vitamin detection.The electrochemical detection method comprises the following steps that S1, an electrode is prepared, specifically, a glassy carbon electrode is sequentially polished for 2-3 minutes by adopting 0.05-1 micron of aluminum oxide powder, and then the glassy carbon electrode is sequentially subjected to ultrasonic cleaning for 1-3 minutes by adopting pure water, absolute ethyl alcohol and pure water; a glassy carbon electrode subjected to ultrasonic treatment is cleaned up, a platinum wire electrode is adopted as a counter electrode, a saturated calomel electrode is adopted as a reference electrode, cyclic voltammetry scanning is performed in a 0.01 mol / L-1 mol / L potassium ferricyanide solution, a 0.01 mol / L-1 mol / L potassium chloride solution and a 0.01 mol / L-1 mol / L phosphoric acid buffer salt solution until the electrodes are stable, the initial electrode potential of an instrument is-1.1 V to-0.35 V, the termination electrode potential is 0.55 V to 1.5 V, and the detection wavelength is 250nm. And performing cyclic voltammetry scanning until the electrode is stable to successfully prepare the electrooxidation modified electrode. The method has the advantages that the high-sensitivity detection of the vitamin B12 is realized by preparing the electrooxidation modified electrode and optimizing a reagent formula, an electrochemical method and parameters, and the CV of the detected vitamin B2 is smaller.

Description

Technical Field

[0001] The invention relates to the technical field of vitamin detection, and in particular to an electrochemical detection method for vitamin B2 detection. Background Art

[0002] Vitamin B2, also known as riboflavin, has a relative molecular mass of 376. Riboflavin is a nutrient necessary for maintaining normal metabolism of the body. It participates in biological oxidation and energy metabolism, maintains the integrity of skin and mucous membranes, participates in drug metabolism, anti-oxidation and visual photosensitivity, and affects adrenaline production and red blood cell formation. In addition, it also has diuretic, lipid-lowering and heart function improvement effects. Secondly, it is the main active gene of the flavin coenzyme necessary for the redox process in the body. It can be widely distributed in human tissues and organs in a free or conjugated form and is called "a bridge connecting single-electron reduction and double-electron reduction in the body". Therefore, it is very important to strengthen the detection of vitamin B2 in the human body. The most common detection methods for vitamin B2 are high performance liquid chromatography, fluorescence, ultraviolet spectrophotometry, atomic absorption spectroscopy and electrochemical method. Among them, the electrochemical method is widely used due to its simple operation and simple equipment.

[0003] Many current methods for electrochemical detection of vitamin B2 have problems such as complex electrode preparation and high detection limit. Summary of the invention

[0004] In view of the shortcomings of the prior art, the object of the present invention is to provide an electrochemical detection method for vitamin B2 detection.

[0005] The technical solution of the present invention is as follows: an electrochemical detection method for vitamin B2 detection, characterized in that it comprises the following steps:

[0006] S1. Electrode preparation: Use 0.05um-1um alumina powder to polish the glassy carbon electrode for 2-3 minutes, and then use pure water, anhydrous ethanol, pure water, and ultrasonic cleaning for 1-3 minutes;

[0007] The ultrasonically treated glassy carbon electrode was cleaned, a platinum wire electrode was used as a counter electrode, a saturated calomel electrode was used as a reference electrode, and cyclic voltammetry was performed in 0.01mol / L-1mol / L potassium ferrocyanide, 0.01mol / L-1mol / L potassium chloride, and 0.01mol / L-1mol / L phosphate buffer solution until the electrode was stable. The initial electrode potential of the instrument was -1.1--0.35V, and the termination electrode potential was 0.55-1.5V. The electro-oxidation modified electrode was successfully prepared by cyclic voltammetry scanning until the electrode was stable.

[0008] S2. Reagent formula: (1) Phosphate buffer solution formula: 0.001mol / L-0.9mol / L potassium dihydrogen phosphate solution and 0.001mol / L-0.9mol / L potassium dihydrogen phosphate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a phosphate solution;

[0009] Or the formula of phosphate buffer solution: 0.001mol / L-0.9mol / L sodium dihydrogen phosphate solution and 0.001mol / L-0.9mol / L sodium dihydrogen phosphate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a phosphate solution;

[0010] The dihydrogen phosphate may be potassium dihydrogen phosphate, sodium dihydrogen phosphate, or a mixture of potassium dihydrogen phosphate and sodium dihydrogen phosphate in any molar ratio;

[0011] The dihydrogen phosphate salt may be dipotassium hydrogen phosphate, disodium hydrogen phosphate, or a mixture of dipotassium hydrogen phosphate and disodium hydrogen phosphate in any molar ratio.

[0012] (2) Citrate buffer solution formula: 0.001mol / L-0.9mol / L citric acid solution and

[0013] 0.001mol / L-0.9mol / L sodium citrate solution is mixed in a ratio of 0.1:9.9-9.9:0.1 to form a citrate buffer solution;

[0014] S3. Electrochemical detection method using DPV.

[0015] Preferably, the scanning speed of the instrument is 20-190 mV / s, and the number of scanning cycles is 35-90 times.

