CN102885710B - Sodium alga acid/chitosan/collagen composite micro-ball with active components and preparation method of composite micro-ball - Google Patents
Sodium alga acid/chitosan/collagen composite micro-ball with active components and preparation method of composite micro-ball Download PDFInfo
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Abstract
The invention discloses a micro-ball with a plurality of cosmetic active components, which is prepared by using collagen, sodium alga acid and chitosan as carrier materials through the following steps of: dissolving collagen in acid medium, mixing with sodium alga acid and active components to form a uniform colloidal solution; enabling the mixture to enter a divalent positive ion solution by using an injector in a liquid drop form, forming a primary micro-ball under the action of gel; arranging the micro-ball in a chitosan solution, further curing; and washing the micro-ball by using de-ionized water to recombine the collagen in the micro-ball, and finally, forming the sodium alga acid/chitosan/collagen composite micro-ball. The micro-ball prepared by using the method provided by the invention has a smooth and flat surface, excellent monodispersity, uniform particle size distribution and fast swelling property, and belongs to a novel cosmetic active component carrier system; and the free-dried micro-ball can be used together with solvents, in this way, the effect of the common action of multi-component various-function cosmetic active components is achieved.
Description
Technical field
The present invention relates to a kind of novel cosmetic load system, be specifically related to alginate/chitosan/collagen composite microsphere of a kind of carrying active composition and preparation method thereof, belong to cosmetics preparing technical field.
Background technology
Arbutin, fruit acid, anisic acid etc. are the current popular whitening of water solublity more safely and effectively raw materials, can reduce Skin pigmentation, dispelling stain and freckle.Ceramide is the one in glycosphingolipid, is a kind of water-soluble grease metallic substance, can improve cuticular water conservation and barrier function, solves xerosis cutis, wrinkle, coarse phenomenon.Somatomedin is the cytokine with stimulating cellular growth activity, and result of study proves: somatomedin reduces with advancing age gradually, and human body shows various senile symptoms.Taking epithelical cell growth factor (EGF) and fibroblast growth factor (FGF) as a class cell growth factor of representative has cosmetic result significantly, can solve skin senilism, sallow, wrinkle, the phenomenon such as lax, there is deep layer repair.Chinese patent CN101732169A report is the formula for function skin care liquid by arbutin, and Japanese Mercian Corp discloses ceramide synthesis accelerators as cosmetics and the patent of skin preparations for extenal use (CN101583345).Chinese patent CN102406556A discloses one and has contributed to keep the bioactive cosmetic composition of epidermal growth factor, and the main component of said composition is epidermal growth factor, Hy drolyzed Skin Powder, hyaluronate sodium etc.Chinese patent CN101548935 discloses a kind of acid fibroblast growth factor in the purposes of preparing in cosmetics, and its production technology is also provided simultaneously.Prior art self stability is not good enough, is subject to the easy inactivation of external interference, for example, under light and the inappropriate condition of surrounding, easily decompose, and in air, is easily oxidized yet, and lacks the stability of storage, therefore in cosmetic applications, has been subject to certain restriction.
Sodium alginate (C
6h
7o
8na) n is mainly made up of the sodium salt of alginic acid, the copolymer that relies on Isosorbide-5-Nitrae-glycosidic bond to be connected and to be made up of GM, MM and the GG fragment of different proportion with b-D-guluronic acid (G unit) by a-L-mannuronic acid (M unit).Sodium alginate is just taken in American Pharmacopeia as far back as 1938, and its gentle sol-gel process, good biocompatibility make sodium alginate be suitable for the microcapsule carrier as release or embedding medicinal, albumen and cell.Chitosan; chemical name is β-(1; 4)-2-amino-2-deoxidation-D-glucosan; it is the product of chitin N-deacetylation; also be the alkaline aminopolysaccharide of unique one that biosphere exists; water insoluble but can be dissolved in low-concentration organic acid or mineral acid, the microsphere of preparing taking chitosan as carrier has control release, improves many-sided advantages such as medicine stability.
At present, use the method that activated protein or medicine microspheres are carried in maximum preparations to have solvent evaporated method, for example patent CN1463696 has reported the Poly-dl-lactide-b-polyethylene glycol microsphere of two emulsion system solvent evaporated method preparation year protein medicaments, and US Patent No. 5288502 has been reported the microsphere that utilizes emulsion solvent evaporation technique preparation to carry polypeptide and albumen; Patent CN1398584A has reported and has utilized emulsion dispersion legal system for bFGF-PLGA sustained-release micro-spheres; US Patent No. 2007275082 has been reported the method for utilizing spray drying method Bidirectional nozzle bag medicine carrying thing, said method generally all will be used organic solvent or cross-linking agent, it has the following disadvantages, the one, the easy stability of destroying coated active substance, the 2nd, easily remain in and in microsphere, affect microsphere performance, the 3rd, the carrier of using is to synthesize family macromolecule and semi-synthetic family macromolecule as main, not good enough in the biocompatibility of human body.
