Acipenser caviar extract as well as preparation method and application thereof

Abstract

The invention belongs to the field of daily chemicals, and discloses a preparation method of a sturgeon caviar extract, which adopts a Soxhlet extraction method and takes ethanol or diethyl ether as a solvent for extraction to obtain the sturgeon caviar extract. According to the sturgeon caviar extract, a Soxhlet extraction method is adopted, ethyl alcohol or diethyl ether is used as a solvent for extraction, the sturgeon caviar extract is obtained, and in the obtained extract, the content of DHA is 3.1-8.1 wt%, the content of vitamin E is 500-1150mu g / g, and the content of phosphatidylcholine is 3-4.8 wt%; the content of effective components is far higher than that of an enzymolysis method. The invention also aims to provide application of the sturgeon caviar extract.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, and in particular to a preparation method of a sturgeon caviar extract and a preparation method and application thereof. Background Art

[0002] The existing technology mostly uses biological proteases to extract active ingredients (DHA, vitamin E, phosphatidylcholine, amino acids, etc.) from sturgeon roe and fish eggs, but the extracted content is low and the extraction is insufficient. Due to the stability and activity of the enzyme, the extraction efficiency of this method is not high and cannot meet the needs of practical applications. In addition, this method may destroy the structure of the active ingredients during the extraction process, further reducing the activity of the extract.

[0003] For example, publication number CN116999379B discloses a sturgeon roe extract and a preparation method and application thereof. The preparation method comprises the following steps: (1) breaking the wall of sturgeon roe and adding enzymes for enzymolysis; the enzymes used for enzymolysis are protease and lipase; the weight ratio of the sturgeon roe, protease and lipase is 50:(1.15-5.15):0.15; the proteases are papain and trypsin; (2) centrifuging and filtering to obtain the sturgeon roe extract.

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

[0005] One of the purposes of the present invention is to provide a method for preparing a sturgeon caviar extract. In the present invention, the sturgeon caviar extract is obtained by using a Soxhlet extraction method and ethanol or ether as a solvent for extraction. The content of DHA in the obtained extract is 3.1-8.1wt%, the content of vitamin E is 500-1150μg / g, and the content of phosphatidylcholine is 3-4.8wt%; the content of the active ingredient is far higher than that of the enzymatic hydrolysis method.

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

[0007] To achieve the above object, the present invention provides a method for preparing a sturgeon caviar extract, wherein the sturgeon caviar extract is obtained by using a Soxhlet extraction method and ethanol or ether as a solvent for extraction.

[0008] Sturgeon caviar, a food rich in active ingredients such as DHA, amino acids, vitamin E, phosphatidylcholine, etc., has always been a favorite extraction object for researchers. The biological protease extraction technology provided by literature, patents and other materials has a low content of active ingredients, which makes the extraction of sturgeon caviar a challenging task. The biological protease extraction technology mainly extracts the active ingredients in sturgeon caviar through the catalytic action of enzymes. However, due to the stability and activity problems of the enzyme, the extraction efficiency of this method is not high and cannot meet the needs of practical applications. In addition, this method may destroy the structure of the active ingredients during the extraction process, further reducing the activity of the extract.

[0009] At the same time, we analyzed the sturgeon caviar extracts on the market and found that most of them are water-soluble extracts. At the same time, we also tried different enzymatic hydrolysis methods to prepare sturgeon caviar extracts. Regardless of whether it is a commercially available product or a product obtained by a variety of enzymatic hydrolysis methods, its performance in terms of antioxidant and anti-wrinkle cannot satisfy us.

[0010] In order to solve this problem, this project developed and tried a new extraction method: Soxhlet extraction. Soxhlet extraction is a traditional organic chemical extraction technology that uses solvents to dissolve and reflux substances to achieve multiple and sufficient extraction of sturgeon roe and fish eggs. This method is simple to operate and has high extraction efficiency. Compared with biological protease extraction technology, Soxhlet extraction has many advantages. First, Soxhlet extraction has higher extraction efficiency and can extract more active ingredients in a short time. Secondly, Soxhlet extraction does not destroy the structure of active ingredients during the extraction process, so the activity of the extract can be guaranteed. Finally, the operation process of Soxhlet extraction is relatively simple and does not require complex equipment and 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° C.; the extraction time is 1-2 hours; the sturgeon roe is placed in a filter bag for extraction; and the mesh size of the filter bag is 80-100 meshes.