[0016] Preferably, the electrochemical parameters are as follows:

[0017] The initial electrode potential of the instrument is -2--1.2V, the termination electrode potential is -0.1--1.5V, the rest time is 2-30s, the potential increment: 0.001-0.05V, the pulse period: 0.01-0.045s, the amplitude: 0.05-0.45V, the enrichment voltage: -0.8--0.2V, and the enrichment time: 30-240s.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] By preparing electro-oxidation modified electrodes, optimizing reagent formulations, electrochemical methods and parameters, the electrochemical performance of vitamin B 12 Highly sensitive detection, smaller CV for detecting vitamin B2. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The disclosure of the present invention is described with reference to the accompanying drawings. It should be understood that the drawings are only for illustrative purposes and are not intended to limit the scope of protection of the present invention. In the drawings, the same reference numerals are used to refer to the same components. Among them:

[0021] Figure 1 A graph showing different concentrations of vitamin B2 tested for the present invention; DETAILED DESCRIPTION

[0022] It is easy to understand that according to the technical solution of the present invention, without changing the essential spirit of the present invention, a person skilled in the art can propose a variety of interchangeable structural modes and implementation modes. Therefore, the following specific implementation modes and drawings are only exemplary descriptions of the technical solution of the present invention, and should not be regarded as the whole of the present invention or as a limitation or restriction to the technical solution of the present invention.

[0023] An electrochemical detection method for vitamin B2 detection, characterized in that it comprises the following steps:

[0024] S1. Electrode preparation: Use 0.05um-1um alumina powder to polish the glassy carbon electrode for 2-3 minutes, and then use pure water, anhydrous ethanol, pure water, and ultrasonic cleaning for 1-3 minutes;

[0025] The ultrasonically treated glassy carbon electrode was cleaned, a platinum wire electrode was used as a counter electrode, a saturated calomel electrode was used as a reference electrode, and cyclic voltammetry was performed in 0.01mol / L-1mol / L potassium ferrocyanide, 0.01mol / L-1mol / L potassium chloride, and 0.01mol / L-1mol / L phosphate buffer solution until the electrode was stable. The initial electrode potential of the instrument was -1.1--0.35V, the scanning speed was 20-190mV / s, the number of scanning circles was 35-90 times, the termination electrode potential was 0.55-1.5V, and the cyclic voltammetry was performed until the electrode was stable, and the electro-oxidation modified electrode was successfully prepared.

[0026] S2. Reagent formula: (1) Phosphate buffer solution formula: 0.001mol / L-0.9mol / L potassium dihydrogen phosphate solution and 0.001mol / L-0.9mol / L potassium dihydrogen phosphate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a phosphate solution;

[0027] Or the formula of phosphate buffer solution: 0.001mol / L-0.9mol / L sodium dihydrogen phosphate solution and 0.001mol / L-0.9mol / L sodium dihydrogen phosphate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a phosphate solution;

[0028] The dihydrogen phosphate may be potassium dihydrogen phosphate, sodium dihydrogen phosphate, or a mixture of potassium dihydrogen phosphate and sodium dihydrogen phosphate in any molar ratio;

[0029] The dihydrogen phosphate salt may be dipotassium hydrogen phosphate, disodium hydrogen phosphate, or a mixture of dipotassium hydrogen phosphate and disodium hydrogen phosphate in any molar ratio.

[0030] (2) Citrate buffer solution formula: 0.001mol / L-0.9mol / L citric acid solution and 0.001mol / L-0.9mol / L sodium citrate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a citrate buffer solution;

[0031] S3. Electrochemical detection method using DPV.

[0032] The initial electrode potential of the instrument is -2--1.2V, the termination electrode potential is -0.1--1.5V, the rest time is 2-30s, the potential increment: 0.001-0.05V, the pulse period: 0.01-0.045s, the amplitude: 0.05-0.45V, the enrichment voltage: -0.8--0.2V, and the enrichment time: 30-240s.

[0033] The following preparation method is adopted:

[0034] (1) Electrode treatment: The glassy carbon electrode was polished with 0.05um alumina powder for 3 minutes, and then ultrasonically cleaned with pure water, anhydrous ethanol, and pure water for 2 minutes. The ultrasonically treated glassy carbon electrode was cleaned, and a platinum wire electrode was used as the counter electrode and a saturated calomel electrode was used as the reference electrode. Cyclic voltammetry was performed in 0.1mol / L potassium ferrocyanide and 0.2mol / L potassium dihydrogen phosphate solution until the electrode was stable. The initial electrode potential of the instrument was -0.55V, the termination electrode potential was 1.1V, the scanning speed was 85mV / s, and the number of scanning cycles was 50. The electro-oxidation modified electrode was successfully prepared when the cyclic voltammetry scan was performed until the electrode was stable.

[0035] (2) Different reagent formula: 0.1 mol / L potassium dihydrogen phosphate solution and 0.1 mol / L potassium hydrogen phosphate solution were mixed in the following proportions to form a phosphate solution.

