CN115737491B - Plant composition and preparation method and application thereof - Google Patents
Plant composition and preparation method and application thereof Download PDFInfo
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- CN115737491B CN115737491B CN202210290593.4A CN202210290593A CN115737491B CN 115737491 B CN115737491 B CN 115737491B CN 202210290593 A CN202210290593 A CN 202210290593A CN 115737491 B CN115737491 B CN 115737491B
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicines Containing Plant Substances (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a plant composition, a preparation method and application thereof, and belongs to the field of plant extraction research. The composition comprises radix Gentianae, radix Sophorae Flavescentis and radix Ophiopogonis. The plant composition can inhibit inflammatory reaction, inhibit oxidative stress reaction, relieve capsaicin stimulation in time, relieve lactic acid stimulation in time and relieve skin sensitive symptoms, and has higher application value.
Description
Technical Field
The invention relates to the field of plant extraction research, in particular to a plant composition, a preparation method and application thereof.
Background
With the continuous high-speed development of modern society, the influence of external environmental factors such as the rapid urban life rhythm, the deterioration of life environment, ultraviolet irradiation, cosmetic abuse and the like and psychological factors is increasing, and more people are facing sensitive skin trouble. Causes of the development of sensitive skin include: impaired skin barrier function, enhanced nerve signaling, increased inflammatory response, skin oxidative stress, etc. When the sensitive skin is stimulated by the outside, the skin is easy to generate erythema and pimple, especially subjective symptoms such as telangiectasis, subjective symptoms such as subjective tingling pain, burning sensation, itching sensation, tightness and the like.
Cosmetic is spread on any part of the surface of the human body by applying, spraying or the like, for the purpose of cleaning, maintaining, beautifying, modifying and changing the appearance, or correcting the smell of the human body, maintaining a good state. In order to meet the demands of usability, efficacy and stability, surfactants, acidic substances, thickeners, emulsifiers, preservatives, spice, pigments and other ingredients are usually added in the production process of cosmetics, and the ingredients have certain irritation and are an important cause of skin stinging and burning. These components have strong penetration and irritation, and can cause skin stinging and burning of normal skin with perfect barrier function of the stratum corneum.
In conclusion, the skin sensitivity symptoms are influenced by the self-state of the skin, the safety of using cosmetics and other factors, and the 'red, swelling, itching and pain' of the sensitive skin is relieved from the aspect of symptoms, and the skin sensitivity symptoms are easy to repeat if the cosmetic stimulus is not antagonized from root.
Chinese patent (CN 108272723A) discloses a plant anti-allergic skin care composition which comprises radix ophiopogonis extract, purslane extract, cotton rose leaf/flower extract, kuh-seng root extract and tamarix chinensis extract, and can play a role in tranquilizing, diminishing inflammation, clearing heat, detumescence and resisting aging by being compounded together, and has good anti-aging, allergy-relieving and skin care effects. The skin care composition is mainly used for relieving sensitive skin symptoms, can not antagonize the stimulus source in cosmetics, and can reduce the skin sensitivity problem caused by the stimulus of the cosmetics.
In view of this, it is highly desirable to develop a botanical composition that simultaneously has the ability to relieve symptoms of sensitive skin and antagonize sources of irritation in cosmetics.
Disclosure of Invention
In order to solve the technical problems, the invention provides a plant composition and a preparation method and application thereof, wherein the plant composition has the effects of inhibiting inflammatory reaction, inhibiting oxidative stress reaction, relieving lactic acid stimulation, relieving capsaicin stimulation and the like and relieving skin sensitivity, and also has the effect of antagonizing the stimulation source of cosmetics.
For this reason, the technical scheme that this application provided is as follows:
in a first aspect, the present invention provides a plant composition.
According to the invention, the plant composition comprises gentian, kuh-seng and ophiopogon root.
According to the invention, the plant composition comprises the following raw materials in parts by mass: 5-10 parts of kuh-seng, 0.5-2 parts of dwarf lilyturf tuber and 0.5-2 parts of gentian.
According to the invention, the plant composition is prepared by a preparation method comprising the following steps:
1) Mixing radix Sophorae Flavescentis, radix Ophiopogonis and radix Gentianae, adding pure water at a ratio of 1:20-1:40 (m/m) based on the total mass of the raw materials, and extracting at 85-95deg.C under stirring for 1-4 hr to obtain extractive solution.
