Multi-fermented oil and application thereof

Abstract

The invention relates to the technical field of daily chemicals, in particular to multi-fermented oil and application thereof. The multi-fermented oil is prepared from Tibetan cordyceps sinensis, eurotium cristatum and saccharomycetes. The multi-fermented oil provided by the invention contains functional components and mild components, and contains rich fatty acids, antioxidant components and other substances with biological activity. A lipid structure in the fermented oil is similar to a natural sebum layer of skin, so that a protective barrier can be formed on the surface of the skin, and water loss is prevented. This is particularly beneficial to dry skin and helps the skin maintain moist and soft. And micromolecular lipid components in the fermented oil, such as unsaturated fatty acid, can more easily permeate into the deep layer of the skin, so that long-acting nourishing and moisturizing effects are provided for skin cells.

Description

Technical Field

[0001] The invention relates to the technical field of daily chemicals, in particular to multiple fermentation oil and application thereof. Background Art

[0002] Geotrichum candidum is a common filamentous fungus that is widely found in nature. It belongs to the genus Geotrichum and is well known in the fermentation and food industries for its unique metabolic properties and applications. Geotrichum candidum has strong enzyme activity, especially lipase and protease, which can break down fats and proteins and release small molecular fatty acids and amino acids.

[0003] Yeast is a type of single-celled fungus that is widely used in food fermentation, medicine and biotechnology. There are many types of yeast, among which Saccharomyces cerevisiae is one of the most common types, named for its wide use in processes such as brewing and bread fermentation. Yeasts are widely found in nature, especially on plant surfaces, in soil, in fruits and in insects. They usually feed on organic matter and grow and reproduce in an environment rich in sugars. In industrial fermentation and research, yeasts are usually propagated and their metabolism studied under laboratory culture conditions.

[0004] Solid State Aerobic Fermentation (SSAF) is a fermentation technology that uses microorganisms to perform aerobic metabolism on a solid matrix (solid material). Unlike traditional liquid fermentation, solid-state fermentation does not require a large amount of liquid culture medium. Instead, it inoculates microorganisms on solid raw materials and allows them to grow and produce metabolites in an aerobic environment.

[0005] Fermented oil refers to oil or lipid substances produced by microbial fermentation. Different from the traditional method of extracting oil from plants, fermented oil is a biotechnology that synthesizes and accumulates oil through the metabolism of specific microorganisms. Summary of the invention

[0006] The invention provides a multiple fermentation oil, comprising the multiple fermentation oil whose preparation raw materials are Tibetan cordyceps sinensis, golden flower fungus and yeast.

[0007] As an embodiment of the present invention, the preparation method of the multiple fermentation oil is as follows:

[0008] (1) Pretreatment: pre-treating the Tibetan Cordyceps sinensis;

[0009] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0010] (3) Multiple fermentation: the product after solid-state aerobic fermentation is mixed with camellia seeds and then inoculated with yeast for multiple fermentation;

[0011] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0012] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1 and then centrifuged at 9000 rpm for 5 min to obtain the multiple fermentation oil.

[0013] As an embodiment of the present invention, the pre-treatment step in step (1) is: wash and dry the Tibetan Cordyceps sinensis, chop it and pass it through a 10-mesh sieve for standby use.

[0014] As an embodiment of the present invention, the golden flower fungus in step (2) is Eurotium cristatum.

[0015] As an embodiment of the present invention, the solid matrix in step (2) is soybeans and mung beans; and the average diameter of the soybeans is about 6 to 8 mm, and the average diameter of the mung beans is about 4 to 6 mm.

[0016] As an embodiment of the present invention, in the step (2), the water content of the solid-state aerobic fermentation is about 30-40 wt %, the fermentation temperature is 30° C., and the fermentation environment humidity is 35-40% RH.

[0017] As an embodiment of the present invention, in the step (3), the camellia seeds and the product after solid-state aerobic fermentation are mixed and then left to stand at 30° C. for 24 hours.

[0018] As an embodiment of the present invention, the yeast is Candida krusei.

[0019] As an embodiment of the present invention, in the step (3), the camellia seeds are dried and ground before being mixed with the product after solid-state aerobic fermentation.

[0020] As an embodiment of the present invention, the multiple fermentation oil is applied in the field of daily chemicals.

[0021] By adopting the above technical solution, the present invention has the following beneficial effects:

[0022] The multi-fermented oil provided by the present invention comprises functional ingredients and mild ingredients, and is rich in fatty acids, antioxidant ingredients and other biologically active substances. The lipid structure in the fermented oil is similar to the natural sebum layer of the skin, and can form a protective barrier on the skin surface to prevent water loss. This is particularly beneficial for dry skin, helping the skin to remain moisturized and soft. The small molecule lipid components in the fermented oil, such as unsaturated fatty acids, can more easily penetrate into the deep layers of the skin, providing long-lasting nourishing and moisturizing effects for skin cells.

