CN105924654A - Method for regenerating human hair keratin by means of L-cysteine - Google Patents
Method for regenerating human hair keratin by means of L-cysteine Download PDFInfo
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- CN105924654A CN105924654A CN201610292984.4A CN201610292984A CN105924654A CN 105924654 A CN105924654 A CN 105924654A CN 201610292984 A CN201610292984 A CN 201610292984A CN 105924654 A CN105924654 A CN 105924654A
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- crinis carbonisatus
- keratin
- cys
- human hair
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08H—DERIVATIVES OF NATURAL MACROMOLECULAR COMPOUNDS
- C08H1/00—Macromolecular products derived from proteins
- C08H1/06—Macromolecular products derived from proteins derived from horn, hoofs, hair, skin or leather
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Abstract
The invention relates to a method for regenerating human hair keratin by means of L-cysteine. The method comprises the steps of cleaning human hair fiber, then conducting degreasing, drying and smashing, adding obtained powder to L-cysteine-urea complex liquid with a bath ratio of (10:1)-(30:1), adjusting PH to be 10-12, conducting sealing, increasing temperature to 65-95 DEG C, conducting oscillation to obtain human hair and cysteine mixed solution, conducting centrifugal filtration, and taking filtrate as human hair keratin solution; conducting dialysis, adjusting PH to be 4.0-4.5, conducting standing after brown keratin is completely separated out, removing supernate, and conducting refrigeration and drying to obtain the human hair keratin. The method is simple and practical, and keratin powder high in molecular weight and extraction rate is prepared through dissolution and acetic acid precipitation of low-cost L-cysteine. The prepared keratin powder can be stored for a long time and can be applied to traumatology. The keratin powder can be dissolved in proper solvent for composite spinning with other polymers to be applied to bioengineering and other fields.
Description
Technical field
The invention belongs to the regeneration field of discarded Crinis Carbonisatus, utilize Cys regeneration Crinis Carbonisatus angle egg particularly to one
White method.
Background technology
The utilization of discarded Crinis Carbonisatus used as biomaterial for the first time from the eighties in 20th century, just caused many researcheres
Attention, owing to Crinis Carbonisatus is grown on human body, cause great interest in bioengineered tissue field, additionally hair color contain
A large amount of melanin, have strong absorption, it is possible to as uvioresistant function additive to ultraviolet.Crinis Carbonisatus and other animal hair
As similar in Pilus Caprae seu Ovis etc., it is mainly composed of alpha-Keratin, its difference is that Crinis Carbonisatus (about 80um) is relative to animal wool (Pilus Caprae seu Ovis
20um) diameter substantially increases, and its top layer substantial amounts of melanin composition plays the biggest inhibition to the reduction effect of reducing agent,
And define melanoprotein in the fibre.
Human hair keratin molecule forms highly cross-linked three-dimensional stability structure by cystine linkage, hydrogen bond, sat linkage and other key effect,
Due to the complicated compactness of its structure, it does not under general condition dissolve, and it is by by force that tradition dissolves the method for human hair keratin
Cystine linkage or the peptide bond of the reagent destruction protein molecules such as alkali, strong acid, reducing agent or oxidant resolve into small peptide, these methods
Generally there is a large amount of mineral acid, alkali, reducing agent, the discharge of the unfriendly reagent of the environment such as oxidant and carbamide, environment is caused
Serious impact.Cys is again β-mercaptoalanine, is a kind of aminoacid with physiological function, is constitutive protein matter
20 several amino acids in the only aminoacid with reproducibility group sulfydryl (-SH).Cys has many good qualities, first
First Cys, intramolecular also has sulfydryl, has certain reproducibility；Secondly Cys does not has toxicity, experiment condition
The gentleest, relatively low to requirement of experiment；Cys low price again, wide material sources, resource is abundant.
