Natural medical collagen sponge made of sharkskin and a preparation method of the natural medical collagen sponge
A collagen sponge and collagen technology, applied in the field of biomedical material preparation, can solve the problems of shark skin collagen extraction and low thermal denaturation temperature of marine biological collagen, and achieve reliable and safer raw material sources and extraction quality Good, the effect of increasing added value
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[0031] The preparation method of the shark skin natural medical collagen sponge provided by the present embodiment comprises the following steps:
[0032] Step 1: Pretreatment of Raw Materials
[0033] Rinse the fresh shark skin with clean water, remove the blood, and freeze it at -22°C to -18°C, separate the black skin layer and the collagen layer of the shark skin, cut the collagen layer into small pieces, and remove the impurities with NaOH , remove lipid with ethanol, then add pre-cooled acetic acid solution, and pulverize with pulverizer;
[0034] Step 2: Enzymolysis
[0035] Suspend the collagen layer treated in step 1 with 0.5M acetic acid solution, add protease at a ratio of 1:1000, and slowly stir and enzymolyze it for 2-3 days at a temperature lower than 10°C to obtain an undenatured collagen solution ;
[0036] Step 3: Purification of the extract
[0037] Place the collagen solution obtained in step 2 in a high-speed centrifuge for high-speed separation; adjust ...
Embodiment 1
[0043] Rinse the skin of the blue shark with clean water to remove excess blood and meat, put it in -20°C for a week of quick freezing, after taking it out, you can easily separate the collagen layer from the black cortex, then take 15g of fish skin, cut it into pieces After forming small pieces, add 450ml of 0.1M NaOH solution, treat for 4hrs, remove impurity proteins, rinse with clean water, add 150ml of 10% ethanol solution, and finally treat for 16hrs, wash with clean water, add 200ml of 0.5M acetic acid solution, After soaking for 20 minutes, crush it with a meat grinder, add 15 mg of pepsin to treat it at a temperature of 10°C, stir and enzymolyze it for 72 hours, remove the undissolved part by centrifugation, adjust the pH of the supernatant to the center with 10M NaOH solution, and then use Salt out with 2.5M NaCl, dissolve the precipitate with 0.5M acetic acid, and dialyze for about 72 hours to obtain undenatured natural shark skin collagen solution, then cross-link wi...
Embodiment 2
[0045]Rinse the skin of the whale shark with clean water to remove excess blood and meat, put it in a -20°C freezer for a week, and then easily separate the collagen layer from the black cortex, then take 20g of fish skin and cut it into pieces After small pieces, add 600ml of 0.1M NaOH solution, treat for 4hrs, remove impurities, rinse with clean water, add 200ml of 10% ethanol solution, and finally, treat for 16hrs, wash with clean water, add 250ml of 0.5M acetic acid solution, soak After 20 minutes, grind it with a meat grinder, add 20 mg of pepsin to treat it at a temperature of 10 ° C, stir for 72 hours, remove the undissolved part by centrifugation, adjust the pH of the supernatant to the center with 10M NaOH solution, and then use 2.5 Salt out with M NaCl, dissolve the precipitate with 0.5M acetic acid, and dialyze for about 72 hours to obtain an undenatured natural shark skin collagen solution, then cross-link with ultraviolet light for 30 minutes, freeze-dry to obtain ...
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