Ginseng fermentation extract as well as preparation method and application thereof in skin care products

Abstract

The invention discloses a ginseng fermentation extract and a preparation method and application thereof in skin care products, and belongs to the technical field of plant extraction.The preparation method comprises the following steps that ginseng powder and deionized water are evenly mixed, the pH is adjusted to 4.5-5.5, then compound enzyme is added, enzymolysis is conducted for 3-5 h at the temperature of 40-50 DEG C under the ultrasonic condition, and enzymatic hydrolysate is obtained; carrying out inactivation treatment on the enzymatic hydrolysate, adding a carbon source, a magnetic fermentation aid, a complex microbial inoculant and deionized water, and carrying out constant-temperature fermentation at 35-40 DEG C for 4-6 days to obtain fermentation liquor; taking out the magnetic fermentation aid from the fermentation liquor under the action of an external magnetic field to obtain a ginseng fermentation product, performing reflux extraction on the ginseng fermentation product twice with an ethanol solution, performing extraction for 2 hours each time, combining extracting solutions, and performing freeze drying to obtain a ginseng fermentation extract; according to the method, high-yield extraction can be achieved within a short time, the content of active ingredients such as polysaccharide, saponin and flavone in the obtained extract is remarkably increased, and pesticide residues are greatly reduced.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of plant extraction, and particularly relates to a ginseng fermentation extract, a preparation method thereof and application thereof in skincare products. BACKGROUND

[0002] Ginseng (scientific name: Panax ginseng C.A. Meyer) belongs to perennial herbaceous plants of the genus Panax in the family Araliaceae, mainly distributed in the mountainous areas of eastern Asia, and is rich in various components, including ginsenosides, volatile oils, proteins, polysaccharides, flavonoids, vitamins, amino acids and trace elements. Related studies have shown that ginsenosides and polysaccharides are the main active components of ginseng, and ginsenosides have significant effects on nervous system diseases and anti-tumor, can inhibit the generation of free radicals and scavenge free radicals, and can resist the damage of free radicals to tissues and cells, thereby enhancing the function of the body's own antioxidant system. However, most of these trace elements exist in the cytoplasm of ginseng cells, and the cell wall of ginseng cells is composed of cellulose and hemicellulose and other substances, and its structure is dense. If it is taken directly, most of the active ingredients in it are difficult to be absorbed by the human body, and the bioavailability is low. Therefore, at present, ginseng extract is usually prepared by extraction process to break through the cell wall barrier and improve the dissolution and bioavailability of active ingredients.

[0003] At present, the common extraction methods of ginseng mainly include water extraction, solvent extraction and enzyme extraction. Among them, water extraction is the most widely used, which is favored due to its green environmental protection and low cost, but it also has the disadvantages of long extraction time and low efficiency. Compared with water extraction, solvent extraction can reduce the dissolution of water-soluble impurities such as starch and protein, but there may be residual organic solvents, which affects the safety of the product. Enzyme extraction is usually carried out at normal temperature, normal pressure and near neutral pH, and enzymes as biological catalysts have the characteristics of non-toxicity, biodegradability and environmental friendliness, but the cost is high, which limits its large-scale application. In addition, the above-mentioned methods also have a common problem: the ginsenosides obtained in the ginseng extract are mainly natural saponins, and the content of rare saponins is very low. Since natural saponins have large polarity, they are not easily absorbed by the human body, and rare saponins usually have higher pharmacological activity. Therefore, how to effectively improve the content of rare saponins in ginseng extract has become a technical problem to be solved at present. SUMMARY

[0004] One of the purposes of the present application is to provide a preparation method of ginseng fermentation extract, in order to solve the problem of low content of rare saponins in ginseng extract in the prior art.

[0005] The second purpose of the present application is to provide a ginseng fermentation extract obtained by the above-mentioned preparation method of ginseng fermentation extract.

[0006] The third object of the present application is to provide the use of the ginseng fermentation extract in skin care products.

