Oil-controlling and soothing plant fermentation product and preparation method and application thereof

Abstract

The present invention belongs to the field of microbial fermentation technology, and discloses a plant fermentation product for oil control and soothing, and its preparation method and application. The raw materials for preparing the plant fermentation product include: plant raw materials, milk, glycerin and fermentation strains; the plant raw materials include at least one of Platycladus orientalis leaves, Sabina juniper leaves, Juniperus chinensis leaves, and Selaginella leaves; the fermentation strains include at least one of Lactobacillus plantarum, Lactobacillus lactis, and Bifidobacterium longum. The present invention organically combines high-pressure homogenization and directional microbial fermentation technology, and uses milk and glycerin in the high-pressure homogenization process, so that plant tissue cells can be fully broken, active ingredients can be fully released and dissolved, and the utilization efficiency of raw materials is improved. The composite stock solution obtained by high-pressure homogenization is used as a subsequent fermentation matrix. Through the biological transformation ability of microorganisms, the prepared plant fermentation product has significant oil control and soothing effects.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and particularly relates to an oil-controlling and soothing plant fermentation product and a preparation method and application thereof. Background Art

[0002] In recent years, people's demand for skin care has become increasingly strong, and research on oil control and anti-inflammatory has gradually deepened. According to the various mechanisms of skin oil control and anti-inflammatory, various raw materials with oil control and anti-inflammatory activity have been continuously introduced. With the continuous development of science, the active ingredients and their effects in natural plants are constantly being explored. For example, Platycladus orientalis, Juniperus chinensis, Juniperus chinensis and Selaginella truncatula are included in the 2021 edition of the "Catalogue of Used Cosmetic Raw Materials". According to reports, these four plants all have a variety of flavonoid components, especially quercetin and quercetin. Pharmacological studies have shown that their extracts have anti-inflammatory, antioxidant and antibacterial effects. At present, solvent extraction is usually used to obtain the active ingredients in these natural plants. These traditional extraction methods have the problems of low yield of active ingredients and high energy consumption, and the effect of the obtained extracts is not obvious; some manufacturers use microwave extraction and supercritical CO2 extraction for extraction, but the equipment requirements are high and the cost is high.

[0003] Therefore, how to increase the active ingredients in plant extracts is a technical problem that needs to be solved urgently in this field. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention provides an oil-controlling and soothing plant fermentation product and its preparation method and application, wherein the plant fermentation product has a high content of active ingredients total flavonoids and quercetin, thereby having significant oil-controlling and soothing effects.

[0005] The first aspect of the present invention provides a plant fermentation product, the raw materials for preparing the plant fermentation product include: plant raw materials, milk, glycerin and fermentation strains; the plant raw materials include at least one of Platycladus orientalis leaves, Sabina leaves, Juniperus leaves, and Selaginella leaves; the fermentation strains include at least one of Lactobacillus plantarum, Lactobacillus lactis, and Bifidobacterium longum.

[0006] According to some embodiments of the present invention, the ratio of the plant raw material, the milk and the glycerol is (0.01-50g): (0.01-50g): 1g. Further, the ratio of the plant raw material, the milk and the glycerol is (0.01-10g): (0.01-10g): 1g. Further, the ratio of the plant raw material, the milk and the glycerol is (0.1-1g): (0.1-1g): 1g.

[0007] According to some embodiments of the present invention, the raw materials for preparing the plant fermentation product also include water.

[0008] According to some embodiments of the present invention, the ratio of the glycerol to the water is 1 g: (1-1000 g). Further, the ratio of the glycerol to the water is 1 g: (1-100 g). Further, the ratio of the glycerol to the water is 1 g: (10-50 g).

[0009] According to some embodiments of the present invention, the plant fermentation product contains active ingredients, including total flavonoids of >1 mg / mL and quercetin of >80 μg / mL. Further, the active ingredients include total flavonoids of 1-3 mg / mL and quercetin of 80-150 μg / mL.

[0010] According to some embodiments of the present invention, the concentration of the fermentation strain is 10 5 -10 10 CFU / mL. Further, the concentration of the fermentation strain is 10 8 -10 10 CFU / mL.

