A caviar extract, its preparation and use

Abstract

The application belongs to the field of daily chemicals, and discloses a preparation method of caviar extract. The caviar extract is obtained by Soxhlet extraction with ethanol or diethyl ether as solvent. In the application, the caviar extract is obtained by Soxhlet extraction with ethanol or diethyl ether as solvent. The content of DHA in the obtained extract is 3.1-8.1 wt%, the content of vitamin E is 500-1150 μg / g, and the content of phosphatidylcholine is 3-4.8 wt%. The content of effective components is much higher than that of enzymatic hydrolysis. Another purpose of the application is to provide the application of the caviar extract.

Description

Technical Field

[0001] This invention relates to the field of cosmetics, specifically to a method for preparing sturgeon caviar extract, the method of preparation, and its application. Background Technology

[0002] Current technologies primarily utilize biological proteases to extract active components (DHA, vitamin E, phosphatidylcholine, amino acids, etc.) from sturgeon roe. However, the extracted content is low and the extraction is incomplete. Due to issues with enzyme stability and activity, this method has low extraction efficiency and cannot meet the needs of practical applications. Furthermore, this method may damage the structure of active components during extraction, further reducing the activity of the extract.

[0003] For example, CN116999379B, the subject of which is a sturgeon roe extract and its preparation method and application, discloses a sturgeon roe extract and its preparation method and application. The preparation method includes the following steps: (1) after breaking the cell wall of sturgeon roe, an enzyme is added for enzymatic hydrolysis; the enzymes used for enzymatic hydrolysis are protease and lipase; the weight ratio of sturgeon roe, protease and lipase is 50:(1.15-5.15):0.15; the protease is papain and trypsin; (2) centrifugation and filtration are performed to obtain the sturgeon roe extract.

[0004] During the research process of this project, papain, alkaline protease, lecithinase, and neutral protease were used to enzymatically extract sturgeon roe. We found that the contents of DHA, vitamin E, and phosphatidylcholine could not meet the ideal requirements. Summary of the Invention

[0005] One of the objectives of this invention is to provide a method for preparing sturgeon caviar extract. In this invention, Soxhlet extraction is used with ethanol or ether as a solvent to extract sturgeon caviar extract. The extract contains 3.1–8.1 wt% DHA, 500–1150 μg / g vitamin E, and 3–4.8 wt% phosphatidylcholine. The content of its active ingredients is far greater than that of enzymatic hydrolysis.

[0006] Another object of the present invention is to provide the application of the sturgeon caviar extract.

[0007] To achieve the above objectives, the present invention provides a method for preparing sturgeon caviar extract, which involves Soxhlet extraction using ethanol or ether as a solvent to obtain sturgeon caviar extract.

[0008] Sturgeon caviar, a food rich in active ingredients such as DHA, amino acids, vitamin E, and phosphatidylcholine, has long been a popular extraction target for researchers. However, the bio-protease extraction technology described in literature and patents yields relatively low levels of these active ingredients, making sturgeon caviar extraction a challenging task. Bio-protease extraction technology primarily utilizes enzymatic catalysis to extract the active ingredients from sturgeon caviar. However, due to issues with enzyme stability and activity, this method has low extraction efficiency, failing to meet practical application requirements. Furthermore, this method may damage the structure of active ingredients during extraction, further reducing the activity of the extract.

[0009] Meanwhile, an analysis of commercially available sturgeon caviar extracts revealed that the vast majority of them are water-soluble. Furthermore, we explored various enzymatic hydrolysis methods to prepare sturgeon caviar extracts, but none of them, whether commercially available or obtained through multiple enzymatic hydrolysis methods, met our expectations regarding their antioxidant and anti-wrinkle properties.

[0010] To address this problem, this project developed and attempted a new extraction method: Soxhlet extraction. Soxhlet extraction is a traditional organic chemical extraction technique that utilizes solvents to dissolve and reflux substances, achieving multiple and thorough extractions of sturgeon roe and eggs. This method is simple to operate and highly efficient. Compared to biological protease extraction techniques, Soxhlet extraction has many advantages. First, it is more efficient, extracting more active ingredients in a shorter time. Second, it does not damage the structure of active ingredients during the extraction process, thus preserving the activity of the extract. Finally, the operation of Soxhlet extraction is simpler, requiring no complex equipment or high costs.

