Preparation method and application of salmon egg extract
Through the methods of cleaning, crushing, degreasing, purification and homogenization of salt solution and water, the problems of single ingredient extraction and active substance in salmon egg extract research were solved, and high purity and retention of multiple nutrients were achieved. The scope of application of salmon egg extract has been expanded.
Patent Information
- Application Number
- CN202411363515.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-27
- Publication Date
- 2024-12-27
AI Technical Summary
The existing research on salmon egg extracts is at the extraction stage of a single component, and some extraction processes lead to the inactivation of natural active substances, which fails to effectively utilize the various nutrients in salmon eggs.
Salmon eggs are washed successively with salt solution and water, and after crushing, solid-liquid separation, degreasing, and then purification and homogenization are carried out, including filtration, ultrafiltration or dialysis, adjusting pH, reducing protein molecular weight and microbial content, and retaining a variety of Nutrients and biological activities.
It improves the purity and biological activity of salmon egg extract, expands its application scenarios, and is suitable for food, health products and cosmetics. It has a simple process and low cost, and is suitable for industrial production.
Smart Images
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Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of biomaterials, and in particular to a preparation method and application of a salmon egg extract. Background Art
[0002] There are many species of salmonids in the world, which grow fast, have a wide range of adaptability, and have nutritious meat, fresh and delicious. At present, domestic salmon roe processing companies generally make caviar and other foods for human consumption after simple blending. However, eating caviar alone cannot effectively utilize the nutrients in salmon roe, which easily leads to the waste of biologically active substances in salmon roe.
[0003] Salmon eggs are crystal clear in appearance and contain a variety of nutrients required for the development of organisms, such as fatty acids, proteins, peptides, amino acids, DNA, RNA, vitamins, minerals, trace elements and other active ingredients. Fatty acids can maintain the relative fluidity of cell membranes to ensure the normal physiological functions of cells; protein is the material basis of all life, an important component of basal cells, and the main raw material for the renewal and repair of organisms; vitamins play an important role in the growth, metabolism and development of organisms, participating in the biochemical reactions of organisms and regulating the metabolic functions of organisms.
[0004] At present, the research on salmon egg extract is still at the stage of extraction of single components, and some extraction processes will cause the inactivation of natural active substances. It is of great significance to find a preparation method for salmon egg extract that can retain multiple nutrients and maintain high biological activity. Summary of the invention
[0005] The main purpose of the present invention is to provide a preparation method and application of salmon egg extract to solve the problem that the current research on salmon egg extract is still at the stage of extraction of a single component and some extraction processes may cause the inactivation of natural active substances.
[0006] In order to achieve the above-mentioned purpose, according to one aspect of the present invention, a method for preparing a salmon egg extract is provided, the method comprising: step S1, first washing the salmon eggs with a salt solution and water in sequence to obtain purified salmon eggs; step S2, crushing the purified salmon eggs and performing solid-liquid separation to obtain a salmon egg homogenate; step S3, defatting the salmon egg homogenate and then performing solid-liquid separation to obtain a crude salmon egg extract; step S4, purifying and homogenizing the crude salmon egg extract in sequence to obtain a salmon egg extract.
[0007] Furthermore, in step S1, the mass volume concentration of the salt solution is 10 to 60 g / L.
[0008] Furthermore, the salt solution includes multiple groups of salt solutions whose mass volume concentrations decrease sequentially from 60g / L to 10g / L, and the salmon eggs are washed sequentially using the multiple groups of salt solutions, and the mass volume concentrations of the multiple groups of salt solutions decrease sequentially.
[0009] Further, the salt solution includes at least one of sodium chloride solution, potassium chloride solution, magnesium chloride solution, calcium chloride solution or ammonium chloride solution.
[0010] Furthermore, the water includes at least one of purified water, deionized water or distilled water.
[0011] Furthermore, the material-liquid ratio of salmon roe to salt solution is 1:1 to 1:2 g / L, and more preferably 1:2 g / L.
[0012] Furthermore, in step S2, solid-liquid separation is performed by centrifugation, and preferably the centrifugal speed is 5000-8500 rpm / min and the time is 8-15 min.
[0013] Furthermore, in step S3, a degreasing agent is added to the salmon egg homogenate for degreasing treatment, and the degreasing agent includes at least one of methanol, ethanol, isopropanol, sodium hydroxide and potassium hydroxide.
[0014] Furthermore, the mass ratio of the degreasing agent to the salmon egg homogenate is 5 to 12:100.
[0015] Furthermore, the degreasing treatment time is 12 to 60 hours, and the degreasing treatment temperature is 15 to 35°C.
[0016] Furthermore, in step S4, the purification treatment includes at least one of filtration, ultrafiltration or dialysis.
[0017] Furthermore, the homogenization treatment is carried out at a pressure of 300 to 500 MPa and for a time of 10 to 30 min.
