Preparation method of vetch extract and eye protection liquid containing vetch extract
By combining traditional extraction and biofermentation methods, acidic ethanol and selenoprotein raw materials are used to solve the problems of complex extraction process of wild pea and insufficient eye protection liquid performance, achieving high clarity, good atomization effect and strong red blood vessel removal ability eye protection liquid significantly improves the bioavailability and stability of the product.
Patent Information
- Application Number
- CN202411460672.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-18
- Publication Date
- 2025-01-03
AI Technical Summary
The existing wild pea extraction process is complex, the purification process is not thorough, the clarity and transparency of eye protection liquid are insufficient, the atomization effect is poor, the irritation is great, the red blood vessel removal ability is weak, the stability is insufficient, and the selenium element is not effectively utilized.
Using a combination of traditional extraction and biofermentation, the pea seeds were washed with ultra-low deuterium water at pH 1.5, soaked in selenoglucose solution, sprayed with L-selenomethionine, and carried out acid-base precipitation treatment. , activated carbon and macroporous resin were decolorized, and selenoid protein raw materials were added to obtain a wild pea extract rich in negative divalent selenium by spray drying, and a clear eye protection liquid was prepared.
Improves the clarity, transparency and atomization effect of eye protection liquid, reduces irritation, enhances the ability and stability of red blood vessels, and gives the product the ability to balance intraocular pressure and repair eye muscle tissue, significantly improving bioavailability and the effect of preventing eye diseases.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of eye protection liquid, in particular to a method for preparing a vetch extract and an eye protection liquid containing the extract. Background Art
[0002] Pea is a common perennial herb, rich in flavonoids, polysaccharides and other chemical substances, and is widely used in anti-oxidation, antibacterial and anti-inflammatory, liver-clearing and eye-improving. At present, the main extraction processes for wild peas include ethanol reflux, acid-base precipitation treatment, resin decolorization extraction and other methods. Patent CN104017099B, a microwave extraction, separation and purification method for pea polysaccharides, discloses that peas are subjected to microwave extraction with ethanol water, and then adsorbed by D3520 macroporous adsorption resin, and finally purified with ethanol to obtain pea polysaccharides with a high extraction rate. However, the pea extract has no good practical application in eye protection products.
[0003] The present invention aims to combine traditional extraction and biological fermentation, improve bioavailability, add a new component "selenium" element to the extract, and achieve the effect of containing negative divalent selenoprotein components in the final eye protection liquid product. It can solve the problems of complex process of wild pea extraction, purification in the refining link, and no need to study "selenoprotein" separately, and at the same time improve the problems of unclear eye protection liquid, insufficient transparency, poor atomization effect, strong irritation, weak red blood streak removal ability, and good stability. Summary of the invention
[0004] 1. Technical issues to be solved
[0005] In view of the deficiencies in the prior art, the present invention provides an eye protection solution containing selenium-enriched Vicia fructus extract.
[0006] (II) Technical solution: A method for preparing a wild pea extract:
[0007] (1) Wash the vetch seeds with ultra-low deuterium water at a pH of 1.5, dry them after washing, and then place them in a solution of seleno-glucose with a mass fraction of 0.2-0.4% for 18-24 hours. After soaking, place the vetch seeds in a seedling tray for seedling cultivation. After the seedling cultivation is completed, transplant them. During the growth period of the vetch, spray the leaves with a solution of L-selenomethionine with a mass fraction of 0.1-0.16% for 5-8 times. When mature, harvest the vetch, dry the roots, wash them with water, dry them, and crush them to obtain coarse vetch powder rich in organic selenium.
[0008] (2) Add the organic selenium-rich wild pea powder into an acidic ethanol solution for extraction, collect the filtrate and the residue after filtering, add sodium hydroxide solution to the filtrate, adjust the pH to neutral, and concentrate to obtain a thick paste.
[0009] (3) Add the thick paste to a saturated calcium hydroxide aqueous solution, stir, then drop dilute sulfuric acid, let stand for 18-24 hours, remove the supernatant, filter, add sodium hydroxide solution to the filtrate, and concentrate to a density of 1.15-1.25 g / cm 3 , let it stand for 18-24 hours, filter and add the precipitate into the hydrochloric acid solution, stir and centrifuge, take the supernatant and obtain the centrifuged liquid.