[0023] Fermented oil is rich in unsaturated fatty acids (such as linoleic acid, linolenic acid, oleic acid, etc.), which help repair the skin's barrier function and restore its natural defense mechanism. Fermented oil can nourish the stratum corneum and help it restore its normal moisture retention and protective function, thereby reducing problems such as rough skin and peeling.

[0024] Oils obtained through microbial fermentation are often rich in powerful antioxidants such as vitamin E, coenzyme Q10, and plant polyphenols. These ingredients can neutralize free radicals, reduce damage to the skin caused by external environments such as ultraviolet rays and pollution, and prevent skin aging caused by oxidative stress.

[0025] The active ingredients in fermented oil can stimulate the repair and regeneration of skin cells, help reduce fine lines and wrinkles, improve skin elasticity, and make the skin firmer and younger. Fermented oil has good anti-inflammatory effects and can effectively relieve skin symptoms such as redness, sensitivity, and itching. Its mild nature is suitable for all skin types, especially sensitive skin. DETAILED DESCRIPTION

[0026] The technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] The invention provides a multiple fermentation oil, comprising the multiple fermentation oil whose preparation raw materials are Tibetan cordyceps sinensis, golden flower fungus and yeast.

[0028] As an embodiment of the present invention, the preparation method of the multiple fermentation oil is as follows:

[0029] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 10-mesh sieve for later use;

[0030] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0031] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0032] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 6-8 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:1; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 30wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 40%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0033] (3) Multiple fermentation: The product after solid-state aerobic fermentation was mixed with camellia seeds and allowed to stand at 30° C. for 24 hours, and then yeast and culture medium X were inoculated for multiple fermentation; the yeast was Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0034] Before mixing with the product after solid-state aerobic fermentation, the camellia seeds were stir-fried at 100° C. for 10 minutes, ground and crushed, and passed through a 100-mesh sieve, and then mixed with the product after solid-state aerobic fermentation, which was recorded as mixture A.

[0035] The yeast was inoculated into the medium X, which was a 5° Bé wort medium. The inoculation amount of the yeast in the medium X was 10 vol%. After the yeast was aerobically cultured in the medium X at 28° C. for 24 h, the mixture of the yeast and the medium X was mixed with the mixture A (mass ratio was 1:2) for multiple fermentation, and the fermentation time was 72 h, aerobically, at 28° C.

[0036] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0037] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0038] As an embodiment of the present invention, the multiple fermentation oil of the present invention is applied in the field of daily chemicals.

[0039] As a preferred embodiment of the present invention, the present invention provides a whitening essential oil, the raw materials for its preparation include:

[0040] Mineral oil 57.37 parts

[0041] 5 servings of multi-fermented oil

[0042] 25 parts of ethylhexyl palmitate

[0043] 0.5 parts of squalane

[0044] 10 parts of Cetyl Ethylhexanoate

[0045] 0.8 parts of ascorbyl tetraisopalmitate

[0046] Tocopherol acetate 0.5 parts

[0047] 0.5 parts of bisabolol

[0048] 0.11 parts of Limnanthes Alba seed oil

[0049] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0050] Macadamia ternifolia seed oil 0.11 parts.

[0051] Cordyceps sinensis contains a lot of mannitol, which is sensitive to humidity and easily absorbs moisture and agglomerates, which will reduce the fermentation effect for solid-state aerobic fermentation. The present invention grinds the Cordyceps sinensis into fine powder before treatment to increase its surface area, which helps to prevent agglomeration. However, it is easy to accumulate after being ground into powder. Therefore, the present invention controls the water content of Cordyceps sinensis during solid-state aerobic fermentation, and adds soybeans and mung beans as a matrix to the solid matrix for ventilation. The temperature and humidity environment are adjusted to obtain a better solid aerobic fermentation effect; however, the subsequent oil production effect is not good, because soybeans can provide nitrogen and carbon sources at the same time. When the nitrogen source is abundant, yeast bacteria focus on growth rather than conversion into storage substances. For this reason, the present invention adds more carbon sources (camellia seeds) when producing oil; at the same time, it is found that the effect is better than directly adding camellia seed oil, because golden flower fungi can consume more amino acids to rapidly reduce the content of protein and amino acids; at the same time, camellia seeds also contain protein, minerals, vitamin E, tea saponin and other ingredients, which can alleviate the discoloration problem (the active ingredients in Cordyceps sinensis oil (such as unsaturated fatty acids, steroids, polysaccharides, etc.) are easily affected by oxygen and undergo oxidation reactions. Oxidation usually causes the color of the oil to gradually darken, from the original light yellow and golden yellow to brown or dark brown).

[0052] The present invention is further explained below in conjunction with specific implementation modes.