According to the invention, the dwarf lilyturf tuber, lightyellow sophora root and gentian coarse powder is sieved by a 10-50 mesh sieve (the 10 mesh sieve and the 50 mesh sieve are taken for standby).
According to the invention, the feed-to-liquid ratio in step 1) may be 1:20 (m/m), 1:21 (m/m), 1:22 (m/m), 1:24 (m/m), 1:26 (m/m), 1:27 (m/m), 1:30 (m/m), 1:32 (m/m), 1:35 (m/m), 1:40 (m/m), and point values between the above values.
According to the invention, the extraction temperature in step 1) may be 85 ℃, 86 ℃, 88 ℃, 90 ℃, 92 ℃, 95 ℃, and point values between the above values.
According to the invention, the extraction time of the step 1) can be 1h, 2h, 3h and 4h.
According to the present invention, the method for preparing a plant composition further comprises:
2) Filtering the extracting solution obtained in the step 1), standing and filtering to obtain filtrate;
3) Decolorizing the filtrate in the step 2) to obtain decolorized solution;
4) Adding protease and amylase into the decolorized solution in the step 3) for enzymolysis to obtain enzymolysis solution;
5) Concentrating the enzymolysis liquid in the step 4) under reduced pressure to obtain concentrated liquid.
According to the invention, the protease is added in the step 4) in an amount of 0.5-1.0% based on the total mass of the raw materials.
According to the present invention, the amylase is added in an amount of 0.005 to 0.010% by mass of the decolorized solution.
According to the invention, the protease in step 4) is a neutral protease.
According to the present invention, the protease may be added in the amount of 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0% based on the total mass of the raw materials in the step 4).
According to the present invention, the amylase may be added in an amount of 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.010% by mass of the decolorized solution in the step 4).
According to the invention, the enzymolysis temperature in the step 4) is 50-55 ℃ and the enzymolysis time is 15-60min.
According to the invention, the enzymatic hydrolysis temperature in step 4) may be 50 ℃, 51 ℃, 52 ℃,53 ℃, 54 ℃, 55 ℃.
According to the invention, the enzymolysis time in the step 4) can be 15min, 16min, 20min, 25min, 30min, 35min, 40min, 45min, 50min, 55min, 60min, and point values between the above values.
According to the invention, the step 4) further comprises enzyme deactivation, wherein the enzyme deactivation temperature is 90-95 ℃ and the enzyme deactivation time is 15-45min.
According to the invention, the decoloring mode of the step 3) is a conventional decoloring mode in the field.
According to the invention, the decoloring mode of the step 3) is as follows: adding 2% of active carbon by mass of the filtrate obtained in the step 2), and decoloring for 1h at 80-85 ℃.
According to the invention, the filter paper in the step 2) is F100 paperboard.
According to the invention, the reduced pressure concentration pressure in the step 5) is as follows: -0.9 MPa-1.0 MPa, at a temperature of: 65-70 ℃.
According to the invention, the step 5) further comprises fine filtration, compounding and sterilization.
According to the invention, the fine filtration mode of the step 5) is that the concentrated solution is filtered by using H70 paperboard.
According to the invention, the compounding mode of the step 5) is to compound 5% of glycerol, 2% of 1, 2-pentanediol and 1% of 1, 2-hexanediol by the mass of the H70 paperboard filtering filtrate.
According to the invention, the sterilization mode of the step 5) is a conventional sterilization mode in the field.
According to the invention, the sterilization mode of the step 5) is sterilization at 90-95 ℃ for 40min.
In a second aspect, the invention provides a method of preparing a plant composition according to the first aspect of the invention.
According to the invention, the preparation method comprises the following steps:
1) Mixing 5-10 parts of radix sophorae flavescentis, 0.5-2 parts of dwarf lilyturf tuber and 0.5-2 parts of gentian, adding pure water according to a material-liquid ratio of 1:20-1:40 (m/m), and carrying out heat preservation and stirring extraction for 1-4h at 85-95 ℃ to obtain an extracting solution;
2) Filtering the extracting solution obtained in the step 1), standing and filtering to obtain filtrate;
3) Decolorizing the filtrate in the step 2) to obtain decolorized solution;
4) Adding protease and amylase into the decolorized solution in the step 3), and performing enzymolysis for 15-60min at 50-55 ℃;
5) Concentrating the enzymolysis liquid in the step 4) under reduced pressure to obtain concentrated liquid.
