TW202114634A - Pearl preparation and preparation method therefor and application thereof - Google Patents

Abstract

Provided by the present invention are a pearl preparation and a preparation method therefor and an application thereof. The pearl preparation is prepared by means of the following method: 1) crushing a pearl using a jet mill; 2) mixing the crushed pearl powder with deionized water to obtain a mixed solution; 3) dissolving the mixed solution using weak acid; 4) putting the solution obtained in step 3) to microbial expansion culture broth for fermentation culture; and 5) freeze-drying the fermentation broth obtained in step 4) to obtain a freeze-dried pearl preparation powder. The pearl preparation has a very significant anti-aging ability, and acts on mitochondria by means of a PI3K/AKT pathway to play an anti-aging effect.

Description

Pearl preparation and its preparation method and use

This application relates to a pearl preparation, its preparation method, and its use.

Aging is a special law of nature, and anti-aging is also a technical problem that mankind has always wanted to overcome. Especially in cosmetics and health products, products with anti-aging functions are very popular among consumers. Many scientific research institutions at home and abroad have carried out comprehensive and in-depth research on the mechanism of aging. Combined with the current development technology, anti-aging mechanisms can be roughly divided into seven categories: regulation of extracellular mechanisms, skin whitening function, improvement of stratum corneum barrier function, promotion of cell proliferation, resistance to apoptosis, anti-inflammatory and anti-oxidant effects. These seven mechanisms interact with each other with different focuses. At present, in studying the mechanism of anti-aging active substances, one or more of them are selected to prove that they have anti-aging effects.

Pearl, or pearl, is a hard, smooth product produced by mollusks (mainly oysters). In the "Compendium of Materia Medica", it is especially written: pearls are sweet, salty, non-toxic in nature, calming the heart; the surface of pearls is moisturized and good color. Apply hands and feet to remove skin and diarrhea; drop phlegm, remove facial spots, and stop diarrhea; remove children’s convulsions and calm souls; stop seminal turbidity, relieve acne, treat poison, and make gloss and white. In view of the above functions of pearls, it is necessary to develop a method that can effectively extract and utilize the anti-aging ingredients in pearls.

Technical purpose

The technical purpose of the present invention is to provide a method that can effectively extract and utilize the anti-aging ingredients in pearls, and provide pearl anti-aging products prepared therefrom, so as to be used in skin anti-aging and anti-inflammatory products.

Technical solutions

In one aspect, the present invention provides a method for preparing pearl preparations, which includes the following steps: 1) Crushing: crush the pearls with a jet mill to obtain ultra-fine pearl powder with a particle size of less than 6 microns, preferably 0.3-1 microns; 2) Mix with water: mix pearl powder with deionized water to obtain a mixed solution; 3) Weak acid dissolution: pass excess carbon dioxide into the bottom of the mixed solution, or add a weak acid solution to dissolve the pearl powder in the mixed solution; 4) Fermentation: After sterilizing the solution obtained in step 3), connect to the microbial expansion broth for fermentation culture; 5) Freeze-drying: freeze-dry the fermentation broth obtained in step 4) to obtain freeze-dried powder of pearl preparations.

According to a specific embodiment of the present invention, before step 1), the method may further include the steps of washing, refining, and spray drying the pearls.

According to a specific embodiment of the present invention, in step 2), the mixing weight ratio of pearl powder and deionized water can be 1:5-20, and the mixing conditions are: 30-70℃ through internal stirrer or stirring blade at 50~ Stir and mix at a speed of 100 rpm.

According to a specific embodiment of the present invention, in step 3), if a weak acid solution is added, the volume ratio of the weak acid solution and the mixed solution is 1:4-10, preferably 1:5, and the weak acid solution is selected from acetic acid, lemon One or more of acid, lactic acid, malic acid, etc.; preferably lactic acid, acetic acid or citric acid.