[0007] The object of the present application can be achieved by the following technical solutions: A preparation method of a ginseng fermentation extract, comprising the following steps: S1, mixing ginseng powder and deionized water uniformly, adjusting pH to 4.5-5.5, then adding a composite enzyme, and under ultrasonic conditions, enzymolysis at 40-50℃ for 3-5h to obtain an enzyme solution; S2, inactivating the enzyme solution, adding a carbon source, a magnetic fermentation aid, a composite microbial agent and deionized water, and constant temperature fermentation at 35-40℃ for 4-6 days to obtain a fermentation liquid; S3, taking out the magnetic fermentation aid in the fermentation liquid under the action of an external magnetic field to obtain a ginseng fermentation product, reflux extracting the ginseng fermentation product with an ethanol solution twice, each time for 2h, combining the extraction liquids, and freeze-drying to obtain a ginseng fermentation extract.

[0008] Further, the mass ratio of the enzyme solution, the carbon source, the magnetic fermentation aid, the composite microbial agent and the deionized water in S2 is 100:6-10:3-5:4-6:30-50.

[0009] Further, the composite microbial agent is composed of Bacillus subtilis bacterial liquid and Lactobacillus phytase bacterial liquid according to a volume ratio of 0.6-1:0.6-1.

[0010] Further, the magnetic fermentation aid is phenylsiloxane modified ferroferric oxide.

[0011] The cell wall structure of ginseng is mainly based on cellulose as the basic skeleton, and there are rich protein components in the cell contents and cell membranes. The ginseng powder is treated by ultrasonic-assisted enzyme method, the cavitation effect of ultrasonic is used on the basis of enzyme fermentation, the plant cell wall lysis is accelerated, and the protein, pectin, tannin, ash and other sticky impurities can be efficiently removed. This treatment process not only can significantly reduce the interference of impurities on the extraction of effective components, but also can promote the efficient dissolution of effective components by destroying the physical structure of cell wall and increasing its permeability, especially effectively releasing ginsenosides, providing substrates and precursor substances for subsequent microbial fermentation.

[0012] Due to the low content of rare saponins in the enzymatic products of ginseng, the present application uses the specific enzymes (such as glucosidase) secreted by microorganisms to carry out deglycosylation reaction on natural saponins, so as to convert natural saponins into rare saponins. The Bacillus subtilis in the composite microbial inoculant can secrete a large amount of extracellular enzymes to hydrolyze and remove the sugar groups on the macromolecular saponin aglycone, thereby starting the conversion process of saponins. The organic acid produced by Lactobacillus phytase creates and maintains a suitable acidic environment for biological conversion, and the enzyme system produced by fermentation can be complementary to Bacillus subtilis to catalyze the deglycosylation reaction at different sites. The synergistic effect of the two makes the saponin conversion path more smooth and the conversion rate higher.

[0013] Due to the long fermentation cycle of microorganisms and the pesticide residues in the fermentation products, the present application introduces a magnetic fermentation aid into the fermentation system. The aid is a phenylsiloxane modified ferroferric oxide, and the surface is rich in benzene ring structure. It has a magnetic effect, which can stimulate the microorganisms in the composite microbial inoculant, catalyze the enzyme activity of the microorganisms, improve the metabolism function and biological activity of the microorganisms, and accelerate the fermentation process. On the other hand, the magnetic fermentation aid can be quickly separated from the fermentation system under the action of an external magnetic field. In addition, the benzene ring structure on its surface can combine with the residual pesticide molecules (such as chloronitrobenzene, pentachloroaniline, and pythium agent) in the ginseng fermentation products through π-π interaction, thereby reducing the pesticide residues in the ginseng fermentation extract.

[0014] Further, the raw materials for preparing the magnetic fermentation aid include ferroferric oxide and phenylsiloxane, and the mass ratio of ferroferric oxide to phenylsiloxane is 1:0.2-0.5.

[0015] Further, the preparation steps of the magnetic fermentation aid are as follows: The ferroferric oxide is ultrasonically dispersed in an ethanol solution, phenylsiloxane is added, and stirring reaction is carried out at 25-30℃ for 12-24h. After the reaction is completed, filtration is performed, the filter cake is dried, and the magnetic fermentation aid is obtained.