[0053] Example 1

[0054] This embodiment provides a multiple fermentation oil, and the preparation method of the multiple fermentation oil is as follows:

[0055] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 10-mesh sieve for later use;

[0056] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0057] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0058] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 6-8 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:1; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 30wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 40%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0059] (3) Multiple fermentation: The product after solid-state aerobic fermentation was mixed with camellia seeds and allowed to stand at 30° C. for 24 hours, and then yeast and culture medium X were inoculated for multiple fermentation; the yeast was Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0060] The ratio of camellia seeds to the product after solid aerobic fermentation is 1:9.

[0061] Before mixing with the product after solid-state aerobic fermentation, the camellia seeds were stir-fried at 100° C. for 10 minutes, ground and crushed, and passed through a 100-mesh sieve, and then mixed with the product after solid-state aerobic fermentation, which was recorded as mixture A.

[0062] The yeast was inoculated into the medium X, which was a 5° Bé wort medium. The inoculation amount of the yeast in the medium X was 10 vol%. After the yeast was aerobically cultured in the medium X at 28° C. for 24 h, the mixture of the yeast and the medium X was mixed with the mixture A (mass ratio was 1:2) for multiple fermentation, and the fermentation time was 72 h, aerobically, at 28° C.

[0063] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0064] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0065] This embodiment also provides a whitening essential oil, the raw materials for its preparation include:

[0066] Mineral oil 57.37 parts

[0067] 5 parts of the multiple fermentation oil described in this example

[0068] 25 parts of ethylhexyl palmitate

[0069] 0.5 parts of squalane

[0070] 10 parts of Cetyl Ethylhexanoate

[0071] 0.8 parts of ascorbyl tetraisopalmitate

[0072] Tocopherol acetate 0.5 parts

[0073] 0.5 parts of bisabolol

[0074] 0.11 parts of Limnanthes Alba seed oil

[0075] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0076] Macadamia ternifolia seed oil 0.11 parts.

[0077] Example 2

[0078] This embodiment provides a multiple fermentation oil, and the preparation method of the multiple fermentation oil is as follows:

[0079] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 100-mesh sieve for later use;

[0080] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0081] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0082] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 6-8 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:1; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 30wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 40%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0083] (3) Multiple fermentation: The product after solid-state aerobic fermentation was mixed with camellia seeds and allowed to stand at 30° C. for 24 hours, and then yeast and culture medium X were inoculated for multiple fermentation; the yeast was Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0084] Before mixing the camellia seeds with the product after solid-state aerobic fermentation, the camellia seeds were stir-fried at 100° C. for 10 minutes, ground and crushed through a 100-mesh sieve, and then mixed with the product after solid-state aerobic fermentation, which was recorded as mixture A. The ratio of camellia seeds to the product after solid-state aerobic fermentation was 1:9.

[0085] The yeast was inoculated into the medium X, which was a 5° Bé wort medium. The inoculation amount of the yeast in the medium X was 10 vol%. After the yeast was aerobically cultured in the medium X at 28° C. for 24 h, the mixture of the yeast and the medium X was mixed with the mixture A (mass ratio was 1:2) for multiple fermentation, and the fermentation time was 72 h, aerobically, at 28° C.

[0086] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0087] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0088] This embodiment also provides a whitening essential oil, the raw materials for its preparation include:

[0089] Mineral oil 57.37 parts

[0090] 5 parts of the multiple fermentation oil described in this example

[0091] 25 parts of ethylhexyl palmitate

[0092] 0.5 parts of squalane

[0093] 10 parts of Cetyl Ethylhexanoate

[0094] 0.8 parts of ascorbyl tetraisopalmitate

[0095] Tocopherol acetate 0.5 parts

[0096] 0.5 parts of bisabolol

[0097] 0.11 parts of Limnanthes Alba seed oil

[0098] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0099] Macadamia ternifolia seed oil 0.11 parts.

[0100] Example 3

[0101] This embodiment provides a multiple fermentation oil, and the preparation method of the multiple fermentation oil is as follows:

[0102] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 10-mesh sieve for later use;

[0103] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0104] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0105] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 4-6 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:2; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 40wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 50%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0106] (3) Multiple fermentation: The product after solid-state aerobic fermentation was mixed with camellia seeds and allowed to stand at 30° C. for 24 hours, and then yeast and culture medium X were inoculated for multiple fermentation; the yeast was Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0107] Before mixing the camellia seeds with the product after solid-state aerobic fermentation, the camellia seeds were stir-fried at 100° C. for 10 minutes, ground and crushed through a 100-mesh sieve, and then mixed with the product after solid-state aerobic fermentation, which was recorded as mixture A. The ratio of camellia seeds to the product after solid-state aerobic fermentation was 1:9.

[0108] The yeast was inoculated into the medium X, which was a 5° Bé wort medium. The inoculation amount of the yeast in the medium X was 10 vol%. After the yeast was aerobically cultured in the medium X at 28° C. for 24 h, the mixture of the yeast and the medium X was mixed with the mixture A (mass ratio was 1:2) for multiple fermentation, and the fermentation time was 72 h, aerobically, at 28° C.