According to a specific embodiment of the present invention, in step 4), the added microbial expansion culture solution accounts for 1-10% by volume of the solution obtained in step 3), and the microorganisms used are selected from the group consisting of Saccharomyces cerevisiae, grape juice yeast, lactic acid bacteria, and subtilis One or more of Bacillus, Bacillus licheniformis, etc.; preferably, Saccharomyces cerevisiae, Lactobacillus rhamnosus, Bacillus subtilis or Bacillus licheniformis; fermentation culture conditions can be: 25~38℃ in the fermentation tank, pH 7.0 ~7.5, 300~500 r/min, ventilation 3-5L/min and culture for 24~72 hours.

According to a specific embodiment of the present invention, in step 5), after stirring the fermentation broth at 85~100°C for 20~50 minutes for inactivation, the sterile filtrate is obtained by pressure filtration through a 0.22μm filter membrane, and sterile filtered The liquid and the freeze-dried protective agent are mixed at a volume/weight ratio of 100:0.5~2. In the above volume/weight ratio, the volume unit is ml and the weight unit is grams, using liquid nitrogen or at -70~-85℃ Pre-freeze for 3-10 hours, then put it in a vacuum freeze-drying bin and freeze-dry for 24 to 72 hours. Finally, freeze-dried powder with a moisture content of 2 to 5 wt% is harvested.

According to a specific embodiment of the present invention, after step 5), the method may further include the step of placing the lyophilized powder in a brown vial or a vacuum-packed plastic bag for storage, preferably between 1~ Store at 10°C.

In another aspect, the present invention provides a pearl preparation prepared by the above preparation method.

In another aspect, the present invention provides a composition comprising at least the aforementioned pearl preparation.

In another aspect, the present invention provides a use of the above-mentioned pearl preparation or the above-mentioned composition as an anti-aging product.

According to a specific embodiment of the present invention, the anti-aging product is a health care product or a cosmetic.

According to a specific embodiment of the present invention, the anti-aging product does not contain preservatives.

Beneficial effect

The method of the present invention has the following advantages:

1. After jet pulverization, the pseudo-agglomeration of pearl powder can be broken up, and then further broken to obtain a small and concentrated small particle size powder with a small particle size distribution range.

2. The acidic solution components in pearls are extracted gently and non-destructively through weak acid dissolution;

3. Retaining part of the calcium in pearls is conducive to improving skin calcium ion channels, and is conducive to pearl anti-aging peptides to cross the skin barrier and act on inflammatory cells and fibroblasts in the basal layer of the skin;

4. It is degraded by microbial decomposition to obtain small molecular peptides, thereby obtaining a richer active mixture under the action of microorganisms. At the same time, because a variety of microbial metabolites contain bacteriostatic peptides, it can ensure that pearl preparations do not need to be added in the later stage preservative;

5. The pearl preparation can effectively inhibit the expression of inflammatory factors bcl-2, caspase-3 and caspase-9, inhibit the production of reactive oxygen species and malondialdehyde, and simultaneously promote the production of glutathione and superoxide dismutase produce. Through the mitochondrial anti-aging pathway, it can effectively repair senescent cells and prolong cell activity. It has great application prospects in anti-aging cosmetics and health products.

Experimental materials: Pearls were purchased from Zhejiang Oushiman Group Deqing Biotechnology Co., Ltd., keratinocytes and fibroblasts were purchased from the cell bank of the Type Culture Collection Committee of the Chinese Academy of Sciences, and the antibodies were all from Cell Signaling Technology, Inc (CST) in the United States.

Preparation Example 1

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.1 to 1 micron;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:10 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 20% (v/v) lactic acid solution to dissolve and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% of the expansion culture solution of Saccharomyces cerevisiae, and place it in a fermentation tank at 28°C, pH 7.0, rotation speed 300r/min, ventilation 3L/min 36 hours;

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the sterile filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight, that is, ml/g), pre-freeze with liquid nitrogen for 2 hours, and then put it in a vacuum freeze-drying bin and freeze-dry for 12 hours . The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 2

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.1~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 20% (v/v) acetic acid solution to dissolve and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% of Saccharomyces cerevisiae expansion culture solution, and place it in a fermentation tank at 28°C, pH 6.8, rotation speed 250r/min, ventilation 4L/min, and incubate for 36 hours ；

