CN110141527B

CN110141527B - Preparation method of nutritional keratin mask

Info

Publication number: CN110141527B
Application number: CN201910512840.9A
Authority: CN
Inventors: 汤佳鹏; 葛彦; 朱俐
Original assignee: Nantong University
Current assignee: Baitide Pharmaceutical Biotechnology Guangdong Co ltd
Priority date: 2019-06-14
Filing date: 2019-06-14
Publication date: 2021-12-28
Anticipated expiration: 2039-06-14
Also published as: CN110141527A

Abstract

The invention discloses a preparation method of a nutritional keratin mask, which comprises the steps of (1) completely dissolving a certain amount of regenerated human keratin in 98% formic acid solution, and uniformly stirring to obtain spinning solution; (2) carrying out electrostatic spinning by adopting the spinning solution to obtain a keratin nano-fiber membrane; (3) washing the keratin nano-fiber membrane to pH7.4 with a commercially available phosphate buffer solution, drying, and activating by a plasma processor; (4) the activated nano-fiber membrane is soaked in a phosphate buffer solution containing skin nutritional factors for negative pressure flash explosion, then grafting reaction is carried out, and centrifugal freeze-drying is carried out. The nutritional keratin mask provided by the invention can be repeatedly used for 3-5 times, has the advantages of environmental protection and resource saving, is safe to use, and has obvious effects of repairing epidermis, refreshing skin, resisting aging and beautifying skin.

Description

Preparation method of nutritional keratin mask

Technical Field

The invention relates to the fields of cosmetics and biomedical engineering, in particular to a preparation method of a nutritional keratin mask.

Background

With the development of global economy and the improvement of the living standard and the overall quality of people, more and more people begin to use cosmetics, especially for daily care of facial skin, and with the development of society, people have higher and higher requirements on the cosmetics, and people want to use cosmetics with better activity and no preservative. In the existing cosmetics, besides cream cosmetics used in daily life, facial masks are also necessary products for daily facial skin care. The facial mask mainly has four types, namely a paste type, a tearing type, a jelly type and a wet tissue type. The wet tissue type is generally a facial mask paper which is packaged in a single piece and is soaked with essence, and the facial mask paper is divided into the following components by the material weight: non-woven fabrics, silk, cotton, terylene, blended fabrics, tencel, biological fiber, etc. The wet tissue type facial mask has the characteristics of simplest use method and uniform distribution of nutrient components, so that the facial mask is the facial mask with the widest application range.

Common masks in the market mainly have the functions of cleaning, moisturizing, whitening, wrinkle removing, aging resisting, grease balancing and the like, a certain amount of microorganisms are inevitably introduced into the masks in the production process through raw materials, ingredients, packaging materials and the like, and in addition, the mask essence contains rich components which are easy to cause microorganism propagation, such as protein, polysaccharide, oligosaccharide, vitamins and the like, so that the bacteria in the masks are easy to propagate in a large quantity, and the deterioration of the masks is caused. Therefore, various preservatives are added in the production process of the mask to inhibit the propagation of microorganisms or kill microorganisms. The preservative used in the cosmetic has the main effects of preventing the growth of microorganisms in the product, keeping the stable property of the skin care product and preventing the deterioration of the skin care product after the use of the product after the cover is opened. Skin care products with more moisture require preservatives.

In addition, with the progress of science and technology, more and more active ingredients are used in mask products, mainly comprising: growth factors, active proteins, active polypeptides, vitamins and some polyphenols have antioxidant activity. These substances are easily deactivated in the wet tissue type mask. Therefore, the development of the mask which has high nutrient factor combination efficiency and is not easy to deteriorate has extremely important significance for the development of mask products.

Keratin has the advantages of biodegradability, high bioactivity and good compatibility, and the extracted keratin from human hair also has low immunological rejection. Keratin sources are almost unlimited with respect to other bulk materials, and thus research applications in the cosmetic field are receiving increasing attention.

