Application of a donkey bone glue characteristic polypeptide in detecting donkey bone glue components in animal hide glue and its products
A technology for animal skin glue and animal detection, which is applied in the fields of peptides, analytical materials, measuring devices, etc., can solve the problems of difficulty in distinguishing animal skin glue and bone glue, and can not distinguish and distinguish, and achieves rapid identification and detection, high sensitivity and characteristic. strong effect
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Embodiment 1
[0028] 1 Materials and Reagents
[0029] Samples to be tested: various pure glues, including donkey bone glue, donkey-hide gelatin, yellow gelatin, and pig skin glue, obtained by decocting donkey bone, donkey skin, cowhide, and pig skin respectively.
[0030] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).
[0031] 2 detection method
[0032] Take 0.05g of the sample to be tested in a 25ml measuring bottle, and use 1% w / vNH 4 HCO 3 The solution (pH 8.0) was dissolved and fixed to volume, passed through a 0.22 μm microporous membrane, 300 μl of the filtrate was accurately measured, 30 μl of 2 mg / ml trypsin solution was added, and the enzymatic hydrolysis was obtained at a constant temperature of 37° C. overnight.
[0033] Take 5 μl of the enzymatic hydrolysis solution and put it into the liquid mass spectrometer for detection. Liquid phase condition: C 18 Reve...
Embodiment 2
[0036] 1 Materials and Reagents
[0037] Sample to be tested: the mixed glue sample is made by adding 5%w / w donkey bone glue to the pure glue sample in Example 1, including donkey-hide gelatin containing 5%w / w donkey bone glue, donkey-hide gelatin containing 5%w / w donkey bone glue Yellow gelatin, pig skin glue 5% w / w donkey bone glue.
[0038] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).
[0039] 2 detection method
[0040] Take 0.05g of the sample to be tested in a 25ml measuring bottle, and use 1% w / vNH 4 HCO 3 The solution (pH 8.0) was dissolved and fixed to volume, passed through a 0.22 μm microporous membrane, 300 μl of the filtrate was accurately measured, 30 μl of 2 mg / ml trypsin solution was added, and the enzymatic hydrolysis was obtained at a constant temperature of 37° C. overnight.
[0041] Take 5 μl of the enzymatic hydrolysis solution and put it...
Embodiment 3
[0044] 1 Materials and Reagents
[0045] Animal glue samples: donkey bone glue and donkey-hide gelatin were obtained by decocting donkey bone and donkey skin respectively.
[0046] Mixed gum samples: donkey-hide gelatin with 5% w / w donkey bone glue, donkey-hide gelatin with 10% w / w donkey bone glue, donkey-hide gelatin with 20% w / w donkey bone glue, donkey-hide gelatin with 40% w / w Made with a precise blend.
[0047] Reagents: ammonium bicarbonate (analytical pure), trypsin (serial pure, purchased from China Institute for the Control of Pharmaceutical and Biological Products).
[0048] 2 detection method
[0049] Take 0.05g of the sample to be tested in a 25ml measuring bottle, and use 1% w / vNH 4 HCO 3 The solution (pH 8.0) was dissolved and fixed to volume, passed through a 0.22 μm microporous membrane, 300 μl of the filtrate was accurately measured, 30 μl of 2 mg / ml trypsin solution was added, and the enzymatic hydrolysis was obtained at a constant temperature of 37° C. ...
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