[0109] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0110] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0111] This embodiment also provides a whitening essential oil, the raw materials for its preparation include:

[0112] Mineral oil 57.37 parts

[0113] 5 parts of the multiple fermentation oil described in this example

[0114] 25 parts of ethylhexyl palmitate

[0115] 0.5 parts of squalane

[0116] 10 parts of Cetyl Ethylhexanoate

[0117] 0.8 parts of ascorbyl tetraisopalmitate

[0118] Tocopherol acetate 0.5 parts

[0119] 0.5 parts of bisabolol

[0120] 0.11 parts of Limnanthes Alba seed oil

[0121] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0122] Macadamia ternifolia seed oil 0.11 parts.

[0123] Example 4

[0124] This embodiment provides a multiple fermentation oil, and the preparation method of the multiple fermentation oil is as follows:

[0125] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 10-mesh sieve for later use;

[0126] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0127] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0128] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 6-8 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:1; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 30wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 40%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0129] (3) Multiple fermentation: The product after solid-state aerobic fermentation and camellia oil were mixed and allowed to stand at 30° C. for 24 hours, and then yeast and culture medium X were inoculated for multiple fermentation; the yeast was Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0130] The camellia oil is mixed with the product after solid-state aerobic fermentation, which is recorded as mixture A. The ratio of camellia oil to the product after solid-state aerobic fermentation is 1:9.

[0131] The yeast was inoculated into the medium X, which was a 5° Bé wort medium. The inoculation amount of the yeast in the medium X was 10 vol%. After the yeast was aerobically cultured in the medium X at 28° C. for 24 h, the mixture of the yeast and the medium X was mixed with the mixture A (mass ratio was 1:2) for multiple fermentation, and the fermentation time was 72 h, aerobically, at 28° C.

[0132] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0133] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0134] This embodiment also provides a whitening essential oil, the raw materials for its preparation include:

[0135] Mineral oil 57.37 parts

[0136] 5 parts of the multiple fermentation oil described in this example

[0137] 25 parts of ethylhexyl palmitate

[0138] 0.5 parts of squalane

[0139] 10 parts of Cetyl Ethylhexanoate

[0140] 0.8 parts of ascorbyl tetraisopalmitate

[0141] Tocopherol acetate 0.5 parts

[0142] 0.5 parts of bisabolol

[0143] 0.11 parts of Limnanthes Alba seed oil

[0144] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0145] Macadamia ternifolia seed oil 0.11 parts.

[0146] Example 5

[0147] This embodiment provides a multiple fermentation oil, and the preparation method of the multiple fermentation oil is as follows:

[0148] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 10-mesh sieve for later use;

[0149] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0150] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0151] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 6-8 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:1; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 30wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 40%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0152] (3) Multiple fermentation: The product after solid-state aerobic fermentation is inoculated with yeast and culture medium X for multiple fermentation; the yeast is Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0153] The yeast was inoculated into the culture medium X, which was a 5° Bé wort culture medium. The inoculation amount of the yeast in the culture medium X was 10 vol%. After the yeast was aerobically cultured in the culture medium X at 28° C. for 24 h, the mixture of the yeast and the culture medium X was mixed with the product after solid aerobic fermentation (mass ratio of 1:1.8) for multiple fermentation, and the fermentation time was 72 h, aerobic, 28° C.

[0154] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0155] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0156] This embodiment also provides a whitening essential oil, the raw materials for its preparation include:

[0157] Mineral oil 57.37 parts

[0158] 5 parts of the multiple fermentation oil described in this example

[0159] 25 parts of ethylhexyl palmitate

[0160] 0.5 parts of squalane

[0161] 10 parts of Cetyl Ethylhexanoate

[0162] 0.8 parts of ascorbyl tetraisopalmitate

[0163] Tocopherol acetate 0.5 parts

[0164] 0.5 parts of bisabolol

[0165] 0.11 parts of Limnanthes Alba seed oil

[0166] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0167] Macadamia ternifolia seed oil 0.11 parts.

[0168] Example 6

[0169] This embodiment provides a multiple fermentation oil, and the preparation method of the multiple fermentation oil is as follows:

[0170] (1) Pretreatment: Wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces and pass it through a 10-mesh sieve for later use;

[0171] (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation;

[0172] The golden flower fungus is Eurotium cristatum, strain number: CICC 2099, China Industrial Microbiological Culture Collection Management Center.

[0173] In step (2), the solid matrix is ​​soybean and mung bean; and the average diameter of the soybean is about 6-8 mm, and the average diameter of the mung bean is about 4-6 mm. The mass ratio of soybean to mung bean is 1:1; the mass ratio of Tibetan Cordyceps sinensis to the solid matrix is ​​1:2; the solid matrix also contains distilled water, and the water content is 30wt%; the solid aerobic fermentation temperature is 30°C, the fermentation environment humidity is 40%RH; the fermentation time is 10d. Turn over 3 times a day during fermentation and keep ventilation.

