CN111349250A - Rabbit hair keratin antibacterial gel and preparation method thereof - Google Patents

Rabbit hair keratin antibacterial gel and preparation method thereof Download PDF

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Publication number
CN111349250A
CN111349250A CN201910601825.1A CN201910601825A CN111349250A CN 111349250 A CN111349250 A CN 111349250A CN 201910601825 A CN201910601825 A CN 201910601825A CN 111349250 A CN111349250 A CN 111349250A
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rabbit hair
keratin
hair keratin
gel
antibacterial
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张毅
葛乃维
张昊
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Tianjin Polytechnic University
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Tianjin Polytechnic University
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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/075—Macromolecular gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
    • A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
    • A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
    • A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M15/00—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment
    • D06M15/01—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment with natural macromolecular compounds or derivatives thereof
    • D06M15/03—Polysaccharides or derivatives thereof
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M15/00—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment
    • D06M15/01—Treating fibres, threads, yarns, fabrics, or fibrous goods made from such materials, with macromolecular compounds; Such treatment combined with mechanical treatment with natural macromolecular compounds or derivatives thereof
    • D06M15/15—Proteins or derivatives thereof
    • D—TEXTILES; PAPER
    • D06—TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06M—TREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M16/00—Biochemical treatment of fibres, threads, yarns, fabrics, or fibrous goods made from such materials, e.g. enzymatic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2389/00—Characterised by the use of proteins; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
    • C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof

Abstract

The rabbit hair keratin antibacterial gel is prepared with rabbit hair keratin extracted from rabbit hair by L-cysteine reducing process, and through heating-cooling process and self-assembling process with intermolecular interaction to form three-dimensional network structure. The preparation method of the keratin gel comprises the following steps: step one, extracting rabbit hair keratin powder, namely preparing the rabbit hair keratin powder from the pretreated rabbit hair fibers through treatment; and step two, preparing rabbit hair protein gel by using the rabbit hair keratin powder prepared in the step one and adopting a heating-cooling method. And step three, selecting carboxymethyl chitosan as an antibacterial agent to prepare the keratin antibacterial gel. The invention makes full use of the waste rabbit hair keratin, changes waste into valuable and also develops the application of the rabbit hair keratin as a gel material in the field of antibacterial biomaterials.

Description

Rabbit hair keratin antibacterial gel and preparation method thereof
Technical Field
The invention relates to a rabbit hair keratin antibacterial gel and a preparation method thereof, in particular to an antibacterial hydrogel material suitable for the field of biomedical materials, which is prepared by chemically crosslinking rabbit hair keratin powder (extracted by an L-cysteine reduction method) extracted before the subject group by a heating and cooling method to form rabbit hair keratin hydrogel and mixing carboxymethyl chitosan for antibiosis.
Background
China is a traditional country for processing and producing rabbit hair, and a large amount of byproducts such as coarse hair, short hair and two-type hair are generated in the rabbit hair processing process. At present, domestic enterprises rarely recycle the byproducts, and most of the byproducts are directly abandoned. If the sulfur-containing waste water is directly incinerated, the high sulfur content of the waste water causes huge pollution to the environment. Although the rabbit hair waste cannot be spun, the rabbit hair waste still belongs to animal protein fibers, wherein the keratin content can reach nine-tenth of the total mass. The problem of how to utilize the keratin in the waste rabbit hair fiber reasonably and to exert its potential value effectively is the subject of increasing attention of researchers.
At present, the research on recycling of waste rabbit hair mainly relates to the extraction of rabbit hair keratin, and the methods are various and can be divided into physical methods and chemical methods. Wherein the physical method (ZUYI, ZUIRui, extraction and application of keratin in waste rabbit hair [ J ] Tianjin textile technology, 2018 (04): 61-64.) is the most original separation method, and disulfide bonds in keratin are broken by heating and pressurizing, high temperature and high pressure or ultrasonic vibration, so that the keratin is changed into a soluble and digestible polypeptide mixture. The method has simple process and low cost, and the obtained keratin has low molecular weight. Chemical methods (see zuki. research on extraction of keratin from waste rabbit hair [ a ]. chinese Animal husbandry Association, seventh term (2017) chinese rabbit industry development symposium [ C ]. chinese Animal husbandry Association, 2017: 4.) are methods of reducing disulfide bonds to thiol groups under alkaline conditions using the reducibility of a reducing agent to form a protein solution or oxidizing disulfide bonds in keratin to sulfonic acid groups using the oxidation of an oxidizing agent to prepare soluble keratin. The yield of keratin extracted by a chemical method is high, but the oxidation and the breakage of a peptide chain are inevitable in the chemical reaction process, so the average molecular weight of the keratin obtained by the oxidation method is small.
