CN116602907A - Fermentation liquor with emulsifying property, product containing fermentation liquor and preparation method and application of fermentation liquor - Google Patents

Fermentation liquor with emulsifying property, product containing fermentation liquor and preparation method and application of fermentation liquor Download PDF

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Publication number
CN116602907A
CN116602907A CN202310735996.XA CN202310735996A CN116602907A CN 116602907 A CN116602907 A CN 116602907A CN 202310735996 A CN202310735996 A CN 202310735996A CN 116602907 A CN116602907 A CN 116602907A
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fermentation
active ingredient
seed
fermentation broth
preparation
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王冬冬
王昌涛
李萌
张佳婵
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Beijing Technology and Business University
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Beijing Technology and Business University
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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]
    • A61K8/9794—Liliopsida [monocotyledons]
    • 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
    • A61K8/9728—Fungi, e.g. yeasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • 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
    • 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
    • A61Q19/004—Aftersun preparations
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
    • 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
    • A61K2800/10—General cosmetic use
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • 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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  • Engineering & Computer Science (AREA)
  • Dermatology (AREA)
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  • Biochemistry (AREA)
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Abstract

The application provides a fermentation broth with emulsifying property, a product containing the fermentation broth, a preparation method and application thereof. The preparation method of the fermentation broth with the emulsifying property comprises the following steps: (1) Mixing 1 part of rice flour, 0.2-2 parts of nitrogen source and 20-150 parts of water to prepare a seed culture medium; inoculating lactobacillus into a seed culture medium, and fermenting and culturing to obtain seed solution A; (2) Inoculating the seed liquid A prepared in the step (1) into a fermentation substrate, and performing fermentation culture and one-time sterilization to prepare a fermentation liquid with emulsifying property; wherein the fermentation substrate comprises 1-5 parts of tremella powder and 20-150 parts of water. The application takes rice, tremella and other substances as fermentation substrates, and prepares the fermentation liquor with emulsifying property through step fermentation culture, can replace an emulsifying agent to be used in the field of cosmetics, improves the stability of the cosmetics, has low preparation cost and high use safety, and also has multiple beauty effects of resisting photodamage, repairing skin, resisting oxidation, resisting inflammation and the like.

Description

Fermentation liquor with emulsifying property, product containing fermentation liquor and preparation method and application of fermentation liquor
Technical Field
The application belongs to the field of cosmetics, and particularly relates to a fermentation broth with emulsifying property, a product containing the fermentation broth, and a preparation method and application of the fermentation broth.
Background
In the field of cosmetics, the emulsifier can combine some insoluble components together to prepare uniform and stable product, so that the formulation of the cosmetics is more flexible. At present, chemical agents are generally adopted as emulsifying agents, the preparation cost is high, the chemical agents have a stimulating effect on skin, and the skin feel of cosmetics can be possibly influenced, so that the problem that the emulsifying agents are carefully considered is solved.
Therefore, there is a need in the art to develop a purely natural substance having emulsifying properties suitable for the cosmetic field, which can improve the stability of cosmetics, ensure the safety of the cosmetics, and have cosmetic effects.
Disclosure of Invention
The application aims to overcome the defects that a chemical emulsifier in the prior art has a stimulation effect on skin and influences the skin feel of cosmetics and the like, and provides a fermentation liquid with emulsifying property, a product containing the fermentation liquid, a preparation method and application thereof. The rice, tremella and other substances are used as fermentation substrates, and fermentation broth with emulsifying property is prepared through step-by-step fermentation culture, so that the fermentation broth can be used for replacing an emulsifying agent in the field of cosmetics, the stability of the cosmetics is improved, the preparation cost is low, the use safety is high, and the fermentation broth has multiple beauty effects of resisting photodamage, repairing skin, resisting oxidation, resisting inflammation and the like. In addition, no organic solvent is added in the preparation process, so that the pollution to the environment is reduced, the preparation condition is mild, and the energy conservation and the environmental protection are realized.
The application adopts the following technical scheme to solve the technical problems:
the application provides a preparation method of fermentation liquor with emulsifying property, which comprises the following steps:
(1) Mixing 1 part of rice flour, 0.2-2 parts of nitrogen source and 20-150 parts of water to prepare a seed culture medium; inoculating lactobacillus into the seed culture medium, and fermenting and culturing to obtain seed solution A;
(2) Inoculating the seed liquid A prepared in the step (1) into a fermentation substrate, and performing fermentation culture and primary sterilization to prepare a fermentation liquid with emulsifying property; wherein the fermentation substrate comprises 1-5 parts of tremella powder and 20-150 parts of water.
In step (1) of some embodiments, the rice flour may be a material obtained by pulverizing rice as conventionally considered by those skilled in the art.
In step (1) of some embodiments, the rice flour may have a particle size of 50 to 150 mesh, preferably 50 to 100 mesh.
In step (1) of some embodiments, the nitrogen source may comprise a nitrogen source conventionally used in the art for providing energy to a fermentation broth, preferably at least one of milk powder, soy peptide and peptone. Wherein the milk powder may be a powder obtained by removing water from animal milk, preferably a powder obtained by removing water from milk, as conventionally known to those skilled in the art.
In step (1) of some embodiments, the nitrogen source is preferably 0.5 to 1 part by weight, more preferably 0.5 to 0.7 part by weight, for example 0.67 part by weight.
In step (1) of some embodiments, the water is preferably 50 to 100 parts by weight.
In step (1) of some embodiments, the seed medium may also include a sterilization procedure prior to use as is conventional in the art.
The method of the sterilization of the seed medium may be conventional in the art, and may generally be a high temperature sterilization method.
When the sterilization of the seed medium is performed by the high temperature sterilization method, the sterilization temperature may be a temperature which is conventional in the art for such operations, preferably 110 to 125 ℃, more preferably 115 to 121 ℃.