[0018] Furthermore, step S4 also includes adjusting the pH after the homogenization process, and adjusting the pH so that the pH of the salmon egg extract is 5.5-7.4.
[0019] Furthermore, the protein molecular weight of the salmon egg extract is 11 to 48 kDa, preferably 11 to 25 kDa, and more preferably 11 to 20 kDa.
[0020] Furthermore, the saturated fatty acid content of the salmon egg extract is 0.6 to 0.9 wt %, preferably 0.7 to 0.9 wt %, and more preferably 0.7 to 0.8 wt %;
[0021] Furthermore, the total colony count of the salmon egg extract is 0 to 300 cfu / g, preferably 0 to 200 cfu / g, and more preferably 0 to 100 cfu / g.
[0022] In order to achieve the above object, according to another aspect of the present invention, there is provided the use of the salmon egg extract obtained by the preparation method of the salmon egg extract in food, health products or cosmetics.
[0023] By applying the technical solution of the present application, the preparation method of the salmon egg extract provided by the present application first uses a salt solution to wash the salmon eggs, which can maintain the osmotic pressure of the salmon eggs while removing impurities and avoiding the leakage of effective ingredients. The salmon eggs are then washed with water and then crushed and solid-liquid separated to obtain a salmon egg homogenate. The salmon egg homogenate is then defatted to remove saturated fatty acids and reduce the content of saturated fatty acids. Finally, it is purified and homogenized to improve the purity of the salmon egg extract while reducing the protein molecular weight and microbial content, thereby effectively expanding the application scenarios of the salmon egg extract.
[0024] In addition, the preparation method of the salmon egg extract provided in the present application is not only simple in process and low in cost, and can be suitable for industrial production, but also the prepared salmon egg extract retains a variety of nutrients and their biological activities in the salmon eggs, reduces the saturated fatty acid content and the molecular weight of the protein, ensures the purity of the product, and effectively reduces the microbial content of the product, and has a high application value in the fields of cosmetics, food, and health care products. DETAILED DESCRIPTION
[0025] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present invention will be described in detail below in conjunction with the embodiments.
[0026] As analyzed in the background technology of this application, the current research on salmon egg extracts is still at the stage of extracting a single component, and some extraction processes will cause the inactivation of natural active substances. Therefore, it is urgent to develop a preparation method for salmon egg extracts that can retain multiple nutrients and maintain high biological activity. In order to solve this problem, this application provides a preparation method and application of salmon egg extracts.
[0027] In a first typical embodiment of the present application, a method for preparing a salmon egg extract is provided, the method comprising: step S1, first washing the salmon eggs with a salt solution and water in turn to obtain purified salmon eggs; step S2, crushing the purified salmon eggs and performing solid-liquid separation to obtain a salmon egg homogenate; step S3, defatting the salmon egg homogenate and then performing solid-liquid separation to obtain a crude salmon egg extract; step S4, purifying and homogenizing the crude salmon egg extract in turn to obtain a salmon egg extract.
[0028] By applying the technical solution of the present application, the preparation method of the salmon egg extract provided by the present application first uses a salt solution to wash the salmon eggs, which can maintain the osmotic pressure of the salmon eggs while removing impurities and avoiding the leakage of effective ingredients. The salmon eggs are then washed with water and then crushed and solid-liquid separated to obtain a salmon egg homogenate. The salmon egg homogenate is then defatted to remove saturated fatty acids and reduce the content of saturated fatty acids. Finally, it is purified and homogenized to improve the purity of the salmon egg extract while reducing the protein molecular weight and microbial content, thereby effectively expanding the application scenarios of the salmon egg extract.
[0029] In addition, the preparation method of the salmon egg extract provided in the present application is not only simple in process and low in cost, and can be suitable for industrial production, but also the prepared salmon egg extract retains a variety of nutrients and their biological activities in the salmon eggs, reduces the saturated fatty acid content and the molecular weight of the protein, ensures the purity of the product, and effectively reduces the microbial content of the product, and has a high application value in the fields of cosmetics, food, and health care products.
[0030] In the above step S1, the specific type of the salt solution is not limited, and any salt solution that can maintain the osmotic pressure of the salmon eggs and prevent the effective ingredients in the salmon eggs from seeping out can be used, including but not limited to any one of sodium chloride solution, potassium chloride solution, magnesium chloride solution, calcium chloride solution or ammonium chloride solution.
[0031] By washing the salmon eggs with a salt solution and water in sequence, the impurities on the surface of the salmon eggs and the dryness of some of the mucous membrane tissue structures remaining in the salmon eggs are significantly reduced, thereby improving the purity of the salmon egg extract.
[0032] In order to further remove impurities on the surface of salmon eggs, the mass volume concentration of the above-mentioned hydrochloric acid solution is preferably 10-60g / L, such as 10g / L, 15g / L, 20g / L, 30g / L, 40g / L, 50g / L, 60g / L or a range consisting of any two values.
