CN112877384A - Preparation method of bacterial cellulose, bacterial cellulose-chitosan composite gel skin-care mask and preparation method thereof - Google Patents
Preparation method of bacterial cellulose, bacterial cellulose-chitosan composite gel skin-care mask and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a preparation method of bacterial cellulose, which comprises the following steps: taking byproduct residues after protein extraction of soybeans as basic raw materials, called soybean residues for short, preparing a soybean residue biological fermentation culture medium by using the soybean residues, adding 10-18% of a gluconacetobacter xylinus seed solution and amygdalase into the soybean residue biological fermentation culture medium, adding 12-25 mL of the gluconacetobacter xylinus seed solution into every 100mL of the soybean residue biological fermentation culture medium, controlling the pH to be 4.8-6.0, controlling the culture temperature to be 28-35 ℃, culturing for 12-18 days, and controlling the dosage of the amygdalase to be 150-450U/mL, thereby obtaining the bacterial membrane cellulose. The invention also discloses a bacterial cellulose-chitosan composite gel skin-care mask and a preparation method thereof. The mask disclosed by the invention is good in tensile strength, high in mechanical strength, good in biocompatibility and capable of inhibiting bacteria and fungi to a certain extent.
Description
Technical Field
The invention relates to a preparation method of bacterial cellulose, a bacterial cellulose-chitosan composite gel skin-care mask and a preparation method thereof, belonging to the technical field of bacterial cellulose.
Background
The traditional non-woven fabric or gelatin facial mask has the following defects: the facial mask does not accord with the bionic design characteristics of human faces, so that the affinity with skin is poor and the water retention is poor; secondly, the mechanical strength is low, the toughness is poor, so that the material is easy to break and peel; thirdly, the material is not environment-friendly, and pollutes the environment after being used and discarded; fourthly, the auxiliary materials or additives such as hyaluronic acid, collagen and the like are added into the mask, so that the production cost is increased and the price is high.
Disclosure of Invention
The invention aims to solve the technical problem that the bacterial cellulose prepared by the method has a superfine nano-scale three-dimensional porous network structure, has high bionic affinity with a human body application part, is not easy to strip, has a stable chemical structure, has a crystallinity index of 70-78 percent, has high crystallinity and is a degradable superfine nano-cellulose material.
Meanwhile, the bacterial cellulose-chitosan composite gel skin-care mask of the bacterial cellulose has the characteristics of good hydrophilicity, antibacterial property, good air permeability, high mechanical strength, good biocompatibility and the like.
Meanwhile, the invention provides a preparation method of the bacterial cellulose-chitosan composite gel skin-care mask of the bacterial cellulose.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
the invention discloses a preparation method of bacterial cellulose, which comprises the following steps:
s01, preparing bacterial cellulose membrane: taking byproduct residues after protein extraction of soybeans as basic raw materials, namely soybean residues for short, preparing a soybean residue biological fermentation culture medium by using the soybean residues, adding 10-18% of a gluconacetobacter xylinus seed solution and amygdalase into the soybean residue biological fermentation culture medium, adding 12-25 mL of the gluconacetobacter xylinus seed solution into every 100mL of the soybean residue biological fermentation culture medium, controlling the pH to be 4.8-6.0, culturing at 28-35 ℃, culturing for 12-18 days, and using the amount of the amygdalase to be 150-450U/mL, so as to obtain a bacterial membrane cellulose;
s02, washing a bacterial cellulose membrane for multiple times by using distilled water, immersing the bacterial cellulose membrane into 0.1-1 mol/mL NaOH solution, soaking in warm water at 70-90 ℃, magnetically stirring at 90-120 rpm/min for at least 2 hours, removing residual bacteria and a culture medium, taking out, cleaning, immersing the bacterial cellulose membrane into 60-90 ℃ deionized water, cleaning for at least 2 hours, repeating for 3-5 times until the bacterial cellulose membrane is white and semitransparent, repeatedly washing with deionized water to be neutral, freeze-drying for 24-48 hours, and crushing to obtain bacterial cellulose.
