CN114668699A

CN114668699A - Preparation method of honeysuckle fermented raw pulp and application of honeysuckle fermented raw pulp in cosmetics

Info

Publication number: CN114668699A
Application number: CN202210568444.XA
Authority: CN
Inventors: 刘青; 周娟; 李西林
Original assignee: Calais Jinan Life Technology Co ltd
Current assignee: Calais Jinan Life Technology Co ltd
Priority date: 2022-05-24
Filing date: 2022-05-24
Publication date: 2022-06-28

Abstract

The invention belongs to the technical field of biological fermentation, and particularly relates to a preparation method of honeysuckle fermented protoplasm and application of the honeysuckle fermented protoplasm in cosmetics, wherein the preparation method comprises the following steps: mixing honeysuckle powder and deionized water in proportion, carrying out high-pressure sterilization treatment to obtain a honeysuckle fermentation initial system, then mixing the honeysuckle fermentation initial system with zymophyte liquid for fermentation, and carrying out high-pressure sterilization and centrifugation to obtain fermentation protoplasm. According to the invention, the honeysuckle is fermented by using the bacillus subtilis, and the prepared honeysuckle fermentation protoplasm has active ingredients for removing DPPH free radicals, also has active ingredients for inhibiting tyrosinase activity and melanin synthesis, and shows better antioxidation and whitening effects.

Description

Preparation method of honeysuckle fermented protoplasm and application of honeysuckle fermented protoplasm in cosmetics

Technical Field

The invention belongs to the technical field of biological fermentation, and particularly relates to a preparation method of honeysuckle fermented raw pulp and application of the honeysuckle fermented raw pulp in cosmetics.

Background

The flos Lonicerae is dried bud or flower with early blossom of Lonicera japonica Thunb of Caprifoliaceae (Caprifoliaceae) and plants of the same genus. The dried flowers are sweet and slightly bitter in taste, slightly cold in nature and nontoxic, and have the effects of promoting the production of body fluid to quench thirst, clearing away heat and toxic materials, resisting bacteria and diminishing inflammation and the like. Researches in the prior art find that the antibacterial agent has an obvious antibacterial effect, the gram-positive bacteria inhibition rate can reach 53.86%, and the gram-negative bacteria inhibition rate can reach 36.14%. In the prior art, honeysuckle is also found to have good antibacterial effect on micrococcus luteus. The flavonoid compounds identified in honeysuckle at present mainly comprise luteolin, loniceraside, luteolin-7-O-D-glucoside, luteolin-7-0-D-galactoside, quercetin-3-0-D-glucoside, hyperin, 5-hydroxy-3, 4, 7-trimethyl flavone and the like. More than 40 organic acid components in honeysuckle have been separated so far, and the organic acid components are key components of honeysuckle for clearing heat and removing toxicity and mainly comprise chlorogenic acid. In addition, there are some triterpenoids, including Lonicera macranthoides saponin A, Lonicera macranthoides saponin B and Akebia quinata saponin D. The volatile oil components include linalool, methyl linoleate, benzyl alcohol, eugenol and other alcohol ester substances.

The traditional extraction process comprises alcohol extraction, water extraction, ultrasonic extraction, microwave extraction and the like, has the limitations of high energy consumption, low efficiency, low purity, introduction of impurities and the like, and causes the functional components of plants to be incompletely extracted and utilized. By using the microbial fermentation process, active ingredients in the plant raw materials can be effectively reserved, the enrichment effect is achieved, and the functional ingredients of the raw materials are better utilized. The microorganism contains certain enzymes, and can degrade cellulose, hemicellulose and other substances in cell walls and intercellular substances of plant raw materials, so that the plant cells are broken, the intercellular spaces are increased, the mass transfer resistance of effective components diffused from the intracellular to the extraction medium is reduced, and the extraction rate of the effective components can be greatly improved. Meanwhile, microorganisms can metabolize to generate new active ingredients by virtue of strong substance conversion capacity and metabolic generation capacity, so that the effect of extracting ingredients from traditional plants is enhanced. Fermentation liquor obtained by fermenting plant raw materials through microorganisms can be directly applied as cosmetics, is a great innovation in the technical development of the cosmetics, and accords with the natural and green development direction of modern cosmetics.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides a preparation method of honeysuckle flower fermentation raw stock and application of the honeysuckle flower fermentation raw stock in cosmetics. Meanwhile, the honeysuckle fermented raw juice prepared by the invention has active ingredients for inhibiting tyrosinase activity and melanin synthesis, and shows a good whitening effect.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for preparing fermented juice from flos Lonicerae comprises mixing flos Lonicerae powder and deionized water at a certain proportion, autoclaving to obtain flos Lonicerae fermentation initial system, mixing with zymophyte liquid, fermenting, autoclaving, and centrifuging to obtain the fermented juice.