[0011] The second aspect of the present invention provides a method for preparing the above-mentioned plant fermentation product, comprising the following steps:

[0012] The plant raw material, the milk and the glycerin are mixed and homogenized under high pressure to obtain a plant composite liquid;

[0013] Mixing the plant complex liquid and the fermentation strain, and fermenting to obtain the plant fermentation product;

[0014] The pressure of the high-pressure homogenization is 20-100 MPa.

[0015] According to some embodiments of the present invention, the pressure of the high-pressure homogenization is 50-100 MPa.

[0016] According to some embodiments of the present invention, the temperature of the high pressure homogenization is 25-50°C, and the time of the high pressure homogenization is 5-40 minutes. Further, the temperature of the high pressure homogenization is 30-50°C, and the time of the high pressure homogenization is 15-20 minutes.

[0017] According to some embodiments of the present invention, before the fermentation, a culture medium is added; the usage ratio of the plant composite liquid, the fermentation strain and the culture medium is (1-500 g): (0.01-10 mL): 1 g. Further, the usage ratio of the plant composite liquid, the fermentation strain and the culture medium is (1-100 g): (0.1-5 mL): 1 g. Further, the usage ratio of the plant composite liquid, the fermentation strain and the culture medium is (20-80 g): (0.1-1 mL): 1 g.

[0018] According to some embodiments of the present invention, the culture medium is MRS medium.

[0019] According to some embodiments of the present invention, the fermentation temperature is 20-45°C, and the fermentation time is 12-120 hours. Further, the fermentation temperature is 30-40°C, and the fermentation time is 72-120 hours.

[0020] According to some embodiments of the present invention, after the fermentation, the method further comprises sterilizing the fermentation broth.

[0021] According to some embodiments of the present invention, the sterilization temperature is 100-150° C., and the sterilization time is 10-30 min.

[0022] According to some embodiments of the present invention, before the high-pressure homogenization, the process further includes mixing water with the plant material, the milk and the glycerin.

[0023] The third aspect of the present invention provides the use of the plant fermentation product in the preparation of cosmetics or external skin preparations.

[0024] A fourth aspect of the present invention provides a cosmetic comprising the above-mentioned plant fermentation product.

[0025] According to some embodiments of the present invention, the cosmetic further comprises an auxiliary material acceptable in cosmetics.

[0026] According to some embodiments of the present invention, the acceptable auxiliary materials in the cosmetics include at least one of an emulsifier, a moisturizer, a thickener, and a preservative.

[0027] According to some embodiments of the present invention, the amount of the plant fermentation product added to the cosmetic is 0.01-80wt%. Further, the amount of the plant fermentation product added to the cosmetic is 0.05-20wt%. Further, the amount of the plant fermentation product added to the cosmetic is 0.1-10wt%.

[0028] According to some embodiments of the present invention, the cosmetic comprises shampoo, shower gel or facial cleanser.

[0029] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0030] The present invention adopts specific plant raw materials, fermentation strains, as well as milk and glycerin as preparation raw materials. The plant fermentation product obtained by fermentation has a high content of active ingredients total flavonoids and quercetin, wherein the content of total flavonoids reaches 1-3 mg / mL, and the content of quercetin reaches 80-150 μg / mL, and has significant oil-controlling and soothing effects.

[0031] The present invention uses milk and glycerin as an auxiliary in the high-pressure homogenization process to fully break up plant tissue cells and improve the dissolution rate of active ingredients. It also organically combines the technology with directional microbial fermentation, uses specific strains for fermentation, and uses the plant composite stock solution obtained by high-pressure homogenization as the subsequent fermentation matrix. Through the biotransformation ability of microorganisms, the content of total flavonoids and quercetin in the plant fermentation product is finally increased, so that the plant fermentation product of the present invention has significant oil-controlling and soothing effects. DETAILED DESCRIPTION

[0032] In order to make the technical scheme of the present invention more clearly understood by those skilled in the art, the following embodiments are listed for illustration. It should be pointed out that the following embodiments do not limit the protection scope of the present invention.