[0016] Further, the phenylsiloxane is at least one of phenyltrimethoxysilane, p-chlorophenyltrimethoxysilane, triethoxyp-tolylsilane, trimethoxy (4-fluorophenyl) silane, 3- (phenylamino) propyltrimethoxysilane, methylphenyldimethoxysilane, and chlorophenyltriethoxysilane, and is preferably phenyltrimethoxysilane.

[0017] Further, the ferroferric oxide is nanoscale ferroferric oxide.

[0018] Further, the preparation steps of the ferroferric oxide are as follows: FeCl3·6H2O, FeCl2·4H2O and deionized water were added into a flask, nitrogen was introduced to remove oxygen, and the temperature was raised to 50-55℃, and then ammonia water was added at a rotation speed of 400-500rpm, and the reaction was stirred for 0.5-1h, after the reaction was completed, the mixture was filtered and the filter cake was dried to obtain the ferroferric oxide.

[0019] Further, the amount ratio of FeCl3·6H2O, FeCl2·4H2O, deionized water and ammonia water was 5.6mmol:11.2mmol:100-150mL:12.5mL, and the mass fraction of ammonia water was 25%.

[0020] Further, the amount ratio of ginseng powder and deionized water in S1 was 1g:10-30mL, and the amount of complex enzyme was 0.15-0.25% of the mass of ginseng powder, and the complex enzyme was composed of α-amylase, cellulase and pectinase in a mass ratio of 2-4:2-4:1-5.

[0021] Further, the ultrasonic power was 20-60W, and the pH regulator was citric acid.

[0022] Further, the inactivation treatment temperature was 85-95℃, and the inactivation time was 25-35min.

[0023] Further, the carbon source in S2 was sucrose and / or glucose.

[0024] Further, the number of bacteria in the Bacillus subtilis and Lactobacillus phytase bacterial liquid was 10 5 -10 8 CFU / mL.

[0025] Further, the Bacillus subtilis was Bacillus subtilis subsp. subtilis, which was an existing strain with a preservation number of CICC 10066 and was preserved in the China Industrial Microbial Culture Collection Center.

[0026] Further, the Lactobacillus phytase was named 581, which was an existing strain with a preservation number of CGMCC No.13121 and was preserved in the China General Microbiological Culture Collection Center.

[0027] Further, the volume fraction of ethanol solution in S3 was 75-85%, and the amount ratio of ginseng fermentation product and ethanol solution in the reflux extraction process was 1g:10mL.

[0028] A ginseng fermentation extract prepared by the above preparation method.

[0029] An application of ginseng fermentation extract in skincare products: Ginseng fermentation extract and its ginsenoside Rd can inhibit the induction of UV-A-mediated matrix metalloproteinase-1 (MMP-1) and MMP-13, thereby exerting a photoaging protection effect. Ginseng polysaccharides can induce the production of type I collagen by clearing residual free genes and have good anti-skin aging activity. Therefore, its application in skincare products can exert anti-aging, whitening, anti-wrinkle and anti-inflammatory effects.

[0030] The beneficial effects of this invention are: 1. The ginseng fermentation extract preparation method of the present invention integrates ultrasound-assisted enzymatic hydrolysis, compound microbial fermentation, and magnetic agent separation and purification, significantly improving extraction efficiency and product quality. This method can achieve high-yield extraction in a short time, resulting in a significant increase in the content of active ingredients such as polysaccharides, saponins, and flavonoids in the extracted extract, while drastically reducing pesticide residues. In particular, the present invention significantly increases the content of rare saponins through the targeted biotransformation effect of compound microbial agents, further enhancing the bioactivity and application value of the extract.

[0031] 2. This invention uses Bacillus subtilis and Lactobacillus phytate in a volume ratio of 0.6-1:0.6-1 to form a compound bacterial agent, and then... 5 -10 8 At a high activity concentration of CFU / mL, a stable and efficient multi-enzyme biological reaction system is formed by utilizing the synergistic effect between different bacterial species. During the 4-6 day fermentation period, ordinary ginsenosides released from the enzymatic hydrolysate are continuously and directionally converted into rare saponins with higher bioavailability and stronger activity.