[0013] In the above preparation method, the sturgeon roe is pre-treated before extraction; the pre-treatment is one or more operations of cleaning and removing impurities, decolorization, deodorization, desodiumization, cleaning and filtration.

[0014] In the above-mentioned preparation method, the pretreatment is washing and removing impurities, decolorization, deodorization, desodiumization, washing and filtering operations performed in sequence;

[0015] The decolorization treatment uses a decolorizing agent to treat the sturgeon roe;

[0016] The deodorizing treatment comprises treating the sturgeon roe with a deodorizing agent;

[0017] The desodium treatment uses a desodium agent to treat the sturgeon roe.

[0018] In the above preparation method, the decolorizing agent is diatomaceous earth; the deodorizing agent is Bacillus subtilis liquid; the desodiumizing agent is a mixed solution 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 is water, and the concentration of hydrogen peroxide is 30wt%;

[0019] The treatment temperature of the desodium treatment is 20℃~25℃, and the treatment time is 0.5h~1.5h;

[0020] More specifically, the specific operating steps of the present invention are:

[0021] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0022] S2, decolorization treatment: adding an appropriate amount of decolorizing agent to the obtained clean sturgeon caviar, stirring and mixing, decolorizing at a certain temperature for a period of time, taking out the sturgeon caviar and washing it with sterile distilled water;

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

[0024] S4, desodium treatment: adding an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stirring and mixing, desodiumizing at a certain temperature for a period of time, taking out the sturgeon caviar and washing it with sterile distilled water;

[0025] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0026] S6, extraction: put the clean caviar into a filter bag, pad the bottom layer with rapid analysis filter paper, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent into the reactor, extract at a certain temperature for a period of time, and pour out the caviar extract in the reactor;

[0027] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, and concentrating and recovering it at an appropriate temperature and pressure until there is no obvious taste of the extraction solvent, and then pouring out the sturgeon caviar concentrate, which is the sturgeon caviar extract; that is, the concentration treatment is to remove ethanol or ether as the treatment end point;

[0028] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0029] In step 2, the decolorizing agent is specifically diatomaceous earth, and the amount thereof added is 0.3%-0.5% of the mass of a single portion of sturgeon caviar, the treatment temperature is 40° C.-50° C., and the treatment time is 0.5 h-1.5 h;

[0030] In step 3, the deodorizing agent is specifically Bacillus subtilis liquid, and the addition amount thereof is 0.4%-0.6% of the mass of a single portion of sturgeon caviar, the treatment temperature is 30°C-42°C, and the treatment time is 0.5h-1.5h;

[0031] In step seven, the processing temperature is 70-85° C. and the processing pressure is 0.06-0.07 MPa.

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

[0033] The sturgeon caviar extract is prepared by any of the above methods.

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

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

[0036] The dosage forms of the above cosmetics are: essence, cream, and water-based solution;

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

[0038] Beneficial Effects

[0039] In the present invention, the Soxhlet extraction method is adopted, and ethanol or ether is used as a solvent for extraction to obtain a sturgeon caviar extract. The content of DHA in the obtained extract is 3.1-8.1wt%, the content of vitamin E is 500-1150μg / g, and the content of phosphatidylcholine is 3-4.8wt%; the content of the effective ingredient is far higher than that of the enzymatic hydrolysis method.

[0040] By adopting the Soxhlet extraction method, the extraction efficiency is higher and more active ingredients can be extracted in a short time. The Soxhlet extraction method does not destroy the structure of the active ingredients during the extraction process, so the activity of the extract can be guaranteed. The operation process of the Soxhlet extraction method is relatively simple and does not require complex equipment and high costs.

[0041] By performing functional verification on extracts obtained by different extraction methods, it was verified that the extract of the present invention exhibited better activity under the premise of the same concentration. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 is a flow chart of the hyaluronidase inhibition assay;

[0043] Figure 2 is a flow chart of the elastase inhibition assay;

[0044] Figure 3 This is a flow chart of the DPPH free radical scavenging test.