Disclosure of Invention

The invention aims to solve the technical problem of providing a preparation method of a nutritional keratin facial mask, and the keratin nano-fiber facial mask with skin nutritional factor enhancement is obtained by utilizing multiple technologies, including an electrostatic spinning technology, a low-temperature plasma treatment technology, a negative pressure flash explosion technology and the like.

The invention provides a preparation method of a nutritional keratin mask, which comprises the following steps:

s1: completely dissolving regenerated human hair keratin in 98 v/v% formic acid solution, and uniformly stirring to obtain spinning solution;

s2: performing electrostatic spinning by using the spinning solution obtained in the step S1 to obtain a keratin nanofiber membrane;

s3: washing the keratin nanofiber membrane with a phosphate buffer solution until the pH value is 7.4, drying, and then treating and activating by a plasma processor;

s4: soaking the keratin nano-fiber membrane obtained in the step S3 in a phosphate buffer solution added with skin nutritional factors for negative pressure flash explosion, and then carrying out grafting reaction;

s5: and (3) centrifugally freeze-drying the keratin nano-fiber membrane subjected to the grafting reaction to obtain the nutritional keratin facial mask.

Preferably, in the step S1, the concentration of the regenerated human hair keratin is 10-30 g/L.

Preferably, in step S2, in the electrospinning process, the used syringe is 10ml, the needle head is a flat head, and the needle head is a 7-gauge needle; the electrostatic spinning conditions comprise that the voltage is 12-20 KV, the distance is 7-10 cm, the sample introduction speed is 0.3-1.0 ml/h, and the temperature is 25-35 ℃.

Preferably, in the step S3, the drying temperature is 37-45 ℃ and the drying time is 2-4 h.

Preferably, in step S3, the plasma processor processing conditions are: the gas is nitrogen or oxygen, the treatment power is 250-300W, the pressure is 50-60 Pa, and the treatment time is 10-15 min.

Preferably, in step S4, the skin nutrient factor has the following composition:

preferably, the phosphate buffer has a pH of 7.4.

Preferably, in step S4, the vacuum degree of the negative pressure flash explosion is 0.100 to 0.024 mBar.

Preferably, in step S4, the soaking bath ratio of the grafting reaction is 1:100 to 1:300, the reaction temperature is 0 to 4 ℃, and the reaction time is 12 to 24 hours.

Preferably, in step S5, the centrifugal gravitational acceleration is 10000g, the time is 10min, the freeze-drying temperature is-30 to-20 ℃, the vacuum degree is 0.100 to 0.024mBar, and the freeze-drying time is 3 to 5 d.

Compared with the prior art, the invention has the beneficial effects that: the invention develops the facial mask which has high combination efficiency of the nutritional factors and is not easy to deteriorate by utilizing the preparation method of the nano-fiber and grafting the skin nutritional factors on the keratin with the advantages of biodegradability, high bioactivity, good compatibility and the like. The preparation of the nutrient keratin mask utilizes multiple technologies, including an electrostatic spinning technology, a low-temperature plasma treatment technology, a negative pressure flash explosion technology and the like, the use of the technologies greatly improves the combination rate of skin nutrient factors and fibers, the loading capacity is larger, the covalent connection is not easy to deteriorate, the prepared mask can better nourish and repair skin cells, the effects of repairing epidermis, refreshing skin, resisting aging and beautifying the skin are achieved, and the use frequency of the mask can be increased. In addition, the mask provided by the invention contains no moisture, and the function of the mask can be realized in a rehydration mode when in use, so that the mask provided by the invention has better stability in a preservation process compared with the mask in the prior art.

Drawings

FIG. 1: the proliferation rate of the mouse L929 fibroblast cells treated by the embodiment and the comparative example is shown in the invention.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.