According to the invention, the coarse powder of gentian, dwarf lilyturf tuber and lightyellow sophora root in the step 1) is sieved by a sieve with 10-50 meshes (the sieve with 10 meshes is taken for standby on a sieve with 50 meshes).
According to the invention, the feed-to-liquid ratio in step 1) may be 1:20 (m/m), 1:21 (m/m), 1:22 (m/m), 1:24 (m/m), 1:26 (m/m), 1:27 (m/m), 1:30 (m/m), 1:32 (m/m), 1:35 (m/m), 1:40 (m/m), and point values between the above values.
According to the invention, the extraction temperature in step 1) may be 85 ℃, 86 ℃, 88 ℃, 90 ℃, 92 ℃, 95 ℃, and point values between the above values.
According to the invention, the extraction time of the step 1) can be 1h, 2h, 3h and 4h.
According to the present invention, the protease may be added in the amount of 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0% based on the total mass of the raw materials in the step 4).
According to the present invention, the amylase may be added in an amount of 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.010% by mass of the decolorized solution in the step 4).
According to the invention, the step 4) further comprises enzyme deactivation, wherein the enzyme deactivation temperature is 90-95 ℃ and the enzyme deactivation time is 15-45min.
According to the invention, the decoloring mode of the step 3) is a conventional decoloring mode in the field.
According to the invention, the decoloring mode of the step 3) is as follows: adding 2% of active carbon by mass of the filtrate obtained in the step 2), and decoloring for 1h at 80-85 ℃.
According to the invention, the filter paper in the step 2) is F100 paperboard.
According to the invention, the reduced pressure concentration pressure in the step 5) is as follows: -0.9mpa— -1.0MPa, at a temperature of: 65-70 ℃.
According to the invention, the step 5) further comprises fine filtration, compounding and sterilization.
According to the invention, the fine filtration mode of the step 5) is that the concentrated solution is filtered by using H70 paperboard.
According to the invention, the compounding mode of the step 5) is to compound 5% of glycerol, 2% of 1, 2-pentanediol and 1% of 1, 2-hexanediol by the mass of the H70 paperboard filtering filtrate.
According to the invention, the sterilization mode of the step 5) is a conventional sterilization mode in the field.
According to the invention, the sterilization mode of the step 5) is sterilization at 90-95 ℃ for 40min.
In a third aspect, the present invention provides the use of a phytocomposition according to the first aspect of the present invention and/or a phytocomposition prepared by a method according to the second aspect of the present invention in the field of topical skin care formulations.
According to the present invention, the botanical composition has the efficacy of inhibiting inflammatory factor release, antioxidant, soothing the stimulus, antagonizing the surfactant.
According to the invention, the external skin care preparation can be cosmetics, including facial masks, cream, gel, emulsion, skin care lotion, facial cleanser, shampoo, hair conditioner or bath foam, and the like, and is not limited by dosage forms.
According to the invention, the gel comprises the following raw materials in parts by weight:
the beneficial effects of the invention are that
The plant composition comprises gentian, kuh-seng and dwarf lilyturf tuber, the proportion of each component is optimized, the compatibility and the synergistic effect are scientific, and the plant composition has the effects of inhibiting inflammatory response, inhibiting oxidative stress response, immediately relieving capsaicin stimulation, immediately relieving lactic acid stimulation, antagonizing surfactant SDS (sodium dodecyl sulfate) stimulation and the like. The plant composition has the capability of antagonizing exogenous stimulus sources of cosmetics on the basis of relieving neurogenic sensitive skin and barrier injury type sensitive skin, and has higher application value in the field of external skin care preparations.
Drawings
Fig. 1 is a graph of the effects of the plant compositions of examples 1-4, comparative examples 1-4 on inhibition of inflammatory factor IL-1 alpha (panel: n=3, using one-way analysis of variance (ANOVA), <0.01, <0.05, < p);
fig. 2 is a graph of the effect of the plant compositions of examples 1-4, comparative examples 1-4 on inhibition of inflammatory factor IL-1 beta (panel: n=3, using one-way analysis of variance (ANOVA), <0.01, <0.05, < p);
FIG. 3 is a graph showing the effect of the plant compositions of examples 1-4 and comparative examples 1-4 on inhibiting inflammatory factor IL-6 (panel: n=3, using one-way analysis of variance (ANOVA)), where p <0.01, p <0.05
FIG. 4 is a graph showing the effect of the plant compositions of examples 1-4 and comparative examples 1-4 on scavenging ABTS free radicals;
FIG. 5 is a graph of the results of the TiVi700 test on the skin of the test area before and after use of samples 1-8;
FIG. 6 is a graph of the effect of samples 1-8 on relieving capsaicin stimulation;
fig. 7 is a graph of the effect of samples 1-8 on relieving lactic acid-induced skin pricking (legend: n=10, p <0.05 compared to the blank with rank sum test);
FIG. 8 is a graph showing the effect of samples 1 to 8 on relieving itching of the skin caused by lactic acid;
FIG. 9 is a graph showing the effect of SDS-stimulation resistance of the plant compositions of examples 1 to 4 and comparative examples 1 to 4.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings and examples, but the present invention is not limited to the following examples.