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

[0046] Figure 5 This is a photo of the sample prepared in Comparative Example 1. DETAILED DESCRIPTION

[0047] The present invention will be further described below in conjunction with the embodiments, but it does not constitute any limitation to the present invention. Any limited number of 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] In order to explain the technical content of the present invention in detail, further description is given below in conjunction with the implementation modes.

[0049] Part I. Examples and Comparative Examples

[0050] Example 1

[0051] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0052] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45°C. After 1 hour of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the amount of which is 0.5% of the mass of a single portion of sturgeon caviar, the treatment temperature is 38°C, after the deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0054] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.2%, calcium chloride, 0.1%, and 0.1% glacial acetic acid, the treatment temperature is 20°C, after treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0055] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0056] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with a rapid analysis filter paper, the specific pore size of the analysis filter paper is 80 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the extraction solvent is specifically 70% ethanol, and the addition amount is 9 times the mass of the sturgeon caviar, the processing temperature is 90°C, after extraction for 1.5 hours, pour out the sturgeon caviar extract in the reactor;

[0057] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 80°C and a treatment pressure of 0.06MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0058] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0059] The obtained samples are Figure 4 Photos shown.

[0060] Example 2

[0061] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0062] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.35% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45°C. After 1.5 hours of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the addition amount is 0.6% of the mass of a single portion of sturgeon caviar, the treatment temperature is 38°C, after the deodorization treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water;

[0064] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.25%, 0.1% calcium chloride, and 0.08% glacial acetic acid, the treatment temperature is 20°C, after treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0065] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0066] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with a rapid analysis filter paper, the specific pore size of the analysis filter paper is 80 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the specific extraction solvent is 70% ethanol, and the addition amount is 9 times the mass of the sturgeon caviar, the processing temperature is 85°C, after extraction for 1 hour, pour out the sturgeon caviar extract in the reactor;

[0067] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 80°C and a treatment pressure of 0.06MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0068] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0069] Example 3

[0070] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0071] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 40°C. After 1 hour of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the amount of which is 0.5% of the mass of a single portion of sturgeon caviar, the treatment temperature is 42°C, after deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0073] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.3%, calcium chloride, 0.15%, and 0.05% glacial acetic acid, the treatment temperature is 20°C, after treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0074] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0075] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with rapid analysis filter paper, the specific pore size of the analysis filter paper is 100 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the specific extraction solvent is 85% ethanol, the addition amount is 9 times the mass of the sturgeon caviar, the processing temperature is 85°C, after extraction for 1.5 hours, pour out the sturgeon caviar extract in the reactor;

[0076] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 80°C and a treatment pressure of 0.06MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0077] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0078] Example 4

[0079] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0080] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45°C. After 1.5 hours of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the amount of which is 0.4% of the mass of a single portion of sturgeon caviar, the treatment temperature is 42°C, after the deodorization treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water;

[0082] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.3%, calcium chloride, 0.2%, and 0.05% glacial acetic acid, the treatment temperature is 25°C, after treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water;

[0083] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0084] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with rapid analysis filter paper, the specific pore size of the analysis filter paper is 100 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the extraction solvent is specifically 90% ethanol, the addition amount is 10 times the mass of the sturgeon caviar, the processing temperature is 85°C, after extraction for 2h, pour out the sturgeon caviar extract in the reactor;

[0085] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 85°C and a treatment pressure of 0.07MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0086] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0087] Example 5

[0088] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0089] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.5% of the mass of a single portion of sturgeon caviar. The treatment temperature is 50°C. After 1.5 hours of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the addition amount is 0.6% of the mass of a single portion of sturgeon caviar, the treatment temperature is 42°C, after deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0091] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.35%, calcium chloride, 0.2%, and 0.15% glacial acetic acid, the treatment temperature is 25°C, after treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water;

[0092] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0093] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with a rapid analysis filter paper, the specific pore size of the analysis filter paper is 80 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the specific extraction solvent is 95% ethanol, and the addition amount is 9 times the mass of the sturgeon caviar, the processing temperature is 85°C, after extraction for 2 hours, pour out the sturgeon caviar extract in the reactor;

