A liquid chromatography-mass spectrometric method for identifying goose-derived components based on characteristic peptides
A goose-derived and peptide technology, applied in the field of protein detection, can solve the problems of difficulty in finding the characteristic peptides of goose, identification of no goose-derived components, close kinship, etc., so as to solve food safety problems, operation and detection. Clear method and accurate identification results
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Embodiment 1
[0041] Prepare a protein lysate containing urea (hereinafter referred to as: uric acid lysate), weigh 1.0 g of a mixed (50%: 50%) sample of chicken and duck cut into minced meat, add 5 mL of urea lysate, and shake to extract 12 hours, centrifuge at 14000rpm for 30min, and control the temperature below 10°C. Take 20 μL of the supernatant, add uric acid lysate to 100 μL, add 4 μL of dithiothreitol solution, and react at a constant temperature of 37°C for 1 hour. Add 20 μL of iodoacetamide solution, react in the dark for 30 min, and transfer to an ultrafiltration tube. Centrifuge at 14000rpm for 30min and keep the temperature below 10°C. Add 100 μL of ammonium bicarbonate aqueous solution, centrifuge at 14,000 rpm for 30 min, repeat washing 1-3 times, add 100 μL of ammonium bicarbonate aqueous solution, 10 μg of trypsin, and react at a constant temperature of 37°C for 12 hours. Centrifuge at 14,000 rpm for 30 minutes, keep the temperature below 10°C, add 100 μL of ammonium bica...
Embodiment 2
[0053] Prepare a protein lysate containing urea, weigh 1.0 g of a mixed sample of pork and goose (50%:50%) cut into minced meat, add 5 mL of urea lysate, shake and extract for 20 hours, centrifuge at 14,000 rpm for 30 min, and control the temperature Below 10°C. Take 40 μL of the supernatant, add urea lysate to 200 μL, add 5 μL of dithiothreitol solution, and react at a constant temperature of 37°C for 1 hour. Add 25 μL of iodoacetamide solution, react in the dark for 30 min, and transfer to an ultrafiltration tube. Centrifuge at 14000rpm for 30min, and keep the temperature below 10°C. Add 100 μL of ammonium bicarbonate aqueous solution, centrifuge at 14,000 rpm for 30 min, repeat washing 1-3 times, add 100 μL of ammonium bicarbonate aqueous solution, 20 μg of trypsin, and react at a constant temperature of 37°C for 16 hours. Centrifuge at 14000rpm for 30min, keep the temperature below 10°C, add 100μL of ammonium bicarbonate aqueous solution, centrifuge at 14000rpm for 30min...
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