When the sterilization of the seed medium is performed by the high temperature sterilization method, the sterilization time may be a time conventional in the art for such an operation, preferably 20 to 60 minutes, more preferably 20 to 40 minutes, for example 30 minutes.
Wherein, the seed culture medium may further comprise a cooling operation, typically to room temperature, following the sterilization operation, as is conventional in the art.
In step (1) of some embodiments, the lactobacillus may comprise lactobacillus helveticus (Lactobacillus helveticus) and/or lactobacillus plantarum (Lactiplantibacillus plantarum).
Wherein the lactobacillus helveticus may comprise lactobacillus helveticus with a deposit number of CICC 20243 purchased from China industry microbiological culture Collection center.
Wherein the lactobacillus plantarum can comprise lactobacillus plantarum with the preservation number of CICC20261 purchased from China industry microbiological culture Collection center.
In step (1) of some embodiments, the lactobacillus may be added as a lactobacillus bacterial solution having a viable count of 10, as is conventional in the art 6 ～10 10 CFU/mL, preferably 10 7 ～10 9 CFU/mL, e.g. 10 8 CFU/mL。
In step (1) of some embodiments, the number of Lactobacillus inoculated per unit volume of the seed medium may be conventional in the art, preferably 10 4 ～10 8 CFU/mL, more preferably 10 5 ～10 6 CFU/mL。
In step (1) of some embodiments, the conditions and methods of the fermentation culture may be conventional in the art, and typically may be stationary culture in a constant temperature incubator.
In step (1) of some embodiments, the fermentation time may be 12 to 24 hours, preferably 12 to 16 hours.
In step (1) of some embodiments, the temperature of the fermentation culture may be 30 to 45 ℃, preferably 37 to 45 ℃.
In step (2) of some embodiments, the tremella powder may be a material conventionally considered by those skilled in the art to be obtained by pulverizing tremella.
In step (2) of some embodiments, the tremella powder may have a particle size of 50-100 mesh.
In step (2) of some embodiments, the weight part of the tremella powder is preferably 2 to 3.5 parts, for example 3.3 parts.
In step (2) of some embodiments, the water is preferably 50 to 100 parts by weight.
In step (2) of some embodiments, the fermentation substrate may further comprise a carbon source and/or a nitrogen source.
Wherein the carbon source may include at least one of glucose, corn and potato.
Wherein, the weight portion of the carbon source can be 0.1 to 0.5 portion, preferably 0.2 to 0.5 portion.
Wherein the nitrogen source may include at least one of milk powder, soybean peptide and black bean powder.
Wherein, the weight portion of the nitrogen source can be 0.1 to 0.5 portion, preferably 0.2 to 0.5 portion.
In step (2) of some embodiments, the fermentation substrate may also comprise a sterilization procedure as is conventional in the art.
Wherein the method of said sterilizing the fermentation substrate may be conventional in the art, and may generally be a high temperature sterilization method.
When the sterilization of the fermentation substrate is performed by the high temperature sterilization method, the sterilization temperature may be a temperature which is conventional in the art for such operations, preferably 110 to 125 ℃, more preferably 115 to 121 ℃.
When the sterilization of the fermentation substrate is performed by the high temperature sterilization method, the sterilization time may be a time conventional in the art for such an operation, preferably 20 to 60 minutes, more preferably 20 to 40 minutes, for example 30 minutes.
In step (2) of some embodiments, the conditions and methods of the fermentation culture may be conventional in the art, and typically may be stationary culture in a constant temperature incubator.
In step (2) of some embodiments, the fermentation time may be 12 to 30 hours, preferably 12 to 20 hours.
In step (2) of some embodiments, the temperature of the fermentation culture may be 30 to 45 ℃, preferably 35 to 40 ℃.
In step (2) of some embodiments, the method of primary sterilization may be conventional in the art, and may generally be a high temperature sterilization method.
In step (2) of some embodiments, when the primary sterilization is performed by the high temperature sterilization method, the temperature of the primary sterilization may be a temperature conventional in the art for such operations, preferably 100 to 121 ℃.
In step (2) of some embodiments, when the high temperature sterilization method is used for the primary sterilization, the time for the primary sterilization may be a time conventional in the art for such an operation, preferably 15 to 35min, more preferably 30min.
In step (2) of some embodiments, the one sterilization operation may be followed by a centrifugation step, as is conventional in the art, to collect the supernatant.
The rotational speed of the centrifugation may be a rotational speed conventional in the art, preferably 3000 to 9000rpm, more preferably 4000 to 6000rpm, for example 5500rpm.
The centrifugation time may be a time conventional in this type of operation in the art, and is preferably 10 to 40 minutes, more preferably 20 to 40 minutes, for example, 30 minutes.
Wherein the centrifugation may be followed by a secondary sterilization and/or mixing with a preservative.
The secondary sterilization method may be a high temperature sterilization method conventionally used in the art.
When the secondary sterilization is performed using the high temperature sterilization method, the temperature of the secondary sterilization may be a temperature conventional in the art for such an operation, and preferably is 100 to 121 ℃.
When the secondary sterilization is performed by the high temperature sterilization method, the time for the secondary sterilization may be a time conventional in the art for such an operation, preferably 20 to 40 minutes, more preferably 30 minutes.
The temperature of the mixing during the mixing with the preservative may be a temperature conventional in the art for such operations, preferably 50-80 ℃, more preferably 70-80 ℃.
In the mixing process with the preservative, the preservative may include p-hydroxyacetophenone and/or 1, 2-hexanediol as is conventional in the art.
When the preservative comprises the p-hydroxyacetophenone and the 1, 2-hexanediol, the mass percentage of the p-hydroxyacetophenone to the supernatant fluid prepared after the centrifugation can be 0.1-1%, and the mass percentage of the 1, 2-hexanediol to the supernatant fluid prepared after the centrifugation can be 0.1-1%; preferably, the p-hydroxyacetophenone accounts for 0.5% of the supernatant obtained after centrifugation, and the 1, 2-hexanediol accounts for 0.5% of the supernatant obtained after centrifugation.