[0033] In order to further improve the purity of the salmon egg extract, the salt solution preferably includes multiple groups of salt solutions with mass volume concentrations decreasing from 60g / L to 10g / L; the salmon eggs are washed in sequence using multiple groups of salt solutions, and the mass volume concentrations of the multiple groups of salt solutions decrease in sequence. In particular, when the material-liquid ratio of salmon eggs to salt solution is 1:1 to 1:2g / L, it is more conducive to improving the washing efficiency, and in particular, when the material-liquid ratio of salmon eggs to salt solution is 1:2g / L, the washing efficiency is higher.
[0034] In some embodiments, salt solutions with different mass volume concentrations (60g / L, 50g / L, 40g / L, 30g / L, 20g / L, 10g / L) are prepared and used to wash salmon eggs in sequence. The salmon eggs are washed in sequence from high to low concentrations, with each washing lasting 10-20 minutes (such as 10 minutes, 12 minutes, 15 minutes, 18 minutes, 20 minutes, etc.), and the water is drained and set aside.
[0035] In the above step S1, the salt solution is washed with water to elute the salt solution, and the water preferably includes but is not limited to any one or more of purified water, deionized water or distilled water.
[0036] In the above step S2, there are many ways to separate the solid and liquid, preferably centrifugation is used for solid-liquid separation. In order to further improve the preparation efficiency of salmon egg homogenate, the centrifugal speed is preferably 5000-8500rpm / min (such as 5000rpm / min, 5500rpm / min, 6000rpm / min, 6500rpm / min, 7000rpm / min, 7500rpm / min, 8000rpm / min, 8500rpm / min, etc.), and the time is 8-15min (such as 8min, 10min, 12min, 15min, etc.).
[0037] In some specific embodiments, in the above step S2, after crushing by a crusher, centrifugation is performed to remove the precipitate, and the obtained supernatant is the salmon egg homogenate.
[0038] In order to reduce the saturated fatty acid content in the salmon egg extract, in the above step S3, the salmon egg homogenate is defatted to remove the saturated fatty acids, and then solid-liquid separation is performed, and the obtained supernatant is the crude salmon egg extract. Specifically, in some embodiments, a degreasing agent is added to the salmon egg homogenate for degreasing, and the degreasing agent is a commonly used degreasing agent in the art, including but not limited to a mixture of any one or more of methanol, ethanol, isopropanol, sodium hydroxide, and potassium hydroxide.
[0039] The saturated fatty acids in salmon eggs are insoluble fats. Excessive intake of saturated fatty acids may interfere with the normal function of the skin, leading to water loss and dry skin. It may also accelerate the degradation of collagen, leading to sagging skin and wrinkles. It may also affect sebum secretion and inflammatory response, aggravating the occurrence of acne. Therefore, the salmon eggs need to be homogenized and defatted.
[0040] In order to further improve the degreasing efficiency, the mass ratio of the degreasing agent to the salmon egg homogenate is preferably 5% to 12%, such as 5%, 8%, 10%, 12% or a range consisting of any two values.
[0041] In order to further reduce the content of saturated fatty acids in salmon egg extract, the preferred defatting treatment temperature is 15-35°C (such as 15°C, 18°C, 20°C, 22°C, 25°C, 28°C, 30°C) and the time is 12-60h (such as 12h, 15h, 20h, 30h, 40h, 50h, 60h, etc.).
[0042] In order to further improve the removal rate of saturated fatty acids, it is preferred to first adjust the pH of the salmon egg homogenate to 6-8 (such as 6, 6.5, 7, 7.5, 8, etc.), and then add a degreasing agent to mix and perform degreasing treatment.
[0043] In the above step S3, the solid-liquid separation method is preferably centrifugation, and the precipitate is removed by centrifugation, and the obtained supernatant is the crude salmon egg extract.
[0044] In the above step S4, purification treatment is performed by conventional methods in the art to reduce the content of microorganisms in the salmon egg extract. The purification treatment method includes but is not limited to any one or more of filtration, ultrafiltration or dialysis.
[0045] In order to further reduce the molecular weight of protein in salmon egg extract, it is preferred that the homogenization pressure is 300-500 MPa and the time is 10-30 min, such as the homogenization pressure is 300 MPa, 350 MPa, 400 MPa, 450 MPa, 500 MPa or a range consisting of any two values, and the homogenization time is 10 min, 15 min, 20 min, 25 min, 30 min or a range consisting of any two values.
[0046] In order to further improve the stability of the salmon egg extract, the above-mentioned step S4 preferably also includes a pH adjustment after the homogenization treatment, and the pH of the salmon egg extract is adjusted to 5.5-7.4 (such as 5.5, 6, 6.5, 7, 7.2, 7.4, etc.), so as to be more conducive to the use of the salmon egg extract in cosmetics, health products or food.