The preparation method of the soybean residue biological fermentation medium comprises the following steps: 20-60% of soybean residue, 10-75 g/L of glucose, 5-15 g/L of corn protein powder, 0.75-3.5 g/L of yeast extract, 0.5-3.5 g/L of sodium citrate and Na2HPO41-5 g/L, and the balance of purified water, adjusting the pH value to 5.0-7.4, and sterilizing at 121 ℃ for 15-20 min.
The preparation method of the gluconacetobacter xylinus seed liquid comprises the following steps: taking 1cm of slant strain culture medium2The gluconacetobacter xylinus strain is inoculated in the following culture medium, 20-40g/L of glucose and Na2HPO41-3 g/L, 3-8 g/L yeast extract and MgSO4·7H2O1-3 g/L, absolute ethyl alcohol 20-40 m L, and water is added to 1L.
The method for extracting the protein from the soybeans comprises the following steps: soaking soybean meal for 60-120 min at 80-100 ℃ by using a mixed solution containing 2-6% of NaOH and 8-15% of urea, stirring at 100-120 rpm/min, cleaning residues, and then sieving by using a 200-300-mesh sieve to obtain the soybean residues.
The invention also discloses a bacterial cellulose-chitosan composite gel skin-care mask containing the bacterial cellulose obtained by the bacterial cellulose preparation method, which comprises 2-4 parts by weight of bacterial cellulose powder, 2-4 parts by weight of chitosan, 0.5-1.5 parts by weight of safflower yellow, 0.5-1.5 parts by weight of ginsenoside, 0.2-0.3 part by weight of transdermal absorption enhancer, 15-25 parts by weight of alkaline component and 75-85 parts by weight of purified water.
The skin penetration enhancer comprises one or more of azone, 1, 2-propylene glycol, volatile oil of Camellia oleifera twig, oleic acid, linoleic acid, glycerol, mannitol, N-methyl-2-pyrrolidone NMP, polyethylene glycol, menthol, ostrich oil and Borneolum Syntheticum.
The alkaline component comprises one or more of NaOH, potassium hydroxide, urea, ammonia water, sodium carbonate, potassium carbonate and sodium bicarbonate.
The invention also discloses a preparation method of the bacterial cellulose-chitosan composite gel skin-care mask, which comprises the following steps: every 100mL of reaction solution contains 75-85% of purified water and 15-25% of alkaline components; adding 2-4 g of bacterial cellulose particles and 2-4 g of chitosan into every 100mL of reaction solution, and carrying out ultrasonic or microwave reaction for 30-60 min at the reaction temperature of 50-80 ℃; completely dissolving bacterial fiber and chitosan, removing bubbles by ultrasonic or magnetic stirring, sieving with a 80-100-mesh sieve to obtain a uniform membrane material mixed solution, performing freeze thawing circulation for 3 times, freezing at-20 ℃ for 12-24 hours each time, and thawing for 6-10 hours at room temperature; and after the first two times of freezing and thawing cycles, washing the membrane material with purified water to pH 7.0, after the third time of freezing and thawing cycles, adding 0.5-1.5 g of safflower yellow, 0.2-0.3 g of ginsenoside and 0.2-0.3 g of percutaneous absorption enhancer respectively, introducing the uniformly mixed solution into a mask mould, and carrying out freeze drying at a pre-freezing temperature of-55 to-65 ℃ for 6-10 hours, at a partition plate temperature of 45-60 ℃ for 24-48 hours to obtain the bacterial cellulose-chitosan composite gel skin-care mask.
The ultrasonic power is 100-150W when bubbles are removed.
The rotating speed of the magnetic stirring is 100-150 rpm/min when bubbles are removed.
In the ultrasonic or microwave reaction, the ultrasonic power is 300-400W, and the microwave power is 150-250W.
The invention has the following beneficial effects:
according to the invention, the bacterial cellulose and chitosan are adopted, the environment-friendly degradable natural nanofiber material is used as a mask base material, and whitening, anti-wrinkle and freckle-removing active ingredients such as safflower yellow and ginsenoside are added to prepare the beauty and skin care mask, so that the beauty and skin care mask has high bionic affinity with a human body application part, is not easy to strip, and has stable chemical structure, good hydrophilicity, antibacterial property, good air permeability, high mechanical strength and good biocompatibility; the matrix material is degradable and does not pollute the environment; the production cost is reduced, the performance-price ratio is higher, and the method is suitable for industrial production.