[0010] (4) The centrifuged liquid is decolorized by activated carbon, and then chromatographically adsorbed by macroporous resins D101, AB-8 and A-722 in sequence, and finally mechanically eluted with ethanol. The ethanol eluate is collected, concentrated, and diluted to a concentration of 0.4-0.6 g / mL by adding ultra-low deuterium-free water. The liquid is filtered through a 0.45 μm filter membrane to obtain a clear liquid, which is then spray-dried to obtain a fine powder of wild pea extract rich in organic selenium.
[0011] Furthermore, in (2), the ratio of acidic ethanol added to the organic selenium-rich wild pea powder is (10-60) g:1L.
[0012] Furthermore, the acidic ethanol is an ethanol solution of HCl, and the molar concentration is (10 -3 -10 -5 )mol / L.
[0013] Furthermore, in (2), the extraction is performed by condensing and reflux at 75-85°C for 6-12h.
[0014] Furthermore, in (3), dilute sulfuric acid is added dropwise to adjust the pH to 2-4; sodium hydroxide solution is added to adjust the pH to 10-12; and the pH of the hydrochloric acid solution is 2-4.
[0015] Furthermore, a method for preparing eye protection liquid using fine powder of wild pea extract is characterized in that the method comprises the following steps: mixing fine powder of wild pea extract, sodium chloride, taurine, cooling agent menthol, distilled water, and selenoprotein raw material, dissolving them with 600 ppm ultra-low deuterium water and hydrogen-rich water, adjusting the pH to 6-8, filtering with a 0.22 μm microporous filter membrane, and sterilizing them with a sterilization device for 10-20 minutes at a sterilization temperature of 115-125° C. to obtain eye protection liquid.
[0016] Furthermore, the ratio of wild pea extract fine powder, sodium chloride, taurine, cooling agent, distilled water, and selenoprotein raw material is (0.02-0.15) g: (8.5-9) g: (0.04-0.12) g: (0.05-0.08) g: 1 L: (0.03-0.2) g.
[0017] Furthermore, the preparation method of the selenoprotein raw material is as follows: placing the filter residue collected in (2) in a fermentation tank, using Escherichia coli as the fermentation strain, using glucose and corn syrup as nutrient fermentation raw materials, and performing bacterial fermentation, filtration, sterilization, hydrolysis, purification, and crystallization to obtain a selenoprotein raw material rich in active selenium.
[0018] (III) Beneficial technical effects: The patent of the present invention combines traditional extraction and biological fermentation, mainly focusing on the screening, processing, and cultivation of wild pea seeds, as well as extraction, refining, and fermentation, which improves the bioavailability and gives the extract a new ingredient, "selenium". The entire series of processes ensures that the final eye protection liquid product contains negative divalent selenoprotein components.
[0019] The acid ethanol is used to reflux extract the wild pea, and the acid-base precipitation process improves the utilization of the alkaloids of the wild pea's effective ingredients; the activated carbon and macroporous resin refining and decolorization process reduces the damage to the eyes caused by colored substances in the eye drops; the spray drying process is convenient and fast, reducing the risk of pollution. The eye drops are used in an atomized manner, which improves the bioavailability and enhances the full absorption rate of the drug.
[0020] The patent solves the problems of complex extraction process of wild pea, purification in the refining step, no need to study "selenoprotein" separately, improved bioavailability, and the problems of unclear eye solution, insufficient transparency, poor atomization effect, high irritation, weak red blood streak removal ability, and good stability.
[0021] The eye protection liquid obtained by the invention has the advantages of clarity, high transparency, good atomization effect, low irritation, strong ability to remove red blood streaks, stable properties, etc. It is also a unique product that can balance intraocular pressure and repair eye muscle tissue cells, has strong ability to balance intraocular pressure, good effect of preventing and repairing eye diseases, fast effect, can quickly relieve eye fatigue and red blood streaks, and has extremely strong prevention and repair ability for myopia, hyperopia, astigmatism, presbyopia, floaters, glaucoma, cataracts and other problems. DETAILED DESCRIPTION
[0022] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0023] Example 1
[0024] (1) Wash the vetch seeds with ultra-low deuterium water at a pH of 1.5, dry them after washing, and then place them in a solution of 0.4% by mass of seleno-glucose for 18 hours. After soaking, place the vetch seeds in a seedling tray for seedling cultivation. After the seedling cultivation is completed, transplant them. During the growth period of the vetch, spray the leaves with a solution of 0.12% by mass of L-selenomethionine for 8 times. When they are mature, harvest them, dry the roots of the vetch, wash them with water, dry them, and crush them to obtain coarse vetch powder rich in organic selenium.