The application also provides a fermentation broth with emulsifying property, which is prepared by the preparation method of the fermentation broth with emulsifying property.
The application also provides an application of the fermentation broth with the emulsifying property in preparing a skin external preparation.
In some embodiments, the fermentation broth with emulsifying property can be used as at least one of photodamage-resistant active ingredient, skin repair active ingredient, oxidation-resistant active ingredient and anti-inflammatory active ingredient in the external preparation for skin.
The application also provides a skin external agent, which comprises a material A and vegetable oil, wherein the material A comprises the fermentation liquor with emulsifying property; the viscosity of the material A is 6000-8000 mPa.S.
In some embodiments, the material a may further comprise water. The water may be sterile deionized water as is conventionally used in the art. The purpose of the water addition is to adjust the viscosity of the material A to 6000 to 8000 mPa.S. In the research and development process, when the viscosity of the material A is lower than 6000 mPa.S, the prepared skin external agent can generate a water-oil separation phenomenon; when the viscosity of the material A is higher than 8000 mPa.S, the skin feel is poor and delamination occurs.
In some embodiments, the vegetable oil may comprise a vegetable oil conventionally used in the art, and may generally comprise at least one of peony seed oil, prinsepia utilis royle oil, macadamia nut seed oil and meadowfoam seed oil.
In some embodiments, the external preparation for skin may further comprise a cosmetic active ingredient, preferably at least one of a moisturizing active ingredient, a whitening active ingredient, an anti-inflammatory active ingredient, an anti-allergic active ingredient, and an anti-oxidation active ingredient.
In some embodiments, the vegetable oil may be 6% -12% by mass of the material a, preferably 8% -10%.
In some embodiments, the skin external preparation may include, but is not limited to, a cream, essence, or toner as is conventional in the art.
The application also provides a preparation method of the skin external agent, which comprises the following steps: mixing the material A with the vegetable oil, and shearing and homogenizing.
In some embodiments, when the cosmetic active ingredient is also included in the skin external agent, the cosmetic active ingredient may be added during the mixing.
In some embodiments, the shear homogenizing speed may be conventional in the art, and may generally be 10000-20000 r/min, preferably 14000-16000 r/min.
In some embodiments, the time for shear homogenization may be conventional in the art and may generally be 3 to 10 minutes, preferably 5 to 8 minutes.
In some embodiments, the room temperature generally refers to 15-40 ℃.
On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the application.
The reagents and materials used in the present application are commercially available.
The application has the positive progress effects that: the application takes rice, tremella and other substances as fermentation substrates, and prepares the fermentation liquor with emulsifying property through step fermentation culture, can replace an emulsifying agent to be used in the field of cosmetics, improves the stability of cosmetics, has low preparation cost and high use safety, and also has multiple beauty effects of resisting photodamage, repairing skin, resisting oxidation, resisting inflammation and the like. In addition, no organic solvent is added in the preparation process, so that the pollution to the environment is reduced, the preparation condition is mild, and the energy conservation and the environmental protection are realized.
Drawings
The application may be better understood by reference to the following description taken in conjunction with the accompanying drawings. The accompanying drawings, which are included to provide a further illustration of the preferred embodiments of the application and together with a further understanding of the principles and advantages of the application, are incorporated in and constitute a part of this specification.
Wherein:
FIG. 1 is a graph showing the stability of the products obtained in examples 1 to 4 and comparative examples 1 to 4;
FIG. 2 is a graph showing comparison of stability of emulsions A1, A2, A3 and A4 prepared in application example 1;
FIG. 3 is a graph showing comparison of stability of emulsions B1, B2, B3 and B4 prepared in application example 2;
FIG. 4 is a graph showing comparison of stability of emulsions C1, C2, C3 and C4 obtained in application example 3;
FIG. 5 is a graph showing comparison of stability of emulsions D1, D2, D3 and D4 obtained in application example 4;
FIG. 6 is a graph showing the degree of influence of the products prepared in examples 1 to 4 and comparative examples 1 to 4 on the expression level of type I collagen in fibroblasts of damaged human skin;
FIG. 7 is a graph showing the effect of the products prepared in examples 1 to 4 and comparative examples 1 to 4 on the cell viability of fibroblasts on damaged human skin;
FIG. 8 is a graph showing the comparison of the protection ability of HaCaT cells by the products prepared in examples 1 to 4 and comparative examples 1 to 4;
FIG. 9 is a graph showing the DPPH radical scavenging ability of the products obtained in examples 1 to 4 and comparative examples 1 to 4;
FIG. 10 is a graph showing the anti-inflammatory power of the products obtained in examples 1 to 4 and comparative examples 1 to 4.
Detailed Description
The application is further illustrated by means of the following examples, which are not intended to limit the scope of the application. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
The experimental methods used in the following examples are conventional methods unless otherwise specified.
In example 1 below, lactobacillus helveticus was purchased from the chinese industrial microbiological bacterial deposit management center under the deposit number cic 20243;
in the following example 2, lactobacillus plantarum was purchased from the chinese industrial microorganism strain collection center under the collection number cic 20261;
in the following application examples, peony seed oil was purchased from Kangpu biotechnology Co., ltd in lotus city;
in the following application examples, prinsepia utilis oil was purchased from vitamin and beneficial peptide (Guangdong) biotechnology Co., ltd;
in the following application examples, macadamia nut seed oil was purchased from Shanghai Naxin Biotech Co., ltd;
in the following application examples, the white pool flower seed oil was purchased from the company of the import and export of the Argania tree (Hangzhou);
in the following examples and comparative examples, milk powder was purchased from Henan Asian Biotechnology Co., ltd.