[0033] Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods.

[0034] Example 1

[0035] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, milk, glycerol and Lactobacillus plantarum liquid.

[0036] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0037] 1) Mix 10 g of Platycladus orientalis leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite solution;

[0038] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0039] Example 2

[0040] The raw materials for preparing plant fermentation products include: juniper leaves, milk, glycerin and plant lactobacillus liquid.

[0041] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0042] 1) After mixing 10 g of juniper leaves, 20 g of milk, 20 g of glycerin, and 360 g of deionized water, the mixture was added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0043] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0044] Example 3

[0045] The raw materials for preparing plant fermentation products include: juniper leaves, milk, glycerin and plant lactobacillus liquid.

[0046] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0047] 1) 10 g of juniper leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water were mixed, and then added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0048] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0049] Example 4

[0050] The raw materials for preparing plant fermentation products include: Selaginella leaves, milk, glycerol and Lactobacillus plantarum liquid.

[0051] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0052] 1) 10 g of Selaginella leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water were mixed, and then added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0053] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 108 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0054] Example 5

[0055] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, Sabina juniper leaves, milk, glycerol and Lactobacillus plantarum liquid.

[0056] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0057] 1) Mix 5 g of Platycladus orientalis leaves, 5 g of Sabina juniper leaves, 20 g of milk, 20 g of glycerin, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite solution;

[0058] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0059] Example 6

[0060] The raw materials for preparing plant fermentation products include: juniper leaves, selaginella leaves, milk, glycerol and Lactobacillus plantarum liquid.

[0061] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0062] 1) Mix 5 g of juniper leaves, 5 g of Selaginella leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite liquid;

[0063] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0064] Example 7

[0065] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, Sabina juniper leaves, Juniperus chinensis leaves, Selaginella leaves, milk, glycerol and Lactobacillus plantarum bacterial liquid.

[0066] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0067] 1) Mix 2.5 g of Platycladus orientalis leaves, 2.5 g of Sabina juniper leaves, 2.5 g of Juniperus chinensis leaves, 2.5 g of Selaginella truncatula leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add them into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite liquid;

[0068] 2) 200g of plant compound solution and 2mL of plant lactobacillus solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 30°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0069] Example 8

[0070] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, milk, glycerol and lactobacillus lactic acid bacterial liquid.

[0071] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0072] 1) Mix 10 g of Platycladus orientalis leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite solution;

[0073] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0074] Example 9

[0075] The raw materials for preparing plant fermentation products include: juniper leaves, milk, glycerin and lactobacillus lactic acid liquid.

[0076] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0077] 1) After mixing 10 g of juniper leaves, 20 g of milk, 20 g of glycerin, and 360 g of deionized water, the mixture was added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0078] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0079] Example 10

[0080] The raw materials for preparing plant fermentation products include: juniper leaves, milk, glycerin and lactobacillus lactic acid liquid.

[0081] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0082] 1) 10 g of juniper leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water were mixed, and then added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0083] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0084] Embodiment 11

[0085] The raw materials for preparing plant fermentation products include: Selaginella leaves, milk, glycerol and lactobacillus lactic acid bacterial liquid.

[0086] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0087] 1) 10 g of Selaginella leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water were mixed, and then added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0088] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0089] Example 12

[0090] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, Sabina juniper leaves, milk, glycerol and Lactobacillus lactis liquid.

[0091] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0092] 1) Mix 5 g of Platycladus orientalis leaves, 5 g of Sabina juniper leaves, 20 g of milk, 20 g of glycerin, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite solution;

[0093] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0094] Embodiment 13

[0095] The raw materials for preparing plant fermentation products include: juniper leaves, selaginella leaves, milk, glycerol and lactobacillus lactis liquid.

[0096] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0097] 1) Mix 5 g of juniper leaves, 5 g of Selaginella leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite liquid;

[0098] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0099] Embodiment 14

[0100] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, Sabina juniper leaves, Juniperus chinensis leaves, Selaginella leaves, milk, glycerol and Lactobacillus lactis liquid.