Furthermore, the honeysuckle powder is prepared by drying honeysuckle for 4-6 hours at 50-70 ℃, crushing and sieving with a 50-100 mesh sieve.

Further, the autoclaving condition is at 121 ℃ for 20-30 minutes at 110-.

Further, the honeysuckle flower powder in the honeysuckle flower fermentation initial system: the proportion of the deionized water is 5-20 g: 100 g.

Further, the zymocyte is Bacillus subtilis, and the preservation information of the strain is as follows: china center for Industrial microbial culture Collection (CICC), the storage unit address: no. 6 building of 24 th hospital of the morning bridge of yangxin district of beijing city, preservation date: no. 11/2008, accession No. 17: cic 23659, category name: the bacillus subtilis subspecies Ski, which is the existing strain in the prior art.

Further, the zymophyte is added into a honeysuckle fermentation initial system in the form of a bacterium liquid, and the concentration of the inoculated bacterium is 10⁵-10⁸CFU/mL。

Further, the proportion relation between the zymophyte liquid and the honeysuckle initial fermentation system is 5-15 mL: 100 g.

Further, in the fermentation step, the temperature is 30-45 ℃, the fermentation time is 40-70 hours, and the rotation speed is 100-200 r/min.

Further, in the centrifugation step, the rotating speed is 6000-12000r/min, the centrifugation radius is 6-12cm, and the centrifugation time is 10-40 minutes.

Further, the pH value of the honeysuckle fermented puree is 4.50-6.50.

Further, the zymophyte is pretreated in advance, and the pretreatment process comprises the following steps:

activating strains: inoculating single colony of zymocyte into nutrient liquid culture medium, and culturing in 30-45 deg.C incubator for 10-30 hr;

and (3) strain purification: streaking and inoculating the activated strain on a nutrient agar plate to obtain a single colony;

expanding culture of strains: inoculating single colony of strain to be used to nutrient liquid culture medium, culturing at 30-45 deg.C in incubator until OD value reaches 0.5-1.0, to obtain suitable inoculation concentration of 10⁵-10⁸CFU/mL。

The honeysuckle fermented raw juice provided by the invention has the effects of oxidation resistance and whitening.

The invention also aims to provide application of the honeysuckle fermented raw juice in cosmetics, wherein the cosmetics comprise the honeysuckle fermented raw juice.

Advantageous effects

The invention discloses a preparation method of honeysuckle fermented protoplasm and application thereof in cosmetics, and the invention at least has the following advantages:

according to the invention, the bacillus subtilis is adopted to ferment the honeysuckle fermentation initial system, so that the functional components and the activity of the honeysuckle in the plants are retained, and the phenomenon of loss of the active components caused by the traditional extraction method is avoided.

The honeysuckle fermentation raw pulp provided by the invention has the effect of removing DPPH free radicals and shows good antioxidation.

The honeysuckle fermentation raw stock provided by the invention is rich in active substances which have the effects of inhibiting the activity of tyrosinase and inhibiting the synthesis of melanin, thereby having good whitening effect on skin.

After the fermentation is finished in the method, chemical components such as essence and the like are not added into the fermentation raw stock, so that the safety of the product to a human body is ensured, and no adverse reaction or side effect is caused to skin in a patch test.

