Electrochemical detection method for high concentration denatured bovine serum albumin
A bovine serum albumin and detection method technology, applied in the field of electrochemical detection of high-concentration denatured bovine serum albumin, can solve the problems of troublesome sample processing, high test cost, and complex data analysis, and achieve high cost and low detection limited and cumbersome operation
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Embodiment 1
[0021] Example 1: Preparation of different concentrations of denatured BSA solutions containing MB
[0022] (1) Use an acidity meter to prepare 0.02mol L of pH 5.0-9.0 -1 Trishydroxymethylaminomethane / 0.1mol·L - 1 NaCl buffer solution, and prepare 10mmol / L MB solution, 10mol / L urea solution and 1mmol / L BSA solution;
[0023] (2) Use a pipette gun to measure 20 μL of MB solution (10 mmol / L) into the sample tube, add different volumes of BSA solution (1 mmol / L) to the sample tube in turn, and then add 1600 μL to each sample tube in turn Urea solution (10mol / L);
[0024] (3) Shake and mix at room temperature to obtain 2000 μL of mixed solution, which contains 100 μmol / L MB and 8 mol / L urea, and the concentrations of BSA are 0, 5, 10, 20, 50 and 100 μmol / L, respectively. Multiple parallel denatured BSA solutions containing MB with different concentrations were stored in the refrigerator and reacted overnight for 15 hours.
Embodiment 2
[0025] Example 2: Linear sweep voltammetry test of different concentrations of denatured BSA solutions containing MB
[0026] The conductive glass ITO electrode cut in advance is immersed in the solution prepared in Example 1 as the working electrode, the titanium sheet is used as the counter electrode, and the saturated calomel electrode is used as the reference electrode, which are respectively connected to 3 electrode joints of the electrochemical workstation, Carry out linear sweep voltammetry test, measure oxidation peak current density linear change curve with denatured BSA solution concentration;
[0027] The result is as figure 1 As shown, with the increase of denatured BSA concentration, the oxidation peak current density of MB increased significantly. Take the oxidation peak current density as the ordinate and the concentration of denatured BSA as the abscissa to draw a standard curve, i.e. pa =23.17+0.41C 变性BSA , there is a good linear relationship between the ox...
Embodiment 3
[0028] Example 3: Practical application of electrochemical detection method based on high-concentration denatured BSA
[0029] To the practical application after the establishment of the method of the linear sweep voltammetry test of the denatured BSA solution of different concentrations containing MB in Example 2, the specific operation steps are as follows:
[0030] The MB solution containing 100 μmol / L denatured BSA prepared in Example 1 was added to 400 μL milk sample, and the linear sweep voltammetry test was carried out according to the linear change curve of the oxidation peak current density of MB with the concentration of denatured BSA established in Example 2 , to measure denatured BSA in milk samples.
[0031] The result is as Figure 4 As shown, the peak current density difference between milk with 100 μmol / L denatured BSA solution and pure milk is 39.14 μA / cm -2 . And as figure 2 As shown in the standard curve, the difference in peak current density between 1...
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