[0101] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0102] 1) Mix 2.5 g of Platycladus orientalis leaves, 2.5 g of Sabina juniper leaves, 2.5 g of Juniperus chinensis leaves, 2.5 g of Selaginella truncatula leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add them into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite liquid;

[0103] 2) 200g of plant compound solution and 2mL of Lactobacillus lactis solution (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 33°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was obtained to obtain the plant fermentation product.

[0104] Embodiment 15

[0105] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, milk, glycerol and Bifidobacterium longum bacterial liquid.

[0106] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0107] 1) Mix 10 g of Platycladus orientalis leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite solution;

[0108] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0109] Example 16

[0110] The raw materials for preparing plant fermentation products include: juniper leaves, milk, glycerol and bifidobacterium longum bacterial liquid.

[0111] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0112] 1) After mixing 10 g of juniper leaves, 20 g of milk, 20 g of glycerin, and 360 g of deionized water, the mixture was added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0113] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0114] Embodiment 17

[0115] The raw materials for preparing plant fermentation products include: juniper leaves, milk, glycerol and bifidobacterium longum bacterial liquid.

[0116] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0117] 1) 10 g of juniper leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water were mixed, and then added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0118] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0119] Embodiment 18

[0120] The raw materials for preparing plant fermentation products include: Selaginella leaves, milk, glycerol and Bifidobacterium longum bacterial liquid.

[0121] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0122] 1) 10 g of Selaginella leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water were mixed, and then added into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 15 min to obtain a plant composite liquid;

[0123] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0124] Embodiment 19

[0125] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, Sabina juniper leaves, milk, glycerol and Bifidobacterium longum bacterial liquid.

[0126] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0127] 1) Mix 5 g of Platycladus orientalis leaves, 5 g of Sabina juniper leaves, 20 g of milk, 20 g of glycerin, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite solution;

[0128] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0129] Embodiment 20

[0130] The raw materials for preparing plant fermentation products include: juniper leaves, selaginella leaves, milk, glycerol and bifidobacterium longum bacterial liquid.

[0131] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0132] 1) Mix 5 g of juniper leaves, 5 g of Selaginella leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add the mixture into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite liquid;

[0133] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0134] Embodiment 21

[0135] The raw materials for preparing plant fermentation products include: Platycladus orientalis leaves, Sabina juniper leaves, Juniperus chinensis leaves, Selaginella leaves, milk, glycerol and Bifidobacterium longum bacterial liquid.

[0136] The method for preparing the plant fermentation product of this embodiment comprises the following steps:

[0137] 1) Mix 2.5 g of Platycladus orientalis leaves, 2.5 g of Sabina juniper leaves, 2.5 g of Juniperus chinensis leaves, 2.5 g of Selaginella truncatula leaves, 20 g of milk, 20 g of glycerol, and 360 g of deionized water, and then add them into a high-pressure homogenizer for high-pressure homogenization at a homogenization temperature of 40° C., a homogenization pressure of 80 MPa, and a homogenization time of 20 min to obtain a plant composite liquid;

[0138] 2) 200g of plant compound liquid and 2mL of Bifidobacterium longum liquid (concentration of 10 8 CFU / mL) and 4g MRS medium were mixed and fermented at 37°C for 72h. After the fermentation, the fermentation liquid was sterilized at 121°C for 20min, and the sterilized fermentation liquid was centrifuged at a speed of 6000rpm / min for 20min. The supernatant after centrifugation was taken to obtain the plant fermentation product.

[0139] Comparative Example 1 (the difference from Example 7 is only that conventional water is used for decoction)

[0140] A method for preparing a composite extract comprises the following steps:

[0141] Mix 2.5 g of Platycladus orientalis leaves, 2.5 g of Sabina leaf, 2.5 g of Juniperus chinensis leaf, 2.5 g of Selaginella leaf, 20 g of milk, 20 g of glycerol and 360 g of deionized water, heat and extract at 100° C. for 1 h, filter and collect the filtrate to obtain a composite extract.