At present, the process for extracting keratin from waste rabbit hair tends to be mature, but the process is not widely applied due to the characteristics of low quality and small molecular weight.
The waste rabbit hair keratin extracted by a chemical method is selected as a raw material in the project to prepare a soft material with special functions, and carboxymethyl chitosan is used as an antibacterial agent to endow the rabbit hair keratin gel with antibacterial performance and research the antibacterial performance of the rabbit hair keratin gel.
Object of the Invention
Aiming at the limitation of the application field of the waste rabbit hair keratin, the invention aims to solve the problems that: provides a method for preparing an antibacterial hydrogel soft material by taking waste rabbit hair keratin as a raw material. The material breaks the application limitation of the waste rabbit hair keratin, and the prepared antibacterial gel has good biocompatibility and obvious antibacterial effect. The preparation method of the antibacterial gel has the characteristics of simple and convenient operation, no toxic or harmful components, mild reaction conditions, easily controlled process, good repeatability and no pollution in the production process.
Disclosure of Invention
The invention aims to overcome the defect that the application field of the existing waste rabbit hair keratin is limited, and provides rabbit hair keratin antibacterial gel and a preparation method thereof.
The invention provides a rabbit hair protein antibacterial gel which is characterized in that rabbit hair keratin extracted from waste rabbit hair is adopted as a raw material, and the keratin gel is prepared by a self-assembly method of forming a three-dimensional network structure by utilizing intermolecular interaction and crosslinking.
The rabbit hair keratin is extracted by a reduction method, and L-cysteine is selected as a reducing agent.
The keratin gel is prepared by a heating-cooling method.
The keratin antibacterial gel adopts carboxymethyl chitosan as an antibacterial agent.
The invention also provides a preparation method of the rabbit hair protein antibacterial gel, which is characterized by comprising the following steps:
step one, extracting rabbit hair keratin powder: taking the pretreated rabbit hair fiber, and cutting the rabbit hair fiber into small sections of about 1 cm for later use. Weighing urea and L-cysteine, dissolving in water, and preparing into a mixed solution. 5mol/L sodium hydroxide solution is prepared, and the mixed solution is gradually dropped to adjust the pH value to 11. Pouring the mixed solution and the pretreated rabbit hair fiber into a 1000ml three-neck round-bottom flask, and stirring for 5h at 85 ℃ in a heat-collecting constant-temperature magnetic stirrer.
And cooling the reacted mixed solution, placing the cooled mixed solution in a centrifuge for centrifugation (8000r/min, 10min), pouring out supernatant for later use, and collecting and drying precipitates. Pouring the supernatant into dialysis bag, dialyzing in distilled water, changing distilled water every 4 hr, and dialyzing for about 3 days until the keratin solution is milky white. Preparing 18 percent by mass of polyethylene glycol 20000 solution, and concentrating for 1 d. And finally, putting the keratin solution into a freeze dryer for vacuum drying until the keratin solution is completely dried. And grinding the dried rabbit hair keratin for 4min by using a high-throughput tissue grinder to obtain white powdery rabbit hair keratin.
Step two, preparing rabbit hair keratin gel: adding the rabbit hair keratin powder into distilled water, adding L-cysteine, stirring with a glass rod until the mixture is dissolved, adding a rotor for stirring to facilitate full reaction, heating in a heat-collecting constant-temperature magnetic stirrer to a fixed temperature, taking out after half an hour, standing at room temperature for cooling, and forming rabbit hair keratin gel.