Example 1
(1) Weighing 3g of rice flour, 2g of milk powder and 300g of deionized water, mixing, sterilizing at 121 ℃ for 30min, and cooling to room temperature after sterilization to obtain a seed culture medium; wherein, the grain diameter of the rice flour is 100 meshes;
inoculating lactobacillus helveticus bacterial liquid with the preservation number of CICC 20243 into the prepared seed culture medium, wherein the viable count of the lactobacillus helveticus bacterial liquid is 10 8 CFU/mL, unit volume of seed culture medium inoculated with RayleighThe number of lactobacillus helveticus is 10 6 Shaking the CFU/mL, and carrying out fermentation culture in a 37 ℃ incubator for 16 hours to obtain a seed solution A;
(2) Uniformly mixing 10g of tremella powder (with the particle size of 100 meshes), 0.6g of glucose, 0.6g of milk powder and 300g of deionized water, sterilizing at a high temperature of 121 ℃ for 30min, and cooling to room temperature after sterilization to obtain a fermentation substrate;
inoculating the seed liquid A prepared in the step (1) into the prepared fermentation substrate, performing fermentation culture in a 40 ℃ incubator, namely standing fermentation culture for 16 hours, and sterilizing the obtained fermentation liquid in a sterilizing pot at 100 ℃ for 30 minutes once after the fermentation culture is finished, and stopping lactobacillus fermentation; cooling to room temperature after primary sterilization, centrifuging for 30min under 5500r/min to obtain supernatant, and performing secondary sterilization at 100deg.C for 30min to avoid contamination of other bacteria during centrifugation, and cooling to room temperature after secondary sterilization to obtain fermentation broth with emulsifying property.
Example 2
Compared with example 1, the fermentation broth having emulsifying property was prepared by substituting lactobacillus helveticus having the accession number of cic 20243 with lactobacillus plantarum having the accession number of cic 20261 in the same amount, except that the fermentation broth was different from the fermentation broth in example 1.
Example 3
The difference from example 1 is only that the addition amount of part of the raw materials is different, specifically: the mass of rice flour in the step (1) is 4g, the mass of tremella powder in the step (2) is 8g, and other condition parameters are the same as those in the example 1.
Example 4
The only difference compared to example 1 is the fermentation incubation time and the fermentation substrate in step (2), in particular:
(1) Weighing 3g of rice flour, 2g of milk powder and 300g of deionized water, mixing, sterilizing at 121 ℃ for 30min, and cooling to room temperature after sterilization to obtain a seed culture medium; wherein, the grain diameter of the rice flour is 100 meshes;
inoculating lactobacillus helveticus bacterial liquid with the preservation number of CICC 20243 into the prepared seed culture medium, wherein the viable count of the lactobacillus helveticus bacterial liquid is 10 8 CFU/mL, number of Lactobacillus helveticus inoculated per unit volume of seed medium was 10 6 Shaking the CFU/mL, and carrying out fermentation culture in a 37 ℃ incubator for 12 hours to obtain a seed solution A;
(2) Uniformly mixing 10g of tremella powder (with the particle size of 100 meshes) and 300g of deionized water, sterilizing at a high temperature of 121 ℃ for 30min, and cooling to room temperature after sterilization to obtain a fermentation substrate;
inoculating the seed liquid A prepared in the step (1) into the prepared fermentation substrate, performing fermentation culture in a 40 ℃ incubator, namely standing fermentation culture for 20 hours, and sterilizing the obtained fermentation liquid in a sterilizing pot at 100 ℃ for 30 minutes once after the fermentation culture is finished, and stopping lactobacillus fermentation; cooling to room temperature after primary sterilization, centrifuging for 30min under 5500r/min to obtain supernatant, and performing secondary sterilization at 100deg.C for 30min to avoid contamination of other bacteria during centrifugation, and cooling to room temperature after secondary sterilization to obtain fermentation broth with emulsifying property.
Application example 1
Adding sterile deionized water into the fermentation broth with emulsifying property prepared in the above example 1, testing the viscosity of the system under the condition of 12r/min of rotation speed by adopting a No. 3 rotor of a digital viscometer NDJ-8S, and stopping adding water when the viscosity of the system is 7000 mPa.S to prepare a material A;
adding vegetable oil (respectively adding peony seed oil, prinsepia utilis royle oil, macadamia nut seed oil and white pool flower seed oil) into the prepared material A, wherein the addition amount of the vegetable oil accounts for 8% of the mass of the material A, carrying out shearing and homogenizing treatment after mixing, wherein the shearing and homogenizing time is 5min, the rotating speed of the shearing and homogenizing is 14000r/min, and the emulsion prepared by adding the four vegetable oils is named emulsion A1, emulsion A2, emulsion A3 and emulsion A4 respectively.
Application example 2
Adding sterile deionized water into the fermentation broth with emulsifying property prepared in the above example 2, testing the viscosity of the system under the condition of 12r/min of rotation speed by adopting a No. 3 rotor of a digital viscometer NDJ-8S, and stopping adding water when the viscosity of the system is 7000 mPa.S to prepare a material A;
adding vegetable oil (respectively adding peony seed oil, prinsepia utilis royle oil, macadamia nut seed oil and white pool flower seed oil) into the prepared material A, wherein the addition amount of the vegetable oil accounts for 8% of the mass of the material A, mixing, performing shearing homogenization treatment, wherein the shearing homogenization time is 5min, the rotating speed of the shearing homogenization is 14000r/min, and the emulsion prepared by adding the four vegetable oils is named emulsion B1, emulsion B2, emulsion B3 and emulsion B4 respectively.