The invention adopts the ultrasonic-freeze thawing cycle technology or the microwave-freeze thawing cycle technology to prepare the bacterial cellulose-chitosan composite gel skin-care mask, the process is simple, the operability is strong, the obtained mask has stable chemical structure, good hydrophilicity and higher cost performance, and is suitable for industrial production.
Drawings
FIG. 1 is a Scanning Electron Microscope (SEM) image of bacterial cellulose prepared by the present invention;
FIG. 2 is an Atomic Force Microscope (AFM) analysis of bacterial cellulose prepared in accordance with the present invention;
fig. 3 is a view showing the biocompatibility of the mask prepared by the present invention.
Fig. 4 is a flow chart of the present invention for preparing a mask.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below with reference to the accompanying drawings and embodiments. The specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.
Example 1:
a method for preparing bacterial cellulose is characterized by comprising the following steps: the method comprises the following steps:
s01, preparing bacterial cellulose membrane: taking the byproduct residue after extracting the protein from the soybean as a basic raw material, namely the soybean residue for short, preparing a soybean residue biological fermentation culture medium by using the soybean residue, adding 15 percent of gluconacetobacter xylinus seed liquid and amygdalase into the soybean residue biological fermentation culture medium, adding 20mL of gluconacetobacter xylinus seed liquid into every 100mL of the soybean residue biological fermentation culture medium, controlling the pH to be 5.5, culturing at the temperature of 30 ℃ for 15 days, and using the amount of the amygdalase to be 300U/mL to obtain a bacterial cellulose membrane material;
s02, washing the bacterial cellulose membrane for multiple times (specifically 3 times), immersing the bacterial cellulose membrane into 0.1mol/mL NaOH solution, soaking in 80 ℃ water bath, magnetically stirring for 2h at 100rpm/min, removing residual bacteria and culture medium, taking out, washing, immersing the bacterial cellulose membrane into 80 ℃ deionized water, washing for 2h, repeating for 3 times until the bacterial cellulose membrane is white and semitransparent, repeatedly washing with deionized water to neutrality, freeze-drying for 24h, and crushing to obtain the bacterial cellulose.
The method for extracting the protein from the soybeans comprises the following steps: soaking soybean powder with a mixed solution containing 2% NaOH and 8% urea at 80 deg.C for 60min, stirring at 100rpm/min, cleaning the residue, and sieving with 200 mesh sieve to obtain the soybean residue.
The bacterial cellulose-chitosan composite gel skin-care mask containing the bacterial cellulose obtained by the preparation method of the bacterial cellulose comprises 4 parts by weight of bacterial cellulose powder, 2 parts by weight of chitosan, 0.5 part by weight of safflower yellow, 0.5 part by weight of ginsenoside, 0.2 part by weight of transdermal absorption enhancer, 5 parts by weight of NaOH, 20 parts by weight of urea and 75 parts by weight of purified water.
The transdermal absorption enhancer comprises azone, 1, 2-propylene glycol and oleic acid, and the weight ratio of the azone to the 1, 2-propylene glycol to the oleic acid is 1:1: 1.
As shown in fig. 4, the preparation method of the bacterial cellulose-chitosan composite gel skin-care mask comprises the following steps: every 100mL of reaction solution contains 75 percent (mass percentage, w/w percent) of purified water, 5 percent of NaOH and 20 percent of urea; adding 4 g of bacterial cellulose particles and 2 g of chitosan into each 100mL of reaction solution, and carrying out ultrasonic or microwave reaction for 30min at the reaction temperature of 50 ℃; completely dissolving bacterial fiber and chitosan, removing bubbles by ultrasonic or magnetic stirring, sieving with 80 mesh sieve to obtain uniform membrane material mixed solution, performing freeze thawing cycle for 3 times, freezing at-20 deg.C for 12 hr each time, and thawing at room temperature for 6 hr; after the first two times of freezing and thawing, washing the membrane material to pH 7.0 by using purified water, after the third time of freezing and thawing, adding 0.5 g of safflower yellow, 0.5 g of ginsenoside and 0.2 g of transdermal absorption enhancer, respectively, introducing the uniformly mixed solution into a mask mould, and carrying out freeze drying at the pre-freezing temperature of-55 ℃, the pre-freezing time of 6 hours, the temperature of a partition board of 45 ℃ and the drying time of 24 hours to obtain the bacterial cellulose-chitosan composite gel skin-care mask.