Summary of the invention
The object of the present invention is to provide a kind of good biocompatibility, not with an organic solvent, cross-linking agent and emulsifying agent, load does not have alginate/chitosan/collagen composite microsphere of multiple efficacies cosmetic active ingredient.
Another object of the present invention is to provide that a kind of technique is simple, cost is low, gentle environmental protection, the load of the active ingredient good stability preparing has alginate/chitosan/collagen composite method for preparing microsphere of multiple efficacies cosmetic active ingredient.
Implementation procedure of the present invention is as follows:
Alginate/chitosan/collagen composite microsphere of carrying active composition, obtained by sodium alginate, chitosan, Ι Collagen Type VI and active fraction preparation, wherein in complex microsphere, sodium alginate, chitosan, Ι Collagen Type VI and active component weight ratio are 3 ~ 10:2 ~ 9:1 ~ 3:2 ~ 5, described active component is selected from whitening active component, moisturizing active component ceramide and activity of fighting against senium composition cell growth factor at least two kinds, and whitening active component is selected from arbutin, fruit acid, anisic acid, aloin.
Alginate/chitosan/collagen composite method for preparing microsphere of above-mentioned carrying active composition, comprises the steps:
(1) Ι Collagen Type VI is dissolved in acid medium, adds sodium alginate and active component to be mixed to form homogeneous colloid solution, and remove bubble;
(2) above-mentioned colloid solution is squeezed to and in bivalent cation solution, forms elementary microsphere;
(3) elementary microsphere is placed in the further curing alginate/chitosan/collagen composite microsphere that obtains carrying active composition of chitosan acid solution.
In above-mentioned steps (1), acid medium is organic or inorganic acid, and its pH is 1-4.In the colloid solution preparing, sodium alginate concentration is 0.1-10 %, and Ι Collagen Type VI concentration is that 0.1-20 %(is preferably 0.5-10 %), activity component concentration is that 0.5-20 %(is preferably 1-10 %).
In above-mentioned steps (2), bivalent cation is selected from Ba
2+, Ca
2+, Co
2+, Zn
2+, Mn
2+, Ni
2+, bivalent cation solution quality percent concentration is 0.1-10 %.Colloid solution is transferred in the glass syringe that is connected with gas injection pump, regulates syringe pump speed that mixture is squeezed in bivalent cation solution, form elementary microsphere by gelatification.
In above-mentioned steps (3), chitosan acid solution is that chitosan is dissolved in formic acid, acetic acid, propanoic acid, lactic acid, malic acid, citric acid, ascorbic acid, oxalic acid, succinic acid, malonic acid, fatty acid, acetone acid, 1,3-propanedicarboxylic acid, tartaric acid, aspartic acid, salicylic acid or glycollic acid solution and is obtained, the concentration of chitosan is 0.1-10 %, and the molecular weight of chitosan is 2-30 ten thousand.
Alginate/chitosan/collagen composite microsphere of above-mentioned carrying active composition can be used in cosmetics, when use, by the swelling rear use in solvent of alginate/chitosan/collagen composite microsphere of carrying active composition, described solvent is butanediol and hyaluronic aqueous solution, pH is 6.0-6.8, and wherein the concentration of butanediol is 2%.
The microsphere features smooth surface rounding that uses the inventive method to make, monodispersity is good, particle size distribution homogeneous, there is Fast-swelling character, its diameter Distribution coefficient can be controlled in 20%, diameter can be in 50-1500 μ m free adjustment, the swellbility > 200% in its 30s solvent.
Diameter Distribution coefficient=[(
di-
d)
2/
n]
1/2/
d} × 100% (1)
Swellbility=[(
m t-
m 0)/
m 0] × 100% (2)
In formula (1),
di is the diameter of each microsphere,
dfor several average diameters, the microsphere quantity that N is calculated diameter, N > 200; In formula (2),
m tthe quality (g) of microsphere in the t moment;
m 0it is the quality (g) of dry rear microsphere.