Application example 3
Adding sterile deionized water into the fermentation broth with emulsifying property prepared in the above example 3, testing the viscosity of the system under the condition of 12r/min of rotation speed by adopting a No. 3 rotor of a digital viscometer NDJ-8S, and stopping adding water when the viscosity of the system is 7000 mPa.S to prepare a material A;
adding vegetable oil (respectively adding peony seed oil, prinsepia utilis royle oil, macadamia nut seed oil and white pool flower seed oil) into the prepared material A, wherein the addition amount of the vegetable oil accounts for 8% of the mass of the material A, mixing, performing shearing homogenization treatment, wherein the shearing homogenization time is 5min, the rotating speed of the shearing homogenization is 14000r/min, and the emulsion prepared by adding the four vegetable oils is named emulsion C1, emulsion C2, emulsion C3 and emulsion C4 respectively.
Application example 4
Adding sterile deionized water into the fermentation broth with emulsifying property prepared in the above example 4, testing the viscosity of the system under the condition of 12r/min of rotation speed by adopting a No. 3 rotor of a digital viscometer NDJ-8S, and stopping adding water when the viscosity of the system is 7000 mPa.S to prepare a material A;
adding vegetable oil (respectively adding peony seed oil, prinsepia utilis royle oil, macadamia nut seed oil and white pool flower seed oil) into the prepared material A, wherein the addition amount of the vegetable oil accounts for 8% of the mass of the material A, mixing, performing shearing homogenization treatment, wherein the shearing homogenization time is 5min, the rotating speed of the shearing homogenization is 14000r/min, and the emulsion prepared by adding the four vegetable oils is named emulsion D1, emulsion D2, emulsion D3 and emulsion D4 respectively.
Comparative example 1
The only difference compared to example 1 is the one-step fermentation process, which comprises the following steps:
weighing 3g of rice powder, 2g of milk powder, 10g of tremella powder and 600g of deionized water, mixing, sterilizing at 121 ℃ for 30min, and cooling to room temperature after sterilization to obtain a fermentation substrate; wherein, the mesh number of the rice powder is 100 meshes, and the mesh number of the tremella powder is 100 meshes;
standing in a 37 ℃ incubator for fermentation culture for 16 hours, sterilizing the obtained fermentation liquor for 30 minutes at the temperature of 100 ℃ after the fermentation culture is finished, and stopping lactobacillus fermentation; cooling to room temperature after sterilization, centrifuging for 30min under 5500r/min to obtain supernatant, performing secondary sterilization at 100deg.C for 30min to avoid contamination by other bacteria during centrifugation, and cooling to room temperature after sterilization.
Comparative example 2
Compared with example 1, the difference is that the fermentation strain is different, the lactobacillus helveticus with the preservation number of CICC 20243 in the step (1) is replaced by the same amount of yeast with the product number of AS2.1392 purchased from food brewing institute of Beijing city, and other condition parameters are the same AS those of example 1;
the yeast is cultured in YPD liquid culture medium for 48 hr before use, and the viable bacteria concentration is 10 8 CFU/mL yeast liquid.
Comparative example 3
The difference compared with example 1 is that the rice flour in step (1) is replaced by the same amount of millet flour, the mesh number of the millet flour is 100 mesh, and other condition parameters are the same as example 1.
Comparative example 4
Compared with example 1, the difference is only that the tremella powder added in step (2) is different, specifically, 2g tremella powder is added, and other condition parameters are the same as those of example 1.
Effect example 1 stability evaluation
The products obtained in examples 1 to 4 and comparative examples 1 to 4 were evaluated for stability, and the products were left at 50℃and normal temperature and-8℃for 4 weeks, respectively, and the sample state was observed after 4 weeks, and the results are shown in FIG. 1 and Table 1.
The stability of the products obtained in the above application examples 1 to 4 was evaluated, and the products were left at 50℃and normal temperature at-8℃for 4 weeks, respectively, and the sample state after 4 weeks was observed, and the results are shown in FIGS. 2 to 5 and Table 2.
TABLE 1
Numbering device 50℃ Normal temperature -8℃
Example 1 Normal state Normal state Normal state
Example 2 Normal state Normal state Normal state
Example 3 Normal state Normal state Normal state
Example 4 Normal state Normal state Normal state
Comparative example 1 Has the advantages of precipitation and non-uniform product state Normal state Normal state
Comparative example 2 Has the advantages of precipitation and non-uniform product state Normal state Normal state
Comparative example 3 Has the advantages of precipitation and non-uniform product state Normal state Normal state
Comparative example 4 Has the advantages of precipitation and non-uniform product state Normal state Normal state
TABLE 2
For the products prepared in examples 1 to 4 and comparative examples 1 to 4, the results show that the products prepared in examples 1 to 4 are uniformly and stably maintained under different conditions, and the products prepared in comparative examples 1 to 4 are precipitated at 50 ℃ to cause non-uniform product conditions. The emulsion prepared by homogeneously mixing the products prepared in examples 1 to 4 with vegetable oil is uniform and stable in state, and has no unstable phenomena such as layering or oil-water separation.
Effect example 2 repair ability (type I collagen expression level)
In the experiment, the restoration capacity of the products prepared in the examples 1-4 and the comparative examples 1-4 on damaged skin is verified by measuring the COL-I content in human skin fibroblasts.