[0094] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 85°C and a treatment pressure of 0.07MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0095] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0096] Example 6

[0097] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0098] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.45% of the mass of a single portion of sturgeon caviar. The treatment temperature is 40°C. After 1 hour of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the addition amount is 0.6% of the mass of a single portion of sturgeon caviar, the treatment temperature is 42°C, after deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0100] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.1%, calcium chloride, 0.2%, and glacial acetic acid 0.2%, the treatment temperature is 20°C, after treatment for 1.5 hours, take out the sturgeon caviar and wash it with sterile distilled water;

[0101] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0102] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with a rapid analysis filter paper, the specific pore size of the analysis filter paper is 80 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the extraction solvent is specifically 95% ethanol, and the addition amount is 9 times the mass of the sturgeon caviar, the processing temperature is 85°C, after extraction for 1.5 hours, pour out the sturgeon caviar extract in the reactor;

[0103] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 85°C and a treatment pressure of 0.07MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0104] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0105] Example 7

[0106] S1. Cleaning and removing impurities: Place fresh caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean caviar;

[0107] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45°C. After 1 hour of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the amount of which is 0.5% of the mass of a single portion of sturgeon caviar, the treatment temperature is 38°C, after the deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0109] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.2%, calcium chloride, 0.1%, and 0.1% glacial acetic acid, the treatment temperature is 20°C, after treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0110] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0111] S6, extraction: put the clean sturgeon caviar into a filter bag, pad the bottom layer with a rapid analysis filter paper, the specific pore size of the analysis filter paper is 80 mesh, and then place the two in a reaction extraction device, add an appropriate amount of extraction solvent to the reactor, the extraction solvent is specifically ether, and the addition amount is 9 times the mass of the sturgeon caviar, the processing temperature is 90°C, after extraction for 1.5 hours, pour out the sturgeon caviar extract in the reactor;

[0112] S7, concentration and recovery; placing the obtained sturgeon caviar extract in a rotary evaporator, at a treatment temperature of 80°C and a treatment pressure of 0.06MPa, and concentrating and recovering until there is no obvious taste of the extraction solvent, pouring out the sturgeon caviar concentrate, i.e., the sturgeon caviar extract;

[0113] S8. Sterilization: sterilize the obtained sturgeon caviar extract.

[0114] Comparative Example 1

[0115] S1. Cleaning and removing impurities: put fresh sturgeon caviar in a container, soak and clean it with 2 / 3 sterile distilled water, control the water content, remove impurities and obtain clean sturgeon caviar;

[0116] S2, decolorization treatment: add an appropriate amount of decolorizer to the obtained clean sturgeon caviar and stir and mix well. The decolorizer is specifically diatomaceous earth. The added amount is 0.3% of the mass of a single portion of sturgeon caviar. The treatment temperature is 45°C. After 1 hour of treatment, 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, the deodorizing agent is specifically Bacillus subtilis liquid, the amount of which is 0.5% of the mass of a single portion of sturgeon caviar, the treatment temperature is 38°C, after the deodorization treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0118] S4, desodium treatment: add an appropriate amount of desodium agent to the desodiumized sturgeon caviar, stir and mix, the desodium agent is specifically a mixture of 30% hydrogen peroxide, 0.2%, calcium chloride, 0.1%, and 0.1% glacial acetic acid, the treatment temperature is 20°C, after treatment for 1 hour, take out the sturgeon caviar and wash it with sterile distilled water;

[0119] S5, cleaning and filtration: the sodium-removed sturgeon caviar is cleaned 2-3 times with sterile distilled water to remove impurities and obtain clean sturgeon caviar;

[0120] S6. Grinding: adding the cleaned sturgeon caviar to sterile water and grinding it with a grinder;

[0121] S7, enzymatic treatment: add alkali solution to the obtained solution to adjust the pH to 7.5-8.0, then add 0.25% hydrolyzing protease (the weight ratio of papain, alkaline protease, lecithinase and neutral protease is 0.32:0.08:0.1:0.2), react at 60°C for 3h, and then centrifuge to obtain the suspension and supernatant;

[0122] S7, sterilization treatment: sterilize the obtained sturgeon caviar extract. Figure 5 Photos shown.