[0174] (3) Multiple fermentation: The product after solid-state aerobic fermentation is mixed with camellia seeds and then inoculated with yeast and culture medium X for multiple fermentation; the yeast is Candida krusei, 31748, China Industrial Microbiological Culture Collection Management Center.

[0175] Before mixing the camellia seeds with the product after solid-state aerobic fermentation, the camellia seeds were stir-fried at 100° C. for 10 minutes, ground and crushed through a 100-mesh sieve, and then mixed with the product after solid-state aerobic fermentation, which was recorded as mixture A. The ratio of camellia seeds to the product after solid-state aerobic fermentation was 1:9.

[0176] The yeast was inoculated into the medium X, which was a 5° Bé wort medium. The inoculation amount of the yeast in the medium X was 10 vol%. After the yeast was aerobically cultured in the medium X at 28° C. for 24 h, the mixture of the yeast and the medium X was mixed with the mixture A (mass ratio was 1:2) for multiple fermentation, and the fermentation time was 72 h, aerobically, at 28° C.

[0177] (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected;

[0178] (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1, and then centrifuged at 9000 rpm for 5 min to obtain the liquid layer to obtain the multiple fermentation oil.

[0179] This embodiment also provides a whitening essential oil, the raw materials for its preparation include:

[0180] Mineral oil 57.37 parts

[0181] 5 parts of the multiple fermentation oil described in this example

[0182] 25 parts of ethylhexyl palmitate

[0183] 0.5 parts of squalane

[0184] 10 parts of Cetyl Ethylhexanoate

[0185] 0.8 parts of ascorbyl tetraisopalmitate

[0186] Tocopherol acetate 0.5 parts

[0187] 0.5 parts of bisabolol

[0188] 0.11 parts of Limnanthes Alba seed oil

[0189] Jojoba (SIMMONDSIA CHINENSIS) seed oil 0.11 parts

[0190] Macadamia ternifolia seed oil 0.11 parts.

[0191] Performance Testing

[0192] Test 1:

[0193] The multiple fermentation oils prepared in Examples 1 to 6 were placed at 0°C, 25°C, and 40°C in an open state for 90 days, and the product properties and color were checked. The test results are shown in Table 1.

[0194] Table 1 Test results of performance test 1

[0195] Example 0℃ 25℃ 40℃ Example 1 No change No change No change Example 2 No change No change No change Example 3 No change No change No change Example 4 Darkening Darkening Darkening Example 5 Darkening Darkening Darkening Example 6 Darkening No change Darkening

[0196] Test 2: Nourishing / Firming / Moisturizing / Repairing / Anti-Wrinkle / Sensitive Skin Efficacy

[0197] Inspection basis: Refer to ALSSOP-008 "Standard Operating Procedure for Skin Oil and Moisture Tester", ALSSOP-013 "Skin Glossy Meter Operating Procedure", T / GDCA 009-2022 "Human Evaluation Method for Cosmetic Repair Efficacy", QB / T 4256-2011 "Guidelines for Evaluation of Cosmetic Moisturizing Efficacy", T / CNMIA0015-2020 "Clinical Evaluation Standard for Soothing Functional Skin Care Products", T / GDCDC 019-2021 "Test Method for Anti-wrinkle Efficacy of Cosmetics", T / ZHCA005-2019 "Test Method for Cosmetics to Affect Skin Elasticity", "Cosmetic Safety Technical Specifications" (2015 Edition), [Cosmetic Efficacy Evaluation (XIII)-Consumer Use Test].

[0198] A certain number of qualified sensitive skin subjects were screened through the lactic acid sting test, and instrumental testing was used to observe the subjects' skin gloss, skin oil content, skin firmness F4, skin elasticity R2, stratum corneum moisture content, transepidermal water loss rate, total wrinkle area, and skin melanin before and after product use. The researchers evaluated the subjects' wrinkle grade, lactic acid stinging score, and self-assessment to evaluate the nourishing, firming, moisturizing, repairing, anti-wrinkle, and sensitive skin applicability effects of the product Cordyceps sinensis Multi-Oil Fermentation Factor for 28 days, providing a basis for the nourishing, firming, moisturizing, repairing, anti-wrinkle, and sensitive skin applicability claims of cosmetics.

[0199] Evaluation items and principles

[0200] 2.1 Lactic acid sting test

[0201] The facial sensitive skin test is evaluated using the following criteria. The coating method is selected for testing. 50 μL of 5% lactic acid solution or water is randomly applied to one side of the nasolabial groove at room temperature. The subjects' sensitive reactions are evaluated on a 4-point scale at 2.5 minutes and 5 minutes, respectively. The two scores are then added together. Those with a (lactic acid-water) score of 3 or more are considered sensitive skin. 0 points means no tingling; 1 point means slight tingling; 2 points means moderate tingling; 3 points means severe tingling.