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the sterile filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), pre-freeze at -80°C for 3 hours, and then put it into a vacuum freeze-drying bin for 12 hours to freeze-dry. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 3

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.1~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 20% (v/v) lactic acid solution to dissolve the ultra-fine pearl powder and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% Lactobacillus expansion culture solution and place it in a fermentation tank at 37°C, pH 7.0, rotating speed 300r/min, and ventilating 3L/min for 36 hours ；

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), pre-freeze at -80°C for 5 hours, and then put it into a vacuum freeze-drying bin for 12 hours to freeze-dry. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 4

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.3~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Pour excess carbon dioxide into the bottom of the mixed solution for 0.5 hours, and then filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% lactic acid bacteria and Saccharomyces cerevisiae (1:1 weight ratio) mixed expansion culture solution, and place it in the fermentation tank at 37°C, pH value 7.0, rotation speed Cultivate for 36 hours under 300r/min and 3L/min ventilation;

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the sterile filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), pre-freeze at -80°C for 3 hours, and then put it into a vacuum freeze-drying bin for 12 hours to freeze-dry. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 5

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.3~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 20% (v/v) acetic acid solution to dissolve and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% lactic acid bacteria expansion culture solution, and place it in a fermentation tank at 37°C, pH 7.0, rotating speed 300r/min, and ventilating 3L/min for 36 hours ；

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), pre-freeze it under liquid nitrogen for 5 hours, and then put it into a vacuum freeze-drying bin for 12 hours to freeze-dry. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 6

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.3~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 20% (v/v) lactic acid solution to dissolve and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% lactic acid bacteria expansion culture solution and place it in a fermenter at 37°C, pH 6.5, rotating speed 300r/min, and ventilating 3L/min for 36 hours ；

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), pre-freeze with liquid nitrogen for 3 hours, and then put it into a vacuum freeze-drying bin for 12 hours to freeze-dry. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 7

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.3~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Pour excess carbon dioxide into the bottom of the mixed solution for 0.5 hours, and then filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% Bacillus subtilis expansion culture solution, and place it in a fermentation tank at 37°C, pH 6.5, rotation speed 300r/min, and 3L/min ventilation. 36 hours;

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), pre-freeze with liquid nitrogen, and then put it into a vacuum freeze-drying bin to freeze-dry for 12 hours. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 8

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.3~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 20% (v/v) acetic acid solution to dissolve and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% Bacillus subtilis expansion culture solution, and place it in a fermentation tank at 37°C, pH 6.8, rotation speed 300r/min, and ventilation 3L/min. 36 hours;

Step 5) After stirring the fermentation broth at 85°C for 20 minutes for inactivation, press filtration through a 0.22 μm filter membrane to obtain a sterile filtrate;

Step 6) Mix the filtrate and the freeze-dried protective agent at a ratio of 100:0.5 (volume/weight), use liquid nitrogen or pre-freeze at -80 ℃, and then put it into a vacuum freeze-drying bin to freeze-dry for 24 hours. The freeze-dried powder is finally obtained;

Step 7) Store the lyophilized powder in a brown sample bottle at 4°C.

Preparation Example 9

Step 1) After the pearls are cleaned, refined and spray dried, they are passed through a jet mill to obtain ultra-fine pearl powder with a particle size of 0.3~1 microns;

Step 2) Mix 1kg of pearl powder and deionized water at a weight ratio of 1:5 at 30°C through an internal stirrer or stirring blade at a speed of 50rpm;

Step 3) Add 15% lactic acid solution to dissolve and filter;

Step 4) After sterilizing the solution obtained after filtering in the previous step, insert 5% Bacillus subtilis expansion culture solution, and place it in a fermentation tank at 37°C, pH 6.8, rotation speed 300r/min, and ventilation 3L/min. 48 hours;

Step 4) After stirring for 20 minutes at 85°C for inactivation, press filtration through a 0.22μm filter membrane to obtain a sterile filtrate;

Step 5) The sterile filtrate and the freeze-dried protective agent are mixed at a ratio of 100:0.5 (volume/weight), pre-freeze at -80°C for 1 hour, and then placed in a vacuum freeze-drying bin for freeze-drying for 72 hours. Finally, the lyophilized powder is obtained.

