Poultry-derived characteristic collagen peptide and application thereof in detection of collagen hydrolysate and products thereof
A technology for sex glue and poultry, applied in specific peptides, biological tests, measuring devices, etc., can solve problems such as insurmountable difficulties, and achieve the effects of high accuracy, high sensitivity and strong characteristics
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Embodiment 1
[0022] 1 Materials and Reagents
[0023] Materials: Gelatin from different animal sources, from chicken, duck, goose, cattle, pig, sheep, respectively, extracted from chicken skin, duck skin, goose skin, cowhide, pig skin, sheepskin.
[0024] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).
[0025] 2 Screening and determination of characteristic collagen peptides
[0026] Collagen products from chickens, ducks, geese and other animals are mainly type I collagen, including COL1A1 and COL1A2 proteins, and have a certain degree of conservation. We can screen a peptide from COL1A2 protein as a characteristic collagen peptide of poultry origin. In order to screen out a stable characteristic collagen peptide for detection from the gene, the following conditions should be met: (a) the three bases adjacent to the mRNA sequence corresponding to the peptide should be argin...
Embodiment 2
[0039] 1 Materials and Reagents
[0040] Materials: Mixed gelatin samples (including: porcine gelatin with 5% chicken gelatin, porcine gelatin with 10% chicken gelatin, porcine gelatin with 20% chicken gelatin, porcine gelatin with 40% chicken gelatin, porcine gelatin with 80% chicken gelatin, pure chicken gelatin) is made from an exact mix of the gelatin from Example 1.
[0041] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).
[0042] 2 detection method
[0043] (1) Take 0.05g gelatin sample in a 25ml measuring bottle, add a small amount of 1% NH 4 HCO 3 Soak the solution for 10 minutes, heat it at 60°C to dissolve it, and use 1% NH 4 HCO 3 Dilute the solution to the mark, shake well, filter with a microporous membrane, accurately measure 0.2ml of the subsequent filtrate, add 20μl of 2mg / ml trypsin solution, and enzymolyze at a constant temperature of 37°C f...
Embodiment 3
[0047] 1 Materials and Reagents
[0048] Materials: chicken gelatin, QQ sugar with bovine gelatin (made by adding), QQ sugar with chicken gelatin (made by adding);
[0049] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).
[0050] 2 detection method
[0051] (1) Take 0.05-0.25g of chicken gelatin, QQ sugar added with cow gelatin, and QQ sugar added with chicken gelatin, put them in 25ml measuring bottles, add a small amount of 1% NH 4 HCO 3 Soak the solution for 10 minutes, heat it at 60°C to dissolve it, and use 1% NH 4 HCO 3 Dilute the solution to the mark, shake well, filter with a microporous membrane, accurately measure 0.2ml of the subsequent filtrate, add 20μl of 2mg / ml trypsin solution, and enzymolyze at a constant temperature of 37°C for 12h.
[0052] (2) Take 5 μl of the enzymatic hydrolysis solution and put it into the liquid mass spectrometer for d...
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