Rapid quantitative detection method for peroxidase in pork
A quantitative detection method and peroxidase technology, applied in the field of food hygiene and safety, can solve the problems of inability to provide maps and quantitative data, poor scientificity and accuracy, and complex inspection procedures, and achieve a simple and easy detection method. Effect
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Embodiment 1
[0027] Example 1: Establishment of a standard curve of horseradish peroxidase concentration-OD value
[0028] Take horseradish peroxidase and prepare horseradish peroxidase standard solutions with concentrations of 0.001, 0.002, 0.004, 0.008, 0.016, 0.032, 0.064, 0.128 μg / mL respectively;
[0029] In the cuvette, add 2mL of sodium dihydrogen phosphate buffer (1g / mL, pH4.5), 0.2mL of carbamide peroxide (0.94g / 100mL), and 0.2mL of horseradish peroxidase standard solution, 3,3 After 0.2mL of',5,5'-tetramethylbenzidine (TMD) (0.15g / 100mL), react at 25°C for 60s, and then add 0.1mL of dilute sulfuric acid solution (2mol / L) to stop the reaction;
[0030] Place each reaction product solution in an ultraviolet spectrophotometer, measure its OD value at an absorption wavelength of 450 nm, measure each concentration 3 times, take the average value, and replace the TMD solution with the same amount of normal saline as a blank control. Draw the standard curve with the OD value as the abscissa a...
Embodiment 2
[0032] Example 2: Time change law of peroxidase in healthy pork tissue
[0033] Take 2 mL of peroxidase extract of healthy pork from 6 sampling sites (Tong Tenderloin A, Pork Belly B, Front Butt C, Front D, Hind E, and Hind F) and add them to the test tube , Respectively add 4-5 drops of 0.2% benzidine solution, mix the solution, and then add 3 drops of 1% hydrogen peroxide solution, be careful not to shake, immediately observe the speed and extent of the color change of the solution within 5 minutes, judge and record As a result, set a distilled water control.
[0034] Judgment criteria: the color change time of the solution within 0-30s is recorded as "++++", the color changing time within 30-60s is recorded as "+++", the color changing time within 60-120s is recorded as "++", the color changes The time between 120-180s is marked as "+", and the one that does not change color is marked as "-". The test results are shown in Table 1;
[0035] Table 1: Time change of peroxidase in p...
Embodiment 3
[0038] Example 3: Method for detecting the concentration of peroxidase in pork to be tested
[0039] (1) Collection of pork samples
[0040] Pork samples are in the order of sirloin (longissimus dorsi), pork belly (abdominal wall muscles), front buttocks (brachial head muscles), front thighs (arm biceps), back buttock (mid-arm muscles), hind legs The six parts of the meat (semitendinosus or semimembranosus) were collected in equal parts of pork samples; the collected weight of each pork sample was 100 grams, and it was stored in a refrigerator at 4°C after collection.
[0041] (2) Preparation of peroxidase extract from pork
[0042] Take the collected pork sample, remove the fat and connective tissue, accurately weigh 10g pork sample, place it in a small beaker, chop and grind it, add 5 times the amount of distilled water of the pork sample and stir well, place on a shaker and shake for 20 minutes , Filter with filter paper, the filtrate is the extract of the peroxidase in the pork t...
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