Step 6) Put the lyophilized powder cans in brown sample bottles and store them at 4°C.

Experimental Example 1 Observation of pearl morphology before and after jet pulverization by scanning electron microscope

The processed and dried sample powders (the sample powders before and after airflow pulverization, respectively) were adhered to a clean copper table with double-sided conductive tape. After ion sputtering coating, they were observed and photographed on a scanning electron microscope. The acceleration voltage is 20kV. The experimental results are shown in Figure 1.

It can be seen from Figure 1 that before the jet pulverization, the powder pseudo agglomerates. After air jet pulverization, the false agglomeration can be broken up, and further smashed to obtain a small and concentrated small particle size powder with a small particle size distribution range.

Experimental Example 2 The particle size distribution of pearl powder after jet pulverization was measured by the laser micro-potentiometer method

Steps:

a) Deionized water for the liquid for dispersing the nano pearl powder sample;

b) Check if there are bubbles in the solution. If there are bubbles, use ultrasound to eliminate bubbles;

c) Check whether the sample pool is clean;

d) The refractive index is selected as 1.61, and the absorptance is selected as 1; it is determined according to GB/T 19077.1-2008.

The result is shown in Figure 2. It can be seen from Figure 2 that the particle size distribution range of the pulverized pearl powder is narrow.

Experimental Example 3 SDS-PAGE electrophoresis of pearl preparations before and after fermentation treatment

SDS-PAGE electrophoresis was used to detect the pearl preparations before and after the fermentation treatment. The results are shown in Figure 3A and 3B. It can be seen from the figures that the molecular weight range of the pearl preparations after the fermentation treatment has been significantly reduced, confirming that the present invention Method to obtain small molecule peptides.

Experimental Example 4 In the UV aging model, the effect of pearl preparations on various oxidation indicators in keratinocytes

Test method: After ultraviolet radiation damages keratinocytes, each group of cells is disrupted by ultrasound on ice, the cell homogenate is collected by centrifugation in a centrifuge, and the absorbance of each group is measured according to the instructions of the Nanjing Jiancheng kit. The BCA protein concentration determination kit was used to determine the protein concentration of each group, and finally the content of ROS, SOD, and MDA in each group was calculated.

As shown in Figure 4A to Figure 4D, after keratinocytes were irradiated by UV, the content of ROS, glutathione peroxidase, malondialdehyde and superoxide dismutase in keratinocytes all changed significantly, and then Adding 0.01% and 0.1% (g/ml) concentrations of pearl preparations was measured after 24 hours of action. The results showed that pearl preparations can significantly improve the oxidation index (influence *p<0.1, significant**p<0.01, very Significant **p<0.001).

Experimental Example 5 In the UV aging model, the effect of pearl preparations on the expression of cell inflammatory factors

Test method: Western blot method to detect Bcl-2, Bax, Caspase-3 and Caspase-9 protein expression levels

When the growth of fibroblasts in the six-well plate reaches 80%-90%, collect the cells of each group, discard the supernatant, add 100 μL per well RIPA Lysis Solution (containing 1% PMSF), and let it stand for 3 minutes. Transfer the liquid to a 1.5ml centrifuge tube, let it stand on ice for 30 minutes and then centrifuge (4 ℃, 12000 r/min-15000 r/min), aspirate the supernatant into another new 1.5 mL centrifuge tube, which is the protein . Mix the protein sample with 5 × SDS Loading Buffer at a ratio of 4:1, and heat it in boiling water for 7 minutes to fully denature the protein. Prepare separation gel and concentrated gel, put the prepared gel into the electrophoresis tank, add freshly prepared electrophoresis buffer to the inner and outer tanks, pull out the sample comb, and add the protein sample. The sample volume is 60 μg. Cut the gel at the end of electrophoresis and transfer (100V, 30 minutes). The primary antibody (1: 500) is blocked, overnight at 4 ℃, and the secondary antibody (1: 10 000) is reacted for 1 hour. ECL luminescent liquid A and B are mixed uniformly in a ratio of 1:1, and dropped onto the PVDF membrane to make it Cover evenly, expose and develop in a dark room, and analyze the results.