The technical scheme adopted by the invention is as follows:

the preparation method of the nutritional keratin mask is characterized by comprising the following steps:

s1: completely dissolving a certain amount of regenerated human hair keratin in 98% (v/v) formic acid solution, and uniformly stirring to obtain spinning solution;

s2: carrying out electrostatic spinning by adopting the spinning solution to obtain a keratin nano-fiber membrane;

s3: washing the keratin nano-fiber membrane with a commercially available phosphate buffer solution until the pH value is 7.4, drying, and then treating and activating by a plasma processor;

s4: soaking the nanofiber membrane obtained in the step three in a phosphate buffer solution added with skin trophic factors for negative pressure flash explosion, and then carrying out grafting reaction;

s5: and (3) centrifugally freeze-drying the nanofiber membrane subjected to the grafting reaction to obtain the nutritional keratin facial mask.

In the above production method of the present invention:

the concentration of the regenerated human hair keratin in the step S1 is preferably 10-30 g/L, and more preferably 20 g/L;

in step S2, in the electrospinning process, the used syringe is preferably 10ml in size, and the needle is preferably a flat head 7-gauge needle; the electrostatic spinning condition is preferably that the voltage is 12-20 KV, more preferably 15KV, the distance is 7-10 cm, more preferably 8cm, the sample introduction speed is 0.3-1.0 ml/h, more preferably 0.5ml/h, the temperature is 25-35 ℃, and more preferably 30 ℃;

in step S3, the pH value of the phosphate buffer is 7.4, the drying temperature is 37-45 ℃, the preferred drying temperature is 37 ℃, and the drying time is 2-4 h, the preferred drying time is 4 h. The plasma treatment conditions were: the gas is nitrogen or oxygen, more preferably oxygen, the treatment power is 250-300W, more preferably 280W, the pressure is 50-60 Pa, more preferably 55Pa, and the treatment time is 10-15 min, more preferably 15 min.

In step S4, the phosphate buffer solution has a pH of 7.4. The composition of the skin trophic factor is preferably: 100-300 g/L of tremella polysaccharide, more preferably 200 g/L; 50-100 g/L of sodium alginate, and more preferably 80 g/L; 1-3 g/L of rose essential oil, and more preferably 2 g/L; 1-3 g/L of lemon essential oil, more preferably 2 g/L; vitamin E1-3 g/L, more preferably 2 g/L; 5-10 g/L of vitamin C, more preferably 8 g/L; 5-15 g/L of propylene glycol, more preferably 10 g/L; 5-15 g/L of glycerol, more preferably 10 g/L; monoglyceride 1-5 g/L, preferably 2 g/L; 1-5 g/L of ceteareth, more preferably 2 g/L. The vacuum degree of the negative pressure flash explosion is preferably 0.100-0.024 mBar, and more preferably 0.024 mBar. The soaking bath ratio of the grafting reaction is preferably 1: 100-1: 300, more preferably 1:200, the reaction temperature is preferably 0-4 ℃, more preferably 4 ℃, and the reaction time is preferably 12-24 h, more preferably 24 h.

In step S5, the centrifugal gravitational acceleration is preferably 10000g, the time is preferably 10min, the freeze-drying temperature is preferably-30 to-20 ℃, the freeze-drying temperature is more preferably-30 ℃, the vacuum degree is preferably 0.100 to 0.024mBar, the freeze-drying time is preferably 3 to 5d, and the freeze-drying time is more preferably 4 d.