[0025] (2) Add 40 g of organic selenium-rich wild pea powder to 1 L of a 10 -3 mol / L HCl in ethanol, heat to 75°C, condense and reflux for 12 hours, collect the filtrate and residue after filtration, add sodium hydroxide solution to the filtrate, adjust the pH to neutral, and concentrate to obtain a thick paste.
[0026] (3) The collected filter residue is placed in a fermentation tank, and Escherichia coli is used as the fermentation bacteria, and glucose and corn syrup are used as nutrient fermentation raw materials. After bacterial fermentation, filtration, sterilization, hydrolysis, purification, and crystallization, a selenoprotein raw material rich in active selenium is obtained.
[0027] (4) Add the thick paste obtained in (2) to a saturated calcium hydroxide aqueous solution, stir, and then add dilute sulfuric acid to adjust the pH to 4. Let stand for 18 hours, remove the supernatant, filter, add sodium hydroxide solution to the filtrate to adjust the pH to 10, and concentrate to a density of 1.2 g / cm 3 , let it stand for 24 hours, filter and add the precipitate into a hydrochloric acid solution with a pH of 2, stir and centrifuge, take the supernatant and obtain the centrifuged liquid.
[0028] (5) The centrifuged liquid is decolorized by activated carbon, and then chromatographically adsorbed by macroporous resins D101, AB-8 and A-722 in sequence, and finally mechanically eluted with ethanol. The ethanol eluate is collected, concentrated, and diluted to a concentration of 0.6 g / mL by adding ultra-low deuterium-free water. The liquid is filtered through a 0.45 μm filter membrane to obtain a clear liquid, which is then spray-dried to obtain a fine powder of wild pea extract rich in organic selenium.
[0029] (6) 0.15 g of wild pea extract powder, 8.5 g of sodium chloride, 0.12 g of taurine, 0.06 g of cooling agent menthol, 1 L of distilled water, and 0.03 g of selenoprotein raw material were mixed, dissolved with 600 ppm ultra-low deuterium water and hydrogen-rich water, the pH was adjusted to 7, filtered with a 0.22 μm microporous filter membrane, and sterilized for 20 min in a sterilization equipment at a sterilization temperature of 120° C. to obtain an eye protection solution.
[0030] Example 2
[0031] (1) Wash the vetch seeds with ultra-low deuterium water at a pH of 1.5, dry them after washing, and then place them in a solution of 0.2% by mass of seleno-glucose for 24 hours. After soaking, place the vetch seeds in a seedling tray for seedling cultivation. After the seedling cultivation is completed, transplant them. During the growth period of the vetch, spray the leaves with a solution of 0.1% by mass of L-selenomethionine for 8 times. When they are mature, harvest them, dry the roots of the vetch, wash them with water, dry them, and crush them to obtain coarse vetch powder rich in organic selenium.
[0032] (2) Add 60 g of organic selenium-rich wild pea powder to 1 L of a 10 -3 mol / L HCl in ethanol, heat to 85°C, condense and reflux for 6 hours, collect the filtrate and residue after filtration, add sodium hydroxide solution to the filtrate, adjust the pH to neutral, and concentrate to obtain a thick paste.
[0033] (3) The collected filter residue is placed in a fermentation tank, and Escherichia coli is used as the fermentation bacteria, and glucose and corn syrup are used as nutrient fermentation raw materials. After bacterial fermentation, filtration, sterilization, hydrolysis, purification, and crystallization, a selenoprotein raw material rich in active selenium is obtained.
