Stable collagen system as well as preparation method and application thereof
A collagen system and a stable technology, applied in skin care preparations, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problem of reducing the compatibility and molecular integration of collagen supplementation with the physiological environment in the body Insufficiency, deviation, and difficulty in effectively functioning, etc., to achieve the effects of easy mass production, simple preparation method, and low cost
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Embodiment 1
[0037] Example 1: Preparation of Type I Collagen and Detection of Physicochemical Properties
[0038] Pre-treatment: chopped beef tendon, cleaned, washed with normal saline, encapsulated, and treated with Co 60 disinfect. The remaining steps are performed under sterile conditions.
[0039] Preparation of high-purity type Ⅰ collagen solution: add 0.5M acetic acid in appropriate amount to beef tendon and tendon, and add pepsin (20g beef tendon with 1 gram of pepsin), enzymolysis at 6°C for 24-48 hours, 15mmol / L EDTA Stop enzymatic digestion. The supernatant was collected by centrifugation to obtain a type I collagen solution, and this process was carried out at 4°C.
[0040] Purification: Add collagen solution and 10% NaCl for salting out at a ratio of 1:1 by volume, and let stand overnight. The precipitate was obtained by centrifugation, dissolved in 0.5M acetic acid, and dialyzed in deionized water to pH 5 to obtain a high-purity type I collagen solution (≧10mg / ml).
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Embodiment 2
[0044] Example 2: Preparation of Type II Collagen and Detection of Physicochemical Properties
[0045] 1. Pre-treatment: cut hyaline cartilage from pig joints, wash with normal saline, degrease with 75% ethanol for 30 minutes, wash with normal saline, encapsulate, and 60 disinfect. The remaining steps are performed under sterile conditions.
[0046] 2. Prepare high-purity type II collagen solution: Add appropriate amount of pre-cooled 0.01M EDTA to the hyaline cartilage, mash it in a homogenizer, collect the precipitate by centrifugation, and then stir overnight with 10 times the volume of 4M, pH 7.5 guanidine hydrochloride Remove proteoglycan; fully wash with water, take the precipitate after centrifugation, add 0.5M acetic acid enzymatic hydrolysis solution, wherein the weight ratio of cartilage to pepsin is 50:1, this process is carried out below 4°C, and stirred for 24 hours. High-speed centrifugation collects the supernatant as type II collagen solution, and this step c...
Embodiment 3
[0053] Embodiment 3: Preparation of low concentration type I collagen system (pH 6)
[0054] a. Preparation of balance substance (90ml): Add 0.5g of glucose, 0.5ml of 1,3 propylene glycol, and 800.1ml of Tween to deionized water, and adjust the pH to 6 with 0.2M NaOH (about 0.16mg).
[0055] b. Measure 10ml of 10mg / ml type I collagen solution.
[0056] c. Mix steps a and b to obtain 100ml of a system containing type I collagen, the concentration of glucose is about 0.5% by weight, the concentration of 1,3 propanediol is 0.5ml / ml, and the concentration of Tween 80 is 0.1ml / ml. The concentration of type collagen was 1 mg / ml.
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