Example 1:

1. completely dissolving 2g of regenerated human hair keratin in 100ml of 98% (v/v) formic acid solution, and uniformly stirring to obtain spinning solution;

2. adopting spinning solution to carry out electrostatic spinning, wherein the specification of a used injector is 10ml, and the specification of a needle head is a flat head 7 needle; the electrostatic spinning conditions are that the keratin nano-fiber membrane is obtained under the voltage of 15KV, the distance of 8cm, the sample injection rate of 0.5ml/h and the temperature of 30 ℃;

3. washing the keratin nanofiber membrane with a commercial phosphate buffer (pH7.4) until the pH is 7.4, drying at 37 ℃ for 4 hours, and then treating and activating the keratin nanofiber membrane by a plasma processor under the following conditions: oxygen is adopted as gas, the treatment power is 280W, the pressure is 55Pa, and the treatment time is 15 min;

4. soaking the nanofiber membrane obtained in the third step in a phosphate buffer solution with the pH of 7.4 and containing 200g/L of tremella polysaccharide, 80g/L of sodium alginate, 2g/L of rose essential oil, 2g/L of lemon essential oil, 2g/L of vitamin E, 8g/L of vitamin C, 10g/L of propylene glycol, 10g/L of glycerol, 2g/L of monoglyceride and 2g/L of ceteareth, performing negative pressure flash explosion, wherein the vacuum degree of the negative pressure flash explosion is 0.024mBar, and then performing grafting reaction, wherein the soaking bath ratio of the grafting reaction is 1:200, the soaking temperature is 4 ℃, and the soaking time is 24 hours;

5. centrifuging the nano-fiber after the grafting reaction is finished, wherein the centrifugal gravitational acceleration is 10000g, the time is 10min, and then freeze-drying the nano-fiber at the temperature of minus 30 ℃, the vacuum degree of 0.024mBar, and the freeze-drying time is 4d, thus obtaining the nutritional keratin facial mask.

Example 2:

1. completely dissolving 1g of regenerated human hair keratin in 100ml of 98% (v/v) formic acid solution, and uniformly stirring to obtain spinning solution;

2. adopting spinning solution to carry out electrostatic spinning, wherein the specification of a used injector is 10ml, and the specification of a needle head is a flat head 7 needle; the electrostatic spinning conditions are that the keratin nano-fiber membrane is obtained under the voltage of 12KV, the distance of 7cm, the sample injection rate of 0.3ml/h and the temperature of 25 ℃;

3. washing the keratin nanofiber membrane with a commercial phosphate buffer (pH7.4) until the pH is 7.4, drying at 45 ℃ for 2h, and then treating and activating the keratin nanofiber membrane by a plasma processor, wherein the conditions of the plasma treatment are as follows: adopting nitrogen as gas, treating at 250W under 50Pa for 10 min;

4. soaking the nanofiber obtained in the third step in a phosphate buffer solution with the pH of 7.4 and containing 100g/L of tremella polysaccharide, 50g/L of sodium alginate, 1g/L of rose essential oil, 1g/L of lemon essential oil, 1g/L of vitamin E, 5g/L of vitamin C, 5g/L of propylene glycol, 5g/L of glycerol, 1g/L of monoglyceride and 1g/L of cetearyl alcohol ether to carry out negative pressure flash explosion, wherein the vacuum degree of the negative pressure flash explosion is 0.100mBar, and then carrying out grafting reaction, wherein the soaking bath ratio of the grafting reaction is 1:100, the soaking temperature is 0 ℃, and the soaking time is 12 hours;

5. centrifuging the nano-fiber after the grafting reaction is finished, wherein the centrifugal gravitational acceleration is 10000g, the time is 10min, and then freeze-drying the nano-fiber at the temperature of-20 ℃, the vacuum degree of 0.100mBar, and the freeze-drying time is 3d, so that the nutritional keratin facial mask is obtained.