Summary of the invention
The technical problem to be solved is to provide one and utilizes the quick environmental-friendly and efficient extraction Crinis Carbonisatus angle of Cys
The method of albumen, and the method can significantly reduce the usage amount of the denaturants such as carbamide, simple to operate, the medicine of use and setting
Standby with low cost, it is easy to accomplish industrialized production；The method with the Crinis Carbonisatus chopped fiber that goes out of use that can not directly be utilized as raw material,
Reach twice laid maximize, high-valued；The brown solid powder prepared has the spy that molecular weight is high, extraction ratio is high
Point.
The one of the present invention utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, including:
(1) after Crinis Carbonisatus fiber being cleaned, defat, dry, pulverize；
(2) by step (1) is pulverized the Crinis Carbonisatus 5g obtained join in Cys solution (when urea content is 0,
For joining in Cys-carbamide complex liquid), bath raio is 10:1～30:1, pH regulator to 10～12, seals, and heats up
To 65～95 DEG C, vibration, obtain Crinis Carbonisatus and keratic mixed solution, centrifugal filtration, take filtrate, obtain human hair keratin
Solution；
(3) by the human hair keratin solution in step (2), dialysis, regulation pH value is 4.0～4.5, to brown keratin
Separate out completely, stand, remove supernatant, obtain the brown keratin of precipitation, freezing, it is dried, obtains human hair keratin powder.
Described step (1) is cleaned as first at H2O2Clean with in the alkaline solution of sodium carbonate, then clean with water；Wherein,
The temperature cleaned is 85-95 DEG C, and bath raio is 30:1～40:1；H2O2With 30%H in the alkaline solution of sodium carbonate2O2Dense
Degree is 4-8mL/L, and the concentration of soda is 5-15g/L.
In described step (1), defat is: with ethanol-acetone mixed solvent defat 12～24h；Wherein, ethanol-acetone mixing
In solvent, ethanol is 1:1～2:1 with the volume ratio of acetone；The degreasing agent submergence Crinis Carbonisatus that skimming processes uses.
In described step (1), pulverizing obtains Crinis Carbonisatus is 2～5mm length.
In described step (2) in Cys solution: Cys consumption is 10～50g/L；The consumption of carbamide is
0～8M.
In described step (2), pH regulator is for regulate by NaOH solution.
In described step (2), vibration is mechanical oscillation, and the time is 8～10h；Centrifugal rotational speed is 5000～9500rpm, temperature
Being 10～20 DEG C, the time is 1-5min.
In described step (3), the time of dialysis is 48～72h；During dialysis, the molecular cut off of dialyzer is 8000～14000,
A deionized water is changed every 4～6h.
In described step (3), the mode of regulation pH is: add acetum under stirring condition.
The time stood in described step (3) is 2～3h；Freezing is :-60 DEG C of refrigerator freezings 6～12h；It is dried and is: cold
Being dried on lyophilizer, condition is: 20～5Pa, and temperature is-60 DEG C～-50 DEG C, and the time is 36～48h.
The human hair keratin powder obtained in described step (3) be placed in 4 DEG C stand-by.
Keratin raw material used by the present invention is solid waste Crinis Carbonisatus, and its chemism is to cut off keratin molecule with method of reducing
-S-S-the key of interchain, regulates the pH of reaction system on this basis to desired value, and selects and have strong Swelling Functions
Carbamide, makes protein macromolecule the most swelling until dissolving, and obtains keratin solution and solid, the notable spy that wherein solution has
Levying, its solid phase powder goods are brown, and its technological process is: → pulverizing → dissolving → keratin solution is cleaned in raw material → oxidation
→ centrifugal, filtration → dialysis → precipitating → lyophilization → keratin powder body.
The keratin solution that the present invention prepares may be used for finishing functions, masking and spinning etc., and keratin powder properties is steady
Fixed, traumatic medicine can be may be used for stored longer under certain condition, it is also possible to be combined with other polymer
Spinning, is applied to bioengineering field etc..