The ultrasonic power for removing bubbles was 100W.
The rotation speed of the magnetic stirrer for removing bubbles was 100 rpm/min.
In the ultrasonic or microwave reaction, the ultrasonic power is 300W, and the microwave power is 150W.
1. The method for measuring the content of the bacterial cellulose comprises the following steps:
accurately weighing 100mg of bacterial cellulose membrane material, washing with distilled water for multiple times, immersing in 0.1mol/mL NaOH solution, soaking in 80 ℃ water bath, magnetically stirring at 100rpm/min for 2h, removing residual thallus and culture medium, taking out, washing, immersing in 80 ℃ deionized water, washing for 2h, repeating for 3 times until the bacterial cellulose membrane is white and semitransparent, washing with deionized water repeatedly to neutrality, freeze-drying for 24h, and weighing again.
Bacterial cellulose content (g/L) ═ constant weight (g) of bacterial cellulose/volume of culture medium (L)
And (3) determining the crystallinity of the bacterial cellulose: after drying the purified bacterial cellulose particles, the diffraction pattern was determined using an X-ray diffractometer. The experimental conditions are as follows: the maximum tube pressure is 40kV, the maximum tube flow is 40mA, a Cu target Ka ray source is adopted, and the scanning range is 5-60 degrees; the scanning step size is 0.013 degree/step. The crystallinity index (Xc) is calculated as follows:
crystallization index ═ I020－Iam/I020
Wherein, I020Represents the diffraction intensity of the (020) crystal plane; i isamRepresents the diffraction intensity at 18.0 °. According to the peak intensity method, the crystallinity index of the bacterial cellulose is calculated to be 72.97%, and the crystallinity is higher.
2. The bacterial cellulose nanometer three-dimensional network structure:
as shown in fig. 1 to fig. 2, the bacterial cellulose obtained in this embodiment has an ultrafine nano-scale three-dimensional porous network structure as can be seen from analysis by a Scanning Electron Microscope (SEM) and an Atomic Force Microscope (AFM).
3. Detecting the mechanical property (tensile test) of the bacterial cellulose-chitosan composite gel skin-care mask:
the mask was cut into 20mm by 20mm (length by width) in size. Tensile tests were carried out at room temperature using a WAW-2000D model universal tester at a tensile speed of 5 mm/min. Each set of samples was run in parallel for 3 experiments. The result shows that the mask has a three-dimensional network structure, and the moisture in the mask is lost after freeze drying, so that the tensile property is enhanced, and the tensile strength can reach 1.42 MPa.
4. The antibacterial experiment of the bacterial cellulose-chitosan composite gel skin care mask comprises the following steps:
test strains: escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae.
Secondly, sterilizing the bacterial cellulose-chitosan composite gel skin-care mask solution:
dissolving 4 g of bacterial cellulose and 2 g of chitosan in 100mL of purified water at 80 ℃, filtering by a 200-mesh sieve, preparing into 10mg/mL, 20mg/mL and 30mg/mL membrane material solutions respectively, placing in an autoclave, and sterilizing for 10min at 105 ℃.
Measuring bacteriostatic effect
Preparing test bacteria liquid: activating test bacteria with suitable slant culture medium, respectively selecting small amount of thallus Porphyrae, and making into product with bacteria content of 10 with sterile water6～107cfu/mL of bacterial suspension is ready for use.