The sources of the raw materials used in the invention are shown in table 1, and the raw materials and the instruments used in the invention are all conventional raw materials or instruments unless otherwise specified.
Table 1 use of raw material sources
| Raw material name | Production area | Suppliers/manufacturers |
| Radix gentianae | Yunnan (Yunnan) province | Beijing Qianzao Chinese herbal pieces Co., ltd |
| Radix Sophorae Flavescentis | Hebei river | Beijing Qianzao Chinese herbal pieces Co., ltd |
| Ophiopogon japonicus | Sichuan (Sichuan) | Beijing Qianzao Chinese herbal pieces Co., ltd |
| Neutral protease | Tianjin | Novozyme (China) Biotechnology Co.,Ltd. |
| Amylase enzyme | Tianjin | Novozyme (China) Biotechnology Co.,Ltd. |
Example 1:
1) Weighing 180g of radix sophorae flavescentis, 30g of dwarf lilyturf tuber and 30g of gentian, adding pure water according to the ratio of 1:20 (m/m), and extracting for 2h at 93 ℃ under heat preservation and stirring to obtain an extracting solution;
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of example 1 is obtained by cooling and filling.
Example 2:
1) 200g of kuh-seng, 20g of dwarf lilyturf tuber and 20g of gentian are weighed, pure water is added according to the ratio of 1:20 (m/m), and the mixture is stirred and extracted for 2 hours at 93 ℃ to obtain an extracting solution.
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of example 2 is obtained by cooling and filling.
Example 3:
1) Weighing 192g of kuh-seng, 24g of dwarf lilyturf tuber and 24g of gentian, adding pure water according to the ratio of 1:20 (m/m), and extracting for 2h at 93 ℃ under heat preservation and stirring to obtain an extracting solution;
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of example 3 is obtained by cooling and filling.
Example 4:
1) 171.4g of kuh-seng, 34.3g of dwarf lilyturf tuber and 34.3g of gentian are weighed, pure water is added according to the ratio of 1:20 (m/m) of feed liquid, and the mixture is stirred and extracted for 2 hours at 93 ℃ under heat preservation, thus obtaining an extracting solution.
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of example 4 is obtained by cooling and filling.
Comparative example 1:
1) Weighing 205.7g of radix sophorae flavescentis and 34.3g of dwarf lilyturf tuber, adding pure water according to the ratio of 1:20 (m/m), and extracting for 2h at 93 ℃ under heat preservation and stirring to obtain an extracting solution;
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of comparative example 1 is obtained after cooling and filling.
Comparative example 2:
1) Weighing 240g of gentian, adding pure water according to the ratio of feed to liquid of 1:20 (m/m), and extracting at 93 ℃ for 2h under heat preservation and stirring to obtain an extract;
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of comparative example 2 is obtained after cooling and filling.
Comparative example 3:
1) Weighing 211.8g of radix sophorae flavescentis, 14.1g of dwarf lilyturf tuber and 14.1g of gentian, adding pure water according to the ratio of 1:20 (m/m), and extracting for 2h at 93 ℃ under heat preservation and stirring to obtain an extracting solution;
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of comparative example 3 is obtained by cooling and filling.