It is carrier that the present invention adopts sodium alginate, chitosan and the collagen of high-biocompatibility and reduced immunogenicity, by self assembly Electrostatic Absorption method, the multiple different efficacies active component in cosmetics is coated and prepares alginate/chitosan/collagen composite microsphere, have the following advantages:
(1) in preparation method, do not add poisonous organic solvent, cross-linking agent and emulsifying agent etc., therefore in final microsphere, there is no residual noxious substance, accomplished zero real pollution;
(2) a kind of microsphere supported bag simultaneously carries multiple different efficacies cosmetic active ingredient, has realized the feature that same cosmetics have both different efficacies, has met the demand of multifunction nursing articles for use;
(3) microsphere and solvent collocation are used, and convenient and swift, instant joining, active component discharges in time, has improved the availability of active component;
(4) preparation method technique is simple, cost is low, gentle environmental protection, and market prospect is wide.
Brief description of the drawings
Fig. 1 microspheres flow chart.
Fig. 2 embodiment 1 microsphere photo.
The optical microscope photograph of microsphere prepared by Fig. 3 embodiment 2.
Fig. 4 embodiment 4 microsphere scanning electron microscope (SEM) photographs.
The microspherulite diameter scattergram of Fig. 5 embodiment 1-4, (A) the particle size distribution figure (particle size distribution coefficient is 17.85%) of the microsphere that prepared by embodiment 1, (B) the particle size distribution figure (particle size distribution coefficient is 18.23%) of the microsphere that prepared by embodiment 2, (C) the particle size distribution figure (particle size distribution coefficient is 18.77%) of the microsphere that prepared by embodiment 3, the particle size distribution (figure particle size distribution coefficient is 19.05%) of the microsphere that (D) prepared by embodiment 4.
The different indexs of Fig. 6 effect checking-test combinations matched group change contrast trendgram.
Detailed description of the invention
Below in conjunction with embodiment, the invention will be further described, and concentration of the present invention is mass percent concentration, measuring instrument: scanning electron microscope JSM5800, laser particle analyzer COULTER LS200.
The method that the present invention prepares bag carries alginate/chitosan/collagen composite microsphere of multiple cosmetic active ingredient, comprise the following steps: purified abortive calfskin type i collagen is dissolved in the acid medium that pH value is 1-4, at 4-10 DEG C, be mixed to form mutually homogeneous colloid solution with sodium alginate and active component, the ultrasonic bubble of removing, wherein: the concentration range of collagen is 0.1-20 %, the concentration range of sodium alginate is 0.1-10%, the concentration range of cosmetic active ingredient (except cell growth factor) is 0.5-20 %, the concentration range of cell growth factor is 0.1-10 μ g/ml.
Above-mentioned mixed colloidal solution is transferred in the glass syringe that is connected with gas injection pump, gas in syringe can be nitrogen, helium, argon, carbon dioxide and airborne any one, gas flow rate in syringe can be controlled by syringe pump, can form like this microsphere of different size.
Mixture is pushed by gas injection pump, and mixture overcomes viscous force and surface tension, enters calcium chloride solution with droplet-like from pin hole, forms elementary microsphere by gelatification, and gelatification time 5-20min, collects microsphere after sucking filtration.Cation in solution can be Ba
2+, Ca
2+, Co
2+, Zn
2+, Mn
2+, Ni
2+in any one, conventionally select Ca
2+carry out gelation, in the present invention, the concentration of bivalent cation is 0.1-10 %.
Above-mentioned elementary microsphere is transferred in the chitosan solution that concentration is 0.1-10% and is further solidified, hardening time 5-30min, the pH value of chitosan solution is between 4.0-6.5, and the acid that is used for dissolving chitosan comprises at least one in formic acid, acetic acid, propanoic acid, lactic acid, malic acid, citric acid, ascorbic acid, oxalic acid, succinic acid, malonic acid, fatty acid, acetone acid, 1,3-propanedicarboxylic acid, tartaric acid, aspartic acid, salicylic acid, glycolic.
With deionized water wash microsphere 2-3 time, the collagen in microsphere is recombinated, and alginate/chitosan/collagen composite microsphere forms.This step is to carry out in room temperature, and washing microsphere is later preserved after lyophilization.Microsphere after lyophilizing can use with solvent collocation, and active component is transported to skin muscular layer safely and effectively.
embodiment 1:particle diameter is the load whitening composition arbutin of 1500 μ m and alginate/chitosan/collagen composite microsphere of high moisturizing composition ceramide
1, as shown in Figure 1, it is in 3.5 acetic acid aqueous solution that 30mg abortive calfskin type i collagen is dissolved in to 6ml pH value, add the sodium alginate of 2.0ml 6%, 10ml concentration is that arbutin and the 10ml concentration of 6 mg/ml is 1mg/ml ceramide mix homogeneously, and ultrasonic 30s removes bubble.