Step three, preparing the rabbit hair keratin antibacterial gel: dissolving carboxymethyl chitosan in distilled water, adding rabbit hair keratin powder, adding L-cysteine, heating in a heat-collecting constant-temperature magnetic stirrer to a fixed temperature, taking out after half an hour, standing at room temperature, and cooling to obtain rabbit hair keratin antibacterial gel.
8mol/L of urea and 0.165mol/L of L-cysteine in the mixed solution in the first step.
In the second step, the mass of the L-cysteine accounts for 1.5 percent of the mass of the protein.
In the third step, the mass of the L-cysteine accounts for 1.5 percent of the mass of the protein.
The invention has the following advantages and effects: the keratin gel provided by the invention belongs to natural polymer hydrogel, has a microstructure which is an obvious three-dimensional cross-linked reticular structure, has good biocompatibility and degradability, and can be used as a carrier for drug delivery in the biomedical field. The ultraviolet-absorbing gel has good ultraviolet-absorbing function and can be used for preparing a sunscreen gel. The prepared antibacterial keratin gel has good antibacterial performance and water absorption and retention performance, can be used as a medical dressing or a drug sustained-release carrier, and is applied to the protection and repair of skin; can also be used as a finishing agent applied to the traditional textile field and used for functional finishing of fabrics. The invention makes full use of the waste rabbit hair keratin, changes waste into valuable and also exploits the application of the rabbit hair keratin as a gel material in the field of antibacterial biological materials.
Effects of the invention
The structural characterization and physicochemical property analysis of the rabbit hair keratin hydrogel are shown in figures 1 to 6.
As shown in figure 1, a rabbit hair protein gel, the macroscopic form of which is a white soft solid state form with certain mechanical properties, is prepared by a self-assembly method of extracting rabbit hair keratin from rabbit hair and forming a three-dimensional network structure by utilizing intermolecular interaction and crosslinking.
Drawings
FIG. 1 is a schematic representation of rabbit hair keratin gel made according to the present invention.
FIG. 2 is a graph showing the results of an electrophoresis experiment of keratin produced by the present invention.
FIG. 3 is SEM pictures of different concentrations of keratin hydrogel prepared by the present invention.
FIG. 4 shows UV spectra of keratin hydrogels prepared according to the present invention at various concentrations.
FIG. 5 is an infrared spectrum of a keratin hydrogel produced according to the present invention.
FIG. 6 is a TG and DTG curve of the keratin hydrogel produced by the present invention.
FIG. 7 is a graph showing the antibacterial effect of 1% carboxymethyl chitosan antibacterial gel
FIG. 8 is a graph showing the antibacterial effect of 3% carboxymethyl chitosan antibacterial gel
FIG. 9 is a graph showing the antibacterial effect of 5% carboxymethyl chitosan antibacterial gel
FIG. 10 is a graph showing the antibacterial effect of waste rabbit hair keratin gel
FIG. 11 is a graph showing the antibacterial effect of the blank group
The specific implementation method comprises the following steps:
the invention is further described with reference to the following examples and accompanying drawings:
the rabbit hair keratin hydrogel designed by the invention is characterized in that the rabbit hair keratin hydrogel comprises 100 parts of rabbit hair keratin powder by mass; 3 parts of L-cysteine; distilled water 500 parts.
The invention designs a preparation method of rabbit hair keratin gel, which adopts the raw material quality composition and the following process steps:
(1) extracting the waste rabbit hair keratin: taking 20 parts of the pretreated rabbit hair fiber, and cutting the rabbit hair fiber into small sections of about 1 cm for later use. 192 parts of urea and 8 parts of L-cysteine are weighed and dissolved in water to prepare 400 parts of mixed solution (8 mol/L of urea and 0.165mol/L of L-cysteine). 5mol/L sodium hydroxide solution is prepared, and the mixed solution is gradually dropped to adjust the pH value to 11. Pouring the mixed solution and the pretreated rabbit hair fiber into a 1000ml three-neck round-bottom flask, and stirring for 5h at 85 ℃ in a heat-collecting constant-temperature magnetic stirrer. And cooling the reacted mixed solution, placing the cooled mixed solution in a centrifuge for centrifugation (8000r/min, 10min), pouring out supernatant for later use, and collecting and drying precipitates. Pouring the supernatant into dialysis bag, dialyzing in distilled water, changing distilled water every 4 hr, and dialyzing for about 3 days until the keratin solution is milky white. Preparing 18 percent by mass of polyethylene glycol 20000 solution, and concentrating for 1 d. And finally, putting the keratin solution into a freeze dryer for vacuum drying until the keratin solution is completely dried. And grinding the dried rabbit hair keratin for 4min by using a high-throughput tissue grinder to obtain white powdery rabbit hair keratin.