Drawings

FIG. 1: a standard curve graph of the removal rate of Trolox with different concentrations to DPPH;

FIG. 2: and (3) a standard curve graph of inhibition rates of different concentrations of kojic acid on tyrosinase.

Detailed Description

Hereinafter, the present invention will be described in detail. Before the description is made, it should be understood that the terms used in the present specification and the appended claims should not be construed as limited to general and dictionary meanings, but interpreted based on the meanings and concepts corresponding to technical aspects of the present invention on the basis of the principle that the inventor is allowed to define terms appropriately for the best explanation. Accordingly, the description proposed herein is just a preferable example for the purpose of illustrations only, not intended to limit the scope of the invention, so it should be understood that other equivalents and modifications could be made thereto without departing from the spirit and scope of the invention.

The following examples are given by way of illustration of embodiments of the invention and are not to be construed as limiting the invention, and it will be understood by those skilled in the art that modifications may be made without departing from the spirit and scope of the invention. Unless otherwise specified, reagents and equipment used in the following examples are commercially available products.

Example 1

A preparation method of honeysuckle fermented protoplasm comprises the following steps:

1. the zymophyte is firstly pretreated, and the pretreatment process comprises the following steps:

(1) activating strains: inoculating single colony of zymocyte into nutrient liquid culture medium, and culturing in 37 deg.C incubator for 16 hr; the medium used conforms to the industry standard YY/T1187-2010.

(2) And (3) strain purification: streaking and inoculating the activated strain on a nutrient agar plate to obtain a single colony; the medium used conforms to the industry standard YY/T1239-2010.

(3) Expanding culture of strains: inoculating single colony of strain to be used into nutrient liquid culture medium, culturing at 37 deg.C in incubator until OD value reaches 0.5-1.0, to obtain suitable inoculation concentration of 10⁵-10⁸CFU/mL. The medium used conforms to the industry standard YY/T1187-2010.

2. Drying honeysuckle for 5 hours at 65 ℃, crushing and sieving with a 100-mesh sieve to obtain honeysuckle powder; honeysuckle powder and deionized water are added according to the weight ratio of 5 g: mixing 100g of the raw materials uniformly, and performing high-pressure sterilization treatment at 121 ℃ for 30min to obtain the initial honeysuckle fermentation system.

3. Fermenting bacteria (concentration is 10) after purification and amplification culture⁵-10⁸CFU/m 1) bacterial liquid 5mL is inoculated into honeysuckle flower fermentation initial system 100g, after culturing for 50 hours in an incubator at 37 ℃, the obtained fermentation liquid is sterilized under high pressure at 121 ℃ for 30min to inactivate the bacteria; filtering residues of the sterilized fermentation liquor by using 8 layers of filter cloth, centrifuging the filtrate for 30min under the conditions of 10000r/min and the centrifugal radius of 9cm, and collecting supernatant fluid, namely the honeysuckle fermentation raw stock provided by the invention.

The honeysuckle fermented puree prepared in the example is viscous liquid in appearance and light yellow to brown in color. The pH value is 4.50-6.50, the viscosity is 50-400cP, the content of soluble solid is 1.0-5.0%, the total number of colonies is less than 50CFU/mL, no pathogenic bacteria are detected, and the requirements related to the management of cosmetic quality tubes are met.

Example 2

(1) activating strains: inoculating a single colony of zymophyte into a nutrient liquid culture medium, and culturing for 30 hours in an incubator at the temperature of 30 ℃; the medium used conforms to the industry standard YY/T1187-2010.

(3) Expanding culture of strains: inoculating single colony of strain to be used into nutrient liquid culture medium, culturing in 30 deg.C incubator until OD value reaches 0.5-1.0, to obtain suitable inoculation concentration of 10⁵-10⁸CFU/mL. The medium used conforms to the industry standard YY/T1187-2010.

2. Drying honeysuckle flower for 6 hours at 50 ℃, crushing and sieving with a 100-mesh sieve to obtain honeysuckle flower powder; honeysuckle powder and deionized water are mixed according to the weight ratio of 15 g: mixing 100g of the raw materials uniformly, and performing autoclaving treatment at 115 deg.C for 25min to obtain initial system of flos Lonicerae fermentation.