[0011] In the above preparation method, the weight of the ethanol or ether is 8 to 10 times the weight of the sturgeon roe.

[0012] In the above preparation method, the extraction temperature is 80-100℃; the extraction time is 1-2 hours; the sturgeon roe is extracted in a filter bag; and the mesh size of the filter bag is 80-100 mesh.

[0013] In the above preparation method, the sturgeon roe is pretreated before extraction; the pretreatment includes one or more of the following operations: cleaning and impurity removal, decolorization, deodorization, sodium removal, and cleaning and filtration.

[0014] In the above preparation method, the pretreatment consists of sequential cleaning and impurity removal, decolorization, deodorization, sodium removal, and cleaning and filtration operations.

[0015] The decolorization process involves treating sturgeon caviar with a decolorizing agent;

[0016] The deodorization process involves treating sturgeon roe with a deodorizing agent;

[0017] The desodium treatment involves using a desodiuming agent to treat sturgeon roe.

[0018] In the above preparation method, the decolorizing agent is diatomaceous earth; the deodorizing agent is Bacillus subtilis bacterial solution; the desodiumning agent is a mixture of 0.1wt% to 0.35wt% hydrogen peroxide, 0.05wt% to 0.2wt% calcium chloride, 0.05wt% to 0.2wt% glacial acetic acid, and the balance being water, with the concentration of hydrogen peroxide being 30wt%.

[0019] Sodium removal treatment temperature: 20℃~25℃, treatment time: 0.5h~1.5h;

[0020] More specifically, the specific operating steps of this invention are as follows:

[0021] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0022] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the clean sturgeon caviar, stir and mix well, and decolorize at a certain temperature for a period of time. Then take out the sturgeon caviar and wash it with sterile distilled water.

[0023] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well, and deodorize at a certain temperature for a period of time. Then take out the sturgeon caviar and wash it with sterile distilled water.

[0024] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well, and after desodium removal treatment at a certain temperature for a period of time, take out the sturgeon caviar and wash it with sterile distilled water.

[0025] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0026] S6. Extraction: Place the cleaned sturgeon caviar into a filter bag, with rapid analysis filter paper at the bottom, and then place both in a reaction extraction device. Add an appropriate amount of extraction solvent to the reactor, extract at a certain temperature for a period of time, and then pour out the sturgeon caviar extract from the reactor.

[0027] S7. Concentration and recovery: The obtained sturgeon caviar extract is placed in a rotary evaporator and concentrated and recovered under appropriate temperature and pressure until there is no obvious smell of the extraction solvent. The sturgeon caviar concentrate is then poured out, which is the sturgeon caviar extract. That is, the concentration process ends with the removal of ethanol or ether.

[0028] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0029] In step two, the decolorizing agent is diatomaceous earth, and the amount added is 0.3%-0.5% of the weight of a single portion of sturgeon caviar. The treatment temperature is 40℃-50℃, and the treatment time is 0.5h-1.5h.

[0030] In step three, the deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.4%-0.6% of the weight of a single portion of sturgeon caviar. The treatment temperature is 30℃-42℃, and the treatment time is 0.5h-1.5h.

[0031] The processing temperature in step seven is 70-85℃, and the processing pressure is 0.06-0.07MPa.

[0032] Meanwhile, the present invention also discloses a sturgeon caviar extract containing 3.1-8.1 wt% DHA, 500-1150 μg / g vitamin E, and 3-4.8 wt% phosphatidylcholine.

[0033] The sturgeon caviar extract described above was prepared using any of the methods described above.

[0034] Finally, the present invention also discloses the use of the sturgeon caviar extract as described above in the preparation of cosmetics, and a cosmetic containing the sturgeon caviar extract as described above.

[0035] Generally, it is recommended to add 1.0–5.0 wt% sturgeon caviar extract to cosmetics;

[0036] The dosage forms of the above-mentioned cosmetics are: serums, creams, and liquids;

[0037] The functions of the above-mentioned cosmetics are: anti-aging, moisturizing, and anti-oxidation.