[0047] In some embodiments, the molecular weight of the protein in the salmon egg extract obtained by the above-mentioned preparation method of the salmon egg extract is 11-48 kDa, preferably 11-25 kDa, and more preferably 11-20 kDa; the saturated fatty acid content of the above-mentioned salmon egg extract is 0.6%-0.9wt%, preferably 0.7%-0.9%, and more preferably 0.7%-0.8%; the total colony count of the above-mentioned salmon egg extract is 0-300 cfu / g, preferably 0-200 cfu / g, and more preferably 0-100 cfu / g.
[0048] Typically but not limiting, the protein molecular weight of the salmon egg extract obtained by the preparation method of the salmon egg extract provided in the present application is 11 kDa, 12 kDa, 13 kDa, 15 kDa, 16 kDa, 17 kDa, 18 kDa, 20 kDa, 22 kDa, 25 kDa, 28 kDa, 30 kDa, 35 kDa, 38 kDa, 40 kDa, 45 kDa, 48 kDa or a range consisting of any two values; the saturated fatty acid mass content of the salmon egg extract is 0.6%, 0.62%, 0.65%, 0.68%, 0.7%, 0.7 2%, 0.74%, 0.75%, 0.78%, 0.8%, 0.85%, 0.9% or a range consisting of any two values; the total colony count of salmon egg extract is 0 cfu / g, 5 cfu / g, 10 cfu / g, 20 cfu / g, 30 cfu / g, 40 cfu / g, 50 cfu / g, 60 cfu / g, 70 cfu / g, 80 cfu / g, 90 cfu / g, 100 cfu / g, 150 cfu / g, 200 cfu / g, 250 cfu / g, 300 cfu / g or a range consisting of any two values.
[0049] In a second typical embodiment of the present application, there is also provided the use of the salmon egg extract prepared in the first typical embodiment in food, health products or cosmetics.
[0050] The preparation method of the salmon egg extract provided in the present application first washes the salmon eggs with a salt solution and water in sequence to remove impurities, then performs crushing treatment, solid-liquid separation and homogenization treatment in sequence, and finally performs purification treatment and homogenization treatment. The obtained salmon egg extract not only has significantly reduced impurity content, microbial content and protein molecular weight, but also retains a variety of natural nutrients in the salmon eggs and the biological activity of the nutrients, so that the salmon egg extract has broad application prospects in the fields of food, health products or cosmetics, thereby improving its economic benefits.
[0051] The beneficial effects of the present application will be further illustrated below in combination with embodiments and comparative examples.
[0052] Example 1
[0053] This embodiment provides a method for preparing a salmon egg extract, which comprises the following steps:
[0054] (1) preparing multiple groups of sodium chloride solutions of different concentrations (60 g / L, 50 g / L, 40 g / L, 30 g / L, 20 g / L, 10 g / L), pouring salmon eggs into a cleaning tank, stirring and cleaning them 6 times in descending order according to the salt concentration at a solid-liquid ratio of 1:2 g / L, each time for 15 minutes, draining the water and pouring them into another cleaning tank, pouring deionized water into the cleaning tank at a solid-liquid ratio of 1:2 g / L, stirring for 10 minutes at room temperature, cleaning a total of 6 times, and draining the water to obtain purified salmon eggs;
[0055] (2) pouring the purified salmon eggs into a crusher for crushing, centrifuging the crushed salmon eggs at a centrifugal speed of 8000 rpm for 10 min, removing the precipitate and taking the supernatant to obtain salmon egg homogenate;
[0056] (3) adjusting the pH of the salmon egg homogenate to 6-8, adding methanol to the salmon egg homogenate at room temperature, shaking and stirring for 24 hours to perform defatting, and centrifuging to obtain the supernatant to obtain a crude salmon egg extract, wherein the amount of methanol is 10 wt% of the salmon egg homogenate;
[0057] (4) The crude salmon egg extract is purified by an ultrafiltration system and subjected to high-pressure homogenization treatment in sequence, and the pH value is adjusted to 5.5-7.4 after the homogenization treatment to obtain the salmon egg extract; wherein the high-pressure homogenization treatment is performed at a pressure of 500 MPa, for 10 min, and for 2 times.
[0058] Comparative Example 1.1
[0059] This comparative example provides a method for preparing a salmon egg extract, which comprises the following steps:
[0060] (1) treating salmon eggs: washing salmon eggs (20 g) with deionized water to remove impurities, and removing water for later use to obtain treated salmon eggs;
[0061] (2) Defatting: Add 6 times the amount of edible alcohol (relative to the mass of the treated sturgeon eggs) to the treated salmon eggs, immerse at a constant temperature of 40°C for 40 min, filter out the alcohol with filter paper, and repeatedly wash with clean water until there is no alcohol smell, thereby obtaining defatted salmon eggs;
[0062] (3) Enzymatic hydrolysis: grind the defatted salmon eggs, add 8 times the amount (relative to the mass of the defatted salmon eggs) of distilled water, add papain until the enzyme activity reaches 300-3000U per 1g of substrate (i.e., defatted salmon eggs), and adjust the pH to 6.5-7.5; divide the above solution into five portions, perform enzymatic hydrolysis at 60°C for 0.5 hour, 1 hour, 2 hours, 3 hours and 4 hours respectively, filter with filter paper to obtain enzymatic hydrolyzate;
[0063] (4) Centrifugation: Centrifuge the enzymatic hydrolysates at 9000 r / min and take the supernatant;
[0064] (5) Freeze drying: The obtained supernatant was freeze dried by a freeze dryer to obtain a salmon egg extract. The salmon egg extract was found to be salmon egg protein peptide after testing.