3. The fermentation bacteria (with the concentration of 10) after purification and propagation culture are treated⁵-10⁸CFU/m 1) bacterial liquid 10mL is inoculated into a honeysuckle fermentation initial system 100g, after culturing for 70 hours in an incubator at 30 ℃, the obtained fermentation liquid is sterilized under high pressure at 115 ℃ for 25min to inactivate the bacteria; filtering residues of the sterilized fermentation liquor by using 8 layers of filter cloth, centrifuging the filtrate for 40min under the conditions of 12000r/min and 12cm of centrifugal radius, and collecting supernatant fluid, namely the honeysuckle flower fermentation raw stock provided by the invention.

Example 3

(1) activating strains: inoculating single colony of zymocyte into nutrient liquid culture medium, and culturing in 45 deg.C incubator for 10 hr; the medium used conforms to the industry standard YY/T1187-2010.

(3) Expanding culture of strains: inoculating single colony of strain to be used into nutrient liquid culture medium, culturing in 45 deg.C incubator until OD value reaches 0.5-1.0, to obtain suitable inoculation concentration of 10⁵-10⁸CFU/mL. The culture medium usedMeets the industry standard YY/T1187-2010.

2. Drying honeysuckle for 4 hours at 70 ℃, crushing and sieving with a 50-mesh sieve to obtain honeysuckle powder; honeysuckle powder and deionized water are mixed according to the weight ratio of 20 g: mixing 100g of the raw materials uniformly, and performing high-pressure sterilization treatment at 110 ℃ for 20min to obtain the initial honeysuckle fermentation system.

3. The fermentation bacteria (with the concentration of 10) after purification and propagation culture are treated⁵-10⁸CFU/m 1) bacterial liquid 15mL is inoculated into a honeysuckle fermentation initial system 100g, after culture is carried out in an incubator at 45 ℃ for 40 hours, the obtained fermentation liquid is sterilized under high pressure at 110 ℃ for 20min, so that the bacteria are inactivated; filtering residues of the sterilized fermentation liquor by using 8 layers of filter cloth, centrifuging the filtrate for 10min under the conditions of 6000r/min and the centrifugal radius of 6cm, and collecting supernatant fluid, namely the honeysuckle flower fermentation raw stock provided by the invention.

Example 4

Antioxidant efficacy test of Lonicera japonica fermentation puree obtained in example 1

1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH for short) is a stable long-life free radical, and the ethanol solution of the free radical is dark purple and has strong absorption near 517 nm. In the presence of free radical scavengers, the light absorption of the DPPH ethanol solution is reduced due to its one-electron pairing. The degree of discoloration of the DPPH ethanol solution is linear with the number of electrons it receives, and thus the ability of the test sample to scavenge free radicals, i.e., the magnitude of antioxidant activity, can be evaluated.

Referring to Table 1, a 10mL test tube was used to set up the sample tube (T), sample background (T)₀) DPPH tube (C) and solvent background (C)₀) For each sample concentration to be tested, 3 parallel tubes are required for each sample tube (T), and 3 parallel tubes are required for DPPH tube (C).

TABLE 1 DPPH radical scavenging test sample test liquid adding meter

The test procedure was as follows:

(1) in the sample tube (T) and sample background（T₀) 1mL of the same concentration of each sample solution was added. And (4) diluting the test sample of the honeysuckle fermentation raw stock by 40 times, and then testing.

(2) All tubes (T, T0, C, C0) were supplemented with deionized water to make up 3mL and mixed well.

(3) 1mL of DPPH ethanol solution was added to the sample tubes (T) and DPPH tube (C), the sample background (T)₀) And solvent background (C)₀) Replace with 95% ethanol, shake gently, and stand at room temperature for 5 minutes.

(4) Each reaction solution was transferred to a 1cm cuvette and absorbance was measured at 517 nm.