The logarithmic phase human skin fibroblasts were inoculated with the cell suspension at a density of 25 ten thousand cells/mL on a 6-well cell culture plate, and 2mL of the cell suspension was added to each well and cultured for 12 hours. Experimental, blank and model groups were set. Wherein, the experimental group and the model group adopt 18mJ/cm 2 The cells were irradiated with UVA for 40min. After irradiation, the supernatant fluid in the experimental group, the blank group and the model group is sucked and removed, 2mL of DMEM solution is added into the experimental group, 2mL of the liquid to be tested is added into the experimental group (the liquid to be tested prepared in the example or the comparative example is prepared into the liquid to be tested with the volume percentage of 2% by adopting DMEM, the liquid to be tested needs to be subjected to a sterile filter membrane with the size of 0.22 μm before being used), the cells are subjected to post-treatment for 24 hours, the supernatant fluid is sucked and removed, the cells are washed for 2 to 3 times by PBS, then the cells are treated by adopting cell lysate, the cell lysate is transferred into a centrifuge tube, and the cell lysate is obtained after centrifugation is carried out for 10 minutes at the temperature of 10000r/min and the temperature of 4 ℃. Taking 20 mu L of cell lysate and detecting the total protein content in a sample by using a BCA kit; measurement of collagen content Experimental procedures were performed according to ELISA kit instructions, and each OD value was measured at 450nm, with standard curve equation y=0.4522x+0.1038, R 2 The expression amount x of the collagen is calculated according to a standard curve, the total protein content b is used for correction to obtain A=x/b, and the relative expression amount of the collagen I is calculated, wherein the relative expression amount of the collagen I is shown in the tableQuantity of arrival = a Experimental group/model group /A Blank group The results are shown in table 3 and fig. 6 (in fig. 6, ### p < 0.001, indicating an extremely significant decrease compared to the blank group; p > 0.05, ns indicates no significant increase compared to the model group; * p < 0.05, indicating an increase in comparison to the model group, p < 0.01, indicating a significant increase in comparison to the model group, p < 0.001, indicating an extremely significant increase in comparison to the model group.
TABLE 3 Table 3
Numbering device Relative expression level of type I collagen
Blank group 1
Model group 0.33±0.033 ###
Example 1 0.846±0.039 ***
Example 2 0.748±0.034 ***
Example 3 0.826±0.017 ***
Example 4 0.741±0.017 ***
Comparative example 1 0.436±0.029 *
Comparative example 2 0.391±0.025 ns
Comparative example 3 0.444±0.033 *
Comparative example 4 0.416±0.031 *
The results show that the products prepared in the examples 1-4 have ideal repairing effect on the fibroblasts of the damaged human skin, and can obviously improve the collagen content in the fibroblasts of the damaged human skin. The products prepared in comparative examples 1 to 4 were significantly inferior to the products prepared in examples 1 to 4, although they had a certain ability to repair skin fibroblasts of the damaged human.
Effect example 3 repair Capacity (cell viability)
1. The experimental steps are as follows:
the products prepared in the above examples and comparative examples were prepared with serum-free DMEM medium as test group test solutions having a volume percentage of 2%, respectively. The test solution needs to pass through a sterile filter membrane of 0.22 μm.
HaCaT cells were cultured in a culture medium containing 10% fetal bovine serum and 1% diabody (1X 10) 5 U/L penicillin, 100mg/L streptomycin) in DMEM medium. Cell growth at 37℃with 5% CO 2 When the cell fusion reached 85% or more in an incubator with saturated humidity, cells in the logarithmic growth phase were digested with 0.05% pancreatin, and the digestion reaction was terminated with DMEM containing serum. Cell counting plate counts, cell suspension concentration was adjusted to 7X 10 4 Each mL of the cell suspension was inoculated into a 96-well plate at a rate of 100. Mu.L per well, 37℃and 5% CO 2 Is incubated for 12h in an incubator. After incubation, the old broth is removed and the incubation is performed with phosphoric acidWashing cells twice with salt buffer solution, adding 100 μl of PBS solution, and irradiating the model group and experimental group with UVB at a dose of 40mJ/cm 2 The irradiation time was 80s, and the negative control group was not irradiated; absorbing and discarding PBS, adding serum-free DMEM culture medium into a model group and a negative control group, adding 100 mu L of the prepared filtered and sterilized experimental group to each hole in the experimental group, and making 6 compound holes for each liquid to be tested; the blank group was cell-free and 100. Mu.L of PBS was added. Then at 37 ℃ and 5% CO 2 Is incubated for 24h in an incubator. Then 10. Mu.L of CCK-8 solution was added to each well, incubated for 3 hours, absorbance was measured at a wavelength of 450nm, and the cell viability of each group was calculated, results are shown in Table 4 and FIG. 7 (in FIG. 7, ### p < 0.001, indicating an extremely significant decrease compared to the negative control group; p > 0.05, ns indicates no significant increase compared to the model group; * p < 0.05, indicating an increase in comparison to the model group, p < 0.01, indicating a significant increase in comparison to the model group, p < 0.001, indicating an extremely significant increase in comparison to the model group.
The cell viability was calculated as follows:
cell viability of experimental or model group= (a experimental or model group-a blank)/(a negative control group-a blank) ×100%.
TABLE 4 Table 4
Cell viability
Negative control group 100％
Model group 54.67±2.05％ ###
Example 1 73.33±2.87％ ***
Example 2 68.85±1.53％ ***
Example 3 70.12±1.8％ ***
Example 4 68.21±1.65％ ***
Comparative example 1 59.52±2.07％ *
Comparative example 2 57.84±0.99％ ns
Comparative example 3 60.62±2.76％ *
Comparative example 4 59.46±1.7％ *
The results show that the products prepared in the examples 1-4 have ideal repairing effect on damaged HaCaT cells, and the survival rate of the damaged HaCaT cells can be remarkably improved. The products prepared in comparative examples 1 to 4 were significantly inferior to those prepared in examples 1 to 4, although they had a certain ability to repair damaged HaCaT cells.
Effect example 4 photodamage resistance
The serum-free DMEM media for the products prepared in examples 1 to 4 and comparative examples 1 to 4 were prepared as experimental group test solutions having a volume percentage of 2%, and the test solutions were subjected to a 0.22 μm sterile filter.