[0065] Comparative Example 1.2
[0066] This comparative example provides a method for preparing a salmon egg extract, which comprises the following steps:
[0067] (1) 2.0 kg of fresh or room temperature thawed salmon eggs were taken, and after impurities were removed, washed three times, homogenized by colloid milling, and freeze-dried, 3 L of 95 wt% ethanol was added for defatting for 15 h, during which the ethanol was changed four times; centrifuged at 3500 rpm for 10 min, the supernatant was removed, and the precipitate was defatted salmon egg protein;
[0068] (2) Defatted salmon egg protein was freeze-dried, and 6 L of distilled water was added and preheated to 50°C; 3 g of neutral protease and 3 g of alkaline protease were added under constant temperature stirring for hydrolysis for 5 h, and the pH was 7.0; then the enzyme activity was inactivated at 100°C for 7 min, the enzymatic solution was cooled to room temperature, centrifuged at 5000 rpm for 5 min, the supernatant was taken, ultrafiltered through an ultrafiltration membrane with a molecular weight cutoff of 10 kD, and the filtrate was freeze-dried to obtain a salmon egg extract. After testing, the salmon egg extract was found to be salmon egg protein peptide.
[0069] Comparative Example 1.3
[0070] This comparative example provides a method for preparing a salmon egg extract, which comprises the following steps:
[0071] (1) 50 g of salmon eggs were loaded into an extraction kettle barrel for de-oiling treatment, wherein the fish oil extraction conditions were: extraction pressure 25 MPa, extraction time 90 min, extraction temperature 40° C., CO2 flow rate 25 L / h, to obtain an extract;
[0072] (2) adding 5% by weight of ethanol to the obtained extract, and extracting under the conditions of extraction pressure of 30 MPa, extraction temperature of 30° C., extraction time of 2 h, and CO2 flow rate of 15 L / h to obtain an extract;
[0073] (3) The extract was distilled under reduced pressure, concentrated and dried to obtain 1.83 g of salmon egg extract. The salmon egg extract was found to be phospholipids with an unsaturated fatty acid content of 39.05 wt %.
[0074] Comparative Example 1.4
[0075] This comparative example provides a method for preparing a salmon egg extract, which comprises the following steps:
[0076] Weigh 2kg of frozen salmon eggs, thaw naturally at room temperature, and boil in boiling water for 10 minutes. Take out the salmon eggs (weighing 1.98kg) and mix with 2.5kg of deionized water to make pulp, adjust its pH to 5.5, then add 2g of phospholipase Lecitaseultra (phospholipase A1), 3g of phospholipase PLA2NAGASE (phospholipase A2) and 10g of papain to the pulp, mix well and stir to react at 50°C. After 6 hours of reaction, heat the reaction mixture to boiling and keep it warm for 15 minutes to fully inactivate the added enzyme preparation, freeze-dry the reaction mixture to a water content of 3wt% and add it to an extraction kettle, extract with supercritical carbon dioxide at a temperature of 50°C, a carbon dioxide flow rate of 100L / h, and a pressure of 33MPa for 4 hours to obtain salmon egg extract. After testing, the total fat content of the salmon egg extract is 0.4wt%.
[0077] Example 2.1
[0078] This embodiment provides a method for preparing a salmon egg extract, which comprises the following steps:
[0079] (1) preparing multiple groups of potassium chloride solutions of different concentrations (60 g / L, 50 g / L, 40 g / L, 30 g / L, 20 g / L, 10 g / L), pouring salmon eggs into a cleaning tank, stirring and cleaning them 6 times in descending order of salt concentration according to a solid-liquid ratio of 1:2 g / L, each time for 15 minutes, draining the water and pouring them into another cleaning tank, pouring deionized water into the cleaning tank according to a solid-liquid ratio of 1:2 g / L, stirring for 15 minutes at room temperature, cleaning a total of 6 times, and draining the water to obtain purified salmon eggs;
[0080] (2) pouring the purified salmon eggs into a crusher for crushing, centrifuging the crushed salmon eggs at a centrifugal speed of 7500 rpm for 10 min, removing the precipitate and taking the supernatant to obtain salmon egg homogenate;
[0081] (3) adjusting the pH of the salmon egg homogenate to 6-8, adding ethanol to the salmon egg homogenate at room temperature, shaking and stirring for 12 hours to perform defatting, and centrifuging to obtain the supernatant to obtain a crude salmon egg extract, wherein the amount of ethanol used is 12 wt% of the salmon egg homogenate;
[0082] (4) The crude salmon egg extract is purified by an ultrafiltration system and subjected to high-pressure homogenization treatment in sequence, and the pH value is adjusted to 5.5-7.4 after the homogenization treatment to obtain the salmon egg extract; wherein the pressure of the high-pressure homogenization treatment is 300 MPa, the time is 10 min, and the number of times is 2 times.