[0032] 3. To shorten the fermentation cycle and improve product purity, this invention introduces a magnetic fermentation aid. This aid has a dual function: firstly, its magnetic effect can stimulate microbial activity, increase enzyme activity and metabolism, thereby accelerating the fermentation process and facilitating rapid separation under the influence of an external magnetic field; secondly, its surface benzene ring structure can adsorb residual pesticide molecules (such as non-chloronitrobenzene) through π-π interactions, effectively reducing pesticide residues in the extract. Detailed Implementation

[0033] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0034] The ginseng powder in the present application is obtained by crushing dry ginseng through a 60-mesh sieve, the dry ginseng is ginseng from Changbai Mountain in Jilin, the bacillus subtilis is bacillus subtilis subsp. subtilis, which is an existing strain, and the preservation number is CICC 10066, preserved in China Industrial Microbial Culture Collection Center; the lactobacillus phytis is named 581, which is an existing strain, and the preservation number is CGMCC No. 13121, preserved in China General Microbiological Culture Collection Center; the enzyme activity of alpha-amylase is 2000 u / g, the enzyme activity of cellulase is 20000 u / g, the enzyme activity of pectinase is 50000 u / g, and other raw materials and reagents are purchased from the market.

[0035] The following will be specifically described in combination with examples.

[0036] Preparation Example 1

[0037] The preparation of the magnetic fermentation aid is as follows: 10 g of ferroferric oxide is ultrasonically dispersed in an ethanol solution, 2 g of phenyltrimethoxysilane is added, stirring is carried out at 25°C for 12 h, after the reaction is completed, suction filtration is carried out, the filter cake is dried, and the magnetic fermentation aid is obtained.

[0038] The preparation steps of ferroferric oxide are as follows: 56 mmol of FeCl3·6H2O, 112 mmol of FeCl2·4H2O and 1 L of deionized water are added to a flask, nitrogen is introduced to remove oxygen, the temperature is raised to 50°C, 125 mL of 25wt% ammonia water is added at a rotation speed of 400 rpm, and the reaction is carried out under stirring for 0.5 h, after the reaction is completed, filtration is carried out, the filter cake is dried, and ferroferric oxide is obtained.

[0039] Preparation Example 2

[0040] The preparation of the magnetic fermentation aid is as follows: 10 g of ferroferric oxide is ultrasonically dispersed in an ethanol solution, 2 g of phenyltrimethoxysilane is added, stirring is carried out at 25°C for 12 h, after the reaction is completed, suction filtration is carried out, the filter cake is dried, and the magnetic fermentation aid is obtained.

[0041] The preparation steps of ferroferric oxide are as follows: 56 mmol of FeCl3·6H2O, 112 mmol of FeCl2·4H2O and 1.3 L of deionized water are added to a flask, nitrogen is introduced to remove oxygen, the temperature is raised to 52°C, 125 mL of 25wt% ammonia water is added at a rotation speed of 450 rpm, and the reaction is carried out under stirring for 0.8 h, after the reaction is completed, filtration is carried out, the filter cake is dried, and ferroferric oxide is obtained.

[0042] Preparation Example 3

[0043] The preparation of the magnetic fermentation aid is as follows: 10g of ferroferric oxide was ultrasonically dispersed in an ethanol solution, 5g of phenyltrimethoxysilane was added, and the reaction was stirred at 30℃ for 24h. After the reaction was completed, the filter cake was dried by suction filtration to obtain the magnetic fermentation aid.

[0044] The ferroferric oxide was prepared by the following steps: 56mmol of FeCl3·6H2O, 112mmol of FeCl2·4H2O and 1.5L of deionized water were added to a flask, nitrogen was introduced to remove oxygen, the temperature was raised to 55℃, 125mL of 25wt% ammonia water was added at a rotation speed of 500rpm, and the reaction was stirred at constant temperature for 1h. After the reaction was completed, the filter cake was dried by suction filtration to obtain the ferroferric oxide.