HaCaT cells were cultured in a culture medium containing 10% fetal bovine serum and 1% diabody (1X 10) 5 U/L penicillin, 100mg/L streptomycin) in DMEM medium. Cell growth at 37℃with 5% CO 2 When the cell fusion reached 85% or more in an incubator with saturated humidity, cells in the logarithmic growth phase were digested with 0.05% pancreatin, and the digestion reaction was terminated with DMEM containing serum. Cell counting plate counts, cell suspension concentration was adjusted to 7X 10 4 Each mL of the cell suspension was inoculated into a 96-well plate at a rate of 100. Mu.L per well, 37℃and 5% CO 2 Is incubated for 12h in an incubator. Removing old culture solution after incubation, washing cells twice by using phosphate buffer solution, sucking and removing the washed PBS solution, adding serum-free DMEM culture medium into a model group and a negative control group, adding 100 mu L of the prepared filtered and sterilized experimental group to-be-detected liquid into each hole of the experimental group, and making 6 compound holes for each to-be-detected liquid; the blank group was cell-free and 100. Mu.L of PBS was added. Then at 37 ℃ and 5% CO 2 Is incubated for 24h in an incubator. After incubation, the model group and the experimental group were subjected to irradiation damage by UVB with an irradiation dose of 40mJ/cm 2 The irradiation time was 80s, and the negative control group was not irradiated; after the completion of the injury, 10. Mu.L of CCK-8 solution was added to each well, incubated for 3 hours, absorbance was measured at a wavelength of 450nm, and the cell viability of each group was calculated, and the results are shown in Table 5 and FIG. 8 (in FIG. 8, ### p < 0.001, indicating an extremely significant decrease compared to the negative control group; p > 0.05, ns indicates no significant increase compared to the model group; * p < 0.05, indicating an increase in comparison to the model group, p < 0.01, indicating a significant increase in comparison to the model group, p < 0.001, indicating an extremely significant increase in comparison to the model group.
The cell viability was calculated as follows:
cell viability in experimental or model group = (a experimental or model group-a blank)/(a negative control group-a blank) ×100%.
TABLE 5
The results show that the products prepared in examples 1-4 have ideal protection effects on HaCaT cells, can prevent the damage of ultraviolet rays to the HaCaT cells to a certain extent, and the protection effects are obviously better than those of the products prepared in comparative examples 1-4.
Effect example 5DPPH radical scavenging experiment
The products prepared in examples 1 to 4 and comparative examples 1 to 4 were subjected to DPPH radical scavenging test.
DPPH is an early synthetic organic radical, commonly used to evaluate the hydrogen donating ability of antioxidants, which is very stable in organic solvents, purple in color, and has a characteristic absorption peak at 517nm, when a radical scavenger is encountered, the lone pair of electrons of DPPH are paired to fade it, i.e., the absorbance at the maximum absorption wavelength becomes small. Therefore, the effect of the sample on DPPH radical scavenging can be evaluated by measuring the change in absorbance.
Liquid to be measured: the products obtained in the above examples or comparative examples.
The DPPH free radical scavenging experiment comprises the following specific experimental steps:
(1) Taking an equal volume (1 mL) of the liquid to be measured and 2X 10 -4 mixing the DPPH solution of mol/L (A) 1 A tube);
(2) Taking equal volume (1 mL) of absolute ethanol and 2X 10 -4 mixing the DPPH solution of mol/L (A) 2 A tube);
(3) Mixing the same volume (1 mL) of absolute ethanol with the liquid to be measured (A) 3 A tube);
(4) After reaction in the dark for 30min, A was measured at 517nm 1 Tube A 2 Tube A 3 Tube absorbance values; the clearance rate calculation formula is: clearance = [ (A) 2 +A 3 )-A 1 ]/A 2 X 100%, measureThe test results are shown in tables 6 and 9 (in fig. 9, p > 0.05, ns indicates no statistical difference from example 1; p < 0.05 indicates a statistical difference from example 1; p < 0.001 indicates a very significant statistical difference from example 1).
TABLE 6
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The results show that the products prepared in examples 1-4 have ideal antioxidant efficacy, and the antioxidant efficacy is significantly better than that of the products prepared in comparative examples 1-4.
Effect example 6 anti-inflammatory Capacity determination
The logarithmic phase HaCaT cells were seeded at a density of 25 ten thousand cells/mL on a 6-well cell culture plate, 2mL of cell suspension was added to each well, and cultured for 12 hours. Experimental, blank and model groups were set. Wherein, the experimental group and the model group adopt 40mJ/cm 2 Is irradiated to cells for 80s. After irradiation, the supernatant fluid in the experimental group, the blank group and the model group is sucked and removed, 2mL of DMEM solution is added into the experimental group, 2mL of the liquid to be tested is added into the experimental group (the liquid to be tested prepared in the example or the comparative example is prepared into the liquid to be tested with the volume percentage of 2% by adopting DMEM, the liquid to be tested needs to be subjected to a sterile filter membrane with the size of 0.22 μm before being used), the cells are subjected to post-treatment for 24 hours, the supernatant fluid is sucked and removed, the cells are washed for 2 to 3 times by PBS, then the cells are treated by adopting cell lysate, the cell lysate is transferred into a centrifuge tube, and the cell lysate is obtained after centrifugation is carried out for 10 minutes at the temperature of 10000r/min and the temperature of 4 ℃. Taking 20 mu L of cell lysate and detecting the total protein content in a sample by using a BCA kit; measurement of inflammatory factors the experimental procedure was followed by measurement of OD values at 450nm according to the ELISA kit instructions.
According to the standard curve (y=0.0018x+0.0652, r 2 = 0.9996) to calculate the expression level a of inflammatory factor using the protein contentB is corrected to obtain c=a/B, relative expression = C/C blank. The results are shown in table 7 and figure 10 (in figure 10, ### p < 0.001, indicating an extremely significant decrease compared to the blank group; * p < 0.05 indicates a decrease in comparison to the model group, p < 0.01 indicates a significant decrease in comparison to the model group, and p < 0.001 indicates an extremely significant decrease in comparison to the model group.