Example 3:

1. completely dissolving 3g of regenerated human hair keratin in 100ml of 98% (v/v) formic acid solution, and uniformly stirring to obtain spinning solution;

2. adopting spinning solution to carry out electrostatic spinning, wherein the specification of a used injector is 10ml, and the specification of a needle head is a flat head 7 needle; the electrostatic spinning conditions are that the keratin nano-fiber membrane is obtained under the voltage of 20KV, the distance of 10cm, the sample injection rate of 1.0ml/h and the temperature of 35 ℃;

3. washing the keratin nanofiber membrane with a commercial phosphate buffer (pH7.4) until the pH is 7.4, drying at 37 ℃ for 4 hours, and then treating and activating by a plasma processor, wherein the conditions of the plasma treatment are as follows: oxygen is adopted as gas, the treatment power is 300W, the pressure is 60Pa, and the treatment time is 15 min;

4. soaking the nanofiber obtained in the third step in a phosphate buffer solution with the pH of 7.4 and containing 300g/L of tremella polysaccharide, 100g/L of sodium alginate, 3g/L of rose essential oil, 3g/L of lemon essential oil, 3g/L of vitamin E, 10g/L of vitamin C, 15g/L of propylene glycol, 15g/L of glycerol, 5g/L of monoglyceride and 5g/L of cetearyl alcohol ether to carry out negative pressure flash explosion, wherein the vacuum degree of the negative pressure flash explosion is 0.024mBar, then carrying out grafting reaction, the soaking bath ratio of the grafting reaction is 1:300, the soaking temperature is 4 ℃, and the soaking time is 24 hours;

5. centrifuging the nanofiber subjected to the grafting reaction at a centrifugal gravity acceleration of 10000g for 10min, and freeze-drying at-30 ℃, a vacuum degree of 0.024mBar for 5d to obtain the nutritional keratin facial mask.

Comparative example 1:

4. soaking the nanofiber membrane obtained in the step three in a phosphate buffer solution with the pH value of 7.4 for negative pressure flash explosion, wherein the vacuum degree of the negative pressure flash explosion is 0.024mBar, and then carrying out grafting reaction, wherein the soaking bath ratio of the grafting reaction is 1:200, the soaking temperature is 4 ℃, and the soaking time is 24 hours;

5. centrifuging the nano-fiber after the grafting reaction is finished, wherein the centrifugal gravitational acceleration is 10000g, the time is 10min, and then freeze-drying the nano-fiber at the temperature of minus 30 ℃, the vacuum degree of 0.024mBar, and the freeze-drying time is 4d, thus obtaining the plasma treated nano-fiber mask.

Comparative example 2:

4. soaking the nanofiber membrane obtained in the third step in a phosphate buffer solution with the pH of 7.4 and containing 200g/L of tremella polysaccharide, 80g/L of sodium alginate, 2g/L of rose essential oil, 2g/L of lemon essential oil, 2g/L of vitamin E, 8g/L of vitamin C, 10g/L of propylene glycol, 10g/L of glycerol, 2g/L of monoglyceride and 2g/L of cetearyl alcohol ether to carry out grafting reaction, wherein the soaking bath ratio of the grafting reaction is 1:200, the soaking temperature is 4 ℃, and the soaking time is 24 hours;

5. centrifuging the nano-fiber after the grafting reaction is finished, wherein the centrifugal gravitational acceleration is 10000g, the time is 10min, and then freeze-drying the nano-fiber at the temperature of minus 30 ℃, the vacuum degree of 0.024mBar, and the freeze-drying time is 4d, thus obtaining the nano-fiber facial mask of the plasma surface grafted skin trophic factor.

Comparative example 3:

3. washing the keratin nanofiber membrane with a commercial phosphate buffer solution (pH7.4) until the pH is 7.4, and drying at 37 ℃ for 4 h;

5. centrifuging the nano-fiber after the grafting reaction is finished, wherein the centrifugal gravitational acceleration is 10000g, the time is 10min, and then freeze-drying the nano-fiber at the temperature of minus 30 ℃, the vacuum degree of 0.024mBar, and the freeze-drying time is 4d, thus obtaining the nano-fiber facial mask with the surface grafted with the skin nutrition factor.