Under the aseptic condition, a flat plate is prepared by using a sterilized culture medium, after cooling and solidification, 0.5mL of test bacterium liquid is respectively added, and the test bacterium liquid is uniformly coated by using an aseptic coater. And (3) clamping the dried filter paper containing the bacterial cellulose and the chitosan by using sterile tweezers, and putting the filter paper on the surface of the culture medium, wherein 3 pieces of filter paper are put in each petri dish, 2 pieces of filter paper are samples, and 1 piece of filter paper is sterile water as a control. Each strain was grouped in 3 parallel groups. Culturing in constant temperature incubator (bacteria 37 deg.C, 24 hr, fungi 28 deg.C, 48 hr), taking out, measuring the diameter of antibacterial zone, and comparing antibacterial effects.
TABLE 1 determination of antibacterial activity of bacterial cellulose-chitosan composite gel skin-care mask by filter paper method
The result shows that the bacterial cellulose-chitosan composite gel skin-care mask has a certain inhibition effect on bacteria and fungi.
5. Biocompatibility experiment of bacterial cellulose:
cell platform: l929 mouse fibroblasts.
The preparation method of the membrane material leaching liquor comprises the following steps: accurately weighing 100mg of bacterial cellulose membrane material, washing with distilled water for multiple times, immersing into 0.1mol/mL NaOH solution, soaking in 80 ℃ water bath, magnetically stirring at 100rpm/min for 2h, removing residual thallus and culture medium, taking out, cleaning, dissolving in 100mL of purified water at 80 ℃, filtering with a 200-mesh sieve, and preparing into 0.001mol/L, 0.01mol/L, 0.1mol/L and 1mol/L membrane material solutions respectively for later use.
The DMEM high-sugar culture medium is a commercial product.
Grouping experiments: an experimental group, wherein bacterial cellulose-chitosan composite gel skin-care mask leaching liquor is given; blank group, DMEM high sugar medium solution; for the control group, the high density polyethylene extract was administered.
The concentrations of the experimental group, the blank group and the control group are all prepared according to 0.001, 0.01, 0.1 and 1mol/L from low to high.
The execution standard is as follows: GB/T16886.5.
The test steps are as follows: taking L929 mouse fibroblast in logarithmic growth phase, adding into DMEM cell culture solution, and adjusting cell density to 1 × 104Individual cells/mL. The prepared cell suspension was inoculated into 96-well culture plates, each set having 6 wells, and 100. mu.L of the cell suspension was inoculated into each well. Placing in 5% CO2The cells were cultured overnight at 37 ℃ and the medium was replaced to continue the culture. After 24 hours, the medium was replaced again, and then 500mL of 0.5mg/mL MTT medium was added for culture. After 4 hours, the culture was terminated and the medium in the wells was aspirated. Then, 600. mu.L of dimethyl sulfoxide was added to each well and shaken on a shaker at a low speed for 10 minutes so that the crystals were completely dissolved. Finally, detection was performed using a full-wavelength microplate reader (OD ═ 570), and the image display mode was Mean ± SD. The cell survival rate is calculated by the formula:
survival rate of cells D/DOD×100％
Wherein D isODIs the absorbance value of the control group, and D is the absorbance value of the experimental group.
As shown in FIG. 3, the results show that the bacterial cellulose membrane still has a cell viability of 90.6% at a concentration of 1mol/L, and meets the national standards according to the cytotoxicity classification standard I. The bacterial cellulose of the embodiment has good biocompatibility and low cytotoxicity.
Example 2:
a method for preparing bacterial cellulose comprises the following steps:
s01, preparing bacterial cellulose membrane: taking the byproduct residue after extracting the protein from the soybean as a basic raw material, short for soybean residue, preparing a soybean residue biological fermentation culture medium by using the soybean residue, adding 10 percent of gluconacetobacter xylinus seed liquid and amygdalase into the soybean residue biological fermentation culture medium, adding 12mL of gluconacetobacter xylinus seed liquid into every 100mL of the soybean residue biological fermentation culture medium, controlling the pH to be 4.8, culturing at the temperature of 28 ℃, culturing for 12 days, and using the amount of the amygdalase to be 150U/mL, thus obtaining a bacterial cellulose membrane material;
s02, washing the bacterial cellulose membrane by distilled water for multiple times, immersing the bacterial cellulose membrane into 1mol/mL NaOH solution, soaking in 70 ℃ water bath, magnetically stirring for 3 hours at 90rpm/min, removing residual thalli and culture medium, taking out, washing, immersing the bacterial cellulose membrane into 60 ℃ deionized water, washing for 2.5 hours, repeating for 3 times until the bacterial cellulose membrane is white and semitransparent, repeatedly washing with deionized water to neutrality, freeze-drying for 24 hours, and crushing to obtain the bacterial cellulose.