[0038] Beneficial effects

[0039] In this invention, Soxhlet extraction is used with ethanol or ether as solvent to extract sturgeon caviar extract. The extract contains 3.1-8.1 wt% DHA, 500-1150 μg / g vitamin E, and 3-4.8 wt% phosphatidylcholine. The content of its active ingredients is far greater than that of enzymatic hydrolysis.

[0040] Soxhlet extraction offers higher extraction efficiency, allowing for the extraction of more active ingredients in a shorter time. The Soxhlet extraction process does not damage the structure of the active ingredients, thus preserving the activity of the extract. Furthermore, the Soxhlet extraction method is relatively simple to operate, requiring no complex equipment or high costs.

[0041] By verifying the function of extracts obtained by different extraction methods, it was confirmed that the extract of the present invention exhibits better activity under the same concentration conditions. Attached Figure Description

[0042] Figure 1 This is a flowchart of the hyaluronidase inhibition test;

[0043] Figure 2 This is a flowchart of the elastase inhibition assay;

[0044] Figure 3 This is a flowchart of the DPPH free radical scavenging assay.

[0045] Figure 4 This is a photograph of the sample prepared in Example 1;

[0046] Figure 5 This is a photograph of the sample prepared in Comparative Example 1. Detailed Implementation

[0047] The present invention will be further described below with reference to embodiments, but this does not constitute any limitation on the present invention. Any limited modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention.

[0048] To illustrate the technical content of the present invention in detail, the following description is provided in conjunction with the embodiments.

[0049] Part One: Examples and Comparative Cases

[0050] Example 1

[0051] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0052] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0053] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 38℃. After deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0054] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide 0.2%, calcium chloride 0.1%, and glacial acetic acid 0.1%. The treatment temperature is 20℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0055] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0056] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 80 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent was added to the reactor. The extraction solvent was 70% ethanol, and the amount added was 9 times the mass of the sturgeon caviar. The treatment temperature was 90℃. After extraction for 1.5 hours, the sturgeon caviar extract was poured out of the reactor.

[0057] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 80℃ and a processing pressure of 0.06MPa until there is no obvious smell of the extraction solvent. Pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0058] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0059] The obtained sample is Figure 4 The photo shown.

[0060] Example 2

[0061] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0062] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.35% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45℃. After treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0063] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.6% of the mass of a single portion of sturgeon caviar. The treatment temperature is 38℃. After the deodorization treatment is carried out for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0064] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide, 0.25% calcium chloride, and 0.08% glacial acetic acid. The treatment temperature is 20℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0065] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0066] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 80 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent was added to the reactor. The extraction solvent was 70% ethanol, and the amount added was 9 times the mass of the sturgeon caviar. The treatment temperature was 85℃. After extraction for 1 hour, the sturgeon caviar extract was poured out of the reactor.

[0067] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 80℃ and a processing pressure of 0.06MPa until there is no obvious smell of the extraction solvent. Pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0068] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0069] Example 3

[0070] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0071] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 40℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0072] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 42℃. After deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0073] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide, 0.3% calcium chloride, and 0.15% glacial acetic acid. The treatment temperature is 20℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0074] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0075] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 100 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent was added to the reactor. The extraction solvent was 85% ethanol, and the amount added was 9 times the mass of the sturgeon caviar. The treatment temperature was 85℃. After extraction for 1.5 hours, the sturgeon caviar extract was poured out of the reactor.

[0076] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 80℃ and a processing pressure of 0.06MPa until there is no obvious smell of the extraction solvent. Pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0077] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0078] Example 4

[0079] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0080] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45℃. After treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0081] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.4% of the mass of a single portion of sturgeon caviar. The treatment temperature is 42℃. After the deodorization treatment is carried out for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0082] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide, 0.3%, calcium chloride, and glacial acetic acid. The treatment temperature is 25℃. After treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0083] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0084] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 100 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent was added to the reactor. The extraction solvent was 90% ethanol, and the amount added was 10 times the mass of the sturgeon caviar. The treatment temperature was 85℃. After extraction for 2 hours, the sturgeon caviar extract was poured out of the reactor.