Fibroblast proliferation assay:

the keratin nanofiber masks of examples and comparative examples were spread on the bottom of a 96-well plate, and mouse L929 fibroblasts were cultured at 1X 10⁵The density of each ml is planted in a 96-well plate, each well is 100 mu l, each group has 5 multiple wells, and the wells are placed in 5% CO₂Culturing in 37 deg.C incubator for 24 hr, removing culture medium by suction, adding 20 μ L MTT (5mg/ml) prepared with PBS into each well, and placing in 5% CO₂And adding 150 mul DMSO into a constant temperature incubator at 37 ℃ for 3h, then oscillating for 10min, placing in a microplate reader at 570nm to read the absorbance OD value, calculating the average value of the OD of each group, and judging the cell proliferation condition.

As can be seen from fig. 1, the keratin nanofiber facial masks of the respective examples can effectively promote the proliferation of fibroblasts, and the effect is superior to that of the comparative example. The results show that the keratin nanofiber facial mask prepared by the invention can effectively activate skin cells and promote metabolism.

The application method of the mask comprises the following steps: when the product is used, the mask can be quickly rehydrated only by opening the package, pouring 20-30ml of purified water or sterile water, standing for 3-5min, and then applying the mask on the skin for normal skin care, and storing at 4 ℃ after use for 3-5 times.

TABLE 1 Total bacterial colony count on repeated use

	Number of times of use	Measurement results (CFU/g)		Number of times of use	Measurement results (CFU/g)
						Example 1	1	<10	Comparative example 1	1	<10
	3	<10		3	8000
							5	<10		5	>10000
Example 2	1	<10	Comparative example 2	1	<10
							3	<10		3	5000
	5	40		5	>10000
						Example 3	1	<10	Comparative example 3	1	<10
	3	<10		3	2000
							5	60		5	>10000

As can be seen from table 1, in the case of repeated use for 3-5 times, the vegetative keratin mask bacteria prepared according to the present invention substantially stopped growing, indicating that the trophic factors thereof were not utilized and deteriorated, whereas the mask bacteria of the comparative example grew in large quantities, indicating that the trophic factors thereof were absorbed and utilized by the bacteria and deteriorated.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.

Claims

1. The preparation method of the nutritional keratin mask is characterized by comprising the following steps:

s2: performing electrostatic spinning by using the spinning solution obtained in the step S1 to obtain a keratin nanofiber membrane; in the electrostatic spinning process, the specification of an injector used is 10ml, and the specification of a needle head is a flat head 7-gauge needle; the electrostatic spinning conditions are that the voltage is 12-20 KV, the distance is 7-10 cm, the sample introduction speed is 0.3-1.0 ml/h, and the temperature is 25-35 ℃;

s3: washing the keratin nanofiber membrane with a phosphate buffer solution until the pH value is 7.4, drying, and then treating and activating by a plasma processor; the plasma processor processing conditions are as follows: the gas is nitrogen or oxygen, the treatment power is 250-300W, the pressure is 50-60 Pa, and the treatment time is 10-15 min;

wherein the skin trophic factor comprises the following components:

the vacuum degree of the negative pressure flash explosion is 0.100-0.024 mBar;

the soaking bath ratio of the grafting reaction is 1: 100-1: 300, the reaction temperature is 0-4 ℃, and the reaction time is 12-24 hours;

2. The method for preparing a nutritional keratin mask as claimed in claim 1, wherein in step S1, the concentration of regenerated human keratin is 10-30 g/L.

3. The method for preparing a nutritional keratin mask as claimed in claim 1, wherein in step S3, the drying temperature is 37-45 ℃ and the drying time is 2-4 h.

4. The method of claim 1, wherein the phosphate buffer has a pH of 7.4.

5. The method for preparing a nutritional keratin mask as claimed in claim 1, wherein in step S5, the centrifugal acceleration is 10000g for 10min, the freeze-drying temperature is-30 to-20 ℃, the vacuum degree is 0.100 to 0.024mBar, and the freeze-drying time is 3 to 5 days.