CN115025025B - Paeonia lactiflora fermentation product and preparation method and application thereof - Google Patents
Paeonia lactiflora fermentation product and preparation method and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- 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
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q9/00—Preparations for removing hair or for aiding hair removal
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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
Abstract
The invention discloses a preparation method of a peony flower fermentation product, which comprises the following steps: providing Paeonia lactiflora flower which is derived from medicinal Paeonia lactiflora and has gallic acid content of not less than 9000ppm, drying, pulverizing, mixing with water, adjusting pH to 4.5-5.0, and sterilizing to obtain fermentation culture medium; providing a saccharomyces cerevisiae strain CICC 1400 for activation and domestication culture; inoculating the zymophyte liquid to a fermentation culture medium, fermenting at 32 ℃ for 24h, cooling to 25 ℃, and fermenting for 48-60h to obtain fermentation liquid; centrifuging, sequentially performing microfiltration and ultrafiltration on the supernatant to obtain filtrate, namely the peony flower fermentation product. According to the method, the source and the quality of the paeoniflora are controlled, the saccharomyces cerevisiae strain CICC 1400 is adopted to prepare the zymophyte liquid, and the fermentation time and the fermentation temperature are strictly limited to ensure the fermentation effect, so that the paeoniflora fermentation product with multiple effects and stable quality is prepared, and the resource utilization of the medicinal paeoniflora is realized.
Description
Technical Field
The invention relates to the technical field of daily cosmetics, and particularly relates to a peony flower fermentation product, and a preparation method and application thereof.
Background
Due to the improvement of the economic living standard and the change of the consumption concept, cosmetic consumers and merchants are increasingly interested in healthy, green and purely natural raw material components. The plant raw materials are processed by utilizing the microbial fermentation technology, so that the release and conversion of active ingredients are promoted through the physiological activity of zymophyte, toxic ingredients are reduced, and the effect is greatly enhanced. Fermentation technology has been applied to some traditional Chinese medicinal materials, for example, CN114540426a discloses a method for fermenting peony roots with lactic acid bacteria, and for example, CN112999127a provides a fermented product of traditional Chinese medicines such as gentian. However, even if the same herb is used in different growing environments, there is a large difference in efficacy or active substance content between different producing areas or batches, and the difference directly or indirectly affects the quality of the fermentation product, but none of the above patents performs quality control on herb raw materials or final products.
The roots of peony, i.e. red peony and white peony, are traditional Chinese medicines and currently relevant planting and processing industries are very large-scale. However, the overground part of the medicinal peony is generally directly discarded, which often causes huge waste. Modern researches find that the flowers of the Chinese herbaceous peony contain polyphenol active matters such as gallic acid and the like, have physiological effects such as anti-inflammation, antioxidation and the like, and have wide application prospects. However, since various researches on paeonia lactiflora focus on roots, the reference data for processing and fermentation of paeonia lactiflora is less, and compared with common traditional Chinese medicinal materials, the fermentation process design and verification are more difficult. At present, no relevant reports on the fermentation of the peony flowers exist.
The invention aims to research a preparation method of a peony fermentation product, so as to fill the blank of the prior art and realize the resource utilization of medicinal peony.
Disclosure of Invention
One of the purposes of the invention is to provide a preparation method of a peony fermentation product, which can recycle peony and obtain the peony fermentation product with various effects and stable quality.
In order to achieve the purpose, the invention adopts the following technical scheme:
the preparation method of the peony flower fermentation product comprises the following steps:
s10, providing peony flowers which are derived from medicinal peonies and have gallic acid content of more than or equal to 9000ppm, drying, crushing, mixing with water, adjusting the pH to 4.5-5.0, and sterilizing to obtain a fermentation culture medium; wherein lactic acid is adopted to adjust the pH value of the mixture of the peony flower powder and pure water, and the pH value is preferably 5.0; sterilizing at 115 deg.C for 15min;
s20, providing a saccharomyces cerevisiae strain CICC 1400, and performing activation and domestication culture to obtain a zymophyte liquid;
s30, inoculating the zymophyte liquid to a fermentation culture medium, fermenting at 32 ℃ for 24 hours, cooling to 25 ℃, and fermenting for 48-60 hours to obtain fermentation liquid;
s40, centrifuging the fermentation liquor, removing thalli and medicinal powder, sequentially performing microfiltration and ultrafiltration treatment on the supernatant, and obtaining filtrate, namely the peony flower fermentation product.
The method controls the source of the peony flower, namely the medicinal peony, limits the content of gallic acid in the peony flower as a raw material to be more than or equal to 9000ppm, simultaneously adopts saccharomyces cerevisiae strain CICC 1400 to prepare zymocyte liquid, strictly limits the fermentation time and temperature after inoculation, and particularly controls the temperature to ensure the best fermentation effect, thereby realizing the resource utilization of the medicinal peony flower and preparing the peony flower fermentation product with various effects and stable quality.
