CN106491416B - Preparation method and application of hibiscus syriacus enzyme - Google Patents
Preparation method and application of hibiscus syriacus enzyme Download PDFInfo
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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Abstract
The invention relates to the technical field of enzyme preparation, and also relates to application of the enzyme in the field of skin care products. A preparation method of hibiscus syriacus enzyme comprises the steps of taking hibiscus flowers, hibiscus leaves and hibiscus stems as raw materials, sterilizing, fermenting with bacillus subtilis for a period of time, adding yeast and lactobacillus, continuing to ferment, adding a nitrogen source, a carbon source and inorganic salt during fermentation, sterilizing after fermentation, filtering, and filtering with an ultrafiltration membrane to obtain the hibiscus enzyme. The preparation method of the invention can obviously improve the content of amino acid and flavone in the product, and has more obvious effects of resisting aging, preserving moisture and recovering skin elasticity.
Description
Technical Field
The invention relates to the technical field of enzyme preparation, and also relates to application of the enzyme in the field of skin care products.
Background
With the healthy development of socioeconomic in China, people pay more attention to personal images, and the healthy skin of human is moist, fine, soft, elastic and glossy and has a natural protective function. Therefore, the antioxidant and anti-aging cosmetics have a far-reaching market prospect.
Hibiscus syriacus is shrub or arborvitae deciduous in Hibiscus of Malvaceae, and its flower, fruit, root, leaf and bark can be used as medicine. The hibiscus flower not only has ornamental value, but also has use and medicinal value, and the essential value of the hibiscus leaf is recorded in China. However, the hibiscus flower has less application in the aspect of beauty treatment, and hibiscus flower ferment has less application in the aspect of anti-aging and anti-free radical.
According to the literature, every 100g of hibiscus flower contains 2.8g of protein with the water content of 92g, 0.3g of fat, 3.26g of saccharides, 133.98kJ of heat, 1.2g of ash, 0.8g of crude fiber, 14mg of calcium, 56mg of phosphorus, 1.2mg of iron, carotene, amino acid, flavonoid, multivitamin and the like, has excellent skin care effects of scavenging free radicals, resisting oxidation, preserving moisture, resisting wrinkles and the like, and is an excellent skin care raw material.
At present, few researches on extraction and separation of active ingredients in hibiscus flower are carried out, organic solvents are more used in the extraction method, the residue of the organic solvents in the extract and treatment make the organic solvents become a defect in the extraction process, the extraction efficiency is not ideal, the waste of raw materials is caused, the anti-aging and skin-care ingredients in the hibiscus flower cannot be fully extracted, and the environment is polluted; the plant fermentation ensures the safety of the fermentation product, does not produce substances harmful to the environment and does not harm human bodies while obtaining the required fermentation product.
The hibiscus flower is fermented by the fermentation process, so that the utilization rate of the hibiscus flower raw material is improved, the waste of the raw material is avoided, the yield of active substances such as amino acid and flavonoid is improved, the molecular weight of hibiscus flower ferment is controlled, and the skin absorption rate of the active substance components is effectively improved.
Disclosure of Invention
The invention aims to solve the technical problem of providing a preparation method of hibiscus syriacus ferment and also providing application of the hibiscus syriacus ferment prepared by the method. The hibiscus syriacus enzyme prepared by the invention can obviously improve the content of amino acid and flavone in the product, and has more obvious effects of resisting aging, preserving moisture and recovering skin elasticity.
The technical problem to be solved by the invention is realized by the following technical scheme:
a preparation method of hibiscus syriacus ferment comprises the following steps:
(1) mixing flos Hibisci, folium Hibisci and stem Hibisci at a ratio of 30-50: 10-20: washing 10-20 weight parts, mashing, drying, and sieving;
(2) sterilizing the material obtained in the step (1) by adopting an ultraviolet lamp irradiation method, wherein the irradiation time is 30-60 min;
(3) putting the material obtained in the step (2) into a fermentation vat, adding sterile water, uniformly stirring, controlling the temperature at 27-37 ℃, and adjusting the pH value to 5-7;
(4) adding 0.7-1.2 parts of bacillus subtilis, 5-10 parts of nitrogen source, 0.5-3 parts of carbon source and 0.01-0.2 part of inorganic salt solution with the mass percent concentration of 1% for fermentation;
(5) adding 0.2-0.4 part of yeast and 1.3-1.8 parts of lactic acid bacteria into the material obtained in the step (4) for continuous fermentation, avoiding light irradiation in the fermentation process, controlling the rotating speed of a shaking table to be 150 plus 200rpm, and fermenting for 7-10 days until no new bubbles are generated in the fermentation barrel;
(6) after the fermentation is finished, carrying out sterilization treatment, and centrifugally filtering out fermentation liquor and filter residues to obtain crude fermentation liquor;
(7) and filtering the crude fermentation liquor by using an ultrafiltration membrane to obtain a hibiscus syriacus enzyme solution.