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

[0124] Table 1 Process parameters

[0125]

[0126]

[0127] Part II Physical and Chemical Performance Test

[0128] The content of effective ingredients in the samples of each embodiment and comparative example was detected, and the effective 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 2;

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

[0133] VE (α-tocopherol);

[0134] Test method: GB 5009.82-2016 Method 1;

[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 refer to 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] Result analysis:

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

[0148] It should be noted that the present invention not only verifies the composite enzyme hydrolysis process of papain, neutral protease, lecithinase and alkaline protease, but we also tried the enzymatic hydrolysis process such as sturgeon roe composite protease (Changzhou Jiaxun Bioengineering Technology Co., Ltd.), and we still found that there is no essential difference between it and the product of Comparative Example 1.

[0149] Among them, the trace elements, vitamins, 16 kinds of amino acids, niacin and other active ingredients in the sturgeon roe extract of Example 1 were detected. Through the detection, it was found that the method is also compatible with the extraction of water-soluble components. The specific test results are shown in Table 5 below;

[0150] Table 5 Water-soluble component detection results

[0151] Serial number Test items unit Test Results method 1 iodine mg / kg 1.98 GB 5009.267-2020 Method 1 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 B 1 > mg / 100mL 1.70 GB 5009.84-2016 Method 1 5 <![CDATA[Vitamin B 2 > mg / 100mL 3.39 GB 5009.85-2016 Method 1 6 <![CDATA[Vitamin B 6 > mg / kg 8.04 GB 5009.154-2023 Method 1 7 <![CDATA[Vitamin D 3 > μg / 100g 309.0 GB 5009.296-2023 Third Law 8 niacin μg / 100g 4480 GB 5009.89-2023 Method 1 9 Taurine mg / 100g 95.0 GB 5009.169-2016 Second Law 10 Biotin μg / 100g 70.5 GB 5009.259-2023 Method 1 11 Pantothenic acid mg / 100g 9.59 GB 5009.210-2023 Method 1

[0152] From the above, it can be seen that the prior art has seriously neglected the advantages of the oil-soluble extract of sturgeon caviar when developing the functions of sturgeon caviar extract. During the research and development of the present invention, we first performed an enzymatic hydrolysis method and found that the effect of the enzymatic hydrolysis method could not meet expectations. After many failed experiments, we concluded that there was an oil film on the surface of the aqueous solution after enzymatic hydrolysis. By chance, we tried to change the extraction method to Soxhlet oil-soluble extraction method. After verification, we found that the effective ingredients of sturgeon caviar in anti-wrinkle, anti-oxidation and other aspects are contained in its oil phase. At the same time, the extract of the present invention is also compatible with water-soluble active ingredients, so this case was filed.

[0153] In order to further verify the application effect of the extract of the present invention, it was confirmed one by one through the functional verification described below.

[0154] Part III Application Formulation and Performance Testing

[0155] Prepare each application sample according to the following recipe:

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

[0157] Test Example 1:

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

[0159] A human skin closed patch test was performed on application samples 1-7 and comparative application sample 1 to evaluate safety.

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

[0161] Table 6 Different skin reaction scores of application samples 1-7 and comparative application sample 1 in patch test

[0162]

[0163]

[0164]

[0165] A patch test was performed on 31 subjects using the above application samples, and the results showed that all of the 31 subjects showed negative results after the patches were removed.

[0166] Test Example 2

[0167] Hyaluronidase inhibition assay

[0168] Experimental principle: Hyaluronic acid is the most abundant component in the extracellular matrix. It regulates the secretion of cytokines and affects cell adhesion, growth, proliferation and differentiation. Therefore, it plays an important role in maintaining skin moisture and elasticity, wound healing and angiogenesis. Hyaluronidase is a specific cleavage enzyme of hyaluronic acid. It has a strong correlation with the release of histamine by mast cells and is a participant in allergic reactions. Inhibiting the activity of hyaluronidase can ensure the normal content and function of hyaluronic acid. Therefore, the hyaluronidase inhibition rate is used as an indicator to evaluate the soothing effect of cosmetics.

