阅读说明：本技术 一种无菌速溶未变性天然胶原冻干品及制备方法 (Sterile instant undenatured natural collagen freeze-dried product and preparation method thereof ) 是由 鲍柳君 张灿 镇晓华 孙秀娟 于 2019-08-14 设计创作，主要内容包括：本发明提供了一种无菌速溶未变性天然胶原冻干品及其制备方法,制备方法步骤包括：(1)调制一定浓度的活性胶原原液,边搅拌边加入阻断剂,搅拌时间为0.5～2小时,反应温度为10～20℃,然后加入速溶剂,搅拌时间为0.5～2小时,反应温度为10～20℃,控制反应pH＝5～7；(2)反应结束后,进行灌装、冻干得速溶未变性天然胶原冻干品。本发明的胶原冻干品保留了天然未变性胶原的结构特性,未经酰化处理即能速溶于水(1～2min)；该胶原冻干品未添加增稠剂,用水溶解后即有稠的肤感,产品更安全；该胶原冻干品采用低剂量电子束辐照灭菌工艺进行终端灭菌,为无菌产品,灭菌后仍然保持胶原的天然活性。(The invention provides a sterile instant undenatured natural collagen freeze-dried product and a preparation method thereof, wherein the preparation method comprises the following steps: (1) preparing an active collagen stock solution with a certain concentration, adding a blocking agent while stirring, wherein the stirring time is 0.5-2 hours, the reaction temperature is 10-20 ℃, then adding a quick-dissolving agent, the stirring time is 0.5-2 hours, the reaction temperature is 10-20 ℃, and controlling the reaction pH to be 5-7; (2) after the reaction is finished, filling and freeze-drying are carried out to obtain the instant undenatured natural collagen freeze-dried product. The freeze-dried collagen product disclosed by the invention keeps the structural characteristics of natural undenatured collagen, and can be quickly dissolved in water (1-2min) without acylation treatment; the freeze-dried collagen product is not added with a thickening agent, and has thick skin feel after being dissolved by water, so that the product is safer; the freeze-dried collagen product is sterilized by a low-dose electron beam irradiation sterilization process, is an aseptic product, and still maintains the natural activity of collagen after sterilization.)

1. A method for preparing a sterile instant undenatured natural collagen freeze-dried product is characterized by comprising the following steps:

(1) preparing an active collagen stock solution with a certain concentration, adding a blocking agent while stirring, wherein the stirring time is 0.5-2 hours, the reaction temperature is 10-20 ℃, then adding a quick-dissolving agent, the stirring time is 0.5-2 hours, the reaction temperature is 10-20 ℃, and controlling the reaction pH to be 5-7;

(2) after the reaction is finished, filling and freeze-drying are carried out to obtain the instant undenatured natural collagen freeze-dried product.

2. The process according to claim 1, wherein the freeze-dried product of sterile, instant undenatured native collagen is prepared by: and performing electron beam sterilization irradiation on the instant undenatured collagen freeze-dried product at a dose of 6 kGy-10 kGy to obtain the sterile instant undenatured natural collagen freeze-dried product.

3. The method for preparing a lyophilized product of sterile instant undenatured native collagen according to claim 1, wherein the mass ratio of the active collagen stock solution to the blocking agent is 25-30%: 25-30%, wherein the mass ratio of the addition amount of the quick solvent to the active collagen stock solution is 25-30%: 25 to 30 percent.

4. The method for preparing a sterile instant undenatured native collagen lyophilizate as claimed in claim 1, wherein said blocking agent is a sugar alcohol compound.

5. The method of claim 1, wherein the range of said dissolution aid is selected from the group consisting of sodium chloride, disodium hydrogen phosphate, sodium citrate, and sodium carbonate.

6. The method for preparing a sterile lyophilized product of native collagen according to claim 4, wherein said sugar alcohol compound is selected from the group consisting of sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol.