2, mixed solution is transferred to and is connected with gas injection pump (model A-99, Razel scientific instrument company) glass syringe in, outside has a pipe that nitrogen cylinder is connected with pump, the air velocity of control pump is the nitrogen of 5 ml/min, in 1 % calcium chloride solution, solidify 6min, elementary microsphere forms.In order to prevent collagenous degeneration, this step is carried out at 4 DEG C.
3, the chitosan that is 300,000 by molecular weight is dissolved in 0.05% acetic acid and obtains the solution that chitosan mass percent concentration is 1.0 %, its pH is 5.5, elementary microsphere in step 2 is transferred in chitosan solution and is further solidified, and the crosslinking curing time is 5min, and this step is carried out in room temperature.
4, with deionized water wash microsphere 2-3 time, alginate/chitosan/collagen composite microsphere forms.Microsphere is carried out to granularmetric analysis, and the microspherulite diameter of preparation is at 1480 ± 50 μ m(Fig. 2,5).Then microsphere is preserved after lyophilization.
5, the microsphere after lyophilizing is placed in to solvent and carries out swellingly, reach 247% in 30s swellbility.Wherein solvent is butanediol and hyaluronic aqueous solution, and pH is 6.5, and the concentration of butanediol is 2%.
6, by by microballoon lyophilized, weigh, the weight ratio that obtains sodium alginate, chitosan, Ι Collagen Type VI and active component is about: 120:105:30:70.
embodiment 2:particle diameter is the load whitening composition arbutin of 500 μ m and alginate/chitosan/collagen composite microsphere of high moisturizing composition ceramide
Carry out according to the condition in embodiment 1, change the air velocity of the control pump in step 2 into 7.5 ml/min by 5 ml/min.Microsphere is carried out to granularmetric analysis, and the microspherulite diameter of preparation is at 505 ± 20 μ m(Fig. 3,5).Microsphere after lyophilizing is placed in to solvent and carries out swellingly, reach 315% in 30s swellbility.
embodiment 3:particle diameter is alginate/chitosan/collagen composite microsphere of three kinds of active component of load of 1500 μ m
Three kinds of active component are respectively whitening composition arbutin, moisturizing composition ceramide, defying age component list skin cell growth factor EGF
Step 1: it is in 3.5 acetic acid solution that purified 30mg abortive calfskin type i collagen is dissolved in to 6ml pH value, the sodium alginate of 2.0ml 8%, 10ml concentration is that arbutin, the 10ml concentration of 6mg/ml is that 1mg/ml ceramide, 2ml concentration are the EGF(epithelical cell growth factor of 1 μ g/ml) mix homogeneously, ultrasonic 30s removes bubble.
All the other steps are identical with embodiment 1.By by microballoon lyophilized, weigh, the weight ratio that obtains sodium alginate, chitosan, Ι Collagen Type VI and active component is about: 160:140:30:70.
Microsphere is carried out to granularmetric analysis, and the microspherulite diameter of preparation is at 1490 ± 50 μ m(Fig. 5).Microsphere after lyophilizing is placed in to solvent and carries out swellingly, reach 268% in 30s swellbility.
embodiment 4:particle diameter is alginate/chitosan/collagen composite microsphere of three kinds of active component of load of 50 μ m
Three kinds of active component refer to respectively whitening composition arbutin, moisturizing composition ceramide, defying age component list skin cell growth factor EGF
Carry out according to the condition in embodiment 3, change the air velocity of the control pump in step 2 into 12 ml/min by 5 ml/min.Microsphere is carried out to granularmetric analysis, and the microspherulite diameter of preparation is at 53 ± 9 μ m(Fig. 5).Microsphere after lyophilizing is placed in to solvent and carries out swellingly, reach 367% in 30s swellbility.
embodiment 5:alginate/chitosan/collagen composite microsphere effect of three kinds of active component arbutin of load, ceramide, epithelical cell growth factor EGF is evaluated
Study subject: 20 ages without skin sensitivity history are the 25-30 young woman in year, 2cm × 2cm region is tested position inside forearm, and a side is trial zone, and a side is check plot
Using method: complex microsphere and solvent collocation are used, spread upon same position inside forearm, concrete using method is, get 2-3 microsphere in the centre of the palm, drip two solvents on microsphere, wait for about 30s, after microsphere is fully swelling, softly break to spheroid with forefinger and middle finger, then be evenly applied in forearm, sooner or later respectively once.Continue to use 30 days.
