Colla corii asini polypeptide and preparation method thereof
A technology of donkey-hide gelatin and middle donkey-hide gelatin, which is applied in the field of analytical chemistry and can solve the disputes over the efficacy of donkey-hide gelatin and other issues
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Embodiment 1
[0034] Embodiment 1 Preparation of a donkey-hide gelatin polypeptide
[0035] Add 20 times the amount of deionized water to the pulverized donkey-hide gelatin powder, stew at 80°C for 30 minutes, cool and refrigerate overnight. The extract was centrifuged at 18,000 rpm at 4°C for 5 min, and the supernatant was filtered to obtain the donkey-hide gelatin extract. Adjust the pH to 7.5 with 10% NaOH solution, in a water bath at 40°C with a concentration of 6×10 4 U / g trypsin was used for enzymatic hydrolysis, and the pH was adjusted every 1 h during the enzymatic hydrolysis to keep the pH at 7.5. After enzymatic hydrolysis for 4 hours, the enzyme activity was inactivated. After cooling, centrifuge at 18,000 rpm for 5 minutes at 4°C. Take 3 mL of the supernatant and pass it through a C18 SPE solid-phase extraction column (pre-activated with 3 mL of methanol and 3 mL of deionized water) for separation and purification. ×3 mL methanol for elution, collect the eluate, blow dry with...
Embodiment 2
[0045] Example 2 Efficacy evaluation of a donkey-hide gelatin polypeptide
[0046] 1 Hydroxyl radical scavenging activity
[0047] The detection of hydroxyl radical scavenging activity refers to the literature Xu Xiaobing. Research on Improving the Antioxidative Performance of Ejiao[D]. Beijing: Beijing University of Chemical Technology, 2018. Add 40 uL of 1.865 mmol / L o-phenanthroline solution to the microplate in sequence , 80uL embodiment 1 crosses the donkey-hide gelatin polypeptide sample solution that 0.45 μ m filter membrane gains, 40 uL concentration is the FeSO of 1.865 mmol / L. 4 ·7H 2 0 solution and 40 uL of 0.03 % (v / v) H 2 0 2 solution. The microtiter plate was kept at 37°C for 60 min, and the absorbance was measured at 536 nm, and the standard solution of ascorbic acid at the same concentration was used as a control.
[0048] 2 Anticoagulant activity
[0049] The anticoagulant activity experiment refers to the literature Jia Junqiang. The effect of ultrasoun...
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