[0083] Example 2.2
[0084] The difference between this embodiment and embodiment 2.1 is that in step (1), when potassium chloride solution is used for cleaning, the cleaning is carried out by stirring in order from low to high salt solution concentrations.
[0085] Example 2.3
[0086] The difference between this embodiment and embodiment 2.1 is that in step (1), only a potassium chloride solution with a mass concentration of 10 g / L is prepared for cleaning, and the cleaning is stirred and cleaned 6 times in sequence according to a solid-liquid ratio of 1:2 g / L.
[0087] Example 2.4
[0088] The difference between this embodiment and embodiment 2.1 is that in step (1), only a potassium chloride solution with a mass concentration of 60 g / L is prepared for cleaning, and the cleaning is stirred and cleaned 6 times in sequence according to a solid-liquid ratio of 1:2 g / L.
[0089] Example 2.5
[0090] The difference between this embodiment and embodiment 2.1 is that in step (1), when potassium chloride solution is used for cleaning, the material-liquid ratio is 1:0.5.
[0091] Example 2.6
[0092] The difference between this embodiment and embodiment 2.1 is that in step (1), when potassium chloride solution is used for cleaning, the material-liquid ratio is 1:1.
[0093] Example 2.7
[0094] The difference between this embodiment and embodiment 2.1 is that in step (1), when potassium chloride solution is used for cleaning, the material-liquid ratio is 1:3.
[0095] Comparative Example 2.1
[0096] The difference between this comparative example and Example 2.1 is that in step (1), no saline solution is used for cleaning.
[0097] Example 3.1
[0098] This embodiment provides a method for preparing a salmon egg extract, which comprises the following steps:
[0099] (1) preparing multiple groups of magnesium chloride solutions of different concentrations (60 g / L, 50 g / L, 40 g / L, 30 g / L, 20 g / L, 10 g / L), pouring salmon eggs into a cleaning tank, stirring and cleaning them 6 times in descending order of salt concentration according to a solid-liquid ratio of 1:2 g / L, each time for 15 min, draining the water and pouring them into another cleaning tank, pouring deionized water into the cleaning tank according to a solid-liquid ratio of 1:1 g / L, stirring for 5 min at room temperature, cleaning 3 times in total, and draining the water to obtain purified salmon eggs;
[0100] (2) pouring the purified salmon eggs into a crusher for crushing, centrifuging the crushed salmon eggs at a centrifugal speed of 8500 rpm for 10 min, removing the precipitate and taking the supernatant to obtain salmon egg homogenate;
[0101] (3) adjusting the pH of the salmon egg homogenate to 6-8, adding ethanol to the salmon egg homogenate at room temperature, shaking and stirring for 60 hours to perform defatting, and centrifuging to obtain the supernatant to obtain a crude salmon egg extract, wherein the amount of ethanol used is 8 wt% of the salmon egg homogenate;
[0102] (4) The crude salmon egg extract is purified by an ultrafiltration system and subjected to high-pressure homogenization treatment in sequence, and the pH value is adjusted to 5.5-7.4 after the homogenization treatment to obtain the salmon egg extract; wherein the pressure of the high-pressure homogenization treatment is 400 MPa, the time is 10 min, and the number of times is 2 times.
[0103] Example 3.2
[0104] The difference between this embodiment and embodiment 3.1 is that in step (3), sodium hydroxide is used instead of ethanol as the degreasing agent, and the amount of sodium hydroxide used is 8 wt % of the amount of salmon eggs used.
[0105] Example 3.3
[0106] The difference between this embodiment and embodiment 3.1 is that in step (3), isopropanol is used instead of ethanol as the degreasing agent, and the amount of isopropanol used is 10 wt % of the amount of salmon eggs used.
[0107] Example 3.4
[0108] The difference between this embodiment and embodiment 3.1 is that in step (3), potassium hydroxide is used instead of ethanol as the degreasing agent, and the amount of potassium hydroxide used is 10 wt % of the amount of salmon eggs used.
[0109] Example 3.5
[0110] The difference between this embodiment and embodiment 3.1 is that in step (3), the oscillating and stirring reaction time is 6 hours.
[0111] Comparative Example 3.1
[0112] The difference between this comparative example and Example 3.1 is that step (3) is not performed, and the salmon egg homogenate obtained in step (2) is directly subjected to ultrafiltration system purification and high-pressure homogenization treatment in step (4).
