A kind of tyrosinase electrochemical biosensor and its application
A biosensor, tyrosinase technology, applied in the field of tyrosinase electrochemical biosensor, to achieve ultra-high selectivity, improve sensitivity and detection limit, and enhance the effect of electron transfer
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Embodiment 1
[0024] Example 1. Preparation of tyrosinase electrochemical biosensor
[0025] Glassy carbon electrode assembly steps:
[0026] (1) Polish the surface of the glassy carbon electrode with aluminum oxide powders with particle sizes of 1, 0.3, and 0.05 μm, and then repeatedly ultrasonically clean it in absolute ethanol and deionized water, and then dry the electrode surface with high-purity nitrogen stand-by. Put in 1mmolL -1 The cyclic voltammetry curve was scanned between -0.1 and +0.6V in the potassium ferricyanide / potassium ferrocyanide solution (molar ratio 1:1). The redox peak potential difference of the curve is less than 75mV, indicating that the redox reaction on the surface of the glassy carbon electrode is a completely reversible reaction, and the electrode polishing is in good condition, and the next experiment can be carried out.
[0027] (2) Construction of mesoporous carbon-cobalt tetroxide-tyrosinase composite material. The mesoporous carbon material was added...
Embodiment 2
[0030] Example 2. Standard solution for detection of phenolic compounds by tyrosinase electrochemical biosensor
[0031] Dissolve catechol, phenol, p-cresol, m-cresol and p-chlorophenol in water respectively to prepare a standard solution with a concentration of 1 mmol / L.
[0032]A certain volume (8μL) of catechol standard solution was continuously added dropwise (once every 40s) to the detection solution 8ml PBS of the three-electrode system composed of tyrosinase-modified glassy carbon electrodes, and the working potential was -0.1V Under the situation of constant potential scanning, the obtained current-time (I-t) relationship curve, because the concentration of catechol in the detection solution can be known at a fixed time point in the I-t curve, the correlation between the current intensity and the concentration of catechol can be obtained curve. Catechol is a natural substrate of tyrosinase. In the presence of oxygen, tyrosinase catalyzes catechol to generate o-quinone...
Embodiment 3
[0034] Example 3. Detection of interfering substances in tyrosinase electrochemical biosensors
[0035] Dissolve common interfering substances such as uric acid, glucose, and hydrogen peroxide in water, and prepare an aqueous solution with a certain concentration. When the concentration of catechol is 2 μM, add uric acid, glucose, and hydrogen peroxide aqueous solution dropwise to the detection solution. , scan the I-t curve, and obtain the electrical signal responses of the above three interfering substances. The results proved that the uric acid concentration of 40μM, the glucose concentration of 2mM and the hydrogen peroxide concentration of 0.1mM did not affect the detection of catechol by the tyrosinase electrochemical biosensor. Use the same detection process to detect inorganic salt ions (concentration is 2mM K + 、Na + , NO 3 - 、H 2 PO 4 - 、HPO 4 2- , Cl - 、Ac - Ions) and organic solvents (acetone, acetonitrile, ethanol) have carried out the interference test...
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