Beneficial effect
Medicine the most used in the present invention and equipment cost are cheap, simple to operate, it is easy to accomplish industrialized production；
Reducing agent the most used in the present invention does not has toxicity, wide material sources, and resource is abundant；
Urea content the most used in the present invention can greatly reduce；
4. the present invention is with the Crinis Carbonisatus chopped fiber that goes out of use that can not directly be utilized as raw material, reach twice laid maximize, high-valued；
5. the distinguishing features such as it is high that the human hair keratin that the present invention extracts has molecular weight, and extraction ratio is high.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments be merely to illustrate the present invention and
It is not used in restriction the scope of the present invention.In addition, it is to be understood that after having read the content that the present invention lectures, people in the art
The present invention can be made various changes or modifications by member, and these equivalent form of values fall within the application appended claims equally and limited
Scope.
Embodiment 1
(1) by Crinis Carbonisatus at H2O2Alkaline solution is cleaned, wherein 30%H2O2Concentration be 8mL/L, the concentration of soda is
10g/L, temperature is 95 DEG C, bath raio 40:1.Crinis Carbonisatus again with ethanol-acetone mixed solvent defat 12h, after then drying
Pulverize；Wherein, the temperature of cleaning is 85 DEG C, and bath raio is 30:1, the degreasing agent submergence Crinis Carbonisatus that skimming processes uses, second
In alcohol-acetone mixed solvent, ethanol is 2:1 with the volume ratio of acetone.
(2) the Crinis Carbonisatus 5g after pulverizing joins in Cys-carbamide composite fluid, wherein bath raio 20:1, L-half
Cystine consumption is 10g/L, and the consumption of carbamide is 8M.With the NaOH regulation pH value of 200g/L to about 12, cover
Plug seal, is warming up to 75 DEG C, after mechanical oscillation 10h, obtains Crinis Carbonisatus and keratic mixed solution.
(3) being centrifuged Crinis Carbonisatus and keratin mixed solution, rotating speed is 9500rmp, and temperature is 25 DEG C, and the time is 5min,
Filter, take filtrate, obtain human hair keratin solution.
(4) to obtained filtrate dialysis 72h, the molecular cut off of dialyzer is 8000～14000, changes every 4-6h
Deionized water.
(5), after having dialysed, in dialysis solution, add acetum while stirring, regulate its pH to 4.0～4.5, make brown
Keratin separates out completely.
(6), after being stood 2h, remove supernatant, take precipitation.
(7) to pellet frozen 12h, then carrying out lyophilization, condition is: air pressure 20Pa, and temperature is-60 DEG C, the time
For 36h, obtain brown keratin pressed powder, be placed on 4 DEG C stand-by.Final keratin molecule amount is at more than 10Kda
Yield reach as high as 25.74%.
Embodiment 2
(1) by Crinis Carbonisatus at H2O2Alkaline solution is cleaned, wherein 30%H2O2Concentration be 8mL/L, the concentration of soda is
10g/L, temperature is 95 DEG C, bath raio 40:1.Crinis Carbonisatus again with ethanol-acetone mixed solvent defat 12h, after then drying
Pulverize.Wherein, the temperature of cleaning is 85 DEG C, and bath raio is 30:1, the degreasing agent submergence Crinis Carbonisatus that skimming processes uses, second
In alcohol-acetone mixed solvent, ethanol is 2:1 with the volume ratio of acetone.
(2) the Crinis Carbonisatus 5g after pulverizing joins in Cys-carbamide composite fluid, wherein bath raio 20:1, L-half
Cystine consumption is 10g/L, and the consumption of carbamide is 2M.With the NaOH regulation pH value of 200g/L to about 12, cover
Plug seal, is warming up to 85 DEG C, after mechanical oscillation 10h, obtains Crinis Carbonisatus and keratic mixed solution.
(3) being centrifuged Crinis Carbonisatus and keratin mixed solution, rotating speed is 9500rmp, and temperature is 25 DEG C, and the time is 5min,
Filter, take filtrate, obtain human hair keratin solution.