As shown in Figure 5, under the action of different concentrations of pearl preparations, the expression of Caspase3 and Caspase3 apoptotic factors were significantly reduced, and the expression of inhibitory apoptosis factor Bcl-2 increased, indicating that the pearl preparations of the present invention passed PI3K/ The AKT pathway has the effect of inhibiting the expression of inflammatory factors.

In summary, the pearl preparation of the present application still has a very significant anti-aging ability at a concentration of 1/10,000, and it acts on the mitochondria through the PI3K/AKT pathway to play an anti-aging effect.

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Figures 1A and 1B show the electrographic images of the pearl powder before and after the jet milling treatment (Figure 1A: before treatment, Figure 1B: after treatment).

Figure 2 shows the particle size distribution of the pearl powder after jet pulverization.

Figures 3A and 3B show the SDS-PAGE electrophoresis of pearl preparations before and after microbial fermentation. Figure 3A: Lane 1 is the test sample before fermentation, Lane 2 is the standard protein; Figure 3B: Lanes 1 and 4 are the standard protein, Lane 2 and 3 are test samples after fermentation.

Figures 4A to 4D show that the pearl preparation prepared according to Preparation Example 1 of the present invention has different concentrations (1*10 ^-4 g/ml and 1*10 ^-3 g/ml) on the oxidation indicators in keratinocytes ( Figure 4A: ROS; Figure 4B: GSH-Px; Figure 4C: MDA; and Figure 4D: SOD) schematic diagram of the impact.

Figure 5 shows a schematic diagram of the effect of the pearl preparation prepared according to Preparation Example 2 of the present invention at different concentrations (1*10 ^-4 g/ml and 1*10 ^-3 g/ml) on the expression of cell inflammatory factors (swimming lanes) 1: blank, lane 2: UV treatment, lane 3: UV+10 ^-4 g/ml, lane 4: UV+10 ^-3 g/ml).

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Claims (10)

A method for preparing pearl preparations, which comprises the following steps: 1) Crushing: crush the pearls with a jet mill to obtain pearl powder, the particle size of which is less than 6 microns; 2) Mixing: Mix pearl powder with deionized water to obtain a mixed solution; 3) Weak acid dissolution: pass carbon dioxide into the bottom of the mixed liquid, or add a weak acid solution to dissolve the pearl powder in the mixed liquid; 4) Fermentation: the solution obtained in step 3) is sterilized and then connected to the microbial expansion broth for fermentation culture; and 5) Freeze-drying: freeze-dry the fermentation broth obtained in step 4) to obtain freeze-dried powder of pearl preparations. The preparation method according to claim 1, wherein, before step 1), the method further includes the steps of washing, refining, and spray drying the pearls. The preparation method according to claim 1, wherein in step 2), the mixing weight ratio of the pearl powder and the deionized water is 1:5-20. The preparation method according to claim 1, wherein, in step 3), the volume ratio of the weak acid solution and the mixed solution is 1:4-10. The preparation method according to claim 1, wherein, in step 4), the microbial expansion culture solution accounts for 1-10% by volume of the solution to be connected. The preparation method according to claim 1, wherein, in step 5), after the fermentation broth is inactivated, the sterile filtrate is obtained by pressure filtration through a filter membrane, and the sterile filtrate and the lyophilized protective agent are determined by volume/weight The ratio is 100:0.5~2 for mixing. In the above volume/weight ratio, the unit of volume is milliliter (ml) and the unit of weight is gram. Use liquid nitrogen or pre-freeze at -70~-85℃ for 3~10 hours Then put it into a vacuum freeze-drying bin and freeze-dry for 24 to 72 hours. The preparation method according to claim 1, wherein, after step 5), the method further includes the step of placing the freeze-dried powder of the pearl preparation in a brown sample bottle or a vacuum-packed plastic bag for storage. A pearl preparation prepared according to the preparation method described in any one of claims 1 to 7. A composition comprising the pearl preparation according to claim 8. A use of the pearl preparation according to claim 8 or the composition according to claim 9 as an anti-aging product.

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