[0142] Comparative Example 2

[0143] The only difference between this comparative example and Example 7 is that the milk and glycerol in step 1) are replaced by deionized water, and the added amounts are the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0144] Comparative Example 3

[0145] The only difference between this comparative example and Example 7 is that the milk in step 1) is replaced by honey, and the added amount is the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0146] Comparative Example 4

[0147] The only difference between this comparative example and Example 7 is that the glycerol in step 1) is replaced by deionized water, and the added amount is the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0148] Comparative Example 5

[0149] The only difference between this comparative example and Example 7 is that the milk in step 1) is replaced by lecithin, and the added amount is the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0150] Comparative Example 6

[0151] The only difference between this comparative example and Example 7 is that glycerol in step 1) is replaced by butanediol, and the added amount is the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0152] Comparative Example 7

[0153] The only difference between this comparative example and Example 7 is that the plant lactobacillus liquid in step 2) is replaced with deionized water, and the added amount is the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0154] Comparative Example 8

[0155] The only difference between this comparative example and Example 7 is that the plant lactobacillus liquid in step 2) is replaced by Monascus liquid, and the added amount is the same. Other raw materials, added amounts and preparation methods are the same as those in Example 7.

[0156] Test Example 1: Determination of total flavonoid content

[0157] This test example tests the total flavonoid content in the plant fermentation products of Examples 1-21 and the plant fermentation products or composite extracts of Comparative Examples 1-8 (hereinafter referred to as "samples"). The specific test method is as follows:

[0158] Adjust the total flavonoid concentration of the sample solution to the measurement range, accurately draw 2.0mL and place it in a dry and clean test tube, add 0.2mL of 5% sodium nitrite solution, shake well and place for 5 minutes, add 0.2mL of 10% aluminum nitrate solution and shake well, place for 6 minutes, add 1.0mL of 5% sodium hydroxide solution, add solvent to the scale, and shake the sample solution. Repeat 3 times for each concentration. Rapidly measure the absorbance value of each tube of the sample solution at a wavelength of 510nm. According to the standard curve of rutin solution, the concentration of total flavonoids is calculated from the absorbance value of the sample solution, and the total flavonoid content in the sample is expressed as rutin.

[0159] Test Example 2: Determination of the content of quercetin

[0160] This test example tests the quercetin content in the plant fermentation products of Examples 1-21 and the plant fermentation products or composite extracts of Comparative Examples 1-8 (hereinafter referred to as "samples"). The specific test method is as follows:

[0161] Preparation of test samples: Take 5 mL of sample, add 5 mL of chromatography-grade acetonitrile, mix well, centrifuge at 6000 rpm / min for 20 min, take the supernatant, and filter with a 0.45 μm filter membrane to obtain the test sample.

[0162] Chromatographic conditions: Use an Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm); use 0.1% phosphoric acid (A)-acetonitrile (B) as the mobile phase for gradient elution (0 min, 15% B; 10 min, 30% B; 19 min, 30% B), flow rate 1.0 mL / min; detection wavelength 254 nm; column temperature 25 ° C.

[0163] The test results of Experimental Example 1 and Experimental Example 2 are shown in Table 1.

[0164] Table 1

[0165]

[0166]

[0167] As shown in Table 1, compared with Comparative Examples 1-8, the total flavonoid content in the samples of Examples 1-21 is greatly improved, which may be due to the fact that the microorganisms can efficiently separate plant tissues, making it easier to dissolve the active ingredients; at the same time, the addition of milk and glycerol in the pretreatment helps to dissolve the low-polarity flavonoid components, thereby increasing the total flavonoid content in the samples. In addition, the content of quercetin is also significantly increased, which may be achieved by the active enzymes present in the microorganisms, which transform other components.