Comparative example 4:
1) Weighing 80g of radix sophorae flavescentis, 80g of radix ophiopogonis and 80g of gentian, adding pure water according to the ratio of 1:20 (m/m), and extracting for 2h at 93 ℃ under heat preservation and stirring to obtain an extracting solution;
2) Standing the extracting solution obtained in the step 1) for 16 hours, and filtering the supernatant F100 paper board to obtain filtrate;
3) Adding 2% of active carbon into the filtrate in the step 2) based on the mass of the filtrate, decoloring for 1 hour at 83 ℃, and filtering by using F100 paper board to obtain decolored liquid;
4) Adding neutral protease accounting for 1% of the mass of the raw materials, and amylase accounting for 0.007% of the mass of the decolorized solution, reacting for 30min at 53 ℃, heating to 93 ℃, and inactivating the enzyme for 30min;
5) Concentrating at 67 deg.C and-0.9 MPa until the solid content of the feed liquid is 4+ -0.5%, and stopping concentrating;
6) Cooling the concentrated solution to 35 ℃, filtering by using H70 paper board, and controlling the turbidity of the feed liquid to be less than or equal to 10FNU to obtain refined filtrate;
7) The fine filtrate is prepared by the following steps: glycerol: 1, 2-pentanediol: 1, 2-hexanediol=100:5:2:1 (m/m) and sterilized at 93 ℃ for 40min, and the plant composition of comparative example 4 is obtained after cooling and filling.
Test example 1: inhibiting inflammatory factor release
Detecting a sample: examples 1 to 4, comparative examples 1 to 4
Detection principle:
lipopolysaccharide (LPS) is one of the components of the cell wall of gram-negative bacteria. When LPS acts on macrophages, inflammatory reactions of the macrophages are induced, so that a large amount of cell inflammatory factors such as interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and the like are generated. The present test evaluates the ability of a botanical composition to inhibit the release of inflammatory factors based on the LPS-induced mouse macrophage (RAW 264.7) inflammatory model, thereby predicting its ability to inhibit further inflammatory responses. The positive control for this test was 0.01%/0.001% dexamethasone; model control was RAW264.7 with no sample added for LPS induction (lps+); the blank is cell culture medium with Phosphate Buffered Saline (PBS) added.
Cell efficacy test concentration selection criteria:
(1) Relative cell viability ± SD > 85% (relative cell viability mean between 85-90% (excluding 90%) was required to have no significant difference (p > 0.05) compared to negative control group);
(2) Cell morphology was not significantly altered from the negative control.
Note that: the negative control was medium.
The detection method comprises the following steps:
cell tests were performed by selecting the series of concentrations of the plant compositions of examples 1 to 4 and comparative examples 1 to 4, and the final concentrations to be tested were selected to be 0.08%, 0.31% and 1.25% based on the toxicity test results of RAW264.7 cells and morphological observation results of the cells, and the test results are shown in FIGS. 1 to 3 and tables 2 to 4.
TABLE 2 inhibition of IL-1 alpha by the plant compositions of examples 1-4, comparative examples 1-4
As shown in FIGS. 1 and 2, at concentrations of 0.08%, 0.31%, 1.25%, the example and comparative example plant compositions each inhibited the release of inflammatory factor IL-1. Alpha. At a test concentration of 0.31%, the inhibition rate of the inflammatory factor IL-1 alpha by comparative example 1 was 27.27%, and the inhibition rate of the inflammatory factor IL-1 alpha by the plant composition of example 1 was 87.44%, which is about 3 times; the inhibition of inflammatory factor IL-1α by the example 3 phytochemicals was about 2-fold compared to comparative example 2. The above test results demonstrate that the synergistic effect among the components of the plant composition of the present application provides a better effect of inhibiting the release of inflammatory factor IL-1 alpha. Compared with comparative example 4, the IL-1 alpha inhibition rate of the plant composition of example 1 is about 1.6 times, which shows that the components of the plant composition have better effect of inhibiting the release of inflammatory factors IL-1 alpha by scientific compatibility under the specific proportion.
TABLE 3 inhibition of IL-1β by the plant compositions of examples 1-4, comparative examples 1-4
As shown in fig. 2 and table 3, at concentrations of 0.08%, 0.31%, 1.25%, the example plant compositions inhibited the release of inflammatory factor IL-1β, which was significantly better than the model control. At a concentration of 0.08%, the compositions of comparative examples 1-4 showed substantially no inhibition of inflammation due to IL-1β; the inhibition of inflammation by IL-1β was slightly increased by the compositions of comparative examples 1-4 as the concentration tested was increased; at a test concentration of 1.25%, the IL-1β inhibition rate of the example 1 plant composition was 42.20%, approximately 3 times that of comparative example 2, and approximately 2.9 times that of comparative example 4. The experimental results show that the plant composition has better effect of inhibiting the release of inflammatory factors IL-1 beta through the combination and scientific compatibility of gentian, kuh-seng and dwarf lilyturf tuber.