TABLE 7
/>
The results show that the products prepared in examples 1-4 can significantly reduce the inflammatory factor content in damaged HaCaT cells, and the anti-inflammatory effect is significantly better than that of the products prepared in comparative examples 1-4.
Finally, it is also noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
While the application has been disclosed by the foregoing description of specific embodiments thereof, it will be appreciated that those skilled in the art may devise various modifications, adaptations, or equivalents of the application within the spirit and scope of the appended claims. Such modifications, improvements, or equivalents are intended to be included within the scope of this application as claimed.

Claims (10)

1. A method for preparing a fermentation broth with emulsifying properties, comprising the steps of:
(1) Mixing 1 part of rice flour, 0.2-2 parts of nitrogen source and 20-150 parts of water to prepare a seed culture medium; inoculating lactobacillus into the seed culture medium, and fermenting and culturing to obtain seed solution A;
(2) Inoculating the seed liquid A prepared in the step (1) into a fermentation substrate, and performing fermentation culture and primary sterilization to prepare a fermentation liquid with emulsifying property; wherein the fermentation substrate comprises 1-5 parts of tremella powder and 20-150 parts of water.
2. The method for producing a fermentation broth having emulsifying properties according to claim 1, wherein the production method satisfies at least one of the following conditions:
in the step (1), the grain size of the rice powder is 50-150 meshes, preferably 50-100 meshes;
in the step (1), the nitrogen source comprises at least one of milk powder, soybean peptide and peptone;
in the step (1), the weight portion of the nitrogen source is 0.5 to 1 part, preferably 0.5 to 0.7 part;
in the step (1), the weight portion of the water is 50-100 portions;
in step (1), the seed medium further comprises a sterilization operation prior to use.
3. The method for producing a fermentation broth having emulsifying properties according to claim 1, wherein the production method satisfies at least one of the following conditions:
in step (1), the lactobacillus comprises lactobacillus helveticus (Lactobacillus helveticus) and/or lactobacillus plantarum (Lactiplantibacillus plantarum); preferably, the lactobacillus helveticus includes lactobacillus helveticus with a deposit number of CICC 20243 purchased from China center for type culture Collection of microorganisms; preferably, the lactobacillus plantarum comprises lactobacillus plantarum with the preservation number of CICC20261 purchased from China center for industry microbiological culture collection center;
in the step (1), the number of the lactobacillus inoculated in the seed culture medium per unit volume is 10 4 ～10 8 CFU/mL, preferably 10 5 ～10 6 CFU/mL；
In the step (1), the fermentation culture time is 12-24 hours, preferably 12-16 hours;
in step (1), the temperature of the fermentation culture is 30 to 45 ℃, preferably 37 to 45 ℃.
4. A method of preparing a fermentation broth having emulsifying properties according to any of claims 1 to 3, wherein the preparation method satisfies at least one of the following conditions:
in the step (2), the particle size of the tremella powder is 50-100 meshes;
in the step (2), the weight part of the tremella powder is 2-3.5 parts, preferably 3.3 parts;
in the step (2), the weight portion of the water is 50-100 portions;
in the step (2), the fermentation substrate further comprises a carbon source and/or a nitrogen source; preferably, the carbon source comprises at least one of glucose, corn and potato; preferably, the carbon source is 0.1 to 0.5 part by weight, more preferably 0.2 to 0.5 part by weight; preferably, the nitrogen source comprises at least one of milk powder, soybean peptide and black bean powder; preferably, the nitrogen source is 0.1 to 0.5 part by weight, more preferably 0.2 to 0.5 part by weight;
in the step (2), the fermentation culture time is 12-30 hours, preferably 12-20 hours;
in the step (2), the temperature of the fermentation culture is 30-45 ℃, preferably 35-40 ℃;
in the step (2), the operation of sterilizing once further comprises the operation of centrifuging and collecting supernatant; preferably, the centrifugation is followed by a secondary sterilization and/or mixing with a preservative.
5. A fermentation broth having emulsifying properties, which is obtained by the process for producing a fermentation broth having emulsifying properties according to any one of claims 1 to 4.
6. Use of the fermentation broth having emulsifying properties according to claim 5 for the preparation of an external preparation for skin;
preferably, the fermentation broth with emulsifying property is also used as at least one of photodamage-resistant active ingredient, skin repair active ingredient, oxidation-resistant active ingredient and anti-inflammatory active ingredient in the skin external agent.
7. A skin external preparation comprising a material a and a vegetable oil, wherein the material a comprises the fermentation broth with emulsifying properties according to claim 5; the viscosity of the material A is 6000-8000 mPa.S.
8. The external skin preparation according to claim 7, wherein the external skin preparation satisfies at least one of the following conditions:
the material A also comprises water;
the vegetable oil comprises at least one of peony seed oil, prinsepia utilis royle oil, macadamia nut seed oil and white pool flower seed oil;
the skin external preparation further comprises a cosmetic active ingredient, preferably at least one of a moisturizing active ingredient, a whitening active ingredient, an anti-inflammatory active ingredient, an anti-allergic active ingredient and an anti-oxidation active ingredient;
the vegetable oil accounts for 6-12% of the mass of the material A, and is preferably 8-10%.
9. A method for producing the external preparation for skin according to claim 7 or 8, comprising the steps of: mixing the material A with the vegetable oil, and shearing and homogenizing.
10. The method for producing a skin external preparation according to claim 9, wherein the production method satisfies at least one of the following conditions:
when the skin external agent further comprises a cosmetically active ingredient, the cosmetically active ingredient is added during the mixing;
the shearing homogenizing speed is 10000-20000 r/min, preferably 14000-16000 r/min;
the time for shearing and homogenizing is 3-10 min, preferably 5-8 min.
CN202310735996.XA 2023-06-20 2023-06-20 Fermentation liquor with emulsifying property, product containing fermentation liquor and preparation method and application of fermentation liquor Pending CN116602907A (en)

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