The method for extracting the protein from the soybeans comprises the following steps: soaking soybean powder with a mixed solution containing 6% NaOH and 15% urea at 100 deg.C for 120min, stirring at 120rpm/min, cleaning the residue, and sieving with 300 mesh sieve to obtain the soybean residue.
The bacterial cellulose-chitosan composite gel skin-care mask containing the bacterial cellulose obtained by the preparation method of the bacterial cellulose comprises 2 weight parts of bacterial cellulose powder, 4 weight parts of chitosan, 1.5 weight parts of safflower yellow, 1.5 weight parts of ginsenoside, 0.3 weight part of transdermal absorption enhancer, 5 weight parts of potassium hydroxide, 5 weight parts of urea, 5 weight parts of ammonia water and 85 weight parts of purified water.
The transdermal absorption enhancer comprises camellia oleifera branch volatile oil, polyethylene glycol, menthol, ostrich oil and borneol in a weight ratio of 1:1:1:1: 2.
The preparation method of the bacterial cellulose-chitosan composite gel skin-care mask comprises the following steps: every 100mL of reaction solution contains 85% of purified water, 5% of potassium hydroxide, 5% of urea and 5% of ammonia water; adding 2 g of bacterial cellulose particles and 4 g of chitosan into each 100mL of reaction solution, and carrying out ultrasonic or microwave reaction for 60min at the reaction temperature of 80 ℃; completely dissolving bacterial fiber and chitosan, removing bubbles by ultrasonic or magnetic stirring, sieving with 100 mesh sieve to obtain uniform membrane material mixed solution, performing freeze thawing cycle for 3 times, freezing at-20 deg.C for 24 hr each time, and thawing at room temperature for 10 hr; after the first two times of freezing and thawing, washing the membrane material to pH 7.0 by using purified water, after the third time of freezing and thawing, adding 1.5 g of safflower yellow, 1.5 g of ginsenoside and 0.3 g of percutaneous absorption enhancer, respectively, introducing the uniformly mixed solution into a mask mould, and carrying out freeze drying at the pre-freezing temperature of-65 ℃, the pre-freezing time of 10 hours, the temperature of a partition board of 60 ℃ and the drying time of 48 hours to obtain the bacterial cellulose-chitosan composite gel skin-care mask.
The ultrasonic power for bubble removal was 150W.
The rotation speed of the magnetic stirrer for removing bubbles was 150 rpm/min.
In the ultrasonic or microwave reaction, the ultrasonic power is 400W, and the microwave power is 250W.
Through detection, the crystallinity index of the bacterial cellulose prepared by the embodiment is 70.15%, and the crystallinity is higher. The bacterial cellulose-chitosan composite gel skin care mask prepared by the embodiment has the tensile strength of 1.54MPa, has a certain inhibition effect on bacteria and fungi, and has good biocompatibility.
Example 3:
a method for preparing bacterial cellulose comprises the following steps:
s01, preparing bacterial cellulose membrane: taking the byproduct residue after extracting the protein from the soybean as a basic raw material, namely the soybean residue for short, preparing a soybean residue biological fermentation culture medium by using the soybean residue, adding 18 percent of gluconacetobacter xylinus seed liquid and amygdalase into the soybean residue biological fermentation culture medium, adding 25mL of gluconacetobacter xylinus seed liquid into every 100mL of the soybean residue biological fermentation culture medium, controlling the pH to be 6.0, culturing at the temperature of 35 ℃, culturing for 18 days, and using the amount of the amygdalase to be 450U/mL, thus obtaining a bacterial cellulose membrane material;
s02, washing the bacterial cellulose membrane for multiple times by using distilled water, immersing the bacterial cellulose membrane into 0.5mol/mL NaOH solution, soaking the bacterial cellulose membrane in water bath at 90 ℃, magnetically stirring the bacterial cellulose membrane for 2 hours at 120rpm/min, removing residual thalli and culture medium, taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane, immersing the bacterial cellulose membrane in deionized water at 90 ℃ for 2 hours, repeating the steps for 4 times until the bacterial cellulose membrane is white and semitransparent, repeatedly washing the bacterial cellulose membrane to be neutral by using deionized water, freezing and drying the bacterial cellulose membrane for 36 hours, and crushing the bacterial cellulose membrane to obtain the bacterial cellulose.