(5) Positive control: the standard Trolox (water-soluble vitamin E) is dissolved to 0.010mg/mL, 0.015mg/mL, 0.020mg/mL, 0.025mg/mL and 0.030mg/mL by using a 95% ethanol solution.

(6) Calculating DPPH free radical clearance rate:

TABLE 2 results of DPPH clearance of Lonicera japonica fermentation puree

As can be seen from Table 2, the DPPH radical scavenging effect of the honeysuckle fermentation raw stock is obvious, and is improved by about 7% compared with the fermentation initial system. The fermentation raw stock with the concentration of 2.5% can remove 57.49% of free radicals, the removal effect of Trolox is better than that of 0.02mg/mL, and the strong oxidation resistance is shown. According to a standard curve of the concentration and the inhibition rate of Trolox and a regression formula (figure 1), the equivalent of Trolox in the honeysuckle fermentation protoplasm is 1.084 mg/mL.

Example 5

Whitening efficacy test of honeysuckle fermentation puree obtained in example 1

Tyrosinase is a key enzyme in skin melanin biosynthesis, acting on dopa to form dopaquinone, which spontaneously undergoes a series of reactions to finally form melanin. Tyrosinase catalyzes the conversion of dopa to dopaquinone in a phosphoric acid solution with a pH of 6.8, and the absorbance can be measured at 475nm of a spectrophotometer. The raw material with tyrosinase activity inhibition effect can reduce the conversion of dopa into dopaquinone, thereby reducing the light absorption value, and the inhibition effect of the raw material on tyrosinase activity is evaluated according to the change of the light absorption value.

Referring to Table 3, a 10mL test tube was used to set up the sample tube (T), sample background (T)₀) Enzyme reaction tube (C) and solvent background (C)₀) 3 parallel tubes are required to be set up for each sample tube (T) of each concentration to be tested for each sample, and 3 parallel tubes are required to be set up for the enzyme reaction tube (C).

TABLE 3 tyrosinase inhibition test sample test liquid adding meter

The test procedure was as follows:

(1) in the sample tube (T) and sample background (T)₀) 1mL of the same concentration of each of the sample solutions, an enzyme reaction tube (C) and a solvent background (C)₀) 1mL of disodium hydrogen phosphate-citric acid buffer (pH6.8) was added.

(2) Adding 0.5mL of tyrosinase solution into each of the sample tube (T) and the enzyme reaction tube (C), and obtaining a sample background (T)₀) Replacing with 0.5mL of disodium hydrogen phosphate-citric acid buffer solution with pH6.8, mixing the sample and tyrosinase thoroughly, and incubating in a 37 deg.C water bath for 10 min.

(3) And sequentially adding 2mL of levodopa solution into each tube, controlling the reaction time of each tube to be 5 minutes, immediately transferring each tube of reaction solution into a cuvette, and measuring the light absorption value at 475 nm.

(4) Positive control: the pH of the buffer solution was adjusted to 100. mu.g/mL, 75. mu.g/mL, 50. mu.g/mL, 25. mu.g/mL, 12.5. mu.g/mL, 6.25. mu.g/mL using disodium hydrogen phosphate-citric acid at pH 6.8.

(5) Calculation of tyrosinase inhibition:

TABLE 4 tyrosinase inhibition rate results of fermented honeysuckle pulp

From table 4, it can be seen that the inhibition effect of the honeysuckle fermented raw pulp on tyrosinase is obvious, and compared with the fermentation initial system, the inhibition effect is improved by about 10%. The fermented primary pulp can inhibit 46.37% of tyrosinase, has the same effect as 12.5 mu g/mL of kojic acid, and shows better whitening effect. According to the standard curve of the concentration and the inhibition rate of the kojic acid and a regression formula (figure 2), the equivalent of the kojic acid in the honeysuckle fermentation raw pulp is 13.52 mu g/mL.

Example 6

Safety test of honeysuckle fermentation puree obtained in example 1

The human body patch test is mainly used for detecting the irritation of the final cosmetic product or raw materials. The honeysuckle flower fermentation protoplasm obtained in the embodiment 1 is subjected to a human body closed patch test, and the potential skin irritation of the honeysuckle flower fermentation protoplasm is evaluated.