(2) Preparation of rabbit hair keratin gel: weighing 20 parts of rabbit hair keratin powder, adding 100 parts of distilled water, preparing keratin gel with different concentrations, adding L-cysteine accounting for 1.5% of the protein mass, stirring with a glass rod until the protein is dissolved, adding a rotor for stirring to facilitate full reaction, putting the mixture into a heat collection type constant temperature magnetic stirrer for heating to a fixed temperature, taking out after half an hour, standing and cooling at room temperature to form the rabbit hair keratin gel, as shown in figure 1.
As shown in FIG. 2 and FIG. 6, the molecular weight of the extracted waste rabbit hair keratin and the physical and chemical properties of the prepared gel were measured and analyzed, and the results are as follows.
1 molecular weight determination of Rabbit Hair Keratin
The relative molecular weight of the keratin extracted from the waste rabbit hair was determined by using the principle that charged particles would move to an electrode of opposite charge in the matrix under the action of an electric field. Finally, observing a protein band and a standard sample band of the sample to determine that the relative molecular weight of the rabbit hair keratin is 11-22 KDa, which shows that most of keratin with high relative molecular weight is decomposed, and the prepared keratin belongs to keratin with small relative molecular weight.
2 scanning Electron microscope SEM
And observing the surface appearance of the keratin gel by adopting a scanning electron microscope, carrying out freeze drying treatment on the prepared keratin gel with different concentrations, and preparing a sample by selecting a complete section of the block gel or cutting the complete section. And (3) putting the sample into a sample chamber, adjusting the contrast, the brightness and the focal distance until the image is clear, sequentially observing the surface appearance of the gel, and taking a picture. The image is shown in fig. 3, and it can be seen that the concentration of keratin is greater and the porous structure of the gel becomes denser, while the concentration of keratin is less and the porous structure of the gel is loose. And the pore diameter of each pore in the porous structure is obviously reduced along with the increase of the concentration of the keratin, which shows that the higher the concentration of the keratin, the more compact the crosslinking between molecules in the prepared gel.
2 ultraviolet spectrophotometric analysis
Taking 10mg of each of 0.130g/ml, 0.149g/ml and 0.166g/ml keratin gel with different keratin concentrations, pouring the 10mg keratin gel into 10ml of 1mol/L sodium hydroxide solution respectively for full dissolution to prepare a solution to be tested, drawing a standard curve, pouring the prepared solution to be tested into a quartz cuvette for testing, measuring the absorbance of the solution, and drawing an absorption spectrum. The measurement result is shown in FIG. 4, the higher the concentration of the keratin gel, the greater the absorbance, the position of the maximum absorption peak at 206nm, and the absorbance of 3.4; as is clear from FIG. 4, the higher the temperature at which keratin is formed, the greater the absorbance, the position of the maximum absorption peak at 207nm, and the absorbance at 3.2. Therefore, the keratin gel has higher absorbance than keratin, has stronger absorption capacity to ultraviolet light, and shows good ultraviolet-proof function, and the higher the concentration and the higher the temperature of the gel, the tighter the structure of the gel is, and the better the absorbance is.
3 Fourier Infrared Spectroscopy (FT-IR)
In order to determine the chemical structure of the rabbit hair keratin hydrogel, a Fourier infrared spectroscopy method is adopted, the vibration characteristics of functional groups and molecular structures contained in samples with different structures are utilized, an infrared absorption spectrum with corresponding characteristics is scanned, and the structure of a substance is determined by searching a standard spectrogram, so that the method is a commonly used method for determining the molecular structure.