TABLE 4 inhibition of IL-6 by the plant compositions of examples 1-4, comparative examples 1-4
As shown in FIG. 3 and Table 4, at concentrations of 0.08%, 0.31%, 1.25%, the example and comparative example plant compositions each inhibited the release of inflammatory factor IL-6. At a test concentration of 0.08%, the inhibition rate of inflammatory factor IL-6 by the example plant composition was 32.88% (average), and the inhibition rate of inflammatory factor IL-6 by the comparative example plant composition was 7.16% (average), which is about 4.5 times that of the comparative example; at a test concentration of 1.25%, the inhibition rate of the inflammatory factor IL-6 by comparative example 1 was 13.38%, and the inhibition rate of the inflammatory factor IL-6 by the plant composition of example 1 was 52.99%, which is about 4 times; the inhibition of inflammatory factor IL-1α by the example 1 phytochemical composition was about 3-fold compared to comparative example 4. The experimental results show that the plant composition has better effect of inhibiting the release of inflammatory factors IL-6 through the combination and scientific compatibility of gentian, kuh-seng and dwarf lilyturf tuber.
In conclusion, the plant composition has the effect of obviously inhibiting the release of inflammatory factors IL-1 alpha, IL-1 beta and IL-6 through the synergistic effect between gentian, kuh-seng and dwarf lilyturf tuber and scientific compatibility.
Test example 2: scavenging free radicals and inhibiting oxidative stress
Detecting a sample: examples 1 to 4, comparative examples 1 to 4
Detection principle:
ABTS free radical scavenging assay as a method of evaluating inhibition of oxidative stress, the inhibition of oxidative stress by a substance can be evaluated. In general, a greater ABTS radical scavenging rate indicates a greater ability of the substance to inhibit oxidative stress. ABTS is oxidized under the action of a proper oxidant to generate stable blue-green cation free radical ABTS+ and when an antioxidant exists, the ABTS+ reacts with the antioxidant component of the ABTS+ to fade the system color. And measuring the absorbance value of the solution at the maximum absorption wavelength, and measuring and evaluating the capacity of the product for inhibiting the oxidative stress reaction.
The detection method comprises the following steps:
the ability of examples 1-4 and comparative examples 1-4 (0.1%, 0.25%, 0.5%, 0.75%, 1.0%, 2.5%) to scavenge ABTS free radicals was examined and the test results are shown in fig. 4. Examples 1-4, comparative examples 1-4 were diluted to the concentrations of 0.1%, 0.25%, 0.5%, 0.75%, 1.0%, 2.5% for testing using 50% ethanol solution.
As shown in FIG. 4, at the concentrations tested in the present application, examples 1-4 and comparative examples 1-4 each had the ability to scavenge ABTS free radicals and inhibit the occurrence of oxidative stress. At the same test concentration, the examples have better ability to scavenge ABTS free radicals than the comparative examples, and the examples have a significant improvement in ABTS free radical scavenging rate over the comparative examples. The experimental result shows that the specific components of the plant composition have better capacity of inhibiting oxidative stress under the scientific compatibility condition.
Test example 3: relieving skin irritation caused by capsaicin
Detecting a sample: samples 1-8
The detection method comprises the following steps:
the specific formulations of the 0.01% capsaicin solution and the 3% gel containing examples 1-4 or comparative examples 1-4 are shown in Table 5. Each sample is used for screening 8 volunteers, a capsaicin-stimulated skin model is established, a filter paper sheet containing 0.01% of capsaicin solution is attached to the nasal and labial sulcus of the volunteers, the filter paper sheet is attached for 3min to take off, after the filter paper sheet is left for 5min, one side of the filter paper sheet is coated with the sample, the other side of the filter paper sheet is not treated, the filter paper sheet is re-timed after the sample is coated, before the sample is used (after capsaicin stimulation), after the sample is used for 0min, 1min, 3min, 5min and 7min, a skin sensitivity imager (TiVi 700) is used for collecting and analyzing skin pictures of a test area, the smaller the average value of TiVi-index is, the better the soothing effect of the sample is indicated, and the soothing effect of the sample on skin stimulation caused by capsaicin is evaluated by comparing the average value of TiVi-index between the samples.