The method for extracting the protein from the soybeans comprises the following steps: soaking soybean powder with a mixed solution containing 4% NaOH and 10% urea at 90 deg.C for 100min, stirring at 110rpm/min, washing the residue, and sieving with 250 mesh sieve to obtain the soybean residue.
The bacterial cellulose-chitosan composite gel skin-care mask containing the bacterial cellulose obtained by the preparation method of the bacterial cellulose comprises 3 weight parts of bacterial cellulose powder, 4 weight parts of chitosan, 1 weight part of safflower yellow, 1 weight part of ginsenoside, 0.2 weight part of transdermal absorption enhancer, 10 weight parts of sodium carbonate, 10 weight parts of sodium bicarbonate and 80 weight parts of purified water.
The transdermal absorption enhancer is N-methyl-2-pyrrolidone NMP.
The preparation method of the bacterial cellulose-chitosan composite gel skin-care mask comprises the following steps: every 100mL of reaction solution contains 80% of purified water, 10% of sodium carbonate and 10% of sodium bicarbonate; adding 3g of bacterial cellulose particles and 4 g of chitosan into each 100mL of reaction solution, and carrying out ultrasonic or microwave reaction for 40min at the reaction temperature of 70 ℃; completely dissolving bacterial fiber and chitosan, removing bubbles by ultrasonic or magnetic stirring, sieving with 100 mesh sieve to obtain uniform membrane material mixed solution, performing freeze thawing cycle for 3 times, freezing at-20 deg.C for 18 hr each time, and thawing at room temperature for 8 hr; after the first two times of freezing and thawing, washing the membrane material with purified water to pH 7.0, after the third time of freezing and thawing, adding 1 g of safflower yellow, 1 g of ginsenoside and 0.2 g of percutaneous absorption enhancer, respectively, introducing the uniformly mixed solution into a mask mould, and carrying out freeze drying at the pre-freezing temperature of-60 ℃ for 8 hours, at the temperature of a partition board of 50 ℃ for 36 hours to obtain the bacterial cellulose-chitosan composite gel skin-care mask.
The ultrasonic power when removing bubbles was 120W.
The rotation speed of the magnetic stirrer for removing bubbles was 120 rpm/min.
In the ultrasonic or microwave reaction, the ultrasonic power is 350W, and the microwave power is 200W.
Through detection, the crystallinity index of the bacterial cellulose prepared by the embodiment is 78.23%, and the crystallinity is higher. The bacterial cellulose-chitosan composite gel skin care mask prepared by the embodiment has the tensile strength of 1.48MPa, has a certain inhibition effect on bacteria and fungi, and has good biocompatibility.
Example 4:
this example differs from example 1 only in that:
the transdermal absorption enhancer is glycerol and mannitol, and the weight ratio of the glycerol to the mannitol is 1: 1.
The alkalinity is potassium carbonate.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (9)
1. A method for preparing bacterial cellulose is characterized by comprising the following steps: the method comprises the following steps:
s01, preparing bacterial cellulose membrane: taking byproduct residues after protein extraction of soybeans as basic raw materials, namely soybean residues for short, preparing a soybean residue biological fermentation culture medium by using the soybean residues, adding 10-18% of a gluconacetobacter xylinus seed solution and amygdalase into the soybean residue biological fermentation culture medium, adding 12-25 mL of the gluconacetobacter xylinus seed solution into every 100mL of the soybean residue biological fermentation culture medium, controlling the pH to be 4.8-6.0, culturing at 28-35 ℃, culturing for 12-18 days, and using the amount of the amygdalase to be 150-450U/mL, so as to obtain a bacterial membrane cellulose;
s02, washing a bacterial cellulose membrane for multiple times by using distilled water, immersing the bacterial cellulose membrane into 0.1-1 mol/mL NaOH solution, soaking in warm water at 70-90 ℃, magnetically stirring at 90-120 rpm/min for at least 2 hours, removing residual bacteria and a culture medium, taking out, cleaning, immersing the bacterial cellulose membrane into 60-90 ℃ deionized water, cleaning for at least 2 hours, repeating for 3-5 times until the bacterial cellulose membrane is white and semitransparent, repeatedly washing with deionized water to be neutral, freeze-drying for 24-48 hours, and crushing to obtain bacterial cellulose.