Furthermore, the hibiscus flower is complete in petals, bright and fresh in color and luster and in full bloom.
Further, the carbon source in the step (4) is one or more of glucose, lactose and sucrose.
Further, in the step (4), the nitrogen source is urea or NH
4One or two of Cl.
Further, the inorganic salt solution in the step (4) is a 1% aqueous solution of: MgSO (MgSO)
4Solution, Na
2HPO
4Solution of, K
2SO
4Solution of MnSO
4One or more of the solutions.
Further, the fermentation time of the bacillus subtilis in the step (4) is 12-28 hours.
Further, the fermentation time in the step (4) is preferably 20 hours.
Further, the sterilization treatment in the step (6) is to place the ultrasonic generator of the ultrasonic cell crushing instrument in the mixed solution, perform cell wall breaking with an ultrasonic power of 400-600w, a frequency of 50-100KHz, a work time of 10-20 seconds, an interval time of 10 seconds, and a total ultrasonic time of 20-30 min.
The interception molecular weight of the ultrafiltration membrane in the step (7) is less than 10000 Da.
Application of the hibiscus syriacus ferment prepared by the preparation method in a personal care composition is provided.
The invention has the following beneficial effects:
1. the hibiscus flower in the full bloom stage is selected, the content of reducing sugar and flavone in petals is the highest, and the quality of the hibiscus enzyme can be effectively improved. Meanwhile, the hibiscus leaf and hibiscus stem are used as fermentation raw materials, because the hibiscus leaf and hibiscus stem also contain abundant amino acids, flavones and other nutrient substances.
2. The bacillus subtilis is added in the fermentation step to provide protease for the subsequent fermentation of the added lactobacillus and saccharomycetes, the fermentation time of the bacillus subtilis is optimized, the survival rate of the bacillus subtilis is improved, the strain dosage is reduced, the yield of the protease is increased, and therefore the content of amino acid and flavone in the hibiscus syriacus ferment is improved.
3. The addition of inorganic salt, carbon source and nitrogen source can prolong the survival time of the strain and increase the active matter content of the ferment. The cell wall is broken by ultrasonic waves, and the skin-activating components and intracellular active products synthesized and secreted in the fermentation process are reserved. Wherein, the inorganic salt is the essential component of the microorganism life substance, and the addition of a proper amount of the inorganic salt can promote the growth of the microorganism and simultaneously promote the microorganism to secrete various enzymes and nutrient substances.
4. The molecular weight range of the product is controlled by the ultrafiltration membrane, so that the quality of the hibiscus syriacus ferment is optimized, and the product is easier to be absorbed and utilized by the skin.
5. The production process utilizes a biological fermentation technology, does not use any organic solvent, and simultaneously avoids the problem of environmental pollution in the fermentation process and the extraction engineering.
Detailed Description
The present invention will be described in detail with reference to examples, which are only preferred embodiments of the present invention and are not intended to limit the present invention.
Example 1
A preparation method of hibiscus syriacus ferment comprises the following steps:
(1) mixing flos Hibisci, folium Hibisci and stem Hibisci according to the weight ratio of 35: 18: 13 parts by weight of the raw materials are washed, smashed, dried and sieved by a 200m sieve; wherein the hibiscus flower is selected from hibiscus flower with complete petals, bright and fresh color and full bloom;
(2) sterilizing the material obtained in the step (1) by adopting an ultraviolet lamp irradiation method, wherein the irradiation time is 30-60 min;
(3) putting the material obtained in the step (2) into a fermentation vat, adding sterile water, uniformly stirring, controlling the temperature at 27-37 ℃, and adjusting the pH value to 5-7;
(4) adding 0.9 weight part of bacillus subtilis, 5 weight parts of nitrogen source (urea), 0.5 weight part of carbon source (glucose) and 1 mass percent of inorganic salt solution (MgSO)
4Solution) 0.01 weight portion for 20 hours;
(5) adding 0.3 weight part of saccharomycetes and 1.5 weight parts of lactic acid bacteria into the material obtained in the step (4) for continuing fermentation, avoiding illumination in the fermentation process, controlling the rotating speed of a shaking table to be 150 plus 200rpm, and fermenting for 7-10 days until no new bubbles are generated in the fermentation barrel;
(6) after the fermentation is finished, carrying out sterilization treatment, and centrifugally filtering out fermentation liquor and filter residues to obtain crude fermentation liquor; wherein the sterilization treatment is specifically to place an ultrasonic generator of the ultrasonic cell crushing instrument in the mixed solution, the ultrasonic power is 400-600w, the frequency is 50-100KHz, the operation is 10-20 seconds, the interval time is 10 seconds, and the total ultrasonic time is 20-30min to break the cell wall.