[0202] 2.2 VISIA-CR facial images

[0203] Facial images were collected using VISIA-CR (Canfield, USA). VISIA-CR is a facial imaging analysis system dedicated to scientific research and clinical trials. The instrument has 6 light source modes: standard light 1, standard light 2, blue light, orange light, cross-polarized light, and parallel polarized light. Both standard light 1 and standard light 2 can be used to directly observe skin characteristics, such as spots, skin color, pore size, glossiness, wrinkles, etc. Standard light 2 balances the light intensity, making it easier to observe and calculate. Under cross-polarized light, two images can be separated, one is the red area image, and the other is the brown spot image.

[0204] 2.3 Skin glossiness

[0205] The skin glossiness is measured using the skin brightness measuring instrument Deflin. The glossiness of the skin surface is reflected by the direct reflection and scattering of light irradiated to the skin surface. Because the skin is different not only in structure and brightness, but also in color, by testing the reflected light and scattered light on the skin surface, different skin glossiness can be accurately and conveniently tested.

[0206] Skin radiance.

[0207] 2.4 Skin oil content

[0208] Using a skin sebum meter, SM 815 collects skin oil content parameters. The principle of this method is based on a photometer with special tape, which is used to detect the sebum level on the skin surface. The instrument collects all the oil on the surface of the stratum corneum and is widely used in the efficacy evaluation of various moisturizing, nourishing and oil control products. The larger the value, the higher the amount of oil in the stratum corneum.

[0209] 2.5 Skin elasticity R2, skin firmness F4

[0210] Use German MPA580, skin elasticity probe MPA580 (probe diameter 2mm) collects skin elasticity parameters. The testing principle of this method is to apply a constant negative pressure to the skin in the vertical direction, then eliminate the negative pressure and measure the displacement change of the skin within a specified time period. In this test, R2 is used as the parameter of skin elasticity and F4 is used as the parameter of skin firmness.

[0211] 2.6 Water content of stratum corneum

[0212] Using the Skin Moisture Test Probe CM 825 collects the moisture content of the stratum corneum. The capacitance method is used to measure the moisture content of the stratum corneum of human skin. The dielectric constant of water and other substances is significantly different. According to the different moisture content of the stratum corneum of the skin, the measured capacitance value of the skin is different, and its parameters can represent the moisture content of the skin.

[0213] 2.7 Transepidermal water loss

[0214] Using the Skin Water Loss Test Probe TM Hex collects the value of transepidermal water loss. The measurement is based on the law of diffusion, and the transepidermal water loss rate is detected through the water evaporation concentration gradient at the skin contact point.

[0215] 2.8 Total wrinkle area

[0216] The total wrinkle area was collected using the American facial image analyzer, VISIA-CR, and the Image-Pro skin comprehensive analysis software. The testing principle of this method is to use the facial acquisition instrument to obtain skin images, use image analysis to quantify wrinkles, and analyze the degree of wrinkle improvement. The smaller the total wrinkle area, the greater the degree of anti-wrinkle improvement.

[0217] 2.9 Wrinkle grade assessment

[0218] Facial wrinkles are evaluated using the following four levels. Human skin wrinkles are divided into four levels: light, moderate, heavy and severe. Level I is light wrinkles, which are not easily noticeable when the facial expression is static, and can only be vaguely seen when the face is making an expression; Level II is moderate wrinkles, which are vaguely visible when the facial expression is static, but clearly visible when the face is moving; Level III is severe wrinkles, which are clearly visible when the facial expression is static, but wrinkles can be significantly reduced when the wrinkles are stretched; Level IV is severe wrinkles, which are still clearly visible even if the wrinkles are stretched with force.

[0219] 2.10 Evaluation of skin reaction grading

[0220] Select at least 30 voluntary subjects according to the subject inclusion criteria, have the subjects directly use the test products according to the trial characteristics and methods indicated on the cosmetic product label, and record the results according to the skin reaction grading standards in the table below.

[0221] Table 2 Classification standards for skin reactions in human trial tests

[0222]

[0223]

[0224] The skin reactions in Examples 1 to 6 were all grade 0.

[0225] 2.11 Skin Melanin

[0226] Skin melanin is collected using the skin melanin test probe Mexameter MX 18. The measurement is based on the principle of spectral absorption, and the amount of skin melanin is determined by measuring the amount of reflection of light of a specific wavelength on human skin.

[0227] 3 Subject screening

[0228] 3.1 Number of subjects: No less than 30 valid cases.