[0085] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 85°C and a processing pressure of 0.07MPa until there is no obvious smell of the extraction solvent. Then pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0086] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0087] Example 5

[0088] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0089] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 50℃. After treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0090] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.6% of the mass of a single portion of sturgeon caviar. The treatment temperature is 42℃. After deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0091] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide, 0.35% calcium chloride, and 0.15% glacial acetic acid. The treatment temperature is 25℃. After treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0092] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0093] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 80 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent was added to the reactor. The extraction solvent was 95% ethanol, and the amount added was 9 times the mass of the sturgeon caviar. The treatment temperature was 85℃. After extraction for 2 hours, the sturgeon caviar extract was poured out of the reactor.

[0094] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 85°C and a processing pressure of 0.07MPa until there is no obvious smell of the extraction solvent. Then pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0095] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0096] Example 6

[0097] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0098] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.45% of the mass of a single portion of sturgeon caviar. The treatment temperature is 40℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0099] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.6% of the mass of a single portion of sturgeon caviar. The treatment temperature is 42℃. After deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0100] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide, 0.1%, calcium chloride, and glacial acetic acid. The treatment temperature is 20℃. After treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water.

[0101] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0102] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 80 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent was added to the reactor. The extraction solvent was 95% ethanol, and the amount added was 9 times the mass of the sturgeon caviar. The treatment temperature was 85℃. After extraction for 1.5 hours, the sturgeon caviar extract was poured out of the reactor.

[0103] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 85°C and a processing pressure of 0.07MPa until there is no obvious smell of the extraction solvent. Then pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0104] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0105] Example 7

[0106] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0107] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0108] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 38℃. After deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0109] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide 0.2%, calcium chloride 0.1%, and glacial acetic acid 0.1%. The treatment temperature is 20℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0110] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0111] S6. Extraction: The cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper at the bottom. The specific pore size of the analytical filter paper was 80 mesh. Then, the two were placed in a reaction extraction device. An appropriate amount of extraction solvent, specifically diethyl ether, was added to the reactor. The amount added was 9 times the mass of the sturgeon caviar. The treatment temperature was 90℃. After extraction for 1.5 hours, the sturgeon caviar extract was poured out of the reactor.

[0112] S7. Concentration and recovery: Place the obtained sturgeon caviar extract in a rotary evaporator and concentrate it at a processing temperature of 80℃ and a processing pressure of 0.06MPa until there is no obvious smell of the extraction solvent. Pour out the sturgeon caviar concentrate, which is the sturgeon caviar extract.

[0113] S8. Sterilization treatment: The obtained sturgeon caviar extract is sterilized.

[0114] Comparative Example 1

[0115] S1. Cleaning and removing impurities: Place the fresh sturgeon caviar in a container and soak it in 2 / 3 sterile distilled water. Drain the water to remove impurities and obtain clean sturgeon caviar.

[0116] S2. Decolorization treatment: Add an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar and stir well. The decolorizing agent is diatomaceous earth, and the amount added is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0117] S3. Deodorization treatment: Add an appropriate amount of deodorizing agent to the decolorized sturgeon caviar, stir and mix well. The deodorizing agent is Bacillus subtilis bacterial solution, and the amount added is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 38℃. After deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0118] S4. Desodium removal treatment: Add an appropriate amount of desodium removal agent to the desodiumed sturgeon caviar, stir and mix well. The desodium removal agent is a mixture of 30% hydrogen peroxide 0.2%, calcium chloride 0.1%, and glacial acetic acid 0.1%. The treatment temperature is 20℃. After treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water.

[0119] S5. Washing and filtration: Wash the desodiumed sturgeon caviar 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar.

[0120] S6. Grinding process: Add the cleaned sturgeon caviar to sterile water and grind it using a grinder.

[0121] S7. Enzymatic hydrolysis: Add alkaline solution to the obtained solution to adjust the pH to 7.5-8.0, then add 0.25% hydrolytic protease (the weight ratio of papain, alkaline protease, lecithinase and neutral protease is 0.32:0.08:0.1:0.2), react at 60℃ for 3 hours, and then centrifuge to collect the suspension and supernatant.