1. Test object

30 persons were selected for the trial according to the criteria for inclusion of the subjects as specified in the technical Specification for cosmetic safety 2015.

2. Test method

The selected area is not more than 50mm²And qualified spot test equipment with the depth of about 1 mm. And (3) test treatment: putting 0.025mL of the honeysuckle fermented raw stock into a chamber of a spot tester, applying the spot tester with the test object to the curved side of the forearm of the subject by using a hypoallergenic adhesive tape, and lightly pressing with the palm to uniformly apply the spot tester to the skin for 24 hours. Blank control treatment is set for each test, no substance is placed in the control spot tester hole, and other processes are the same as those of the test group.

The skin reactions were observed according to the standard of table 1 30min (after disappearance of the indentation), 24h and 48h after removal of the test article plaque test device, respectively, and the observation results were recorded.

Skin closed patch test skin reaction degree grading criteria are as follows:

"-" = negative reaction;

"±" = suspicious reaction, only weak erythema;

"+" = weak positive reaction (erythema reaction); erythema, infiltration, edema, and possibly papules;

"++" = strong positive reaction (herpetic response); erythema, infiltration, edema, pimples, herpes; inverse direction

Should be able to exceed the test area;

"+ + + +" = very strong positive reaction (fusogenic herpes response); obvious erythema, severe infiltration, edema,

Herpes fusiformis; the reaction goes beyond the test area.

The judgment standard of the human body patch test is as follows: the number of plus or minus adverse reactions in 30 subjects is more than 5, or the number of plus or minus adverse reactions is more than 2, or any 1 plus or minus skin adverse reaction is judged to have adverse reactions to human body, otherwise, the subjects are judged to have no adverse reactions to human body.

TABLE 5 Patch test results of fermented Lonicera Japonica flos pulp obtained in example 1

As can be seen from table 5: the honeysuckle fermentation raw pulp obtained in the embodiment 1 has no adverse reaction, which shows that the honeysuckle fermentation raw pulp provided by the invention has safety and does not bring adverse reaction to human body.

Example 7

Application of honeysuckle fermented protoplasm in cosmetics (skin lotion)

The honeysuckle fermented raw juice skin-moistening emulsion comprises the following components in parts by weight:

phase A: 2 parts of simethicone, 3 parts of cetyl alcohol, 1 part of behenyl alcohol, 3 parts of caprylic/capric triglyceride, 2 parts of glycerol stearic acid and 202 parts of behenyl polyether;

phase B: 75 parts of deionized water, 2 parts of 1, 3-butanediol, 4 parts of glycerol, 0.1 part of xanthan gum, 100.05 parts of carbomer U, 0.05 part of disodium EDTA and 5 parts of honeysuckle fermented protoplasm obtained in the embodiment 1;

and C phase: 0.3 part of pentanediol, 0.5 part of p-hydroxyacetophenone and 0.2 part of essence.

The processing technology is as follows:

adding the raw materials of the component A into a container A respectively, heating to 80-85 ℃, and homogenizing and stirring until no insoluble substances exist in the system;

adding the components of the phase B into a container B, heating to 90 ℃, and homogenizing and stirring until no insoluble substances exist in the system;

adding the mixture of the phase A into a container B, and homogenizing for 5 minutes;

after homogenizing, stirring and cooling to 45 ℃, and adding each component of the phase C;

cooling to room temperature, and filling to obtain the product.

Example 8

Application of honeysuckle fermented protoplasm in cosmetics (skin lotion)

phase A: 3 parts of simethicone, 2 parts of cetyl alcohol, 2 parts of behenyl alcohol, 3 parts of caprylic/capric triglyceride, 2 parts of glycerol stearic acid and 201 parts of behenyl polyether;

phase B: 65 parts of deionized water, 3 parts of 1, 3-butanediol, 3 parts of glycerol, 0.1 part of xanthan gum, 100.05 parts of carbomer U, 0.05 part of disodium EDTA and 15 parts of honeysuckle fermented protoplasm obtained in the embodiment 2;

The processing technology is as follows:

cooling to room temperature, and filling to obtain the product.