Testing time: since the 1st day, carry out every three days skin test one time, test process is by dermatologist's complete monitoring.
Test index: comprise that cuticlar water is right, melanin and skin elasticity.
The German CK of the right employing of the cuticlar water CM825 of company type moisture tester contact continuous measurement mode is carried out.Skin color adopts the German CK MEXAMETER MX16 of company melanin analyzer to measure.Melanin value is measured by instrument, and average measurement is averaged for 3 times, and measuring range is 0 ~ 999, measures numerical value higher, illustrates that in skin, melanin content is higher.Skin elasticity adopts SME575 elasticity tester to observe, and average measurement is averaged for 3 times, uses R
2value representation (stretching/resilience ratio).
Concrete measurement result is in table 1, variation tendency is shown in Fig. 6, result shows, after the active component microsphere (wait and carry solvent) of continuous three kinds of different efficacies of working load, the aggregative indicator of skin all has significant change, right average 46.8 points that improved of cuticlar water, melanin has also reduced by 49.3 points than matched group, and skin elasticity has increased by 16.3% than matched group simultaneously.The active component microsphere of therefore, load different efficacies can meet the needs of multifunction nursing articles for use completely.
Claims (9)
1. a preparation method for alginate/chitosan/collagen composite microsphere of carrying active composition, is characterized in that comprising the steps:
(1) Ι Collagen Type VI is dissolved in acid medium, adds sodium alginate and active component to be mixed to form homogeneous colloid solution, and remove bubble;
(2) above-mentioned colloid solution is squeezed to and in bivalent cation solution, forms elementary microsphere;
(3) elementary microsphere is placed in the further curing alginate/chitosan/collagen composite microsphere that obtains carrying active composition of chitosan acid solution;
In complex microsphere, sodium alginate, chitosan, Ι Collagen Type VI and active component weight ratio are 3 ~ 10:2 ~ 9:1 ~ 3:2 ~ 5, described active component is selected from whitening active component, moisturizing active component ceramide and activity of fighting against senium composition cell growth factor at least two kinds, and whitening active component is selected from arbutin, fruit acid, anisic acid, aloin.
2. the preparation method of alginate/chitosan/collagen composite microsphere of carrying active composition according to claim 1, is characterized in that: the particle diameter of described complex microsphere is 50-1500 μ m.
3. the preparation method of alginate/chitosan/collagen composite microsphere of carrying active composition according to claim 1, is characterized in that: in step (1), acid medium is organic or inorganic acid, and its pH is 1-4.
4. the preparation method of alginate/chitosan/collagen composite microsphere of carrying active composition according to claim 1, it is characterized in that: in the colloid solution preparing in step (1), sodium alginate concentration is 0.1-10 %, Ι Collagen Type VI concentration is 0.1-20 %, and activity component concentration is 0.5-20 %.
5. the preparation method of alginate/chitosan/collagen composite microsphere of carrying active composition according to claim 1, is characterized in that: in step (2), described bivalent cation is selected from Ba
2+, Ca
2+, Co
2+, Zn
2+, Mn
2+, Ni
2+, bivalent cation solution quality percent concentration is 0.1-10 %.
6. the preparation method of alginate/chitosan/collagen composite microsphere of carrying active composition according to claim 5, it is characterized in that: in step (2), colloid solution is transferred in the glass syringe that is connected with gas injection pump, regulate syringe pump speed that mixture is squeezed in bivalent cation solution, form elementary microsphere by gelatification.
7. the preparation method of alginate/chitosan/collagen composite microsphere of carrying active composition according to claim 1, it is characterized in that: in step (3), chitosan acid solution is that chitosan is dissolved in formic acid, acetic acid, propanoic acid, lactic acid, malic acid, citric acid, ascorbic acid, oxalic acid, succinic acid, malonic acid, acetone acid, 1,3-propanedicarboxylic acid, tartaric acid, aspartic acid, salicylic acid or glycollic acid solution and is obtained, and the concentration of chitosan is 0.1-10 %.
8. alginate/chitosan/collagen composite microsphere of the carrying active composition preparing according to method described in claim 1 is in the application of preparing in cosmetics.
9. application according to claim 8, it is characterized in that: when use, by the swelling rear use in solvent of alginate/chitosan/collagen composite microsphere of carrying active composition, described solvent is butanediol and hyaluronic aqueous solution, pH is 6.0-6.8, and wherein the concentration of butanediol is 2%.
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