[0168] Experimental Example 3: Cell scratch test

[0169] This test example evaluates the cell repair ability and the effect on cell migration of the plant fermented products of Examples 1-21 and the plant fermented products or composite extracts of Comparative Examples 1-8 (hereinafter referred to as "samples"). The specific test method is as follows:

[0170] Use a pen to evenly draw horizontal lines on the back of the 6-well plate, with an interval of about 0.5-1cm between the lines. Fill the wells with human immortalized epidermal cells (HaCaT). The next day, when the cells fill the wells, use a gun tip to scratch the cells in the wells according to the horizontal lines pre-painted on the 6-well plate, wash the cells 3 times with PBS, remove the scratched cells, add cell culture medium containing 1% (w / w) sample, culture in a 37°C, 5% CO2 incubator for 24 hours, take pictures, and count the 24-hour scratch healing ratio. Replace the sample with deionized water as a blank control group. The evaluation results are shown in Table 2.

[0171] Table 2

[0172]

[0173]

[0174] As shown in Table 2, compared with Comparative Examples 1-8, the samples of Examples 1-21 can more effectively improve the cell migration and repair ability, which indicates that the plant fermentation obtained by the preparation method of the present invention has a good repair effect on epidermal cells. The leaves of Platycladus orientalis, Sabina juniper, Juniperus chinensis, and Selaginella truncatula have not been treated with milk, glycerol, and microbial fermentation, and the products obtained may contain components that affect cell vitality, resulting in a significant reduction in the repair effect of the products on epidermal cells.

[0175] Experimental Example 4: Effects on IL-6 and TNF-α levels in HaCaT cells after UV irradiation

[0176] Human epidermal keratinocytes were seeded into 6-well plates and incubated overnight in an incubator at 37°C and 5% CO2. When the cell plating rate in the 6-well plate reached 50%-60%, cell culture medium was added to the blank group without other treatments; cell culture medium was added to the model group, and cell culture medium containing 1% (w / w) sample was added to the test sample group. After pretreatment for 1 hour, UVB (medium-wave ultraviolet) irradiation was performed for 40 minutes, and the radiation intensity was 30mJ / cm 2 After treatment, the cells were cultured in an incubator at 37°C and 5% CO2 for 24 hours. After the culture, the IL-6 and TNF-α contents were determined according to the Human IL-6 ELISA Kit (Biyuntian) and Human TNF-α ELISA Kit (Biyuntian). The test results are shown in Table 3.

[0177] Table 3

[0178]

[0179]

[0180] The results in Table 3 show that compared with Comparative Examples 1-8, the plant fermentation products prepared in Examples 1-21 can more effectively reduce the levels of inflammatory factors IL-6 and TNF-α produced by HaCaT cells after UV irradiation, showing anti-inflammatory and soothing activity.

[0181] Test Example 5: Effects on Staphylococcus aureus and Malassezia

[0182] Preparation of bacterial suspension: Staphylococcus aureus (ATCC6538) and Malassezia furfur (ATCC14521) were selected as test bacteria. Malassezia furfur is a conditionally pathogenic fungus that reproduces in large numbers under the condition of vigorous secretion of sebaceous glands and can cause folliculitis, tinea versicolor, etc. It is known to be closely related to dandruff production and scalp health; Staphylococcus aureus is also a common pathogenic bacteria of the skin, which can usually cause papules or pustules on the skin. The test bacteria were prepared into bacterial suspensions with PBS at a concentration of 5×10 6 CFU / mL.

[0183] Bacterial inhibition zone test: The Oxford cup method was used for the bacterial inhibition zone test. 100 μL of bacterial suspension was absorbed on the surface of the solidified culture medium, spread evenly with a sterile coating stick, covered with a flat dish, and placed at 20°C for 10 minutes; finally, the sterile Oxford cup was gently inverted on the surface of the culture medium, and sample solutions of different mass fractions were added until the liquid surface was parallel to the surface of the Oxford cup, and placed in an incubator at 37°C for 18 hours. Refer to the enterprise standard "Microbial Antibacterial Evaluation Value In Vitro Inhibition of Bacteria and Fungi Test Operation Guide (Drug Sensitivity Test)" for the criteria for determining the results of the bacterial inhibition zone: the diameter of the bacterial inhibition zone > 20 mm is extremely sensitive, expressed as ++++; 15-20 mm is highly sensitive, expressed as +++; 10-15 mm is moderately sensitive, expressed as ++; < 10 mm is low sensitivity, expressed as +; 0 mm is no antibacterial effect, expressed as -. The test results are shown in Table 4.