[0045] Comparative Example 1

[0046] The magnetic fermentation aid was prepared as in Preparation Example 2, except that the “phenyltrimethoxysilane” in Preparation Example 2 was replaced with an equal amount of “γ-aminopropyltriethoxysilane”.

[0047] Comparative Example 2

[0048] This comparative example is ferroferric oxide, and the preparation process is the same as that of Preparation Example 2.

[0049] Example 1

[0050] A preparation method of a ginseng fermentation extract includes the following steps: S1, 100g of ginseng powder and 1L of deionized water were mixed uniformly, the pH was adjusted to 4.5 with citric acid, and then 0.15g of a composite enzyme composed of α-amylase, cellulase and pectinase in a mass ratio of 2:2:1 was added. The enzyme was hydrolyzed under ultrasonic conditions at a power of 20W and at 40℃ for 3h to obtain an enzyme hydrolysate; S2, 100g of the enzyme hydrolysate was inactivated at 85℃ for 35min, 6g of sucrose, 3g of the magnetic fermentation aid of Preparation Example 1, 4g of a composite bacterial agent and 30g of deionized water were added, and the mixture was fermented at a constant temperature of 35℃ for 4 days to obtain a fermentation liquor; S3, the magnetic fermentation aid in the fermentation liquor was removed under the action of an external magnetic field to obtain a ginseng fermentation product. The ginseng fermentation product was refluxed with a 75% ethanol solution twice, the amount of the ginseng fermentation product and the ethanol solution was 1g:10mL, each time for 2h, the extract was combined and freeze-dried to obtain a ginseng fermentation extract.

[0051] The composite bacterial agent was composed of Bacillus subtilis bacterial liquid and Lactobacillus phytase bacterial liquid in a volume ratio of 1:1.

[0052] The preparation process of the Bacillus subtilis bacterial liquid was as follows: Bacillus subtilis was inoculated in LB medium and activated at 30℃ to a viable cell count of 5.23 x 10 6 CFU / mL.

[0053] Lactobacillus plantarum bacterial solution was prepared as follows: Lactobacillus plantarum was inoculated in MRS medium and activated at 37℃ to a viable cell count of 3.15 x 10 6 CFU / mL.

[0054] Example 2

[0055] A preparation method of a ginseng fermentation extract includes the following steps: S1, 100g of ginseng powder and 2L of deionized water were mixed uniformly, the pH was adjusted to 5.0 with citric acid, and then 0.2g of a composite enzyme was added, the composite enzyme was composed of alpha-amylase, cellulase and pectinase in a mass ratio of 3:3:2, and under the condition of ultrasonic power of 40W, enzymolysis was carried out at 45℃ for 4h to obtain an enzymolysis solution; S2, 100g of the enzymolysis solution was inactivated at 90℃ for 30min, 8g of sucrose, 4g of the magnetic fermentation aid prepared in Preparation Example 1, 5g of a composite bacterial agent and 40g of deionized water were added, and constant temperature fermentation was carried out at 37℃ for 5 days to obtain a fermentation solution; S3, the magnetic fermentation aid in the fermentation solution was taken out under the action of an external magnetic field to obtain a ginseng fermentation product, the ginseng fermentation product was refluxed with an 80% ethanol solution twice, the amount ratio of the ginseng fermentation product to the ethanol solution was 1g:10mL during the reflux extraction process, each extraction was carried out for 2h, the extraction solutions were combined, and freeze-drying was carried out to obtain a ginseng fermentation extract.

[0056] The composite bacterial agent was composed of Bacillus subtilis bacterial solution and Lactobacillus plantarum bacterial solution in a volume ratio of 0.6:1, and the preparation processes of the Bacillus subtilis bacterial solution and the Lactobacillus plantarum bacterial solution were the same as those in Example 1.