As a preferable embodiment of the preparation method of the peony flower fermentation product, in step S10, the pulverized peony flower powder is mixed with water in a ratio of 1.
As a preferable scheme of the preparation method of the peony flower fermentation product, in step S20, a saccharomyces cerevisiae strain CICC 1400 is inoculated into a YPD culture medium and is shake-cultured at 32 ℃ for 36h for activation;
wherein the YPD culture medium consists of 1% of yeast extract, 1% of peptone and 2% of glucose.
As a preferable scheme of the preparation method of the peony flower fermentation product, in step S20, the activated bacterial liquid is inoculated into a domesticated culture medium, and shake culture is performed at 32 ℃ for 36 hours to prepare a zymogen liquid;
wherein the acclimatization culture medium comprises 0.25% of yeast extract, 0.25% of peptone, 0.5% of glucose and 5% of peony flower powder.
As a preferable embodiment of the preparation method of the peony flower fermentation product, in step S30, the inoculum size of the zymocyte liquid is 8%.
As a preferable scheme of the preparation method of the peony flower fermentation product, the particle size of the crushed peony flower is below 100 meshes.
As a preferable scheme of the preparation method of the peony flower fermentation product, the aperture of the filter membrane adopted by microfiltration is 0.22 μm.
As a preferred scheme of the preparation method of the peony flower fermentation product, the ultrafiltration membrane is a 5000KD ultrafiltration membrane.
The second purpose of the invention is to provide a peony flower fermentation product prepared by the preparation method, wherein the content of gallic acid in the peony flower fermentation product is 1/16 to 1/20 of the content of gallic acid in peony flower raw materials.
The invention also aims to provide the application of the peony flower fermentation product prepared by the preparation method in cosmetics. The cosmetic such as facial mask liquid, cream, essence, lotion and the like can have good anti-inflammatory, antioxidant, repairing and other effects by adding the peony flower fermentation product into the cosmetic.
The invention has the beneficial effects that:
1. the method controls the source and quality of raw materials, adopts a fermentation process, and strictly limits the fermentation temperature, so that a peony flower fermentation product with stable quality and high activity can be prepared, the peony flower fermentation product has good anti-inflammatory, antioxidant and repairing effects, and the whole preparation process is simple, controllable and easy to realize;
2. the invention effectively utilizes the waste peony flower in the peony industry, prepares products with high added value and has great social and economic benefits.
Drawings
FIG. 1 is a graph of the results of an in vitro DPPH free radical scavenging assay according to an embodiment of the present invention.
FIG. 2 is a graph showing the results of macrophage cytotoxicity test in the examples of the present invention.
FIG. 3 is a graph showing the results of a macrophage relaxation test in accordance with an embodiment of the present invention.
FIG. 4 is a graph showing the results of a keratinocyte Reactive Oxygen Species (ROS) assay.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments.
Unless otherwise specified, various starting materials of the present invention are commercially available or prepared according to conventional methods in the art.
In the following examples and comparative examples, high performance liquid chromatography (ref: wu Zhongwang, et al. Different drying methods were used to influence and evaluate 6 chemical components in white peony root [ J ]. Natural products research and development, 2016, 28 (11): 7.).
The fermentation process was carried out under aerobic conditions (150 rpm).
Example 1
Raw materials: the peony flower is from medicinal peony planted in Bozhou of Anhui, and the gallic acid content is 13538ppm.
The preparation method of the peony fermentation product in this example is as follows:
s10, cleaning the peony flowers, drying in an oven at 55 ℃, crushing, and sieving with a 100-mesh sieve.
S20, weighing 30g of powder, mixing the powder with 450mL of pure water, adjusting the pH to 5.0 by adopting lactic acid, and sterilizing for 15min at 115 ℃ to be used as a fermentation culture medium for later use.
S30, inoculating a saccharomyces cerevisiae strain CICC 1400 (purchased from Guangdong province microbial strain collection center) to 100ml of YPD culture medium (1% yeast extract, 1% peptone and 2% glucose) for activation by shaking culture at 32 ℃ for 36 h; 2mL of the activated bacterial liquid was inoculated into 100mL of an acclimatization medium (0.25% yeast extract, 0.25% peptone, 0.5% glucose, 5% peony flower powder), and shake-cultured at 32 ℃ for 36 hours to obtain a fermentation bacterial liquid.
S40, inoculating 36mL of bacterial liquid to a fermentation medium, fermenting at 32 ℃ for 24 hours, cooling to 25 ℃, and fermenting for 48 hours.
