Enzyme-colorimetric method for detecting residual hydrogen peroxide in foods
A technology of hydrogen peroxide and peroxidase, which is applied in the direction of material analysis by observing the influence on chemical indicators, and analysis by making materials undergo chemical reactions, which can solve the problems of complex sample pretreatment and long detection time
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Embodiment 1
[0021] Accurately weigh the calibrated hydrogen peroxide solution to prepare a 1000mg / L standard solution. Prepare a 100mg / L standard intermediate solution. Prepare 20 mL of hydrogen peroxide standard solution containing 0, 1, 2, 5, 10, and 20 mg / L respectively, transfer it to a 25 mL colorimetric tube, add 2 mL of peroxidase reagent solution, and keep the temperature in a water bath at 37 ° C for 20 min. Cool to room temperature. Dilute to 25 mL with secondary water, and measure the absorbance of the resulting solution at a wavelength of 470 nm. Take the absorbance as the Y axis and the hydrogen peroxide concentration as the X axis to make a standard curve to obtain the standard curve regression equation.
Embodiment 2
[0023] Analysis steps: pipette 20mL of fresh milk in a bag of a certain brand into a 50mL volumetric flask, add 5mL of zinc sulfate solution and 5mL of potassium ferrocyanide solution in sequence, mix well, add water to make up to 50mL, let stand for 20min, and filter with filter paper to obtain Filtrate, then adjust the pH value to 7.6 with 0.01% sodium hydroxide solution, filter through filter paper, the filtrate is the test solution, pipette 20mL test solution into a 25mL volumetric flask, add 2mL peroxidase reagent solution, shake well, Constant temperature in ℃ water bath for 20min, the solution was light red, cooled to room temperature, fixed to 25mL, measured the absorbance at 470nm, according to the standard curve equation of hydrogen peroxide in Example 1, calculated that the sample contained 5.78mg / L of hydrogen peroxide . The quantification limit of this method is 2.0mg / L.
Embodiment 3
[0025] Analysis steps: Take beef louvers from a bazaar, cut them into small pieces, crush them with a food grinder, weigh 10.0g (accurate to 0.001g) into a 50mL volumetric flask, add 5mL zinc sulfate solution and 5mL ferrocyanide in turn Potassium chloride solution, mix well, add water to 50mL, let stand for 20min, filter paper to obtain filtrate, then adjust pH value to 7.6 with 0.01% sodium hydroxide solution, filter paper, filtrate is test solution, pipette 20mL test solution to Into a 25mL volumetric flask, add 2mL of peroxidase reagent solution, shake well, keep the temperature in a 37°C water bath for 20min, the solution turns pink, cool to room temperature, set the volume to 25mL, measure the absorbance at 470nm, according to Example 1 The standard curve equation of hydrogen peroxide is calculated to contain hydrogen peroxide 9.54mg / kg in this sample. The limit of quantification of this method is 5.0 mg / kg.
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