[0057] Example 3

[0058] A preparation method of a ginseng fermentation extract includes the following steps: S1, 100g of ginseng powder and 3L of deionized water were mixed uniformly, the pH was adjusted to 5.5 with citric acid, and then 0.25g of a composite enzyme was added, the composite enzyme was composed of alpha-amylase, cellulase and pectinase in a mass ratio of 4:4:5, and under the condition of ultrasonic power of 60W, enzymolysis was carried out at 50℃ for 5h to obtain an enzymolysis solution; S2, 100g of the enzymolysis solution was inactivated at 95℃ for 35min, 10g of sucrose, 5g of the magnetic fermentation aid prepared in Preparation Example 1, 6g of a composite bacterial agent and 50g of deionized water were added, and constant temperature fermentation was carried out at 40℃ for 6 days to obtain a fermentation solution; S3, the magnetic fermentation aid in the fermentation broth is taken out under the action of an external magnetic field to obtain ginseng fermentation product, the ginseng fermentation product is refluxed with 85% ethanol solution twice, the ratio of the ginseng fermentation product to the ethanol solution is 1g:10mL, each time for 2h, the extract is combined and freeze-dried to obtain ginseng fermentation extract.

[0059] The complex bacterial agent is composed of Bacillus subtilis bacterial liquid and Lactobacillus phytase bacterial liquid according to a volume ratio of 1:0.6, and the preparation process of the Bacillus subtilis bacterial liquid and the Lactobacillus phytase bacterial liquid is the same as that in Example 1.

[0060] Example 4

[0061] A preparation method of ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is replaced by the product obtained in Preparation Example 2.

[0062] Example 5

[0063] A preparation method of ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is replaced by the product obtained in Preparation Example 3.

[0064] Example 6

[0065] A preparation method of ginseng fermentation extract, compared with Example 2, the only difference is that the magnetic fermentation aid in Example 2 is replaced by the product obtained in Preparation Example 2.

[0066] Example 7

[0067] A preparation method of ginseng fermentation extract, compared with Example 2, the only difference is that the magnetic fermentation aid in Example 2 is replaced by the product obtained in Preparation Example 3.

[0068] Example 8

[0069] A preparation method of ginseng fermentation extract, compared with Example 3, the only difference is that the magnetic fermentation aid in Example 3 is replaced by the product obtained in Preparation Example 2.

[0070] Example 9

[0071] A preparation method of ginseng fermentation extract, compared with Example 3, the only difference is that the magnetic fermentation aid in Example 3 is replaced by the product obtained in Preparation Example 3.

[0072] Comparative Example 1

[0073] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is replaced by the product obtained in Comparative Example 1.

[0074] Comparative Example 2

[0075] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is replaced by the product obtained in Comparative Example 2.

[0076] Comparative Example 3

[0077] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is removed.

[0078] Comparative Example 4

[0079] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the "complex microbial agent" in Example 1 is replaced by an equal amount of Bacillus subtilis bacterial solution, and the preparation process of the Bacillus subtilis bacterial solution is the same as that of Example 1.

[0080] Comparative Example 5

[0081] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the "complex microbial agent" in Example 1 is replaced by an equal amount of Lactobacillus phytase bacterial solution, and the preparation process of the Lactobacillus phytase bacterial solution is the same as that of Example 1.

[0082] Comparative Example 6

[0083] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is removed, and the "complex microbial agent" in Example 1 is replaced by an equal amount of Bacillus subtilis bacterial solution, and the preparation process of the Bacillus subtilis bacterial solution is the same as that of Example 1.

[0084] Comparative Example 7

[0085] A preparation method of a ginseng fermentation extract, compared with Example 1, the only difference is that the magnetic fermentation aid in Example 1 is removed, and the "complex microbial agent" in Example 1 is replaced by an equal amount of Lactobacillus phytase bacterial solution, and the preparation process of the Lactobacillus phytase bacterial solution is the same as that of Example 1.

