Method for rapidly extracting and detecting content of tyramine in cheese
A technology of cheese and tyramide, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome and time-consuming steps such as hollow fiber membrane infiltration and sealing organic solvent removal, and is not conducive to the development of the field of rapid food detection. Processing time and detection time, reduced matrix effects, simple effects of preparation
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Embodiment 1
[0058] The enrichment process of tyramine by the electric membrane extraction device includes:
[0059] Step 1. Add 10mL of 5% (mass concentration) trichloroacetic acid and 30mL of 0.1mol / L hydrochloric acid solution into a 50mL centrifuge tube respectively, and add 0.2mg of tyramine to obtain a tyramine solution with a mass concentration of 5mg / L.
[0060] Step 2, take 30mL 5mg / L tyramine solution as the donor phase solution, 0.6mL10% (mass concentration) trichloroacetic acid as the receiving phase solution, select a polyacrylamide gel film with a thickness of 5mm for 45V, 5min electric film Extraction experiments to obtain extracts;
[0061] Step 3: Take 0.1 mL of the extract, add 5 μL of monoamine oxidase, 50 μL of horseradish peroxidase and 100 μL of chromogenic solution, mix well, and react for 15 minutes to obtain the solution to be tested;
[0062] Step 5: Measure the absorbance value at 550 nm of the solution to be tested, and calculate the tyramine content in the ext...
Embodiment 2
[0067] A rapid extraction and quantitative detection method for tyramine in cheese, comprising the following steps:
[0068] Step 1. Take 5g of shredded cheese in a 50mL centrifuge tube, add 10mL of 5% (mass concentration) trichloroacetic acid, then add 30mL of 0.1mol / L hydrochloric acid solution, and mix to obtain a sample suspension;
[0069] Step 2. Ultrasonicate the sample suspension in an ultrasonic water bath for 10 minutes, centrifuge at 9000 rpm for 5 minutes, and take the supernatant and filter it through a 0.45 μm suction filter membrane to obtain the filtrate;
[0070] Step 3, use an electric membrane extraction device to extract the filtrate, take 30mL of the filtrate as the donor phase solution, and 0.6mL of 10% (mass concentration) trichloroacetic acid as the receiving phase solution, select a polyacrylamide gel membrane with a thickness of 5mm for 45V, 5min electro-membrane extraction experiment to obtain the extract;
[0071] Step 4: Take 0.1 mL of the extract...
Embodiment 3
[0082] A rapid extraction and quantitative detection method for tyramine in cheese, comprising the following steps:
[0083] Step 1. According to the steps described in Example 1, a polyacrylamide gel film with a thickness of 4mm was obtained, and after the electromembrane extraction experiment of 35V and 5min, the enrichment factor R of the extract was 12.5;
[0084] Step 2: Put 5g of shredded cheese into a 50mL centrifuge tube, add 10mL of 5% (mass concentration) trichloroacetic acid, then add 30mL of 0.1mol / L hydrochloric acid solution, mix well to obtain a sample suspension; ultrasonicate in an ultrasonic water bath for 10min , centrifuged at 9000rpm for 5min, and the supernatant was filtered through a 0.45μm suction membrane to obtain the filtrate;
[0085] Step 3, use an electric membrane extraction device to extract the filtrate, take 30mL of the filtrate as the donor phase solution, 0.6mL10% (mass concentration) trichloroacetic acid as the receiving phase solution, sel...
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