Table 5 gel formulation
The preparation process for samples 1-8 is as follows:
(1) Heating the phase A raw material to 80 ℃ and uniformly mixing for later use;
(2) Stirring and cooling, adding the B phase and C phase raw materials at 60 ℃, and uniformly stirring;
(3) Stirring and cooling, adding the phase D raw material at 40 ℃, and uniformly stirring;
(4) Stirring and cooling, and discharging at 38 ℃.
As shown in FIGS. 5-6, samples 1-8 each were able to relieve capsaicin-induced skin irritation, wherein samples 1-4 containing 3% of the examples 1-4 of the present application were significantly better in relieving capsaicin-induced skin irritation than samples 5-8 containing 3% of the comparative examples 1-4. The plant composition has excellent effect of instantly relieving skin irritation caused by capsaicin under a specific proportion through synergistic effect among gentian, kuh-seng and dwarf lilyturf tuber, and can relieve neurogenic sensitive skin.
Test example 4: relieving skin irritation caused by lactic acid
Detecting a sample: samples 1-8
The detection method comprises the following steps:
lactic acid test the effect of the samples on the soothing effect of skin irritation caused by lactic acid was evaluated by asking the volunteers for the scores of the skin's tingling and itching sensation, scoring in a 4-point method (0 for no sensation, 1 for mild sensation, 2 for moderate sensation, 3 for heavy sensation), and comparing the size of the scores between the samples. 10 volunteers are screened from each sample, a lactic acid stimulation skin model is established, a filter paper sheet containing 10% lactic acid aqueous solution is attached to the nasal and labial sulcus of the volunteers, when the feeling score of the nasal and labial sulcus of the two sides of the volunteers is more than or equal to 2, the filter paper sheet is removed, one side of the sample is smeared, the other side of the sample is not treated (blank control), the sample is smeared and counted again, and before the sample is used (after lactic acid stimulation), after the sample is used, the skin of the testing areas on the two sides of the volunteers are inquired about the tingling and itching of the skin of the testing areas on the two sides of the volunteers in 0min, 0.5min, 2.5min, 5min, 7.5min, 15min, 20min and 30min, so that the skin stimulation relieving effect of the sample on lactic acid is evaluated.
As shown in fig. 7-8, samples 1-4 exhibited significant soothing effects on skin irritation caused by lactic acid during the 0-15min test period; samples 5-8 showed substantially no effect of soothing lactic acid stimulation compared to the blank. In the test period of 15-30min, the value is smoothed afterwards due to the self-repairing ability of the skin. The lactic acid test experiment result shows that the plant composition has excellent effect of immediately relieving skin stinging and itching caused by lactic acid under specific proportion through synergistic effect among gentian, kuh-seng and dwarf lilyturf tuber, scientific compatibility, and can relieve barrier-damaged sensitive skin.
Test example 5: antagonizing the stimulation of cosmetic stimulus sources
Detecting a sample: examples 1 to 4, comparative examples 1 to 4
Detection principle:
sodium Dodecyl Sulfate (SDS) acts on erythrocytes to cause a change in cell membrane permeability, so that hemoglobin exudes, and the exuded hemoglobin has a maximum absorption peak at a wavelength of 530 nm. After the sample is added to the SDS-containing solution, if the hemolysis of the erythrocytes is reduced, the added sample is considered to be capable of inhibiting the hemolysis of the erythrocytes caused by SDS and playing a role in protecting cell membranes. And (3) adopting a erythrocyte hemolysis coagulation test, mixing the sample with SDS, then incubating the mixture with the erythrocyte suspension, and evaluating the inhibition effect of the sample on erythrocyte hemolysis caused by SDS by comparing the change of erythrocyte hemolysis so as to evaluate the resistance effect of the sample on cell membrane stimulation caused by a cosmetic stimulus.
The detection method comprises the following steps:
taking a centrifuge tube, adding a test substance, PBS, RBC suspension and SDS according to the table 6, and uniformly mixing, wherein the final concentration of a sample system is the test concentration; placing in a shaking table, incubating for 10min, and centrifuging; observing the phenomenon, collecting supernatant, and measuring OD 530 The method comprises the steps of carrying out a first treatment on the surface of the The hemolysis rate was calculated, and the erythrocyte hemolysis inhibition rate was calculated from the hemolysis rate, wherein the sample concentration was 0.10%, 0.20%, and 0.50%.