[0086] The ginseng fermentation extracts obtained in Examples 1-9 and Comparative Examples 1-7 were respectively passed through a 60-mesh sieve, 2 times the mass of chromatographic methanol was added, dissolved thoroughly, and passed through a 0.22 μm filter membrane. The filtrate was used for HPLC detection, and the detection process was as follows: (1) Polysaccharide content test The phenol-sulfuric acid method was used to test the polysaccharide content using glucose as a standard. The glucose mass concentration was used as the horizontal coordinate, the absorbance was used as the vertical coordinate, the absorbance was measured at 490 nm, the standard curve was drawn as y = 7.95657x-0.00716, R 2 = 0.9993, and the polysaccharide content was expressed by the mass of glucose per gram of dry matter; (2) Total flavonoid content test The aluminum chloride colorimetric method was used to test the total flavonoid content using rutin as a standard. The rutin mass concentration was used as the horizontal coordinate, the absorbance was used as the vertical coordinate, the absorbance was measured at 500 nm, the standard curve was drawn as y = 2.8174x-0.00745, R 2 = 0.9991, and the total flavonoid content was expressed by the mass of rutin per gram of dry matter; (3) Total saponin content test The vanillin-glacial acetic acid method was used to test the total saponin content using ginsenoside Re as a standard. The ginsenoside Re mass concentration was used as the horizontal coordinate, the absorbance was used as the vertical coordinate, the absorbance was measured at 465 nm, the standard curve was drawn as y = 2.975x+0.01902, R 2 = 0.9995, and the total saponin content was expressed by the mass of saponin Re per gram of dry matter; (4) Rare saponin content test HPLC was used to identify the ginsenoside components of the sample solution, and a PerkinElmer C 18 chromatographic column (250mmx4.6mm, 5μm) was used, the mobile phase was acetonitrile (A)-0.05% phosphate aqueous solution (B), the detection wavelength was 203 nm, the flow rate was 1.0mL / min, the injection volume was 10μL, and the column temperature was 35℃, and gradient elution was performed. According to the ginsenoside standard curve, the content of rare saponin in the sample was quantitatively calculated, which was expressed by the mass of rare saponin per gram of extract, and rare ginsenoside Rg3 and rare ginsenoside Ck were taken as representatives; (5) Pesticide residue The test sample solution was prepared by the method of solution preparation in document 2.1.1, and the chromatographic peak area was tested under the chromatographic conditions of item 2.2, and the technical fludioxonil content was calculated according to the standard curve equation of item 2.4; The test results are shown in Table 1:

[0087] Table 1

[0088] As can be seen from the recorded data in Table 1, the total polysaccharide content of the ginseng fermentation extract obtained in Examples 1-3 is 422.7-454.6 mg / g, the total saponin content is 178.4-182.2 mg / g, the total flavone content is 7.6-8.1 mg / g, the content of rare ginsenoside Rg3 is 3.5-4.2 mg / g, the content of rare ginsenoside Ck is 1.6-2.1 mg / g, and the content of fluoro fungicide is ≤0.2 ug / L, wherein the total polysaccharide and total saponin contents in Example 2 are high, the content of rare ginsenoside in Example 3 is high, and no fluoro fungicide is detected; As can be seen from the test results of the group of Example 1, Example 4 and Example 5, the group of Example 2, Example 6 and Example 7, and the group of Example 3, Example 8 and Example 9, under the condition that other conditions are unchanged, the effective component contents of the ginseng fermentation products prepared by using the magnetic fermentation aid provided in Preparation Example 1, Preparation Example 2 and Preparation Example 3 are not much different, and the product quality obtained in Preparation Example 2 is relatively better; As can be seen from the experimental results of Example 1, Comparative Example 1 and Comparative Example 2, compared with the use of aminosiloxane modified ferroferric oxide and unmodified ferroferric oxide, the fluoro fungicide content of the product obtained by adding the magnetic fermentation aid of the application is significantly reduced; As can be seen from the experimental results of Example 1 and Comparative Example 3, compared with no addition of the magnetic fermentation aid, the total polysaccharide, total saponin, total flavone and rare ginsenoside contents of the product obtained by adding the magnetic fermentation aid of the application are significantly increased, and the fluoro fungicide content is significantly reduced; As can be seen from the experimental results of Example 1, Comparative Example 4 and Comparative Example 5, compared with the use of Lactobacillus phytase or Bacillus subtilis alone, the active component content of the product obtained by using the composite bacterial solution obtained by compounding the two as the fermentation microorganism is higher; As can be seen from the experimental results of Example 1, Comparative Example 6 and Comparative Example 7, by introducing the magnetic fermentation aid into the fermentation system and using the composite bacterial solution obtained by compounding Lactobacillus phytase and Bacillus subtilis as the fermentation microorganism, high-yield extraction is achieved in a relatively short time, the polysaccharide, saponin and flavone active component contents of the obtained extract are significantly increased, and the pesticide residue is greatly reduced.