(4) to obtained filtrate dialysis 72h, the molecular cut off of dialyzer is 8000～14000, changes every 4-6h
Deionized water.
(5), after having dialysed, in dialysis solution, add acetum while stirring, regulate its pH to 4.0～4.5, make brown
Keratin separates out completely.
(6), after being stood 2h, remove supernatant, take precipitation.
(7) to pellet frozen 12h, then carrying out lyophilization, air pressure 15Pa, temperature is-50 DEG C, and the time is 48h,
To brown keratin pressed powder, be placed on 4 DEG C stand-by.Final keratin molecule amount is the highest in the yield of more than 10Kda
Up to 38.42%.
Embodiment 3
(1) by Crinis Carbonisatus at H2O2Alkaline solution is cleaned, wherein 30%H2O2Concentration be 8mL/L, the concentration of soda is
10g/L, temperature is 95 DEG C, bath raio 40:1.Crinis Carbonisatus again with ethanol-acetone mixed solvent defat 12h, after then drying
Pulverize.Wherein, the temperature of cleaning is 85 DEG C, and bath raio is 30:1, the degreasing agent submergence Crinis Carbonisatus that skimming processes uses, second
In alcohol-acetone mixed solvent, ethanol is 2:1 with the volume ratio of acetone.
(2) the Crinis Carbonisatus 5g after pulverizing joins in Cys-carbamide composite fluid, wherein bath raio 20:1, L-half
Cystine consumption is 15g/L, and the consumption of carbamide is 8M.With the NaOH regulation pH value of 200g/L to about 12, lid
Upper plug seal, is warming up to 85 DEG C, after mechanical oscillation 10h, obtains Crinis Carbonisatus and keratic mixed solution.
(3) being centrifuged Crinis Carbonisatus and keratin mixed solution, rotating speed is 9500rmp, and temperature is 25 DEG C, and the time is 5min,
Filter, take filtrate, obtain human hair keratin solution.
(4) to obtained filtrate dialysis 72h, the molecular cut off of dialyzer is 8000～14000, changes every 4-6h
Deionized water.
(5), after having dialysed, in dialysis solution, add acetum while stirring, regulate its pH to 4.0～4.5, make brown
Keratin separates out completely.
(6), after being stood 2h, remove supernatant, take precipitation.
(7) to pellet frozen 12h, then carrying out lyophilization, air pressure 10Pa, temperature is-55 DEG C, and the time is 42h,
To brown keratin pressed powder, be placed on 4 DEG C stand-by.Final keratin molecule amount is the highest in the yield of more than 10Kda
Up to 51.80%.
Embodiment 4
(1) by Crinis Carbonisatus at H2O2Alkaline solution is cleaned, wherein 30%H2O2Concentration be 8mL/L, the concentration of soda is
10g/L, temperature is 95 DEG C, bath raio 40:1.Crinis Carbonisatus again with ethanol-acetone mixed solvent defat 12h, after then drying
Pulverize.Wherein, the temperature of cleaning is 85 DEG C, and bath raio is 30:1, the degreasing agent submergence Crinis Carbonisatus that skimming processes uses, second
In alcohol-acetone mixed solvent, ethanol is 2:1 with the volume ratio of acetone.
(2) the Crinis Carbonisatus 5g after pulverizing joins in Cys-carbamide composite fluid, wherein bath raio 20:1, L-half
Cystine consumption is 10g/L, and the consumption of carbamide is 8M.With the NaOH regulation pH value of 200g/L to about 10, cover
Plug seal, is warming up to 95 DEG C, after mechanical oscillation 10h, obtains Crinis Carbonisatus and keratic mixed solution.
(3) being centrifuged Crinis Carbonisatus and keratin mixed solution, rotating speed is 9500rmp, and temperature is 25 DEG C, and the time is 5min,
Filter, take filtrate, obtain human hair keratin solution.