[0184] Table 4

[0185]

[0186] From the test results of Table 4, it can be seen that the samples of Examples 1-21 can inhibit Staphylococcus aureus and Malassezia furfur at a mass fraction of 5% or 1%. Compared with the samples of Comparative Examples 1-8, the inhibition of Staphylococcus aureus and Malassezia furfur at the same concentration has a stronger effect. The samples of Examples 1-21 have a good inhibitory effect on pathogenic bacteria, indicating that they can be used to maintain a normal and healthy skin microecology.

[0187] Test Example 6: Evaluation of red blood cell hemolysis

[0188] This test example evaluates the hemolytic activity of red blood cells on the plant fermented products of Examples 1-21 above, and the plant fermented products or composite extracts of Comparative Examples 1-8 (hereinafter referred to as "samples"). The basic principle of the red blood cell hemolysis experiment is to evaluate the damage of chemicals to eye tissue cells by measuring the amount of hemoglobin dissolution and the degree of denaturation. This experiment is in accordance with the experimental test methods and grading standards of the European Center for Validation of Alternative Methods. HD50 is the sample concentration when 50% of the red blood cells are hemolyzed, DI is the protein denaturation index, and L / D is the ratio of HD50 to DI. The evaluation criteria are shown in Table 5, and the evaluation results are shown in Table 6.

[0189] Table 5

[0190] L / D Grading L / D>100 Non-irritating 10<L / D≤100 Slightly irritating 1<L / D≤10 Mild irritant 0.1<L / D≤1 Toxic irritation L / D≤0.1 Severe irritation

[0191] Table 6

[0192]

[0193]

[0194] It can be seen from Table 6 that, except for Comparative Example 8 which is slightly irritating, the materials before and after fermentation and used for fermentation are non-irritating, and the plant fermented products or composite extracts of Examples 1-21 and Comparative Examples 1-7 are relatively mild raw materials.

[0195] Application Example 1

[0196] This application example provides a shampoo, which includes the plant fermentation product of Example 7. The formula thereof is shown in Table 7 in terms of mass percentage. The preparation method of the shampoo of Application Example 1 is prepared according to conventional preparation methods in the art.

[0197] Table 7

[0198]

[0199]

[0200] Application Comparative Example 1

[0201] The difference between Application Example 1 and Application Example 1 is that in Application Example 1, the plant fermentation product of Example 7 is replaced by the composite extract of Comparative Example 1, and the other ingredients, dosages and preparation methods are the same as those of Application Example 1.

[0202] Test Example 7: Evaluation of the anti-dandruff and soothing effects of shampoo

[0203] 60 volunteers were selected and randomly divided into 2 groups, aged between 20 and 50 years old, half male and half female. Group 1 used 5g of shampoo of Application Example 1 on the 1st, 3rd, 5th and 7th days respectively; Group 2 used 5g of shampoo of Comparative Example 1 on the 1st, 3rd, 5th and 7th days respectively. The subjects were revisited on the 8th day to assess the effects, and the subjects scored according to their feelings of use. The scoring standard uses a 10-point system, and the higher the score, the better the feeling of use. Improvement of scales (0 points for no improvement, 10 points for obvious improvement); improvement of itching (0 points for no improvement, 10 points for obvious improvement); overall feeling of use (0 points for very poor, 10 points for very good). The test results are shown in Table 8.