2. The method for preparing bacterial cellulose according to claim 1, wherein: the method for extracting the protein from the soybeans comprises the following steps: soaking soybean meal for 60-120 min at 80-100 ℃ by using a mixed solution containing 2-6% of NaOH and 8-15% of urea, stirring at 100-120 rpm/min, cleaning residues, and then sieving by using a 200-300-mesh sieve to obtain the soybean residues.
3. A bacterial cellulose-chitosan composite gel skin care mask containing the bacterial cellulose obtained by the bacterial cellulose preparation method of claim 1, which is characterized in that: the antibacterial cellulose powder comprises 2-4 parts by weight of bacterial cellulose powder, 2-4 parts by weight of chitosan, 0.5-1.5 parts by weight of safflor yellow, 0.5-1.5 parts by weight of ginsenoside, 0.2-0.3 part by weight of transdermal absorption enhancer, 15-25 parts by weight of alkaline component and 75-85 parts by weight of purified water.
4. The bacterial cellulose-chitosan composite gel skin care mask as claimed in claim 3, wherein: the skin penetration enhancer comprises one or more of azone, 1, 2-propylene glycol, volatile oil of Camellia oleifera twig, oleic acid, linoleic acid, glycerol, mannitol, N-methyl-2-pyrrolidone NMP, polyethylene glycol, menthol, ostrich oil and Borneolum Syntheticum.
5. The bacterial cellulose-chitosan composite gel skin care mask as claimed in claim 3, wherein: the alkaline component comprises one or more of NaOH, potassium hydroxide, urea, ammonia water, sodium carbonate, potassium carbonate and sodium bicarbonate.
6. The method for preparing the bacterial cellulose-chitosan composite gel skin care mask as claimed in any one of claims 3 to 5, wherein: the method comprises the following steps: every 100mL of reaction solution contains 75-85% of purified water and 15-25% of alkaline components; adding 2-4 g of bacterial cellulose particles and 2-4 g of chitosan into every 100mL of reaction solution, and carrying out ultrasonic or microwave reaction for 30-60 min at the reaction temperature of 50-80 ℃; completely dissolving bacterial fiber and chitosan, removing bubbles by ultrasonic or magnetic stirring, sieving with a 80-100-mesh sieve to obtain a uniform membrane material mixed solution, performing freeze thawing circulation for 3 times, freezing at-20 ℃ for 12-24 hours each time, and thawing for 6-10 hours at room temperature; and after the first two times of freezing and thawing cycles, washing the membrane material with purified water to pH 7.0, after the third time of freezing and thawing cycles, adding 0.5-1.5 g of safflower yellow, 0.2-0.3 g of ginsenoside and 0.2-0.3 g of percutaneous absorption enhancer respectively, introducing the uniformly mixed solution into a mask mould, and carrying out freeze drying at a pre-freezing temperature of-55 to-65 ℃ for 6-10 hours, at a partition plate temperature of 45-60 ℃ for 24-48 hours to obtain the bacterial cellulose-chitosan composite gel skin-care mask.
7. The method of claim 6, wherein: the ultrasonic power is 100-150W when bubbles are removed.
8. The method of claim 6, wherein: the rotating speed of the magnetic stirring is 100-150 rpm/min when bubbles are removed.
9. The method of claim 6, wherein: in the ultrasonic or microwave reaction, the ultrasonic power is 300-400W, and the microwave power is 150-250W.
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