Freeze-drying keratin hydrogel, grinding into powder for later use, weighing 0.13mg potassium bromide powder as a standard sample, weighing 0.12mg potassium bromide and 0.01mg keratin powder, mixing to obtain a first sample, weighing 0.12mg potassium bromide and 0.01mg freeze-dried keratin hydrogel powder, mixing to obtain a second sample, grinding respectively until the particle size is less than 2um, sieving, tabletting in a tabletting machine, performing infrared transmission on the prepared slice, and performing infrared transmission at 400-4000cm-1Collecting spectrum data within the range, scanning for 20 times, and resolution up to 0.4cm-1. The results are shown in FIG. 5, where curve a is the IR spectrum of keratin powder and curve b is the IR spectrum of keratin hydrogel. At 2938cm-1, the absorption peak of C-H stretching vibration in-CH 3 in the protein molecule appears, and at 930cm-1 and 588cm-1, the absorption peak appears as cystine acid in keratinThe characteristic absorption peaks of the residue C-S bond and the residue S-S bond can obviously show that the extracted keratin powder contains a large amount of cystine C-S residues. Meanwhile, the absorption peaks at 1666cm-1, 1529cm-1 and 1236 cm-1 are respectively the characteristic absorption peaks of an amide I band, an amide II band and an amide III band of the keratin hydrogel, the amide I band is a stretching vibration absorption peak of C ═ O, and the absorption peaks are generally 1690-1633 cm-1(ii) a The absorption peak of the amide II band for the bending vibration of N-H and the stretching vibration of C-N is generally in the range of 1540-1520cm-1, and the absorption peak of the amide III band for the bending vibration of C-N and the bending vibration of N-H is generally in the range of 1220-1300 cm-1. By comparison, the infrared spectra of the keratin powder and the keratin hydrogel substantially match.
The amide I peak position of the keratin powder is 1673cm-1The peak position of the amide I band of the keratin hydrogel is 1666cm-1The displacement occurs mainly because the keratin molecular chain and water molecule form hydrogen bond in the gel forming process, and the amide I band generates blue shift under the coupling action of the hydrogen bond and H-O-H bending. According to the report of the literature, the absorption peak of the amide II band is 1539cm-1The results show that the keratin has an α -helix structure and an absorption peak at 1508cm-1The keratin has β -fold structure, and it can be seen from FIG. 4-5 that the amide II band peak of keratin powder is 1536cm-1The amide II peak position of the keratin hydrogel is 1529cm-1The shift change value is-7, and the negative change of the shift indicates that the α -helix structure content of the keratin is reduced, the keratin is converted into an irregular coil structure and a β -fold structure, and the secondary structure of the keratin is changed.
4 Heat stability Performance test
As shown in FIG. 6, thermogravimetric analysis was performed on the keratin hydrogel using a TG 209-F3 type thermogravimetric analyzer to test its thermal stability, a freeze-dried keratin gel powder sample of 10mg was taken, the heating rate was 10K/min, the temperature range was 30-600 ℃, and nitrogen was introduced for protection during the experiment. The results are shown in fig. 8, and it can be seen that the thermal decomposition of the keratin hydrogel can be divided into two stages, the first stage is 50-150 ℃, the thermal weight loss rate of the keratin hydrogel is small, and the stage may be caused by the volatilization of trace moisture and a small amount of residual substances in the hydrogel. The second stage is 200-350 ℃, the keratin hydrogel is cracked, the initial decomposition temperature is about 225 ℃, the heat loss weight of the keratin hydrogel is obviously changed when the temperature is higher than 250 ℃, and the TG curve shows that when the heat loss weight of the keratin hydrogel is 50%, the decomposition temperature is 342.6 ℃, the maximum temperature of the weight loss slope is 302 ℃, and the stability of the keratin hydrogel is reduced compared with that of high-molecular keratin gel. Is used as a drug carrier, and is beneficial to the metabolism of the carrier in the body.
Specific examples of the present invention are given below. The specific examples are intended only to illustrate the invention in further detail and are not intended to limit the claims of the invention.
Example 1
Taking the concentration of carboxymethyl chitosan as a variable, weighing 1 part of carboxymethyl chitosan powder into a beaker, adding distilled water to 100 parts, stirring by using a glass rod until the carboxymethyl chitosan powder is completely dissolved, and preparing into a carboxymethyl chitosan solution with the mass fraction of 1%. Adding 15 parts of rabbit hair keratin powder into the solution, adding L-cysteine accounting for 1.5% of the protein mass, stirring with a glass rod until the keratin powder is completely suspended in the carboxymethyl chitosan solution, and performing antibacterial performance tests of 3 concentration gradients on the sample, wherein the concentration gradient is 10 respectively-2、10-3、10-4The effect is shown in fig. 7.