[0204] Test Example 8: Evaluation of the Oil Control Effect of Shampoo

[0205] Recruit 20 volunteers with vigorous scalp oil secretion, aged between 20-30 years old. If the volunteers are female, avoid the menstrual period, have a regular life and work schedule, and try to avoid strenuous exercise, sun exposure, staying up late, etc. Do not wash your hair the night before the test. Before the test, the volunteers sit quietly in a room with a temperature of 20-22℃ and a humidity of 40%-65% for 30 minutes, and cannot drink water or beverages during this period. The test site is exposed, placed in the test state, and stay relaxed. The test time points are before sample use, immediately after sample use, 6 hours, and 8 hours. The test site is the 5cm area from the forehead to the top of the head, and a 1cm×1cm square is drawn on the test area with a skin pen. When using the sample, use half of the head to wash your hair. Divide the head along the hairline in the middle, half of which is the blank control, and wash your hair with clean water. The other half is the sample area, and the test sample is washed with the test sample. Bend the tape into a ring and make it contact with the scalp. The lipid compounds (oils) in the scalp will change the transparency of the test tape. Then flatten the test tape and put it in the slider of the scalp oil tester, and push the slider into the instrument. The transmittance distribution curve on the tape can be tested by the photoelectric tube in the instrument. The peak value of the transmittance curve distribution represents the content of lipid compounds, i.e., oils, in the scalp. During the test, the annular top of the tape is exactly the part that contacts the scalp. The calculation of the mean difference of scalp oil is that the mean difference of scalp oil between the blank control group and the sample group at different time periods = the scalp oil content of the blank control group - the scalp oil content of the sample group. The test results are shown in Table 9.

[0206] Table 8

[0207] sample Anti-dandruff effect Anti-itching effect Overall use experience Example 7 8.6 8.0 8.5 Comparative Example 1 6.2 6.4 7.3

[0208] Table 9

[0209]

[0210] As shown in Table 8, compared with the shampoo of Comparative Example 1, the shampoo of Example 1 with the plant fermentation product of Example 7 added thereto had obvious anti-dandruff and anti-itching effects on the volunteers after use, and the overall feeling of use was also greatly improved. This indicates that the plant fermentation product of Example 7 can not only inhibit the production of IL-6 and TNF-α, but also inhibit Staphylococcus aureus and Malassezia furfur, and the anti-dandruff and anti-itching effects of the shampoo are related to the anti-inflammatory and antibacterial effects of the raw materials.

[0211] In addition, it can be seen from Table 9 that the shampoo to which the plant fermentation product of Example 7 is added can slow down the secretion of scalp oil and achieve an oil control effect.

[0212] The preferred embodiments of the present invention are specifically described above, but the invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A plant fermentation product, characterized in that: The raw materials for preparing the plant fermentation product include: plant raw materials, milk, glycerin and fermentation strains; the plant raw materials are at least one of Platycladus orientalis leaves, Sabina juniper leaves, Juniperus chinensis leaves and Selaginella leaves; the fermentation strains are at least one of Lactobacillus plantarum, Lactobacillus lactis and Bifidobacterium longum; The usage ratio of the plant raw material, the milk and the glycerol is (0.01-50g): (0.01-50g): 1g; The plant fermentation product is prepared by a preparation method comprising the following steps: The plant raw material, the milk and the glycerin are mixed and homogenized under high pressure to obtain a plant composite liquid; Mixing the plant complex liquid and the fermentation strain, and fermenting to obtain the plant fermentation product; The pressure of the high-pressure homogenization is 20-100 MPa.

2. The plant fermentation product according to claim 1, characterized in that The plant fermentation product contains active ingredients, which include total flavonoids in an amount greater than 1 mg / mL and quercetin in an amount greater than 80 μg / mL.

3. The plant fermentation product according to claim 1, characterized in that The concentration of the fermentation strain is 10 5 -10 10 CFU / mL.

4. The plant fermentation product according to claim 1, characterized in that Before the fermentation, a culture medium is added; the usage ratio of the plant complex liquid, the fermentation strain and the culture medium is (1-500 g): (0.01-10 mL): 1 g.

5. The plant fermentation product according to claim 1, characterized in that The fermentation temperature is 20-45°C, and the fermentation time is 12-120h.

6. Use of the plant fermentation product according to any one of claims 1 to 5 in the preparation of cosmetics or external skin preparations.

7. A cosmetic, characterized in that: The invention comprises the plant fermentation product according to any one of claims 1 to 5.

8. The cosmetic according to claim 7, characterized in that: The cosmetics include shampoo, shower gel or facial cleanser.

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