(7) And filtering the crude fermentation liquor by using an ultrafiltration membrane with the interception molecular weight of less than 10000Da to obtain the hibiscus syriacus ferment solution.
Comparative example 1
Comparative example 1 was prepared by the same procedure as example 1, except that no inorganic salt solution was added during comparative example 1.
Comparative example 2
The preparation process of comparative example 2 is the same as that of example 1, but in the process of comparative example 2, bacillus subtilis, lactic acid bacteria and yeast are added simultaneously, namely, the steps (4) and (5) are combined, and the three bacteria are added simultaneously and fermented simultaneously.
Table 1 shows the results of the amino acid and flavone contents test of the ferments of examples and comparative examples.
| Test number | Amino acid content (mg/g) | Flavone content (mg/g) |
| Example 1 | 1.23 | 0.608 |
| Comparative example 1 | 0.85 | 0.481 |
| Comparative example 2 | 1.02 | 0.340 |
It can be seen that the amino acid content and the flavone content of the ferment seed obtained in example 1 are obviously higher than those of comparative example 1 and comparative example 2, and it can be seen that the amino acid content and the flavone content of the ferment seed can be obviously improved by adding the inorganic salt in the fermentation process, adding the bacillus subtilis in the fermentation process, fermenting for a period of time, and adding the yeast and the lactic acid bacteria.
Example 2
A cream added with hibiscus enzyme prepared in example 1 is prepared by the following method:
1. mixing an oil phase: weighing glyceryl stearate/PEG-100 stearate, caprylic/capric triglyceride, and shea butter, and heating to 75-80 deg.C;
2. mixing the water phase: mixing and dispersing glycerol and carbomer, adding pure water, and stirring to add 75-80 deg.C;
3. pouring the oil phase into the water phase, homogenizing with a homogenizer for 5min at a homogenizing rate of 4000r/min
4. After homogenizing, stirring and cooling to 60 ℃, and adding a pH regulator to regulate the pH value to 6.0;
5. and cooling to 25-30 ℃, adding the hibiscus enzyme obtained in the example 1, and continuously stirring uniformly to obtain the skin care cream containing the hibiscus enzyme.
Comparative example 3 was prepared in the same manner as in example 2, except that the hibiscus enzyme was replaced with a crude extract of hibiscus enzyme (i.e., the crude fermentation broth obtained in step (6) of example 1); the preparation method of comparative example 4 is the same as example 2, in which hibiscus flower ferment is replaced by hibiscus flower extract extracted by conventional ultrasonic extraction technology; comparative example 5 was prepared in the same manner as in example 2, wherein commercial composition having anti-aging and moisturizing alleges was used instead of hibiscus ferment;
TABLE 2 formulation compositions (unit: parts by weight) of example 2, comparative example 3, comparative example 4, and comparative example 5
| Ingredients of raw materials | Example 2 | Comparative example 3 | Comparative example 4 | Comparative example 5 |
| PEG-100 stearate/glyceryl stearate | 2 | 2 | 2 | 2 |
| Caprylic/capric triglyceride | 5 | 5 | 5 | 5 |
| Shea butter | 2 | 2 | 2 | 2 |
| Carbomer | 0.5 | 0.5 | 0.5 | 0.5 |
| Glycerol | 5 | 5 | 5 | 5 |
| Preservative | 0.6 | 0.6 | 0.6 | 0.6 |
| pH | 0.15 | 0.16 | 0.17 | 0.17 |
| Hibiscus syriacus flower extract | \ | \ | 2 | \ |
| Hibiscus syriacus enzyme | 2 | \ | \ | \ |
| Commercially available anti-aging compositions | \ | \ | \ | 2 |
| Rough extraction of hibiscus enzyme | \ | 2 | \ | \ |
| Water (W) | 82.75 | 82.74 | 82.73 | 82.73 |
And (3) performing efficacy evaluation test on the obtained samples, selecting 25-35-year-old female volunteers, 40 female volunteers in each group, using the female volunteers in the morning and the evening, analyzing the skin of the test subject by using a multifunctional skin tester after each female volunteer continuously uses the female volunteers for 28 days, and performing data analysis to obtain the efficacy test results shown in the table 3.
