阅读说明：本技术 从动物软骨中提取非变性ii型胶原蛋白的制备方法 (Preparation method for extracting non-denatured type II collagen from animal cartilage ) 是由 赵成军 李佳 于 2020-06-11 设计创作，主要内容包括：本发明公开了一种从动物软骨中提取非变性II型胶原蛋白(复合骨胶原)的制备方法,此工艺可以生产出对人体极为有用的生物营养素。该工艺包括以下步骤：浸泡—清洗—超声清洗—破碎—盐溶液浸泡—过滤—乙醇浸泡—过滤—真空干燥—涡旋粉碎—包装。利用本发明生产工艺提取的非变性II型胶原蛋白(复合骨胶原)的分子结构完整,能很好的保留营养成分的生物活性,脂肪含量低,胶原蛋白含量高,大分子粘多糖含量达20％以上,并含有钙、镁、磷等矿物质营养元素,所得产品无令人不悦的气味,利于消化、吸收和利用,具有良好的市场前景。(The present invention discloses a preparation method for extracting non-denatured type II collagen (composite bone collagen) from animal cartilage, and said process can produce biological nutrient which is very useful for human body. The process comprises the following steps: soaking, cleaning, ultrasonic cleaning, crushing, soaking in a salt solution, filtering, soaking in ethanol, filtering, vacuum drying, vortex crushing and packaging. The non-denatured type II collagen (composite ossein) extracted by the production process has complete molecular structure, can well retain the biological activity of nutrient components, has low fat content and high collagen content, has the macromolecular mucopolysaccharide content of more than 20 percent, contains mineral nutrient elements such as calcium, magnesium, phosphorus and the like, does not have unpleasant smell, is beneficial to digestion, absorption and utilization, and has good market prospect.)

1. A method for extracting non-denatured type II collagen (composite collagen) from animal cartilage comprises the following steps:

1) soaking: soaking animal cartilage in water for 1-4 hours;

2) cleaning: putting the cartilage obtained in the step 1) into a tank, adding water, stirring and cleaning to remove blood and impurities;

3) ultrasonic cleaning: putting the clean cartilage obtained in the step 2) into an ultrasonic cleaning pool, and mixing the materials in a weight ratio of 1: adding purified water according to the proportion of 1.5 for ultrasonic cleaning for 0.5-1 hour, and collecting clean cartilage;

4) crushing: freezing the clean cartilage obtained in the step 3) at the temperature of between 20 ℃ below zero and 1 ℃ below zero, and cutting the cartilage into cartilage particles with the length of less than 1cm by using a crusher;

5) soaking in a salt solution: soaking the cartilage particles obtained in the step 4) and a salt solution according to the ratio (w/w) of 1: 0.1-9 for 0.5-4 hours;

6) and (3) filtering: filtering the solution obtained in the step 5), removing liquid, and reserving a solid obtained by filtering;

7) ethanol soaking: adding ethanol into the solid obtained in the step 6), feeding the solid and the ethanol according to the ratio (w/w) of 1: 0.1-30, stirring and soaking for 0.5-4 hours, and repeating for 2-3 times;

8) and (3) filtering: after the step 7), removing ethanol, and reserving a solid obtained by filtering;

9) and (3) vacuum drying: putting the solid obtained in the step 8) into a low-temperature vacuum drier, and drying until the water content in the product is less than 10% (weight ratio);

10) vortex crushing: putting the dried solid obtained in the step 9) into a vortex crusher for crushing to obtain a product with the mesh number larger than 50 meshes;

11) packaging: sieving the product obtained in the step 10), sealing and packaging.

2. The method for preparing according to claim 1, wherein the cartilage is cartilage of terrestrial mammals, marine and freshwater fishes or poultry cartilage, which is dried cartilage or fresh cartilage.

3. The preparation method according to claim 1, wherein the cleaning process in step 3) uses an ultrasonic cleaning device, which can remove grease and impurities with high efficiency.

4. The method according to claim 1, wherein the clean cartilage of step 4) is frozen and then crushed by a crusher.

5. The method of claim 1, wherein the salt solution of step 5) is selected from the group consisting of: sodium chloride solution, potassium chloride solution, calcium chloride solution or the combination thereof, wherein the mass concentration of the salt solution is 1-25%.

6. The preparation method according to claim 1, wherein ethanol is used for soaking in the step 7), the mass content concentration of the ethanol is required to be more than 96%, and the ethanol can be recycled after soaking.

7. The method of claim 1, wherein the vacuum dryer in the step 9) is a drying apparatus capable of continuously monitoring temperature and humidity.

8. Non-denatured type II collagen (complex collagen) obtained by the production method according to any one of claims 1 to 7, preferably having a glycosaminoglycan content of more than 30 wt%.