Example 2
Weighing 3 parts of carboxymethyl chitosan powder into a beaker, adding distilled water to 100 parts, stirring by using a glass rod until the carboxymethyl chitosan powder is completely dissolved, and preparing the carboxymethyl chitosan solution with the mass fraction of 3%. Adding 15 parts of rabbit hair keratin powder into the solution, adding L-cysteine accounting for 1.5% of the protein mass, stirring with a glass rod until the keratin powder is completely suspended in the carboxymethyl chitosan solution, and performing 3 concentration gradient antibacterial property tests on the sample, wherein the concentration gradient is 10 respectively-2、10-3、 10-4The effect is shown in fig. 8.
Example 3
Weighing 5 parts of carboxymethyl chitosan powder into a beaker, adding distilled water to 100 parts, stirring with a glass rod,until the solution is completely dissolved, preparing a carboxymethyl chitosan solution with the mass fraction of 5%. Adding 15 parts of rabbit hair keratin powder into the solution, adding L-cysteine accounting for 1.5% of the protein mass, stirring with a glass rod until the keratin powder is completely suspended in the carboxymethyl chitosan solution, and performing 3 concentration gradient antibacterial property tests on the sample, wherein the concentration gradient is 10 respectively-2、10-3、 10-4The effect is shown in fig. 9.
Comparative example 1
Selecting keratin gel with the same mass as the above, performing antibacterial test under the same conditions, and selecting 3 concentration gradient antibacterial performance tests, 10-2、10-3、10-4The results are shown in FIG. 10.
Comparative example 2
Performing blank experiment under the above conditions, selecting 3 concentration gradient antibacterial performance tests, 10 each-2、10-3、10-4The results are shown in FIG. 11
To summarize:
in conclusion, the keratin hydrogel prepared by adding the carboxymethyl chitosan has a certain antibacterial effect, and a control test group shows that the antibacterial rate of the keratin hydrogel is negative, which indicates that the keratin hydrogel has no antibacterial property, and can provide nutrients for escherichia coli and promote the growth of bacteria. The antibacterial hydrogel prepared from the keratin/carboxymethyl chitosan according to different proportions has certain difference in antibacterial effect. The antibacterial effect can be shown when the antibacterial rate of the escherichia coli reaches more than 70%, and the antibacterial rate reaches 87.02% when the CMCS content is 1% through calculation of the antibacterial rate of each experimental group, so that the antibacterial effect is realized. When the content of CMCS is 5%, the bacteriostasis rate reaches 99.27%.
It is obvious to those skilled in the art that the present invention is not limited to the above embodiments, and it is within the scope of the present invention to adopt various insubstantial modifications of the method concept and technical solution of the present invention, or to directly apply the concept and technical solution of the present invention to other occasions without any modification.

Claims (7)

1. The rabbit hair keratin antibacterial gel is characterized in that the raw materials are prepared by a self-assembly method of extracting rabbit hair keratin from rabbit hair and forming a three-dimensional network structure by utilizing intermolecular interaction and crosslinking.
2. The rabbit hair keratin antibacterial gel according to claim 1, wherein the rabbit hair keratin is extracted by an L-cysteine reduction method.
3. The rabbit hair keratin antibacterial gel according to claim 1, wherein the keratin gel is prepared by a heating-cooling method.
4. A preparation method of rabbit hair keratin antibacterial gel is characterized by comprising the following steps:
step one, extracting rabbit hair keratin powder: taking the pretreated rabbit hair fiber, and shearing the rabbit hair fiber into small sections of about 1 cm for later use. Weighing urea and L-cysteine, dissolving in water, and preparing into a mixed solution. 5mol/L sodium hydroxide solution is prepared, and the mixed solution is gradually dropped to adjust the pH value to 11. Pouring the mixed solution and the pretreated rabbit hair fiber into a 1000ml three-neck round-bottom flask, and stirring for 5h at 85 ℃ in a heat-collecting constant-temperature magnetic stirrer.