Table 3 results of efficacy test of example 2, comparative example 3, comparative example 4, and comparative example 5
| Contrast item | Blank group | Example 2 | Comparative example 3 | Comparative example 4 | Comparative example 5 |
| Effective rate for repairing fine wrinkles around eyes | 0% | 95% | 75% | 72.5% | 65% |
| Mean rate of increase in skin moisture around the eyes | 12.5% | 29.5% | 19.5% | 22.3% | 25.3% |
| Increased elasticity of skin around the eyes | 0.23% | 7.83% | 5.23% | 6.02% | 6.58% |
As can be seen from the comparison of the results in Table 3, the hibiscus ferment of the present invention has more effective anti-aging, moisturizing, and skin elasticity-restoring effects than the hibiscus extract and the commercial products. And the hibiscus syriacus ferment filtered by the ultrafiltration membrane has more obvious effect compared with the crude filtrate.
Claims (10)
1. A preparation method of hibiscus syriacus enzyme is characterized by comprising the following preparation steps:
(1) mixing flos Hibisci, folium Hibisci and stem Hibisci at a ratio of 30-50: 10-20: washing 10-20 weight parts, mashing, drying, and sieving;
(2) sterilizing the material obtained in the step (1) by adopting an ultraviolet lamp irradiation method, wherein the irradiation time is 30-60 min;
(3) putting the material obtained in the step (2) into a fermentation vat, adding sterile water, uniformly stirring, controlling the temperature at 27-37 ℃, and adjusting the pH value to 5-7;
(4) adding 0.7-1.2 parts of bacillus subtilis, 5-10 parts of nitrogen source, 0.5-3 parts of carbon source and 0.01-0.2 part of inorganic salt solution with the mass percent concentration of 1% for fermentation;
(5) adding 0.2-0.4 part of yeast and 1.3-1.8 parts of lactic acid bacteria into the material obtained in the step (4) for continuous fermentation, avoiding light irradiation in the fermentation process, controlling the rotating speed of a shaking table to be 150 plus 200rpm, and fermenting for 7-10 days until no new bubbles are generated in the fermentation barrel;
(6) after the fermentation is finished, carrying out sterilization treatment, and centrifugally filtering out fermentation liquor and filter residues to obtain crude fermentation liquor;
(7) and filtering the crude fermentation liquor by using an ultrafiltration membrane to obtain a hibiscus syriacus enzyme solution.
2. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: the hibiscus flower is the hibiscus flower with complete petals, bright and fresh color and full bloom.
3. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: and (4) the carbon source in the step (4) is one or more of glucose, lactose and sucrose.
4. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: step (a)4) The nitrogen source is urea and NH
4One or two of Cl.
5. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: the inorganic salt solution in the step (4) is a 1% aqueous solution of: MgSO (MgSO)
4Solution, Na
2HPO
4Solution, K
2SO
4Solution, MnSO
4One or more of the solutions.
6. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: the fermentation time of the bacillus subtilis in the step (4) is 12-28 hours.
7. The method for preparing hibiscus ferment of claim 6, wherein the hibiscus ferment is prepared by the following steps: the fermentation time in the step (4) is 20 hours.
8. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: and (6) the sterilization treatment is to place an ultrasonic generator of the ultrasonic cell crushing instrument in the mixed solution, wherein the ultrasonic power is 400-600w, the frequency is 50-100KHz, the operation lasts for 10-20 seconds, the interval time is 10 seconds, and the total ultrasonic time is 20-30min to break the cell wall.
9. The method for preparing hibiscus ferment according to claim 1, wherein the hibiscus ferment is prepared by the following steps: the interception molecular weight of the ultrafiltration membrane in the step (7) is less than 10000 Da.
10. Use of hibiscus ferment prepared by the preparation method according to any one of claims 1 to 9 for preparing a personal care composition.
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| CN107137280A (en) * | 2017-05-27 | 2017-09-08 | 中南林业科技大学 | A kind of preparation method of the crease-resistant moisturizing rose of Sharon petal eye mask of self-heating |
| CN108277242A (en) * | 2018-01-10 | 2018-07-13 | 精晶药业股份有限公司 | A method of extracting amino acid from plant |
| CN110269250A (en) * | 2019-07-19 | 2019-09-24 | 蓬莱深奥生物科技研究所 | A kind of wall of sea cucumber Stichopus japonicus ferment and preparation method thereof |
| CN110477093A (en) * | 2019-09-19 | 2019-11-22 | 张学海 | A kind of salt-free fruits and vegetables fermentation process of paddy embryo saccharification |
| CN112891283B (en) * | 2021-02-01 | 2022-04-22 | 浙江万里学院 | Dendrobium officinale fermentation extract, skin care product and preparation method thereof |
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