[0229] 3.2 Subject conditions

[0230] 3.2.1 Selection criteria:

[0231] 3.2.1.1 Aged between 30 and 60;

[0232] 3.2.1.2 Chinese healthy male and female subjects, without allergic diseases, and without a history of allergy to cosmetics or other topical preparations;

[0233] 3.2.1.3 In the lactic acid sting test, the difference between the self-assessed scores on both sides of the nasolabial groove is greater than 3 points;

[0234] 3.2.1.4 The subjects have no skin lesions, eczema, etc.;

[0235] 3.2.1.5 The subject has no severe chronic wasting disease (asthma, diabetes, etc.);

[0236] 3.2.1.6 The facial skin should be free of birthmarks, scratches, white spots, pigmented moles, etc. that may affect the test;

[0237] 3.2.1.7 Those who have not participated in other clinical trials;

[0238] 3.2.1.8 The test area should be kept dry during the test, and no liquid substances, other cosmetics, drugs or health products that may affect the results should be used;

[0239] 3.2.1.9 Can read Chinese and understand it accurately, and has signed the informed consent form for the trial study.

[0240] 3.2.2 Exclusion conditions:

[0241] 3.2.2.1 Currently pregnant or breastfeeding;

[0242] 3.2.2.2 History of skin cancer in the past 5 years;

[0243] 3.2.2.3 Subjects with severe bacterial, viral or fungal skin infections;

[0244] 3.2.2.4 The test subject has obvious sunburn, scars, pigmented moles, excessive hair growth or other conditions that may affect the test results;

[0245] 3.2.2.5 Have a history of immunosuppression or immunodeficiency disease (including HIV or AIDS), or currently use immunosuppressive drugs or radiation;

[0246] 3.2.2.6 Suffering from uncontrollable diseases such as asthma, epilepsy, diabetes, hypertension, hyperthyroidism or hypothyroidism;

[0247] 3.2.2.7 Participating in other clinical trials (such as hormone replacement therapy HRT) within 3 months, or taking oral hormones that may affect the results, or planning to take HRT during the trial;

[0248] 3.2.2.8 Currently using any medication that may affect skin condition or reaction, such as antihistamines, antibiotics, insulin, anti-inflammatory drugs, steroids, corticosteroids, aspirin, thyroid medication, etc.;

[0249] 3.2.2.9 Have undergone chemotherapy or other radiation therapy, or received any prescribed medication or treatment that may affect the skin condition;

[0250] 3.2.2.10 Those who have been diagnosed with allergies or other known or suspected allergic reactions (systemic, inhalation or local); those who have a history of sensitivity to some ingredients in the formula;

[0251] 3.2.2.11 People who are receiving dermatological treatment, or subjects who have taken hydroxy acids, whitening and anti-aging drugs within one month;

[0252] 3.2.2.12 Other diseases or conditions that, according to the investigator's judgment, reduce the likelihood of enrollment or complicate enrollment, such as frequent changes in work environment, unstable living environment, etc., which are likely to cause loss to follow-up; alcohol and / or psychoactive substance abusers and drug addicts.

[0253] 3.2.3 Informed Consent:

[0254] Before participating in any research related to benefits and risks, the subjects must sign an informed consent form. The original informed consent form shall be kept and archived by the project technicians.

[0255] 3.2.4 Confidentiality Agreement:

[0256] All subjects participating in the study are identified by numbers. In the research report, only the gender and age of the subjects are disclosed.

[0257] 3.2.5 Exit criteria:

[0258] During the trial, if the subject experiences adverse reactions, is lost to follow-up without reason, or violates the research protocol (such as using other cosmetics or drugs that may affect the research results), or other special reasons, after evaluation and confirmation by the research leader, if the subject is no longer suitable to continue the trial, the subject will be asked to withdraw.

[0259] The total number of subjects participating in the experiment: 30, the total number of subjects in the effective experiment: 30, including 16 males and 14 females, with an average age of 42.80±3.91 years old.

[0260] 4.1 Test cycle: 28 days of continuous use

[0261] 4.2 Test environment temperature: 20℃~22℃, humidity: 40%~60%.

[0262] 4.4 Protocol requirements: single-blind, randomized enrollment, before-and-after comparison of the product itself

[0263] 4.5 Test site: face

[0264] 4.6 Testing Process

[0265] 4.6.1D0 (initial value, before using the product): Qualified sensitive skin subjects clean their faces under the guidance of the staff, and wipe them clean with a dry, absorbent paper towel. Sit quietly in a room that meets the requirements for at least 30 minutes, without drinking water or beverages, with the test area exposed, stay relaxed, and avoid touching the test area. After 30 minutes, the researchers will conduct instrument tests on the evaluation items.

[0266] 4.6.2 After the initial value determination is completed, the test samples will be distributed to the subjects, and the samples will be used according to the Sample Usage Instructions starting from the same day.

[0267] 4.6.3H1 (after using the product for 1 decimal point) and D28 (after using the product for 28 days), the subjects came for testing. The testing requirements and procedures were the same as D0 (initial values). The researchers conducted instrument testing and data analysis on the evaluation items, and the subjects also answered a questionnaire.