S50, taking 5000g of fermentation liquor, centrifuging for 10min, sequentially carrying out microfiltration on supernate through a 0.22-micrometer filter membrane and ultrafiltration through a 5000KD ultrafiltration membrane, and obtaining filtrate which is a peony flower fermentation product.
The content of gallic acid in the peony flower fermentation product is 721ppm.
Example 2
Raw materials: the peony flower is medicinal peony planted in Yuyao from Zhejiang, and the gallic acid content is 9840ppm.
The preparation method of the peony flower fermentation product in this example is the same as that in example 1.
The content of gallic acid detected in the peony flower fermentation product is 583ppm.
Example 3
Raw materials: the flos Paeoniae is from Paeonia lactiflora pall planted in Henan Hengyang, and has gallic acid content of 9112ppm.
The preparation method of the peony flower fermentation product in this example is the same as that in example 1.
The content of gallic acid detected in the peony flower fermentation product is 559ppm.
Comparative example 1
The source of the starting material was the same as in example 1.
The preparation method of the peony aqueous extract in the comparative example is as follows:
1) Cleaning peony flower, oven drying at 55 deg.C, pulverizing, and sieving with 100 mesh sieve.
2) Weighing 30g of powder according to the proportion of 1
3) Heating at 80 deg.C for 1.5h;
4) Centrifuging 5000g of extractive solution for 10min, filtering supernatant,
5) Concentrating under reduced pressure at 50 deg.C to reduce volume by about 1/2
6) Microfiltering the concentrated solution to 0.22 μm to obtain water extract of flos Paeoniae.
The gallic acid content in the water extract of Paeonia lactiflora Pallas was 602ppm.
Comparative example 2
Raw materials: the peony flower for viewing and admiring peony is purchased from south of Guangzhou Ling, and the gallic acid content is 2085mg/kg.
The preparation method of the peony fermentation product in this comparative example is the same as in example 1.
The gallic acid content in the peony flower fermentation product was detected to be 112ppm.
Comparative example 3
The source of the starting material was the same as in example 1.
The preparation method of the peony flower fermentation product in the comparative example is basically the same as that in example 1, except that: and S40, fermenting at constant temperature, namely fermenting at 32 ℃ for 72h.
The content of gallic acid detected by the peony flower fermentation product is 427ppm.
Comparative example 4
The source of the starting material was the same as in example 1.
The preparation method of the peony flower fermentation product in the comparative example is basically the same as that in example 1, except that: and S40, fermenting at constant temperature, namely fermenting at 25 ℃ for 72h.
The content of gallic acid detected by the peony flower fermentation product is 381ppm.
Comparative example 5
The source of the starting material was the same as in example 1.
The preparation method of the peony flower fermentation product in the comparative example is basically the same as that in example 1, except that: the strain used in step S30 was modified to Saccharomyces cerevisiae ATCC 26603 (purchased from Guangdong province culture collection).
The content of gallic acid detected by the peony flower fermentation product is 305ppm.
Comparative example 6
The source of the starting material was the same as in example 1.
The preparation method of the peony flower fermentation product in the comparative example is basically the same as that in example 1, except that: the strain used in step S30 was Saccharomyces diastaticus ACCC 20204 (available from Guangdong province culture Collection).
The content of gallic acid detected in the peony flower fermentation product was 189ppm.
Application example-facial mask liquid
A cosmetic (mask solution) added with a peony flower fermentation product is provided, and a formula of the mask solution is shown in Table 1.
TABLE 1 facial mask liquid formulation
Preparing facial mask liquid according to the formula:
1. accurately weighing the phase A raw material, and uniformly stirring and dispersing;
2. adding phase B, and stirring uniformly;
3. adding the phase C in sequence, and stirring uniformly while heating.
In vitro free radical scavenging assay
The test method was referenced to T/SHRH 006-2018, and the results are shown in FIG. 1.
As shown in figure 1, the free radical clearance of 10% diluent in the embodiment is about 50%, which is obviously higher than that of each control example, and the peony flower fermentation product has strong antioxidation, small difference among the embodiments and stable product quality; the gallic acid content of comparative example 1 is higher than that of examples 2 and 3, but the antioxidant effect is far less than that of examples 2 and 3, indicating that more abundant antioxidant substances are produced or retained during the fermentation process.
Cytotoxicity assay
The test method is briefly described as follows: macrophage Raw 264.7 was inoculated in a 96-well plate and cultured for 24h. The culture medium was aspirated from each well and samples of different concentrations were added. Exposing for 24h, washing the cells with PBS, adding 10% CCK-8, incubating for 1h, and measuring the OD450 value. Blank wells containing medium only and no cells, control cultured cells without any sample, and OD measured in blank wells and control 450 The value is obtained. The relative cell activities were calculated according to formula (1) and the results are shown in FIG. 2.