[0036] Serial number Dipotassium hydrogen phosphate Potassium dihydrogen phosphate PH 1 0 10 4.46 2 0.1 9.9 5.18 3 1 9 5.93 4 3 7 6.61 5 4 6 6.87 6 6 4 7.33 7 7 3 7.54 8 9 1 8.29 9 9.9 0.1 9.19 10 10 0 9.24

[0037] The results of testing 50umol / L of vitamin B2 are as follows:

[0038]

[0039]

[0040] (3) Different DPV parameters: initial electrode potential of the instrument was -1.4 V, final electrode potential was 1.3 V, and rest time was 8 s.

[0041] Potential increment amplitude cycle No. 1 0.012 0.1 0.015 No. 2 0.015 0.15 0.015 No. 3 0.02 0.2 0.015 No. 4 0.012 0.15 0.02 No. 5 0.015 0.2 0.02 No. 6 0.02 0.1 0.02 No.7 0.012 0.2 0.025 No. 8 0.015 0.1 0.025 No. 9 0.02 0.15 0.025

[0042] The results of testing 50umol / L of vitamin B2 are as follows:

[0043] Serial number Current 1 Current 2 Average SD CV 1 120200 122600 121400 1697.056275 1.40％ 2 159100 147200 153150 8414.570696 5.49％ 3 206100 216000 211050 7000.357134 3.32％ 4 152800 165500 159150 8980.256121 5.64％ 5 199100 165100 182100 24041.63056 13.20％ 6 100300 83620 91960 11794.54111 12.83％ 7 38470 44120 41295 3995.153314 9.67％ 8 128900 133500 131200 3252.691193 2.48％ 9 142900 147100 145000 2969.848481 2.05％

[0044] (4) The following buffer solution formula is used: 0.05 mol / L potassium dihydrogen phosphate solution and 0.05 mol / L potassium hydrogen phosphate solution are mixed in a ratio of 6:4 to form a phosphate solution.

[0045] DPV parameters: instrument initial electrode potential -1.4V, termination electrode potential 0.3V, rest time 5s, enrichment voltage: -0.85V, enrichment time: 165s, potential increment: 0.08V, pulse period: 0.05s, amplitude: 0.35V.

[0046] Different concentrations of vitamin B2 were tested as follows:

[0047]

[0048] The technical scope of the present invention is not limited to the contents in the above description. Those skilled in the art can make various deformations and modifications to the above embodiments without departing from the technical idea of ​​the present invention, and these deformations and modifications should all fall within the protection scope of the present invention.

Claims

1. An electrochemical detection method for vitamin B2 detection, characterized in that: The following steps are involved: S1. Electrode preparation: Use 0.05um-1um alumina powder to polish the glassy carbon electrode for 2-3 minutes, and then use pure water, anhydrous ethanol, pure water, and ultrasonic cleaning for 1-3 minutes; The ultrasonically treated glassy carbon electrode was cleaned, a platinum wire electrode was used as a counter electrode, a saturated calomel electrode was used as a reference electrode, and cyclic voltammetry was performed in 0.01mol / L-1mol / L potassium ferrocyanide, 0.01mol / L-1mol / L potassium chloride, and 0.01mol / L-1mol / L phosphate buffer solution until the electrode was stable. The initial electrode potential of the instrument was -1.1--0.35V, and the termination electrode potential was 0.55-1.5V. The electro-oxidation modified electrode was successfully prepared by cyclic voltammetry scanning until the electrode was stable. S2. Reagent formula: (1) Phosphate buffer solution formula: 0.001mol / L-0.9mol / L potassium dihydrogen phosphate solution and 0.001mol / L-0.9mol / L potassium dihydrogen phosphate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a phosphate solution; Or the formula of phosphate buffer solution: 0.001mol / L-0.9mol / L sodium dihydrogen phosphate solution and 0.001mol / L-0.9mol / L sodium dihydrogen phosphate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a phosphate solution; The dihydrogen phosphate may be potassium dihydrogen phosphate, sodium dihydrogen phosphate, or a mixture of potassium dihydrogen phosphate and sodium dihydrogen phosphate in any molar ratio; The dihydrogen phosphate salt may be dipotassium hydrogen phosphate, disodium hydrogen phosphate, or a mixture of dipotassium hydrogen phosphate and disodium hydrogen phosphate in any molar ratio. (2) Citrate buffer solution formula: 0.001mol / L-0.9mol / L citric acid solution and 0.001mol / L-0.9mol / L sodium citrate solution are mixed in a ratio of 0.1:9.9-9.9:0.1 to form a citrate buffer solution; S3. Electrochemical detection method using DPV.

2. An electrochemical detection method for vitamin B2 detection according to claim 1, characterized in that: The scanning speed of the instrument is 20-190 mV / s, and the number of scanning cycles is 35-90 times.

3. An electrochemical detection method for vitamin B2 detection according to claim 1, characterized in that: The electrochemical parameters are as follows: The initial electrode potential of the instrument is -2--1.2V, the termination electrode potential is -0.1--1.5V, the rest time is 2-30s, the potential increment: 0.001-0.05V, the pulse period: 0.01-0.045s, the amplitude: 0.05-0.45V, the enrichment voltage: -0.8--0.2V, and the enrichment time: 30-240s.

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