7. The process for preparing a lyophilized product of sterile instant undenatured native collagen according to claim 1, wherein in step (1), according to the efficacy of the product, the corresponding bioactive substance is added.

8. A sterile instant native collagen lyophilized product, characterized in that it is prepared by the method of any one of claims 1 to 7.

Technical Field

The invention relates to a sterile instant undenatured natural collagen freeze-dried product and a preparation method thereof.

Background

Collagen, or collagen, is widely distributed in the bodies of various animals, is the main structural protein of the animals and accounts for about 30% of the total protein in the bodies. Currently, 28 types of collagen are found in vertebrates, of which type I collagen is the most abundant, accounting for about 90% of the total amount of collagen in animals. The collagen is widely present in connective tissues such as skin, bone, tendon and the like of mammals such as pigs, cows, sheep and the like, and provides possibility for large-scale application of the collagen. The unique triple helix structure of the collagen endows the collagen with a plurality of excellent properties, such as better mechanical property, low antigenicity and good biocompatibility, is beneficial to the survival and growth of cells, and also has the excellent properties of moisturizing property and obviously promoting the growth and the propagation of epidermal cells. Therefore, the natural collagen as a raw material of an excellent biological material can be widely applied to the fields of tissue engineering, biomedicine, plastic, skin care and the like.

At present, the natural collagen which is not denatured is obtained mainly by an acid dissolution method and an acid dissolution enzymolysis method. The collagen extracted by these two methods is acid-soluble collagen and can be stably present only under acidic conditions. When freeze-dried into a solid, the collagen is only redissolved in an acidic solution and has a long dissolution time, and thus cannot be used as a cosmetic material.

Chinese patent No. 200410040487.2 discloses that acid-soluble collagen is subjected to acylation reaction, washed, precipitated, and freeze-dried to obtain water-soluble undenatured natural collagen. Chinese patent publication No. CN105078783A discloses the preparation of a quick-dissolving collagen lyophilized pellet from acetylated collagen. All of the solutions disclosed in the above documents require acylation of collagen, and although the natural structure of collagen is not changed, the molecular composition of single collagen chains is changed, so that the stability of the collagen structure after acylation is lowered, and the lyophilized product thereof can be rapidly dissolved in an aqueous solution.

The collagen is easy to breed bacteria in a solution environment, and a preservative must be added for long-term storage. However, it has been shown that preservatives can cause abnormal skin proliferation, canceration; related components are chemically changed under the irradiation of sunlight, so that people can feel light, and the skin becomes red, black and painful; causing allergy to people and causing symptoms of red and oxygen; or cause skin burn.

Other active ingredients are added into the collagen compound product, if the collagen compound product is stored in a solution environment for a long time, the titer of the active ingredients is possibly reduced or even disappears, and the effectiveness of the product cannot be ensured.

To solve the above problems, the collagen product can be freeze-dried for storage. However, two problems arise, namely, the acid-soluble collagen is a slow process when dissolved in water if the acylation reaction does not occur, the waiting time is long (generally about 12 hours), and the use is influenced by the excessively low pH value. Secondly, the water content of the collagen product is still maintained at 10-20% after freeze drying. Because the production process is not sterile, the risk of bacteria and mold growth exists along with long-term storage.

Disclosure of Invention

The invention aims to solve the technical problem firstly to provide a preparation method of an aseptic instant undenatured natural collagen freeze-dried product, which can obtain the natural undenatured collagen freeze-dried product, can ensure that the freeze-dried product is very easy to redissolve, can be directly redissolved in drinking water environment and is convenient to use. Therefore, the invention adopts the following technical scheme:

a method for preparing a sterile instant undenatured natural collagen freeze-dried product is characterized by comprising the following steps:

(1) preparing an active collagen stock solution with a certain concentration, adding a blocking agent while stirring, wherein the stirring time is 0.5-2 hours, the reaction temperature is 10-20 ℃, then adding a quick solvent, the stirring time is 0.5-2 hours, the reaction temperature is 10-20 ℃, and controlling the reaction pH to be 5-7;

