Sturgeon cartilage type II non-denatured collagen as well as preparation method and application thereof
A non-denaturing and cartilage technology, which is applied in the field of sturgeon cartilage type II non-denaturing collagen and its preparation, can solve the problems of difficult to guarantee product quality and great influence on collagen activity, and achieve the effects of high extraction rate and improved application value.
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Embodiment 1
[0042] After thawing, the sturgeon cartilage was rinsed with pre-cooled distilled water, and then cut into small pieces of 5mm×5mm×5mm. The prepared cartilage was mixed with 0.02mol / L NaOH solution (1:10, w / v), stirred continuously for 48h, and the alkali solution was replaced every 16h, and then rinsed with pre-cooled deionized water until the pH was neutral. Then soak in 1mol / L guanidine hydrochloride-0.05mol / LTris-HCl (pH 7.5, 1:10, w / v) buffer solution and stir for 24h. After repeated rinsing with pre-cooled distilled water, the sample was placed in 0.5 mol / L acetic acid containing 0.7% pepsin at a solid-liquid ratio of 1:20, and stirred continuously for 48 hours. The mixture was centrifuged at 9000 x g for 30 min at 4°C to obtain a supernatant. Add finely ground sodium chloride powder to the supernatant to a final salt concentration of 0.9 mol / L to precipitate collagen. The precipitate was collected by centrifugation at 9000×g, 4° C. for 30 min, and dissolved in 0.5 mol...
Embodiment 2
[0044]After thawing, the sturgeon cartilage was rinsed with pre-cooled distilled water, and then cut into small pieces of 6mm×5mm×5mm. The prepared cartilage was mixed with 0.05 mol / L NaOH solution (1:15, w / v), stirred continuously for 36 h, the alkaline solution was replaced every 12 h, and then rinsed with pre-cooled deionized water until the pH was neutral. Then soak in 1mol / L guanidine hydrochloride-0.05mol / LTris-HCl (pH 7.5, 1:10, w / v) buffer solution and stir for 24h. After repeated rinsing with pre-cooled distilled water, the sample was placed in 0.9 mol / L acid ethyl alcohol containing 0.1% pepsin, the ratio of solid to liquid was 1:10, and stirred continuously for 48 hours. The mixture was centrifuged at 9000 x g for 25 min at 4°C to obtain a supernatant. Add finely ground sodium chloride powder to the supernatant to a final salt concentration of 1 mol / L to precipitate collagen. The precipitate was collected by centrifugation at 9000×g, 4°C for 25 min, and dissolved ...
Embodiment 3
[0046] After thawing, the sturgeon cartilage was rinsed with pre-cooled distilled water, and then cut into small pieces of 5mm×4mm×4mm. The prepared cartilage was mixed with 0.1 mol / L NaOH solution (1:20, w / v), stirred continuously for 24 h, the alkaline solution was replaced every 8 h, and then rinsed with pre-cooled deionized water until the pH was neutral. Then soak in 1mol / L guanidine hydrochloride-0.05mol / L Tris-HCl (pH 7.5, 1:10, w / v) buffer solution, and stir for 24h. After repeated rinsing with pre-cooled distilled water, the sample was placed in 0.5 mol / L acetic acid containing 0.4% pepsin at a solid-liquid ratio of 1:15, and stirred continuously for 48 hours. The mixture was centrifuged at 9000 x g for 20 min at 4°C to obtain a supernatant. Add finely ground sodium chloride powder to the supernatant to a final salt concentration of 1.2 mol / L to precipitate collagen. The precipitate was collected by centrifugation at 9000×g, 4°C for 20 min, and dissolved in 0.5 mol / ...
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