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CN-121991199-A - Human keratin extraction method

CN121991199ACN 121991199 ACN121991199 ACN 121991199ACN-121991199-A

Abstract

The invention discloses a method for extracting protein from human hair fibers, and relates to the technical field of application of human hair fibers. The method comprises cleaning human hair fiber, drying, cutting, adding into NaOH solution, treating in sterilizing pot, filtering, and cleaning insoluble substance with distilled water until pH is close to 7. And then the insoluble matter after the treatment is put into a mixed solution containing Na 2 SO 3 and urea for treatment, and the insoluble matter obtained by filtering after the treatment is washed by distilled water until the pH value is close to 7. And finally, adding the obtained insoluble substances into a keratinase solution for dissolution, filtering, dialyzing and drying to obtain pale yellow human hair keratin powder. The human keratin extracted by the method has the advantages of large molecular weight, stable structure and high purity, can be used for manufacturing various human soft tissue implants or outer membranes, contents and the like of composite implants, can also be manufactured into soft tissue injection filling materials, has extremely wide application prospect and has great market development space.

Inventors

  • XU RUQIANG
  • DONG JIANSHEN
  • GAO YIZHE
  • ZHU YIFENG
  • LIU XIAOPING

Assignees

  • 长沙佰傲生物医药有限公司

Dates

Publication Date
20260508
Application Date
20241105

Claims (6)

  1. 1. A method for extracting human keratin, comprising the steps of: (1) Fully cleaning and drying the selected human hair fibers, cutting the human hair fibers to 2cm, adding the human hair fiber scraps into a NaOH solution with the concentration of 0.1mol/L according to the bath ratio of 1:20, and stirring to uniformly mix the solution with the human hair fiber scraps; (2) Placing the system into a sterilizing pot, sterilizing at 121 ℃ for 10 minutes, preserving heat at 70 ℃ for 1 hour, filtering by a nylon filter screen with 300 meshes after the treatment is finished, and repeatedly cleaning the filtrate by distilled water until the pH value is close to 7; (3) Adding the obtained filtration in the process (2) into a reduction system containing 0.5mol/L Na 2 SO 3 and 0.5mol/L urea according to a bath ratio of 1:15, performing treatment at 56 ℃ for 2 hours, filtering the treated mixture with 300-mesh nylon filter cloth, and repeatedly cleaning insoluble substances with distilled water until the pH value is close to 7; (4) Adding the insoluble substance obtained in the step (3) into 0.5% keratinase solution (PBS pH 7.0), stirring at 37deg.C for reacting for 8 hr, filtering with 300 mesh nylon filter cloth after the treatment, and repeatedly cleaning the insoluble substance with distilled water; (5) Placing the filtrate into a dialysis bag for dialysis for 48h, changing distilled water every 12h, pouring the solution obtained by dialysis into a beaker, and drying in a vacuum drying oven to obtain pale yellow keratin powder.
  2. 2. The method of claim 1, wherein the alkali solution treatment in step 1 (2) is performed at high temperature and high pressure, thereby destroying the hair surface structure and reducing the alkali treatment time.
  3. 3. The method of claim 1, wherein the steps (3) and (4) combine a reduction method and an enzyme method to reduce the strength of the single process.
  4. 4. The method for extracting keratin from human hair fiber according to claim 1, wherein the human hair keratin obtained by the method has the advantages of no excessive degradation, high purity, and more complete molecular structure and biological function, and the molecular weight of the harvested human keratin reaches 70 KD.
  5. 5. The macromolecular human-derived keratin obtained in claim 4 has high purity, more complete molecular structure and biological function, and better supporting performance and higher stability when used for manufacturing the outer membrane and the content of a soft tissue implant or a composite implant.
  6. 6. The macromolecular human keratin obtained in the method of claim 4 has high purity, and can be made into soft tissue injection filling material with good stability, difficult degradation, long retention effect and better filling effect.

Description

Human keratin extraction method Technical Field The invention relates to the technical field of keratin extraction, in particular to a method for extracting keratin from human hair fibers Background Human keratin materials have high sulfur content chemistry, exceptional biocompatibility, self-assembly propensity and natural cell recognition capability, biodegradability, stability persistence and sufficient source, among others, and are also significantly superior to other materials in promoting cell adhesion and proliferation. These characteristics have led to an increasing emphasis on human hair keratin materials in the field of research in the cosmetic and plastic sector, tissue engineering and even in the whole biomedical field. The human keratin is an excellent antigen-free biological material, has good tissue compatibility, does not cause immune rejection reaction or interfere with normal organism immune response, is suitable for manufacturing the outer membranes, the contents and the like of various soft tissue implants or composite implants, can be also manufactured into soft tissue injection filling materials, has extremely wide application prospect and has great market development space. Currently, three main types of extraction methods of keratin include a reduction method, an enzyme method and an acid-base method. The reduction method is a method adopted by most people at present, but has lower efficiency, the enzyme method is environment-friendly and green, but has higher cost and difficult industrial production, and the acid-base method has better effect, but the prepared keratin has lower molecular weight. None of the human keratin obtained by these methods is suitable for the manufacture of soft tissue implants or composite straight-through adventitia and contents, nor is it suitable for the manufacture of soft tissue injection filling materials. The method combines the experience of former people in the aspect of keratin extraction technology, and is continuously innovated and improved, firstly, human hair fibers are put into alkali solution with certain concentration, and treated at high temperature and high pressure to destroy the surface structure of the human hair fibers, so as to reduce the treatment time, then insoluble matters obtained by filtration are treated by adopting a reduction system, finally, the solution treated by the reduction system is filtered, the insoluble matters obtained by filtration are added into keratinase solution for dissolution, and the light yellow human keratin powder is obtained by dialysis and drying of the filtrate after filtration. The method provided by the invention has the advantages that the human keratin is extracted, the treatment time is reduced, the treatment difficulty is reduced, excessive degradation in the keratin extraction process is avoided, the molecular weight of the obtained keratin can reach 70KD, the molecular weight (3000-7000D) of the keratin extracted by a far-reaching conventional method is high, the keratin has more complete molecular structure and biological function, the outer membrane and the content of the soft tissue implant or the composite implant are manufactured, the supporting performance is better, the stability is stronger, the soft tissue injection filling material is prepared, the stability is good, the degradation is difficult, the retention effect is long, and the better filling effect can be achieved. Disclosure of Invention The invention aims to overcome at least one of the defects in the prior art, and provides a method for extracting human keratin, which combines high-temperature high-pressure alkali liquor treatment, reduction treatment and enzyme dissolution to obtain high-molecular-weight human keratin, improves the purity of the human keratin and the extraction efficiency of the human keratin, does not use harmful substances in the extraction process, and is suitable for being applied to the biomedical field. Is especially suitable for manufacturing the adventitia and the content of the soft tissue implant or the composite implant and the soft tissue injection filling material. In a first aspect, an embodiment of the present invention provides a method for extracting human keratin, which is implemented by the following technical scheme, and is characterized by comprising the following steps: fully cleaning and drying the selected human hair fibers, cutting the human hair fibers to 2cm, adding the human hair fiber scraps into a NaOH solution with the concentration of 0.1mol/L according to the bath ratio of 1:20, and stirring to uniformly mix the solution with the human hair fiber scraps; Placing the system into a sterilizing pot, sterilizing at 121 ℃ for 10 minutes, preserving heat at 70 ℃ for 1 hour, filtering by a nylon filter screen with 300 meshes after the treatment is finished, and repeatedly cleaning the filtrate by distilled water until the pH value is close to 7; Adding the obtained filtration in t