The calculation formula of the erythrocyte hemolysis rate is as follows:
the calculation formula of the erythrocyte hemolysis inhibition rate is as follows:
TABLE 6 reaction system (mu L)
Note that: in table 6 "+" represents "added" and "-" represents "not added", the test sample in the model control was deionized water.
As shown in fig. 9, the plant compositions of examples 1 to 4 had inhibitory effects on red blood cell solutions, and the inhibition effect on red blood cell hemolysis of the plant composition of example at the same concentration was significantly better than that of the plant composition of comparative example, and the inhibition rate was about 2 times that of the plant composition of comparative example. The plant composition can effectively antagonize SDS stimulation in a specific proportion range through the synergistic interaction among gentian, kuh-seng and dwarf lilyturf tuber.
In conclusion, the plant composition comprises gentian, kuh-seng and dwarf lilyturf tuber, the proportion of each component is optimized, the compatibility and the synergistic effect are scientific, and the plant composition has the effects of inhibiting inflammatory response, inhibiting oxidative stress response, immediately relieving capsaicin stimulation, immediately relieving lactic acid stimulation, antagonizing surfactant SDS stimulation and the like. The plant composition has the effects of relieving neurogenic sensitive skin and barrier injury type sensitive skin, and antagonizing exogenous stimulation of cosmetics.
The foregoing examples of the present invention are merely illustrative of the present invention and are not intended to limit the embodiments of the present invention, and other variations or modifications of various forms may be made by those skilled in the art based on the foregoing description, and it is not intended to be exhaustive of all embodiments, and all obvious variations or modifications that come within the scope of the invention are defined by the following claims.
Claims (7)
1. The plant composition is characterized by comprising the following raw materials in parts by mass: 5-10 parts of kuh-seng, 0.5-2 parts of dwarf lilyturf tuber and 0.5-2 parts of gentian;
the plant composition is prepared by a preparation method comprising the following steps:
1) Mixing radix Sophorae Flavescentis, radix Ophiopogonis and radix Gentianae, adding pure water at a ratio of 1:20-1:40m/m, and extracting at 85-95deg.C under stirring for 1-4 hr to obtain extractive solution;
the plant composition preparation method further comprises the following steps:
2) Filtering the extracting solution obtained in the step 1), standing and filtering to obtain filtrate;
3) Decolorizing the filtrate in the step 2) to obtain decolorized solution;
4) Adding protease and amylase into the decolorized solution in the step 3) for enzymolysis to obtain enzymolysis solution;
5) Concentrating the enzymolysis liquid in the step 4) under reduced pressure to obtain concentrated liquid.
2. The plant composition according to claim 1, wherein the protease is added in the amount of 0.5-1% based on the total mass of the raw materials in the step 4).
3. The plant composition according to claim 1, wherein the amylase is added in an amount of 0.005-0.010% by mass of the decolorized solution.
4. A plant composition according to any one of claims 1-3, wherein the enzymatic hydrolysis conditions in step 4) are enzymatic hydrolysis temperatures of 50-55 ℃ and enzymatic hydrolysis times of 15-60min.
5. A method of preparing a plant composition, comprising the steps of:
1) Weighing 5-10 parts of kuh-seng, 0.5-2 parts of dwarf lilyturf tuber and 0.5-2 parts of gentian, mixing, adding pure water according to the total mass of the raw materials and the feed liquid ratio of 1:20-1:40m/m, and carrying out heat preservation and stirring extraction for 1-4h at 85-95 ℃ to obtain an extracting solution;
2) Filtering the extracting solution obtained in the step 1), standing and filtering to obtain filtrate;
3) Decolorizing the filtrate in the step 2) to obtain decolorized solution;
4) Adding protease and amylase into the decolorized solution in the step 3), and performing enzymolysis for 15-60min at 50-55 ℃;
5) Concentrating the enzymolysis liquid in the step 4) under reduced pressure to obtain concentrated liquid;
in the step 4), the addition amount of protease is 0.5-1% based on the total mass of the raw materials, and the addition amount of amylase is 0.005-0.010% based on the mass of the decolorized solution.
6. Use of a botanical composition as claimed in any one of claims 1 to 4 or prepared by a method as claimed in claim 5 in the manufacture of a topical skin care formulation.
7. The use according to claim 6, wherein the plant composition is used for preparing a skin care preparation for external use having the efficacy of inhibiting inflammatory factor release, resisting oxidation, relieving irritation, antagonizing surfactant.
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