[0034] (4) Add the thick paste obtained in (2) to a saturated calcium hydroxide aqueous solution, stir, and then add dilute sulfuric acid to adjust the pH to 2. Let stand for 24 hours, remove the supernatant, filter, add sodium hydroxide solution to the filtrate to adjust the pH to 12, and concentrate to a density of 1.15 g / cm 3 , let it stand for 24 hours, filter and add the precipitate into a hydrochloric acid solution with a pH of 3, stir and centrifuge, take the supernatant and obtain the centrifuged liquid.
[0035] (5) The centrifuged liquid is decolorized by activated carbon, and then chromatographically adsorbed by macroporous resins D101, AB-8 and A-722 in sequence, and finally mechanically eluted with ethanol. The ethanol eluate is collected, concentrated, and diluted to a concentration of 0.4 g / mL by adding ultra-low deuterium-free water. The liquid is filtered through a 0.45 μm filter membrane to obtain a clear liquid, which is then spray-dried to obtain a fine powder of wild pea extract rich in organic selenium.
[0036] (6) 0.02 g of wild pea extract powder, 9 g of sodium chloride, 0.06 g of taurine, 0.08 g of cooling agent menthol, 1 L of distilled water, and 0.2 g of selenoprotein raw material were mixed, dissolved with 600 ppm ultra-low deuterium water and hydrogen-rich water, adjusted to pH 7, filtered with a 0.22 μm microporous filter membrane, and sterilized for 10 min in a sterilization equipment at a sterilization temperature of 125° C. to obtain an eye protection solution.
[0037] Example 3
[0038] (1) Wash the vetch seeds with ultra-low deuterium water at a pH of 1.5, dry them after washing, and then place them in a solution of 0.3% by mass of seleno-glucose for 24 hours. After soaking, place the vetch seeds in a seedling tray for seedling cultivation. After the seedling cultivation is completed, transplant them. During the growth period of the vetch, spray the leaves with a solution of 0.16% by mass of L-selenomethionine for 5 times. When they are mature, harvest them, dry the roots of the vetch, wash them with water, dry them, and crush them to obtain coarse vetch powder rich in organic selenium.
[0039] (2) Add 10 g of organic selenium-rich wild pea powder to 1 L of a 10 -5 mol / L HCl in ethanol, heat to 80°C, condense and reflux for 12 hours, collect the filtrate and residue after filtration, add sodium hydroxide solution to the filtrate, adjust the pH to neutral, and concentrate to obtain a thick paste.
[0040] (3) The collected filter residue is placed in a fermentation tank, and Escherichia coli is used as the fermentation bacteria, and glucose and corn syrup are used as nutrient fermentation raw materials. After bacterial fermentation, filtration, sterilization, hydrolysis, purification, and crystallization, a selenoprotein raw material rich in active selenium is obtained.
[0041] (4) Add the thick paste obtained in (2) to a saturated calcium hydroxide aqueous solution, stir, and then add dilute sulfuric acid to adjust the pH to 3. Let stand for 18 hours, remove the supernatant, filter, add sodium hydroxide solution to the filtrate to adjust the pH to 10, and concentrate to a density of 1.25 g / cm 3 , let it stand for 18 hours, filter and add the precipitate into a hydrochloric acid solution with a pH of 4, stir and centrifuge, take the supernatant and obtain the centrifuged liquid.
[0042] (5) The centrifuged liquid is decolorized by activated carbon, and then chromatographically adsorbed by macroporous resins D101, AB-8 and A-722 in sequence, and finally mechanically eluted with ethanol. The ethanol eluate is collected, concentrated, and diluted to a concentration of 0.4 g / mL by adding ultra-low deuterium-free water. The liquid is filtered through a 0.45 μm filter membrane to obtain a clear liquid, which is then spray-dried to obtain a fine powder of wild pea extract rich in organic selenium.
[0043] (6) 0.08 g of wild pea extract powder, 9 g of sodium chloride, 0.04 g of taurine, 0.05 g of cooling agent menthol, 1 L of distilled water, and 0.12 g of selenoprotein raw material were mixed, dissolved with 600 ppm ultra-low deuterium water and hydrogen-rich water, adjusted to pH 7, filtered with a 0.22 μm microporous filter membrane, and sterilized for 10 min in a sterilization equipment at a sterilization temperature of 125° C. to obtain an eye protection solution.