[0089] It should be noted that, in this document, relational terms such as first and second and the like can be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises", "comprising", or any other variations 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 can include other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0090] While embodiments of the application have been shown and described, it is to be understood that the application is not limited to the details of the embodiments described, since numerous changes, modifications, substitutions and variations can be made thereto without departing from the spirit and scope of the application as defined by the appended claims and their equivalents.

Claims

1. A method for preparing ginseng fermentation extract, characterized in that, Includes the following steps: S1. Mix ginseng powder and deionized water evenly, adjust the pH to 4.5-5.5, add compound enzyme, and enzymatically hydrolyze at 40-50℃ for 3-5 hours under ultrasonic conditions to obtain enzymatic hydrolysate; S2. Inactivate the enzyme hydrolysate, add carbon source, magnetic fermentation aid, compound bacterial agent and deionized water, and ferment at 35-40℃ for 4-6 days to obtain fermentation broth; S3. The magnetic fermentation aid in the fermentation broth was removed under the action of an external magnetic field to obtain ginseng fermentation product. The ginseng fermentation product was extracted twice by reflux with ethanol solution for 2 hours each time. The extracts were combined and freeze-dried to obtain ginseng fermentation extract. The magnetic fermentation aid is phenylsiloxane-modified iron(III) oxide.

2. The method for preparing ginseng fermentation extract according to claim 1, characterized in that, The mass ratio of enzymatic hydrolysate, carbon source, magnetic fermentation aid, compound microbial agent and deionized water in S2 is 100:6-10:3-5:4-6:30-50.

3. The method for preparing a ginseng fermentation extract according to claim 1, characterized in that, The compound microbial agent is composed of Bacillus subtilis bacterial solution and Lactobacillus phytate bacterial solution in a volume ratio of 0.6-1:0.6-1.

4. The method for preparing ginseng fermentation extract according to claim 1, characterized in that, The raw materials for preparing the magnetic fermentation aid include iron oxide and phenylsiloxane, with the mass ratio of iron oxide to phenylsiloxane being 1:0.2-0.

5.

5. A method for preparing ginseng fermentation extract according to claim 1 or 4, characterized in that, The preparation steps of the magnetic fermentation aid are as follows: Iron oxide was ultrasonically dispersed in an ethanol solution, phenylsiloxane was added, and the mixture was stirred at 25-30℃ for 12-24 hours. After the reaction was completed, the mixture was filtered, and the filter cake was dried to obtain a magnetic fermentation aid.

6. The method for preparing a ginseng fermentation extract according to claim 4, characterized in that, The phenylsiloxane is at least one of phenyltrimethoxysilane, p-chlorophenyltrimethoxysilane, triethoxy-p-phenylmethylsilane, trimethoxy(4-fluorophenyl)silane, 3-(phenylamino)propyltrimethoxysilane, methylphenyldimethoxysilane, and chlorophenyltriethoxysilane.

7. The method for preparing a ginseng fermentation extract according to claim 4, characterized in that, The iron oxide is nano-sized iron oxide.

8. The method for preparing a ginseng fermentation extract according to claim 1, characterized in that, In S1, the ratio of ginseng powder to deionized water is 1g:10-30mL, and the amount of compound enzyme is 0.15-0.25% of the mass of ginseng powder. The compound enzyme is composed of α-amylase, cellulase and pectinase in a mass ratio of 2-4:2-4:1-5.

9. A ginseng fermentation extract, characterized in that, It is prepared by the preparation method according to any one of claims 1-8.

10. The application of the ginseng fermentation extract as described in claim 9 in a skin care product.

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