[0113] Example 4.1
[0114] This embodiment provides a method for preparing a salmon egg extract, which comprises the following steps:
[0115] (1) preparing multiple groups of calcium chloride solutions of different concentrations (60 g / L, 50 g / L, 40 g / L, 30 g / L, 20 g / L, 10 g / L), pouring salmon eggs into a cleaning tank, stirring and cleaning them 6 times in descending order according to the salt concentration at a solid-liquid ratio of 1:2 g / L, each time for 15 min, draining the water and pouring them into another cleaning tank, pouring deionized water into the cleaning tank at a solid-liquid ratio of 1:1 g / L, stirring for 20 min at room temperature, cleaning a total of 6 times, and draining the water to obtain purified salmon eggs;
[0116] (2) pouring the purified salmon eggs into a crusher for crushing, centrifuging the crushed salmon eggs at a centrifugal speed of 7500 rpm for 10 min, removing the precipitate and taking the supernatant to obtain salmon egg homogenate;
[0117] (3) adjusting the pH of the salmon egg homogenate to 6-8, adding sodium hydroxide to the salmon egg homogenate at room temperature, shaking and stirring for 12 hours to perform defatting, and centrifuging to obtain the supernatant to obtain a crude salmon egg extract, wherein the amount of sodium hydroxide used is 12 wt% of the salmon egg homogenate;
[0118] (4) The crude salmon egg extract is purified by an ultrafiltration system and subjected to high-pressure homogenization treatment in sequence, and the pH value is adjusted to 5.5-7.4 after the homogenization treatment to obtain the salmon egg extract; wherein the high-pressure homogenization treatment is performed at a pressure of 450 MPa, for 10 min, and for 2 times.
[0119] Example 4.2
[0120] The difference between this embodiment and embodiment 4.1 is that in step (4), a filtration replacement ultrafiltration system is used for purification, and the specification of the filter paper is a circular filter paper with a diameter of 9 cm.
[0121] Example 4.3
[0122] The difference between this embodiment and embodiment 4.1 is that in step (4), a dialysis replacement ultrafiltration system is used for purification, and the dialysis bag used is a regenerated cellulose dialysis bag (25 mm, 3.5 kDa, dry type) of Biyuntian.
[0123] Comparative Example 4.1
[0124] The difference between this comparative example and Example 4.1 is that in step (4), no ultrafiltration system purification treatment is performed.
[0125] Example 5.1
[0126] This embodiment provides a method for preparing a salmon egg extract, which comprises the following steps:
[0127] (1) preparing multiple groups of ammonium chloride solutions of different concentrations (60 g / L, 50 g / L, 40 g / L, 30 g / L, 20 g / L, 10 g / L), pouring salmon eggs into a cleaning tank, stirring and cleaning them 6 times in descending order according to the salt concentration at a solid-liquid ratio of 1:2 g / L, each time for 15 min, draining the water and pouring them into another cleaning tank, pouring deionized water into the cleaning tank at a solid-liquid ratio of 1:5 g / L, stirring for 20 min at room temperature, cleaning a total of 6 times, and draining the water to obtain purified salmon eggs;
[0128] (2) pouring the purified salmon eggs into a crusher for crushing, centrifuging the crushed salmon eggs at a centrifugal speed of 5000 rpm for 10 min, removing the precipitate and taking the supernatant to obtain salmon egg homogenate;
[0129] (3) adjusting the pH of the salmon egg homogenate to 6-8, adding ethanol to the salmon egg homogenate at room temperature, shaking and stirring for 12 hours to perform defatting, and centrifuging to obtain the supernatant to obtain a crude salmon egg extract, wherein the amount of ethanol used is 5 wt% of the salmon egg homogenate;
[0130] (4) The crude salmon egg extract is purified by an ultrafiltration system and subjected to high-pressure homogenization treatment in sequence, and the pH value is adjusted to 5.5-7.4 after the homogenization treatment to obtain the salmon egg extract; wherein the pressure of the high-pressure homogenization treatment is 400 MPa, the time is 10 min, and the number of times is 2 times.
[0131] Example 5.2
[0132] The difference between this embodiment and embodiment 5.1 is that in step (4), the pressure of the high-pressure homogenization treatment is 100 MPa.
[0133] Example 5.3
[0134] The difference between this embodiment and embodiment 5.1 is that in step (4), homogenization is carried out under normal pressure.
[0135] Comparative Example 5.1
[0136] The difference between this embodiment and embodiment 5.1 is that in step (4), no homogenization treatment is performed.
[0137] Test example
[0138] The extraction rate, saturated fatty acid content, microbial colony count and protein molecular weight of the salmon egg extract in the above examples and comparative examples were measured respectively, and the results are shown in Tables 1 to 5 below.
[0139] Wherein, (1) the salmon egg extraction rate is calculated according to the following formula:
[0140] The extraction rate of salmon egg extract = (salmon egg extract / salmon egg) x 100%.
[0141] (2) The saturated fatty acid content was determined in accordance with the National Standard of the People’s Republic of China GB / T 2223-2008.