As shown in FIG. 2, the cell activity of 3 examples was more than 90% at 1% and 5% and 10% concentrations, indicating higher safety. Whereas control 1, prepared by extraction with water, exhibited significant cytotoxicity at concentrations of 5% and 10%; in addition, comparative examples 4 and 6 also exhibited some cytotoxicity at 10% concentration, indicating that changing fermentation conditions and fermentation species had a greater effect on the metabolic activity of the fermentation process.
Cell relaxation assay
The assay was performed with reference to T/SHRH033-2020, and the results are shown in FIG. 3.
As shown in fig. 3, compared with the negative control, the 5% peony flower fermentation product significantly reduces the secretion of inflammatory factor TNF- α, and the effect is better than the control, which indicates that the peony flower fermentation product has a good soothing effect.
Cell antioxidant assay
The test method was performed with reference to T/SHRH032-2020, and the results are shown in FIG. 4.
As shown in fig. 4, when keratinocytes were treated with uv light, which resulted in an increase in cellular Reactive Oxygen Species (ROS) (the higher the fluorescence intensity represents the higher the ROS content), the examples all significantly reduced ROS content, indicating that peony flower fermentation product can reduce ROS produced by cells due to uv light, and thus protect them.
Consumer use testing
Sensitive muscle groups were screened by lactic acid sting and 14 female volunteers were recruited together between 25 and 43 years of age. The facial mask liquid prepared by the application example is delivered to volunteers for use, the facial mask liquid is required to be used once a day and is used for 3 weeks continuously for return visit, and 13 volunteers show that the skin condition is obviously improved, and the frequency of redness or stabbing pain of facial skin is reduced.
In conclusion, the fermentation method provided by the invention can prepare the peony flower fermentation product with stable quality and high activity, has good anti-inflammatory, antioxidant and repairing effects, is simple and controllable in the whole process, and is easy to realize; the patent effectively utilizes the waste in the peony flower industry, prepares products with high added value and has great social and economic benefits.
The above examples are only intended to illustrate the detailed process of the present invention, and the present invention is not limited to the above detailed process, i.e., it is not intended that the present invention necessarily depends on the above detailed process for its implementation. It is understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention and the addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
Claims (10)
1. The preparation method of the peony flower fermentation product is characterized by comprising the following steps:
s10, providing peony flowers which are derived from medicinal peony and have gallic acid content of more than or equal to 9000ppm, drying, crushing, mixing with water, adjusting pH to 4.5-5.0, and sterilizing to obtain a fermentation culture medium;
s20, providing a saccharomyces cerevisiae strain CICC 1400, and performing activation and domestication culture to obtain a zymocyte liquid;
s30, inoculating the zymophyte liquid to a fermentation culture medium, fermenting at 32 ℃ for 24 hours, cooling to 25 ℃, and fermenting for 48-60 hours to obtain fermentation liquid;
s40, centrifuging the fermentation liquor, removing thalli and medicinal powder, sequentially performing microfiltration and ultrafiltration treatment on the supernatant, and obtaining filtrate, namely the peony flower fermentation product.
2. The method of preparing a fermented product of peony flower according to claim 1, wherein in step S10, the pulverized peony flower powder is mixed with water at a ratio of 1.
3. The method for preparing a fermented product of peony flower according to claim 1, wherein in step S20, saccharomyces cerevisiae strain cic 1400 is inoculated into YPD medium, and shake-cultured at 32 ℃ for 36h for activation.
4. The method for preparing a fermented product of peony flower according to claim 1, wherein in step S20, the activated bacterial liquid is inoculated into an acclimatization medium, and shake-cultured at 32 ℃ for 36h to obtain a fermented bacterial liquid;
wherein the acclimatization medium contains 0.25% of yeast extract, 0.25% of peptone, 0.5% of glucose and 5% of peony flower powder.
5. The method of claim 1, wherein the inoculum size of the fermentation broth is 8% in step S30.
6. The method of preparing a fermented product of peony flower according to claim 1, wherein the particle size of pulverized peony flower is 100 mesh or less.
7. The method for preparing a fermented product of peony flower according to claim 1, wherein the pore size of the filter membrane used for microfiltration is 0.22 μm.
8. The method for preparing a peony flower fermentation product according to claim 1, wherein the ultrafiltration membrane used in the ultrafiltration is a 5000KD ultrafiltration membrane.
9. A peony flower fermentation product, which is characterized by being prepared by the preparation method of any one of claims 1-8, wherein the content of gallic acid in the peony flower fermentation product is 1/16 to 1/20 of the content of gallic acid in peony flower raw material.
10. Use of the fermented product of peony flower prepared by the method according to any one of claims 1-8 in cosmetics.
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