[0122] S7. Sterilization: The obtained sturgeon caviar extract was sterilized. The resulting sample was... Figure 5 The photo shown.

[0123] The relevant process parameters for the above embodiments and comparative examples are shown in Table 1 below;

[0124] Table 1 Process Parameters

[0125]

[0126]

[0127] Part Two: Physicochemical Performance Testing

[0128] The content of active ingredients in the samples of each embodiment and comparative example was detected. The active ingredients were: DHA (cis-4,7,10,13,16,19-docosahexaenoic acid), VE (α-tocopherol), and PC (phosphatidylcholine).

[0129] Testing equipment and methods:

[0130] DHA (cis-4,7,10,13,16,19-docosahexaenoic acid);

[0131] Test method: GB 5009.168-2016 Method II;

[0132] Equipment: Gas chromatograph (Agilent 7820A);

[0133] VE (α-tocopherol);

[0134] Test method: GB 5009.82-2016 Method I;

[0135] Equipment: Liquid Chromatograph (Agilent 1260);

[0136] PC:

[0137] Test method: GB 5009.272-2016;

[0138] Equipment: Liquid Chromatograph (Agilent 1260)

[0139] The test results are shown in Tables 2 to 4.

[0140] Table 2. DHA (cis-4,7,10,13,16,19-docosahexaenoic acid) content

[0141]

[0142] Table 3. VE (α-tocopherol) content

[0143]

[0144] Table 4 PC (phosphatidylcholine) content

[0145]

[0146] Results analysis:

[0147] As can be seen from the above experiments, the concentrations of DHA (cis-4,7,10,13,16,19-docosahexaenoic acid), VE (α-tocopherol), and PC (phosphatidylcholine) in the product obtained by Soxhlet extraction are significantly better than those in the product obtained by enzymatic hydrolysis.

[0148] It should be noted that this invention not only verified the enzymatic hydrolysis process of papain, neutral protease, lecithinase and alkaline protease, but we also tried the enzymatic hydrolysis process of sturgeon caviar complex protease (Changzhou Jiaxun Bioengineering Technology Co., Ltd.), and we still found that it was not essentially different from the product of Comparative Example 1.

[0149] Among them, the trace elements, vitamins, 16 kinds of amino acids, nicotinic acid and other active ingredients in the sturgeon caviar extract of Example 1 were detected. The detection results showed that the method is also compatible with the extraction of water-soluble components. The specific detection results are shown in Table 5 below.

[0150] Table 5 Results of water-soluble component detection

[0151] Serial Number Test Project unit Test Results method 1 iodine mg / kg 1.98 GB 5009.267-2020 First Law 2 selenium mg / kg 0.785 GB 5009.93-2017 Third Law 3 16 amino acids g / 100g 2.1 GB 5009.124-2016 4 <![CDATA[Vitamin B1]]> mg / 100mL 1.70 GB 5009.84-2016 First Law 5 <![CDATA[Vitamin B2]]> mg / 100mL 3.39 GB 5009.85-2016 First Law 6 <![CDATA[Vitamin B6]]> mg / kg 8.04 GB 5009.154-2023 First Law 7 <![CDATA[Vitamin D3]]> μg / 100g 309.0 GB 5009.296-2023 Third Law 8 niacin μg / 100g 4480 GB 5009.89-2023 First Law 9 Taurine mg / 100g 95.0 GB 5009.169-2016 Second Law 10 Biotin μg / 100g 70.5 GB 5009.259-2023 First Law 11 pantothenic acid mg / 100g 9.59 GB 5009.210-2023 First Law

[0152] In summary, existing technologies for developing the functions of sturgeon caviar extracts have severely overlooked the advantages of oil-soluble extracts of sturgeon caviar. During the development of this invention, we initially attempted enzymatic hydrolysis, but found that the results were unsatisfactory. After numerous failed experiments, we discovered an oil film on the surface of the aqueous solution after enzymatic hydrolysis. By chance, we tried changing the extraction method to Soxhlet oil-soluble extraction. Verification revealed that the anti-wrinkle and antioxidant active ingredients of sturgeon caviar are contained in its oil phase. Furthermore, the extract of this invention also contains water-soluble active ingredients, hence this proposal.