[0169] Hyaluronidase can hydrolyze sodium hyaluronate to generate N-acetylglucosamine, which will condense with p-dimethylaminobenzaldehyde when heated under alkaline conditions to produce a color reaction, and the absorbance value can be measured at 585nm on a spectrophotometer. Samples with hyaluronidase activity inhibition can reduce the generation of chromogenic substances, thereby reducing the absorbance value. The inhibitory effect of the sample on hyaluronidase activity can be evaluated based on the change in absorbance value.

[0170] Test Steps 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] Result analysis: By testing Examples 1-7, the inhibition rates of hyaluronic acid enzyme inhibition 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 has the best inhibition effect.

[0175] Test Example 3

[0176] Elastase inhibition test

[0177] Experimental principle: Elastin plays a key role in the youthfulness of the skin, and the lack of elastin is the main cause of skin aging. Inhibiting the activity of elastase can effectively slow down the decomposition of elastin and maintain the elasticity of the skin. Elastase can catalyze the hydrolysis of hydroxyl groups contained in the polypeptide bonds of various amino acids. Therefore, when it reacts with the substrate N-succinyl-alanine-alanine-alanine-p-nitroaniline, it will produce colored substances, and its absorbance can be measured at 410nm in a spectrophotometer. According to the change in absorbance, the inhibitory effect of the test substance on the activity of elastase is calculated, and whether the test substance has firming and anti-wrinkle effects can be evaluated.

[0178] Test Steps Reference Figure 2 ;

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

[0180] Table 8 Elastase inhibition rate results statistical table

[0181]

[0182] Result analysis: by 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 has 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 for short) is a stable long-lived free radical. Its ethanol solution is dark purple and has strong absorption near 517nm. When there is a free radical scavenger, the light absorption of DPPH ethanol solution is weakened due to pairing with its single electron. The degree of fading of DPPH ethanol solution is linearly related to the number of electrons it accepts, which can be used to evaluate the ability of the test sample to scavenge free radicals, that is, the size of the antioxidant activity.

[0186] Test Steps 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] Result analysis: by 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 has the best effect.

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

Claims

1. A method for preparing a sturgeon caviar extract, characterized in that: The sturgeon caviar extract is obtained by adopting the Soxhlet extraction method with ethanol or ether as solvent.

2. The preparation method according to claim 1, characterized in that: The weight of the ethanol or ether is 8 to 10 times the weight of the sturgeon roe.

3. The preparation method according to claim 1, characterized in that: The extraction temperature is 80-100° C.; the extraction time is 1-2 hours; the sturgeon roe is placed in a filter bag for extraction; and the mesh number of the filter bag is 80-100 meshes.

4. The preparation method according to claim 1, characterized in that: Before extraction, the sturgeon roe is pre-treated; the pre-treatment is one or more operations of cleaning and removing impurities, decolorization, deodorization, de-sodiumization, cleaning and filtration.

5. The preparation method according to claim 4, characterized in that: The pre-treatment is washing and impurity removal, decolorization, deodorization, sodium removal, washing and filtering operations performed in sequence; The decolorization treatment uses a decolorizing agent to treat the sturgeon roe; The deodorizing treatment is to treat the sturgeon roe with a deodorizing agent; The desodium treatment uses a desodium agent to treat the sturgeon roe.

6. The preparation method according to claim 5, characterized in that: The decolorizing agent is diatomaceous earth; the deodorizing agent is Bacillus subtilis liquid; the desodiumizing 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 is water, and the concentration of hydrogen peroxide is 30wt%; The treatment temperature of the desodium treatment is 20℃~25℃, and the treatment time is 0.5h~1.5h; The extraction operation is followed by concentration and sterilization; the concentration treatment ends with the removal of ethanol or ether.

7. A sturgeon caviar extract, characterized in that Contains 3.1-8.1wt% DHA, 500-1150μg / g vitamin E, and 3-4.8wt% phosphatidylcholine.

8. The sturgeon caviar extract according to claim 7, characterized in that The method is prepared by any one of claims 1 to 6.

9. Use of the sturgeon caviar extract according to claim 7 or 8 for preparing cosmetics.

10. A cosmetic, characterized in that: Contains the sturgeon caviar extract according to claim 7 or 8.

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