[0268] SPSS software was used to perform descriptive statistics on each measurement value, including quantity, mean, and standard deviation. Before and after comparison: Shapiro-Wilk Test was used to test the significance of the normal distribution of the data improvement value. If Sig. (bilateral)>0.01, it was normally distributed; t test was performed, and the significant difference level α was 0.01. If Sig. (bilateral) <0.01, it was non-normally distributed, and rank sum test was performed, and the significant difference level α was 0.05.

[0269] Table 3 Descriptive statistical results of some performance tests

[0270]

[0271] in:

[0272] Average improvement rate of skin glossiness=(average glossiness at D28-average glossiness at D0) / average glossiness at D0*100%.

[0273] Skin oil content improvement rate (%) = (average skin oil content at D0 - average skin oil content at D28) / average skin oil content at D0 * 100%.

[0274] Average reduction rate of skin firmness F4 (%) = (average skin firmness F4 value at D0 - average skin firmness F4 value at D28) / average skin firmness F4 value at D0 * 100%.

[0275] Skin elasticity R2 average increase rate (%) = (skin elasticity R2 average value of D28 - skin elasticity R2 average value of D0) / skin elasticity R2 average value of D0 * 100%.

[0276] Table 4 Descriptive statistical results of some performance tests

[0277]

[0278] in:

[0279] Average increase rate of stratum corneum moisture content - H1 (%) = (average stratum corneum moisture content of H1 - average stratum corneum moisture content of D0) / average stratum corneum moisture content of D0 * 100%.

[0280] Average increase rate of stratum corneum moisture content - D28 (%) = (average stratum corneum moisture content of D28 - average glossiness of D0) / average stratum corneum moisture content of D0 * 100%.

[0281] Average reduction rate of transepidermal water loss rate - H1 (%) = (average transepidermal water loss rate test value of H1 - average transepidermal water loss rate test value of D0) / average transepidermal water loss rate test value of D0 * 100%.

[0282] Average reduction rate of transepidermal water loss rate - D28 (%) = (average value of transepidermal water loss rate test value of D28 - average value of transepidermal water loss rate test value of D0) / average value of transepidermal water loss rate test value of D0 * 100%.

[0283] Table 5 Descriptive statistical results of some performance tests

[0284]

[0285] in:

[0286] Average reduction rate of total wrinkle area (%) = (average total wrinkle area of ​​D0 - average total wrinkle area of ​​D28) / average total wrinkle area of ​​D0 * 100%.

[0287] Average skin melanin reduction rate (%) = (average skin melanin value at D0 - average skin melanin value at D28) / average skin melanin value at D0 * 100%.

[0288] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A multi-fermented oil, characterized in that: The raw materials for preparing the multiple fermentation oil are Tibetan cordyceps, golden flower fungus and yeast.

2. The multi-fermented oil according to claim 1, characterized in that The preparation method is as follows: (1) Pre-treatment: pre-treating the Tibetan Cordyceps sinensis; (2) Solid-state aerobic fermentation: The pre-treated Tibetan Cordyceps sinensis is placed in a solid matrix and inoculated with golden flower fungi for solid-state aerobic fermentation; (3) Multiple fermentation: the product after solid-state aerobic fermentation is mixed with camellia seeds and then inoculated with yeast for multiple fermentation; (4) Post-processing: After solid-liquid separation of the fermented product, the liquid portion is collected; (5) Purification: The liquid portion was centrifuged at 9000 rpm for 5 min to obtain the oil phase, which was fully mixed with anhydrous sodium sulfate at a mass ratio of 10:1 and then centrifuged at 9000 rpm for 5 min to obtain the multiple fermentation oil.

3. The multi-fermented oil according to claim 2, characterized in that The pre-treatment step in step (1) is as follows: wash and dry the Tibetan Cordyceps sinensis, chop it into small pieces, and pass it through a 10-mesh sieve for standby use.

4. The multi-fermented oil according to claim 2, characterized in that The golden flower fungus in step (2) is Eurotium cristatum.

5. The multi-fermented oil according to claim 2, characterized in that In the step (2), the solid matrix is ​​soybeans and mung beans; and the average diameter of the soybeans is about 6 to 8 mm, and the average diameter of the mung beans is about 4 to 6 mm.

6. The multi-fermented oil according to claim 2, characterized in that In the step (2), the water content of the solid-state aerobic fermentation is about 30-40 wt%, the fermentation temperature is 30° C., and the fermentation environment humidity is 35-40% RH.

7. The multi-fermented oil according to claim 2, characterized in that In the step (3), the camellia seeds and the product after solid-state aerobic fermentation are mixed and then left to stand at 30° C. for 24 hours.

8. The multi-fermented oil according to claim 2, characterized in that The yeast is Candida krusei.

9. The multi-fermented oil according to claim 2, characterized in that In the step (3), the camellia seeds are dried and ground before being mixed with the product after solid-state aerobic fermentation.

10. The multi-fermented oil according to any one of claims 1 to 9, characterized in that Applied in daily chemical field.

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