(2) after the reaction is finished, filling and freeze-drying are carried out to obtain the instant undenatured natural collagen freeze-dried product

On the basis of the technical scheme, the instant undenatured collagen freeze-dried product is subjected to electron beam sterilization irradiation with the dose of 6 kGy-10 kGy, and the sterile instant undenatured natural collagen freeze-dried product is obtained. The low dose does not denature or crosslink the collagen, but can make the product reach a sterile level and be safer.

Furthermore, the mass ratio of the active collagen stock solution to the blocking agent is 25-30%: 25-30%, wherein the mass ratio of the addition amount of the quick solvent to the active collagen stock solution is 25-30%: 25 to 30 percent.

The blocking agent is a sugar alcohol compound. The sugar alcohol compounds are selected from sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol. These sugar alcohol compounds have the characteristics of small molecular weight and hydroxyl group.

The selection range of the quick solvent comprises sodium chloride, disodium hydrogen phosphate, sodium citrate and sodium carbonate. The low concentration of salt solution aids in the dissolution of collagen.

Further, in the step (1), corresponding bioactive substances can be added according to different product efficacies.

Further, during filling and freeze-drying, the freeze-dried powder can be frozen into a patch, a spherical shape, a columnar shape, a heart shape and the like according to different requirements.

The invention also provides a sterile instant undenatured natural collagen freeze-dried product which is prepared by adopting the method.

The collagen adopted by the invention is natural undenatured collagen, and the collagen in a freeze-dried product is not subjected to acylation reaction, so that the most original triple-helix structure of the natural collagen is ensured, and the single-chain component of the collagen is not changed. The natural collagen has a triple helix structure, has a molecular weight of 300kDa, is a rod-shaped biological macromolecule, can be subjected to self-assembly in a solution to form microfibers, then is in an aggregation state structure, and can be directly freeze-dried to form a sponge-like porous network structure. According to the invention, the hydroxyl sugar alcohol compound with smaller molecular weight is added as a blocking agent, so that the self-assembly behavior of natural collagen molecules can be blocked, the structure of the natural collagen after freeze-drying is changed, and the water absorption is facilitated.

Sugar alcohols are now approved by the international food and health organization as food products that are not required to be used in limited quantities, and therefore are very safe ingredients, and sugar alcohol compounds can be used as moisturizers. The collagen structure stability after acylation reaction is reduced, the freeze-dried product is not full in shape, and other components such as mannitol (30-56%), trehalose (10-15%) and the like are added in combination to serve as fillers to change the freeze-dried balls into pompon. At the same concentration, the viscosity of the collagen after acylation reaction is reduced compared with the natural undenatured collagen, so that a thickening agent such as pullulan (10-15%) is added to increase the viscosity of the final product.

In the invention, because the natural undenatured collagen can form a spongy structure, the aim of blocking the self-assembly behavior of collagen molecules can be achieved only by adding a certain amount of blocking agent, and the plump state of the product during dissolution can be achieved without adding a filling agent and a thickening agent.

The collagen freeze-dried product can be dissolved by drinking water and can be directly contacted with human tissues.

In conclusion, the invention establishes a sterile instant undenatured natural collagen freeze-dried product and a preparation method thereof. The freeze-dried collagen product keeps the structural characteristics of natural undenatured collagen, is not subjected to acylation treatment, is subjected to terminal sterilization by adopting a low-dose electron beam irradiation sterilization process, can be quickly dissolved in water (1-2min), keeps the natural activity of collagen invariance, can be used without adding a thickening agent, is safer, and is an aseptic product.

Drawings

FIG. 1 is a graph showing the results of electrophoresis in examples 1 to 6.

FIG. 2 is a graph showing the results of electrophoresis in examples 7 to 8.

Detailed Description