[0153] To further verify the application effect of the extract of the present invention, it will be confirmed one by one through the functional verification in the following text.

[0154] Part Three: Application Formulation and Performance Testing

[0155] Prepare application samples according to the following formulations:

[0156] Sturgeon caviar extract of each embodiment and comparative example: 2.5 wt%, butylene glycol: 50 wt%, water: 47.5%; application samples 1 to 7 corresponding to each embodiment and comparative example, and comparative application sample 1 were obtained.

[0157] Experimental Example 1:

[0158] Safety tests of samples 1-7 and comparative sample 1

[0159] Human skin closed patch tests were conducted on application samples 1-7 and control application sample 1 to evaluate safety.

[0160] The human skin occlusive patch test was conducted according to the test requirements for occlusive patches in the 2015 edition of the "Cosmetic Safety Technical Specifications". The test was conducted on application samples 1-7 and control application sample 1; the negative control was pure water. A total of 31 subjects were included, with an average age of 33.32±7.09 years. The test results are shown in Table 6.

[0161] Table 6 shows the different skin reaction ratings for patch tests of samples 1-7 and comparative sample 1.

[0162]

[0163]

[0164]

[0165] The results of patch testing on 31 subjects using the above-mentioned application sample showed that all 31 subjects had negative results after the patch was removed.

[0166] Experimental Example 2

[0167] Hyaluronidase inhibition test

[0168] Experimental Principle: Hyaluronic acid is the most abundant component of the extracellular matrix, regulating cytokine secretion and influencing cell adhesion, growth, proliferation, and differentiation. Therefore, it plays a crucial role in maintaining skin hydration and elasticity, wound healing, and angiogenesis. Hyaluronidase is a specific lysin of hyaluronic acid and is strongly correlated with histamine release from mast cells, thus participating in allergic reactions. Inhibiting hyaluronidase activity ensures normal hyaluronic acid content and function; therefore, the hyaluronidase inhibition rate is used as an indicator to evaluate the soothing efficacy of cosmetics.

[0169] Hyaluronidase hydrolyzes sodium hyaluronate to produce N-acetylglucosamine. This substance, upon heating under alkaline conditions, condenses with p-dimethylaminobenzaldehyde, producing a colorimetric reaction. The absorbance can be measured at 585 nm using a spectrophotometer. Samples exhibiting hyaluronidase activity inhibition can reduce the formation of chromogenic substances, thereby lowering the absorbance. The inhibitory effect of the sample on hyaluronidase activity can be evaluated based on changes in absorbance.

[0170] Test procedure reference Figure 1 ;

[0171] The test results are shown in Table 7 below;

[0172] Table 7 Statistical table of hyaluronidase inhibition rate results

[0173]

[0174] Results Analysis: Through testing Examples 1-7, the inhibition rates of hyaluronidase inhibitors in Comparative Example 1 were 57.46%, 39.37%, 44.79%, 41.72%, 20.25%, 26.28%, 31.70%, and 13.29%, respectively, indicating that the sample has the ability to inhibit hyaluronidase activity. Among them, Example 1 showed the best inhibitory effect.

[0175] Experimental Example 3

[0176] elastase inhibition test

[0177] Experimental Principle: Elastin plays a crucial role in youthful skin, and its deficiency is a major cause of skin aging. Inhibiting elastase activity effectively slows down elastin breakdown and maintains skin elasticity. Elastase catalyzes the hydrolysis of hydroxyl groups in polypeptide bonds of various amino acids. Therefore, when it reacts with the substrate N-succinyl-alanine-alanine-alanine-p-nitroaniline, a colored substance is produced, the absorbance of which can be measured at 410 nm using a spectrophotometer. Based on the change in absorbance, the inhibitory effect of the test substance on elastase activity can be calculated, and the firming and anti-wrinkle effects of the test substance can be evaluated.

[0178] Test procedure reference Figure 2 ;

[0179] The test results are shown in Table 8 below;

[0180] Table 8. Statistical table of elastase inhibition rate results

[0181]

[0182] Results Analysis: Through testing Examples 1-7, the elastase inhibition rates of Comparative Example 1 were 48.78%, 42.03%, 36.50%, 30.89%, 18.70%, 24.47%, 19.27%, and 7.72%, respectively, indicating that the sample has firming and anti-wrinkle effects. Among them, Example 1 showed the best firming and anti-wrinkle effects.