Example 9

Application of honeysuckle fermented protoplasm in cosmetics (skin lotion)

phase A: 3 parts of simethicone, 3 parts of cetyl alcohol, 2 parts of behenyl alcohol, 2 parts of caprylic/capric triglyceride, 1 part of glycerol stearic acid and 202 parts of behenyl polyether;

phase B: 55 parts of deionized water, 2 parts of 1, 3-butanediol, 4 parts of glycerol, 0.1 part of xanthan gum, 100.05 parts of carbomer U, 0.05 part of disodium EDTA and 25 parts of honeysuckle fermented raw pulp obtained in example 3;

The processing technology comprises the following steps:

cooling to room temperature, and filling to obtain the product.

The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.

Claims

1. A preparation method of honeysuckle fermented puree is characterized by comprising the following steps:

(1) mixing honeysuckle powder and deionized water in proportion, and then carrying out high-pressure sterilization treatment to obtain a honeysuckle fermentation initial system;

(2) mixing the honeysuckle fermentation initial system obtained in the step (1) with a zymophyte liquid, and fermenting;

(3) after fermentation, sterilizing under high pressure, filtering, centrifuging the filtrate, and collecting the supernatant to obtain the fermented raw stock.

2. The method for preparing fermented raw juice of honeysuckle flowers according to claim 1, wherein in the step (1), the honeysuckle flower powder is prepared by drying honeysuckle flowers for 4-6 hours at 50-70 ℃, crushing and sieving with a 50-100 mesh sieve.

3. The method for preparing fermented raw juice of honeysuckle flowers according to claim 1, wherein the autoclaving conditions in step (1) are at 121 ℃ for 20-30 minutes.

4. The method for preparing fermented puree of honeysuckle flower according to claim 1, wherein in the step (2), the honeysuckle flower powder in the initial system of honeysuckle flower fermentation: the proportion of the deionized water is 5-20 g: 100 g.

5. The method for preparing fermented raw juice of honeysuckle flowers according to claim 1, wherein in the step (2), the zymocyte is bacillus subtilis, and the preservation information of the strain is as follows: china Industrial microorganism culture preservation management center, preservation unit address: no. 6 building of 24 th hospital of the morning bridge of yangxin district of beijing city, preservation date: no. 11/17 in 2008, accession No.: cic 23659, classification name: bacillus subtilis subspecies Spreniae.

6. The method for preparing fermented honeysuckle puree according to claim 1, wherein in the step (2), the zymophyte is added to a honeysuckle fermentation initial system in a bacterial liquid form, and the concentration of an inoculation bacterium is 10⁵-10⁸CFU/mL。

7. The method for preparing fermented raw juice of honeysuckle flowers according to claim 6, wherein in the step (2), the ratio of the zymophyte liquid to the initial system of honeysuckle flower fermentation is 5-15 mL: 100g of the total weight of the feed; in the fermentation step, the temperature is 30-45 ℃, the fermentation time is 40-70 hours, and the rotation speed is 100-.

8. The method for preparing the primary fermented pulp of honeysuckle according to claim 1, wherein in the step (3), the rotation speed is 6000-12000r/min, the centrifugation radius is 6-12cm, and the centrifugation time is 10-40 minutes; the pH value of the honeysuckle fermented protoplasm is 4.50-6.50.

9. The method for preparing honeysuckle fermented puree according to claim 1, wherein in the step (2), the zymophyte is pretreated in advance, and the pretreatment process comprises the following steps:

activating strains: inoculating single colony of zymophyte into nutrient liquid culture medium, and culturing in 30-45 deg.C incubator for 10-30 hr;

10. An application of honeysuckle fermented protoplasm in cosmetics is characterized in that the cosmetics comprise the honeysuckle fermented protoplasm, and the honeysuckle fermented protoplasm is prepared according to the preparation method of any one of claims 1 to 9.