[0044] Example 4
[0045] (1) Wash the vetch seeds with ultra-low deuterium water at a pH of 1.5, dry them after washing, and then place them in a solution of 0.4% by mass of seleno-glucose for 24 hours. After soaking, place the vetch seeds in a seedling tray for seedling cultivation. After the seedling cultivation is completed, transplant them. During the growth period of the vetch, spray the leaves with a solution of 0.1% by mass of L-selenomethionine for 8 times. When they are mature, harvest them, dry the roots of the vetch, wash them with water, dry them, and crush them to obtain coarse vetch powder rich in organic selenium.
[0046] (2) Add 30 g of organic selenium-rich wild pea powder to 1 L of a 10 -3 mol / L HCl in ethanol, heat to 80°C, condense and reflux for 12 hours, collect the filtrate and residue after filtration, add sodium hydroxide solution to the filtrate, adjust the pH to neutral, and concentrate to obtain a thick paste.
[0047] (3) The collected filter residue is placed in a fermentation tank, and Escherichia coli is used as the fermentation bacteria, and glucose and corn syrup are used as nutrient fermentation raw materials. After bacterial fermentation, filtration, sterilization, hydrolysis, purification, and crystallization, a selenoprotein raw material rich in active selenium is obtained.
[0048] (4) Add the thick paste obtained in (2) to a saturated calcium hydroxide aqueous solution, stir, and then add dilute sulfuric acid to adjust the pH to 4. Let stand for 24 hours, remove the supernatant, filter, add sodium hydroxide solution to the filtrate to adjust the pH to 11, and concentrate to a density of 1.2 g / cm 3 , let it stand for 24 hours, filter and add the precipitate into a hydrochloric acid solution with a pH of 2, stir and centrifuge, take the supernatant and obtain the centrifuged liquid.
[0049] (5) The centrifuged liquid is decolorized by activated carbon, and then chromatographically adsorbed by macroporous resins D101, AB-8 and A-722 in sequence, and finally mechanically eluted with ethanol. The ethanol eluate is collected, concentrated, and diluted to a concentration of 0.5 g / mL by adding ultra-low deuterium-free water. The liquid is filtered through a 0.45 μm filter membrane to obtain a clear liquid, which is then spray-dried to obtain a fine powder of wild pea extract rich in organic selenium.
[0050] (6) 0.05 g of wild pea extract powder, 9 g of sodium chloride, 0.1 g of taurine, 0.06 g of cooling agent menthol, 1 L of distilled water, and 0.1 g of selenoprotein raw material were mixed, dissolved with 600 ppm ultra-low deuterium water and hydrogen-rich water, the pH was adjusted to 7, filtered with a 0.22 μm microporous filter membrane, and sterilized for 20 min in a sterilization equipment at a sterilization temperature of 120° C. to obtain an eye protection solution.
[0051] Eye protection liquid test method: The experimental animals were provided by Xi'an Dilupu Biomedical Co., Ltd. 3 New Zealand rabbits (number A, B, C), female, weighing 2.5-3.0 kg, certificate number: 202002379320, ordinary grade, production license number: SCXK (Shaanxi) 2019-002. Feed was provided by the Experimental Animal Center of the School of Medicine of Xi'an Jiaotong University. Animal room environment: ordinary environment, temperature 25°C, relative humidity: 60%.
[0052] Test method: Take 0.1mL eye protection solution and drop it into the conjunctival sac of one eye of the rabbit, and make the rabbit eye passively closed for 4s, and rinse with normal saline after 30 days. The other eye is dripped with the same volume of normal saline as the control group. After 1h, 24h, 48h, 72h, 7d, 14d and 21d after eye drops, observe the damage and recovery of the conjunctiva, iris and cornea of the rabbit with naked eyes. If no irritation reaction occurs within 72h, or the eye irritation reaction is completely restored on the 7th or 14th day, the test can be terminated in advance. If necessary, use 2% sodium fluorescein solution or slit lamp and magnifying glass to check the changes of cornea and iris. Evaluate according to the scoring criteria of eye irritation test in the Technical Specifications for Disinfection. The test results are shown in the table below.
[0053]
[0054] The eye protection solutions prepared in the test examples were found to have no irritation to rabbit eyes.