Technical Field

The invention relates to a method for extracting nutrient substances from animal cartilage tissues, in particular to a method for extracting non-denatured type II collagen (composite ossein) from mammal, marine and freshwater fish cartilage and poultry cartilage.

Background

Collagen is an important component of tissues such as skin, bone, and joints, and is a main component of extracellular matrix of multicellular animals. At present, 27 different types of collagen have been found from mammals, in which non-denatured type II collagen (complex collagen) composed of three identical α chains is mainly present in cartilage tissue and vitreous eye, and thus non-denatured type II collagen (complex collagen) is considered as a structural functional component of cartilage.

Most of the currently marketed collagens are those subjected to over-hydrolysis, the main components of the collagens are amino acids and polypeptides, and the molecular weight distribution is within the range of 200-20000 daltons. The polypeptide has certain functions of keeping skin moisture and increasing immunity, but cannot prevent and treat osteoporosis, degenerative arthritis, osteoarthritis, rheumatoid arthritis and rheumatoid arthritis.

Disclosure of Invention

The invention aims to provide a preparation method for extracting non-denatured type II collagen (composite collagen) from animal cartilage, and the type II collagen (composite collagen) produced by the production process has a complete structure, is non-denatured protein and has a good market prospect.

Non-denatured type II collagen (composite collagen) is extracted from animal cartilage, and contains polypeptide component and macromolecular mucopolysaccharide component which is not possessed by commercial collagen powder, so that the product has the characteristics and advantages different from common collagen.

The preparation method of the invention comprises the following steps: a preparation method for extracting non-denatured type II collagen (composite collagen) from animal cartilage is characterized by sequentially comprising the following steps of:

1) soaking: soaking animal cartilage in water for 1-4 hours;

2) cleaning: putting the cartilage obtained in the step 1) into a tank, adding water, stirring and cleaning to remove blood and impurities;

3) ultrasonic cleaning: putting the clean cartilage obtained in the step 2) into an ultrasonic cleaning pool, and mixing the materials in a weight ratio of 1: adding purified water according to the proportion of 1.5 for ultrasonic cleaning for 0.5-1 hour, and collecting clean cartilage;

4) crushing: freezing the clean cartilage obtained in the step 3) at the temperature of between 20 ℃ below zero and 1 ℃ below zero, and cutting the cartilage into cartilage particles with the length of less than 1cm by using a crusher;

5) soaking in a salt solution: soaking the cartilage particles obtained in the step 4) and a salt solution according to the ratio (w/w) of 1: 0.1-9 for 0.5-4 hours;

6) and (3) filtering: filtering the solution obtained in the step 5), removing liquid, and reserving a solid obtained by filtering;

7) ethanol soaking: adding ethanol into the solid obtained in the step 6), feeding the solid and the ethanol according to the ratio (w/w) of 1: 0.1-30, stirring and soaking for 0.5-4 hours, and repeating for 2-3 times;

8) and (3) filtering: after the step 7), removing ethanol, and reserving a solid obtained by filtering;

9) and (3) vacuum drying: putting the solid obtained in the step 8) into a low-temperature vacuum drier, and drying until the water content in the product is less than 10% (weight ratio);

10) vortex crushing: putting the dried solid obtained in the step 9) into a vortex crusher for crushing to obtain a product with the mesh number larger than 50 meshes;

11) packaging: sieving the product obtained in the step 10), sealing and packaging.

The animal cartilage is preferably fresh cartilage.

The present invention also provides non-denatured type II collagen (complex collagen) obtained by the extraction method of the present invention.

With the above-described production method of the present invention, cartilage of mammals, marine and freshwater fishes, cartilage of poultry (e.g., chicken) is used as the raw material, and the salt solution treatment step, the low-temperature crushing step and the vacuum drying step require determination that the concentration of the salt solution is not more than 35 wt% (preferably 1 wt% to 25 wt%), and the crushing and drying temperature is not more than 50 ℃ in order to ensure that the structure of type II collagen (composite collagen) is not damaged, to obtain a non-denatured type II collagen product having a glycosaminoglycan content of more than 30 wt% (e.g., 30 wt% to 45 wt%, such as 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt%, 40 wt%, or 45 wt%).

Drawings

FIG. 1 is a flow chart of the process for preparing non-denatured type II collagen from animal cartilage according to the present invention;

FIG. 2 shows the result of measuring the glycosaminoglycan content in non-denatured type II collagen (complex collagen) extracted by the method of the present invention;

FIG. 3 is a graph of non-denatured type II collagen (complex collagen) and hydrolyzed type II collagen extracted by the method of the present invention; and

FIG. 4 is gel electrophoresis of non-denatured type II collagen (complex collagen) extracted by the method of the present invention.

Detailed Description

The method for extracting non-denatured type II collagen (complex collagen) from chicken breast cartilage according to the present invention will be described in detail with reference to the following examples.