And cooling the reacted mixed solution, placing the cooled mixed solution in a centrifuge for centrifugation (8000r/min, 10min), pouring out supernatant for later use, and collecting and drying precipitates. Pouring the supernatant into dialysis bag, dialyzing in distilled water, changing distilled water every 4 hr, and dialyzing for about 3 days until the keratin solution is milky white. Preparing 18 percent polyethylene glycol 20000 solution by mass percent, and concentrating for 1 d. And finally, putting the keratin solution into a freeze dryer for vacuum drying until the keratin solution is completely dried. And grinding the dried rabbit hair keratin for 4min by using a high-throughput tissue grinder to obtain white powdery rabbit hair keratin.
Step two, preparing rabbit hair keratin gel: adding the rabbit hair keratin powder into distilled water, adding L-cysteine, stirring with a glass rod until the rabbit hair keratin powder is dissolved, adding a rotor for stirring to facilitate full reaction, putting the mixture into a heat-collecting constant-temperature magnetic stirrer for heating to a fixed temperature, taking out after half an hour, standing at room temperature for cooling, and forming rabbit hair keratin gel.
Step three, preparing the rabbit hair keratin antibacterial gel: dissolving carboxymethyl chitosan in distilled water, adding rabbit hair keratin powder, adding L-cysteine, heating in a heat-collecting constant-temperature magnetic stirrer to a fixed temperature, taking out after half an hour, standing at room temperature, and cooling to obtain rabbit hair keratin antibacterial gel.
5. The method for preparing rabbit hair keratin antibacterial gel as claimed in claim 4, wherein the mixed solution in the first step is 8mol/L urea and 0.165 mol/L-cysteine.
6. The method for preparing rabbit hair keratin antibacterial gel as claimed in claim 4, wherein in step two, the L-cysteine accounts for 1.5% of the protein.
7. The method for preparing rabbit hair keratin antibacterial gel as described in claim 4, characterized in that, in the third step, 5 parts of carboxymethyl chitosan powder, 100 parts of distilled water and 15 parts of rabbit hair keratin powder.
CN201910601825.1A 2019-07-08 2019-07-08 Rabbit hair keratin antibacterial gel and preparation method thereof Pending CN111349250A (en)

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CN113354840A (en) * 2021-06-21 2021-09-07 内蒙古工业大学 Keratin hydrogel and preparation method thereof, and keratin sponge scaffold and preparation method and application thereof
CN114618010A (en) * 2020-12-09 2022-06-14 四川大学 Multifunctional keratin-based hydrogel and preparation method thereof
CN114733489A (en) * 2022-03-23 2022-07-12 广东省科学院生物与医学工程研究所 Formaldehyde-removing gel taking waste feathers as raw materials and preparation method thereof
CN115400735A (en) * 2022-09-15 2022-11-29 广东省科学院生物与医学工程研究所 Honeycomb gel material prepared from waste feathers, preparation method thereof and application of honeycomb gel material in removing organic dye

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114618010A (en) * 2020-12-09 2022-06-14 四川大学 Multifunctional keratin-based hydrogel and preparation method thereof
CN113354840A (en) * 2021-06-21 2021-09-07 内蒙古工业大学 Keratin hydrogel and preparation method thereof, and keratin sponge scaffold and preparation method and application thereof
CN114733489A (en) * 2022-03-23 2022-07-12 广东省科学院生物与医学工程研究所 Formaldehyde-removing gel taking waste feathers as raw materials and preparation method thereof
CN114733489B (en) * 2022-03-23 2023-07-25 广东省科学院生物与医学工程研究所 Formaldehyde-removing gel with waste feathers as raw materials and preparation method thereof
CN115400735A (en) * 2022-09-15 2022-11-29 广东省科学院生物与医学工程研究所 Honeycomb gel material prepared from waste feathers, preparation method thereof and application of honeycomb gel material in removing organic dye
CN115400735B (en) * 2022-09-15 2024-01-30 广东省科学院生物与医学工程研究所 Honeycomb gel material prepared from waste feathers, preparation method thereof and application of honeycomb gel material in removal of organic dye

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