(4) to obtained filtrate dialysis 72h, the molecular cut off of dialyzer is 8000～14000, changes every 4-6h
Deionized water.
(5), after having dialysed, in dialysis solution, add acetum while stirring, regulate its pH to 4.0～4.5, make brown
Keratin separates out completely.
(6), after being stood 2h, remove supernatant, take precipitation.
(7) to pellet frozen 12h, then carrying out lyophilization, air pressure 5Pa, temperature is-60 DEG C, and the time is 36h,
To brown keratin pressed powder, be placed on 4 DEG C stand-by.Final keratin molecule amount is the highest in the yield of more than 10Kda
Up to 12.78%.
Claims (9)
1. utilize Cys to regenerate the keratic method of Crinis Carbonisatus, including:
(1) after Crinis Carbonisatus fiber being cleaned, defat, dry, pulverize；
(2) joining in Cys solution by shredding the Crinis Carbonisatus obtained in step (1), bath raio is 10:1～30:1, pH
Regulation, to 10～12, seals, is warming up to 65～95 DEG C, and vibration obtains Crinis Carbonisatus and keratic mixed solution, centrifugal filtration,
Take filtrate, obtain human hair keratin solution；
(3) by the human hair keratin solution in step (2), dialysis, regulation pH value is 4.0～4.5, complete to brown keratin
Separate out, stand, remove supernatant, freezing, it is dried, obtains human hair keratin.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
State in step (1) and clean as first at H2O2Clean with in the alkaline solution of sodium carbonate, then clean with water；Wherein, clean
Temperature be 85-95 DEG C, bath raio is 30:1～40:1；H2O2With 30%H in the alkaline solution of sodium carbonate2O2Concentration be
4-8mL/L, the concentration of soda is 5-15g/L.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
Stating defat in step (1) is: with ethanol-acetone mixed solvent defat 12～24h；Wherein, in ethanol-acetone mixed solvent
Ethanol is 1:1～2:1 with the volume ratio of acetone.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
State in step (2) in Cys solution: Cys consumption is 10～50g/L；The consumption of carbamide is 0～8M.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
Stating pH regulator in step (2) is to regulate by 200g/L NaOH solution.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
Stating vibration in step (2) is mechanical oscillation, and the time is 8～10h；Centrifugal rotational speed is 2500-5000rpm, temperature be 10～
20 DEG C, the time is 1-5min.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
The time of dialysis in step (3) of stating is 48～72h；The molecular cut off of dialyzer is 8000～14000.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
The mode of regulation pH in step (3) of stating is: add acetum under stirring condition.
One the most according to claim 1 utilizes Cys to regenerate the keratic method of Crinis Carbonisatus, it is characterised in that institute
The time of standing in step (3) of stating is 2～3h；Freezing is :-60 DEG C of refrigerator freezings 6～12h；It is dried as lyophilization, bar
Part is: air pressure 20～5Pa, and temperature is-60 DEG C～-50 DEG C, and the time is 36～48h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610292984.4A CN105924654A (en) | 2016-05-05 | 2016-05-05 | Method for regenerating human hair keratin by means of L-cysteine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610292984.4A CN105924654A (en) | 2016-05-05 | 2016-05-05 | Method for regenerating human hair keratin by means of L-cysteine |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102839441A (en) * | 2012-09-25 | 2012-12-26 | 天津工业大学 | High-elasticity keratin regenerated fiber and preparation method thereof |
| CN104861663A (en) * | 2015-04-28 | 2015-08-26 | 东华大学 | Method for regenerating wool keratin by L-cysteine |
-
2016
- 2016-05-05 CN CN201610292984.4A patent/CN105924654A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102839441A (en) * | 2012-09-25 | 2012-12-26 | 天津工业大学 | High-elasticity keratin regenerated fiber and preparation method thereof |
| CN104861663A (en) * | 2015-04-28 | 2015-08-26 | 东华大学 | Method for regenerating wool keratin by L-cysteine |
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