[0183] Test Example 4

[0184] DPPH free radical scavenging test

[0185] Experimental Principle: 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable, long-lived free radical. Its ethanol solution is deep purple and exhibits strong absorption around 517 nm. In the presence of a free radical scavenger, the light absorption of the DPPH ethanol solution decreases due to the pairing of unpaired electrons with the DPPH. The degree of fading of the DPPH ethanol solution is linearly related to the number of electrons it accepts, thus allowing evaluation of the test sample's ability to scavenge free radicals, i.e., its antioxidant activity.

[0186] Test procedure reference Figure 3 ;

[0187] The test results are shown in Table 9 below;

[0188] Table 9. Statistical Results of Free Radical Scavenging Rate

[0189]

[0190]

[0191] Results Analysis: Through testing Examples 1-7, the free radical scavenging rates of Comparative Example 1 were 89.73%, 72.43%, 74.81%, 67.57%, 39.13%, 54.35%, 45.58%, and 9.08%, respectively, indicating that the sample has the ability to scavenge free radicals. Among them, Example 1 showed the best effect.

[0192] In summary, the sturgeon caviar extract obtained in Example 1 has significant anti-wrinkle, firming, antioxidant, and soothing effects.

Claims

1. A method for preparing sturgeon caviar extract, characterized in that, Cleaned sturgeon caviar was placed in a filter bag, with a rapid analytical filter paper (80 mesh) placed underneath. Both were then placed in a reaction extraction apparatus. An extraction solvent (70% ethanol, 9 times the mass of the sturgeon caviar) was added to the reactor for Soxhlet extraction. The extraction temperature was 90℃, and extraction was carried out for 1.5 hours. The sturgeon caviar extract was then poured out of the reactor. The extract was then concentrated and sterilized to obtain the sturgeon caviar extract. Before extraction, the sturgeon caviar undergoes pretreatment; the pretreatment includes one or more of the following operations: cleaning and impurity removal, decolorization, deodorization, sodium removal, and cleaning and filtration. The sturgeon caviar extract contains 8.07 wt% DHA, 1136.61 μg / g vitamin E, 4.77 wt% phosphatidylcholine, 2.1 g / 100g amino acids, 1.70 mg / 100 mL vitamin B1, 3.39 mg / 100 mL vitamin B2, 8.04 mg / 100 mL vitamin B6, 309.0 μg / 100g vitamin D3, 4480 μg / 100g niacin, 95.0 mg / 100g taurine, 70.5 μg / 100g biotin, and 9.59 mg / 100g pantothenic acid. The sturgeon caviar extract has the ability to inhibit hyaluronidase activity, scavenge free radicals, and has firming and anti-wrinkle effects.

2. The preparation method according to claim 1, characterized in that, The pretreatment consists of sequential cleaning and impurity removal, decolorization, deodorization, sodium removal, and cleaning and filtration operations. The decolorization process involves treating sturgeon caviar with a decolorizing agent; The deodorization process involves treating sturgeon roe with a deodorizing agent; The desodium treatment involves treating sturgeon caviar with a desodiuming agent.

3. The preparation method according to claim 2, characterized in that, The decolorizing agent is diatomaceous earth; the deodorizing agent is Bacillus subtilis bacterial solution; the desodiumning agent is a mixture of 0.1wt%–0.35wt% hydrogen peroxide, 0.05wt%–0.2wt% calcium chloride, 0.05wt%–0.2wt% glacial acetic acid, and the balance being water, with the hydrogen peroxide concentration being 30wt%. The sodium removal treatment temperature is 20℃~25℃, and the treatment time is 0.5h~1.5h. The concentration process ends with the removal of ethanol.

4. A sturgeon caviar extract prepared by the method as described in any one of claims 1 to 3.

5. The use of the sturgeon caviar extract as described in claim 4 in the preparation of cosmetics.

6. A cosmetic product, characterized in that, It contains the sturgeon caviar extract as described in claim 4.

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