[0055] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.
Claims
1. A method for preparing a Vicia fructus extract, characterized in that: The preparation method comprises the following steps: (1) Washing the vetch seeds with ultra-low deuterium water at a pH of 1.5, drying them after washing, and then placing them in a seleno-glucose solution for 18-24 hours. After soaking, placing the vetch seeds in a seedling tray for seedling cultivation. After the seedling cultivation is completed, transplanting, spraying the leaves with a solution of L-selenomethionine 5-8 times during the growth period of the vetch, harvesting them when they are mature, drying the roots of the vetch, washing them with water, drying them, and crushing them to obtain a coarse vetch powder rich in organic selenium; (2) adding the organic selenium-rich wild pea powder to an acidic ethanol solution for extraction, collecting the filtrate and the filter residue after filtering, adding sodium hydroxide solution to the filtrate, adjusting the pH to neutral, and concentrating to obtain a thick paste; (3) Add the thick paste to a saturated calcium hydroxide aqueous solution, stir, then drop dilute sulfuric acid, let stand for 18-24 hours, remove the supernatant, filter, add sodium hydroxide solution to the filtrate, and concentrate to a density of 1.15-1.25 g / cm 3 , let stand for 18-24 hours, filter and add the precipitate into hydrochloric acid solution, stir and centrifuge, take the supernatant to obtain centrifuged liquid; (4) The centrifuged liquid is decolorized by activated carbon, and then chromatographically adsorbed by macroporous resins D101, AB-8 and A-722 in sequence, and finally mechanically eluted with ethanol. The ethanol eluate is collected, concentrated, and diluted to a concentration of 0.4-0.6 g / mL by adding ultra-low deuterium-free water. The liquid is filtered through a 0.45 μm filter membrane to obtain a clear liquid, which is then spray-dried to obtain a fine powder of wild pea extract rich in organic selenium.
2. The method for preparing the Vicia fructus extract according to claim 1, characterized in that: In the above (2), the ratio of adding acidic ethanol to the organic selenium-rich wild pea coarse powder is (10-60) g:1 L.
3. The method for preparing the Vicia fructus extract according to claim 2, characterized in that: The acidic ethanol is an ethanol solution of HCl, the molar concentration of which is (10 -3 -10 -5 )mol / L.
4. The method for preparing the Vicia fructus extract according to claim 1, characterized in that: In the step (2), the extraction is performed by condensing and reflux at 75-85°C for 6-12 hours.
5. The method for preparing the Vicia fructus extract according to claim 1, characterized in that: In the above (3), dilute sulfuric acid is added dropwise to adjust the pH to 2-4; sodium hydroxide solution is added to adjust the pH to 10-12; and the pH of the hydrochloric acid solution is 2-4.
6. A method for preparing eye protection solution using the fine powder of Vicia fructus extract obtained by the preparation method according to any one of claims 1 to 5, characterized in that: The method is as follows: mix the fine powder of wild pea extract, sodium chloride, taurine, cooling agent menthol, distilled water, and selenoprotein raw materials, dissolve them with 600ppm ultra-low deuterium water and hydrogen-rich water, adjust the pH to 6-8, filter with a 0.22μm microporous filter membrane, and then sterilize with a sterilization device for 10-20 minutes at a sterilization temperature of 115-125°C to obtain eye protection solution.
7. The method for preparing eye protection solution using fine powder of Vicia fructus extract according to claim 5, characterized in that: The ratio of the wild pea extract fine powder, sodium chloride, taurine, cooling agent, distilled water and selenoprotein raw material is (0.02-0.15) g: (8.5-9) g: (0.04-0.12) g: (0.05-0.08) g: 1 L: (0.03-0.2) g.
8. The method for preparing eye protection solution using fine powder of Vicia fructus extract according to claim 5, characterized in that: The preparation method of the selenoprotein raw material is as follows: placing the filter residue collected in (2) in a fermentation tank, using Escherichia coli as the fermentation strain, using glucose and corn syrup as the nutrient fermentation raw materials, and subjecting the fermentation, filtration, sterilization, hydrolysis, purification and crystallization to obtain a selenoprotein raw material rich in active selenium.