[0142] (3) The microbial colony count shall be determined in accordance with the microbiological testing methods in Chapter 5 of the Technical Specifications for Safety of Cosmetics (2015 Edition).
[0143] (4) The protein molecular weight was determined by electrophoresis and MALDI-TOF-MS according to General Chapter 0541 of the Pharmacopoeia of the People’s Republic of China (2020 edition).
[0144] Table 1
[0145]
[0146] Table 2
[0147]
[0148]
[0149] Table 4
[0150]
[0151] Table 5
[0152]
[0153] From the above description, it can be seen that the above embodiments of the present invention achieve the following technical effects:
[0154] The preparation method of the salmon egg extract provided in the present application first uses a salt solution to wash the salmon eggs, which can maintain the osmotic pressure of the salmon eggs while removing impurities and avoiding the leakage of effective ingredients. The salmon eggs are then washed with water and then crushed and solid-liquid separated to obtain a salmon egg homogenate. The salmon egg homogenate is then defatted to remove saturated fatty acids and reduce the content of saturated fatty acids. Finally, it is purified and homogenized to improve the purity of the salmon egg extract while reducing the protein molecular weight and microbial content, thereby effectively expanding the application scenarios of the salmon egg extract.
[0155] In addition, the preparation method of the salmon egg extract provided in the present application is not only simple in process and low in cost, and can be suitable for industrial production, but also the prepared salmon egg extract retains a variety of nutrients and their biological activities in the salmon eggs, reduces the saturated fatty acid content and the molecular weight of the protein, ensures the purity of the product, and effectively reduces the microbial content of the product, and has a high application value in the fields of cosmetics, food, and health care products.
[0156] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.
Claims
1. A method for preparing a salmon egg extract, characterized in that: The preparation method comprises: Step S1, first washing the salmon eggs with a salt solution and then with water to obtain purified salmon eggs; Step S2, crushing the purified salmon eggs and performing solid-liquid separation to obtain salmon egg homogenate; Step S3, defatting the salmon egg homogenate and then performing solid-liquid separation to obtain a crude salmon egg extract; Step S4, purifying and homogenizing the crude salmon egg extract in sequence to obtain the salmon egg extract.
2. The preparation method according to claim 1, characterized in that: In step S1, the mass volume concentration of the salt solution is 10 to 60 g / L; Preferably, the salt solution comprises a plurality of groups of salt solutions whose mass volume concentration decreases successively from 60 g / L to 10 g / L; the salmon eggs are washed successively with the plurality of groups of the salt solutions, and the mass volume concentration of the plurality of groups of the salt solutions decreases successively.
3. The preparation method according to claim 1, characterized in that: In step S1, the salt solution includes at least one of sodium chloride solution, potassium chloride solution, magnesium chloride solution, calcium chloride solution or ammonium chloride; and / or, the water comprises at least one of purified water, deionized water or distilled water; And / or, the material-liquid ratio of the salmon roe to the salt solution is 1:1 to 1:2 g / L, more preferably 1:2 g / L.
4. The preparation method according to claim 1, characterized in that: In the step S2, the solid-liquid separation is performed by centrifugation, and preferably the centrifugation speed is 5000-8500 rpm / min and the time is 8-15 min.
5. The preparation method according to claim 1, characterized in that: The step S3, adding a degreasing agent to the salmon egg homogenate to perform the degreasing treatment, wherein the degreasing agent includes at least one of methanol, ethanol, isopropanol, sodium hydroxide, and potassium hydroxide; Preferably, the mass ratio of the degreasing agent to the salmon egg homogenate is 5 to 12:100; Preferably, the degreasing treatment is carried out for 12 to 60 hours at a temperature of 15 to 35°C.
6. The preparation method according to claim 1, characterized in that: In step S4, the purification treatment comprises at least one of filtration, ultrafiltration or dialysis.
7. The preparation method according to claim 1, characterized in that: The homogenization treatment is performed under a pressure of 300 to 500 MPa and for a time of 10 to 30 minutes.
8. The preparation method according to any one of claims 1 to 7, characterized in that The step S4 further includes adjusting the pH after the homogenization process, and the pH of the salmon egg extract is adjusted to be 5.5 to 7.
4.
9. The preparation method according to any one of claims 1 to 7, characterized in that: The protein molecular weight of the salmon egg extract is 11-48 kDa, preferably 11-25 kDa, and more preferably 11-20 kDa; and / or, the saturated fatty acid content of the salmon egg extract is 0.6 to 0.9 wt %, preferably 0.7 to 0.9 wt %, and more preferably 0.7 to 0.8 wt %; And / or, the total colony count of the salmon egg extract is 0 to 300 cfu / g, preferably 0 to 200 cfu / g, and more preferably 0 to 100 cfu / g.
10. Use of the salmon egg extract obtained by the preparation method according to any one of claims 1 to 9 in food, health products or cosmetics.