Sunflower seed oil preparation method and application of scar repair protein polypeptide

Abstract

The present application relates to the technical field of plant oil preparation, and particularly relates to a sunflower seed oil preparation method and a scar repair protein polypeptide for application. The present application discloses a sunflower seed oil preparation method, which comprises the following steps: firstly, sunflower seeds are dried, impurities are removed, the sunflower seeds are shelled and crushed to obtain sunflower seed powder; secondly, an extracting agent is used to obtain a crude extract through ultrasonic treatment, filtration and static stratification; thirdly, a lyotropic liquid crystal agent is used for decolorization to obtain a secondary extract; fourthly, under the alkaline condition of oxalyl chloride solution and triethylamine solution, a purified membrane is used to remove free fatty acids, and then the remaining impurities and solvent are removed through rotary evaporation to obtain sunflower seed oil; fifthly, the sunflower seed oil is embedded by a layered adsorbent to obtain a sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide compound; and finally, the sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide compound is mixed with collagen protein polypeptide to form a scar repair protein polypeptide, so that the protection, slow release, hydrophilicity and biocompatibility of the sunflower seed oil are realized.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of plant oil preparation, and particularly relates to a sunflower seed oil preparation method and application of scar repair protein polypeptide. BACKGROUND

[0002] Sunflower seed oil is extracted from the fruit of sunflower, and the main effective component is triglyceride. Generally, the sunflower seed oil is extracted by pressing method and leaching method. The impurities that may affect the quality of the sunflower seed oil include natural non-oil components and external pollutants. The natural non-oil components include sunflower shell, seed coat debris, protein and carbohydrates in the kernel, phospholipids, free fatty acids, pigments, waxes, sterols, water, etc. The external pollutants include sand and microorganisms. The external pollutants are generally removed by screening and cleaning. For the natural non-oil components, the sunflower seed oil is refined by deacidification, decolorization, deodorization and degumming steps after the sunflower seed oil is extracted, and finally the sunflower seed oil mainly composed of triglyceride is formed.

[0003] A preparation process of strong-flavor sunflower seed oil is disclosed in Chinese Patent No. CN118126769A. First, sunflower seeds are fried, then soaked in a phospholipid and water solution, ground, centrifuged to obtain crude oil and slurry; second, the slurry is subjected to two-step enzymatic hydrolysis to obtain an enzymatic hydrolysate; finally, the crude oil and the enzymatic hydrolysate are mixed, heated, centrifuged, and degummed to obtain sunflower seed oil. After the crude oil and the enzymatic hydrolysate are mixed, heating and stirring are performed at 70-180℃ for 5-90min. In the low-temperature section of 70-100℃, the double bonds of unsaturated fatty acids such as oleic acid and linoleic acid in the mixed solution are prone to react with oxygen to generate peroxides, accompanied by a slight rancid smell. In the medium-high temperature section of 100-180℃, the peroxides will further decompose into aldehydes, ketones, acids and other secondary products, with a pungent odor and increased toxicity.

[0004] A preparation method of sunflower seed oil is disclosed in Chinese Patent No. CN119656070A. First, sunflower seed oil is pressed out by pressing method, and then suspended solids and impurities are filtered out to obtain crude oil. Hydrated phospholipids are removed by hot water at 60-95℃, non-hydrated phospholipids are removed by citric acid, and neutralization is performed by an alkaline solution. Second, oil-water separation is achieved by multiple hot water rinsing at 80-85℃. Finally, sunflower seed oil is obtained after drying, bleaching and decolorization, and vacuum deodorization. In this method, the residual heat of 60-95℃ may cause acid hydrolysis of triglycerides during the removal of non-hydrated phospholipids by citric acid. In addition, in the deodorization process, the triglycerides may be significantly damaged due to thermal cracking reaction under the conditions of 230-240℃ and vacuum.

[0005] A refining method for controlling the odor of sunflower seed oil is disclosed in Chinese Patent Publication No. CN116120987B. By neutralization deacidification process, decolorization process and deodorization process, the sunflower seed oil can achieve zero reverse and remove odor substances at the same time. In the decolorization process, bentonite, attapulgite and their compound soil are used. These clay adsorbents adsorb pigments and other substances in the decolorization process based on their surface adsorption sites and pore structure. However, their adsorption is not absolutely selective. In addition to pigment molecules, triglycerides may also be adsorbed onto the surface or pores of the adsorbent. Especially when the molecular size of triglyceride matches the pore size of the adsorbent, or there is a certain van der Waals force and electrostatic attraction between the triglyceride and the adsorbent surface, it is easy to be adsorbed and cause loss. In the deodorization process, under the conditions of vacuum degree 0-500 Pa, deodorization time 50-70 min, temperature 190-220℃, steam dosage more than 2wt% of oil weight, etc., the triglyceride may produce certain loss. Because the longer the deodorization time, the longer the contact time between steam and oil, the amount of small oil droplets carried into the condenser will increase. At the same time, although vacuum can reduce oxygen concentration and slow down the oxidative decomposition of triglyceride, it cannot completely avoid thermal decomposition. At 190-220℃, close to the initial decomposition temperature of triglyceride, the decomposition rate is low in a short time, but long time may accumulate slight loss.

[0006] Chinese Patent Publication No. CN105505576A discloses a refining method for crude blended oil containing sunflower seed oil, which is refined through pretreatment, degumming, deacidification, decolorization, deodorization, and cooling filtration. In the degumming process, under the conditions of 4-6% boiling water or 40-55% mass concentration of citric acid solution, 60-80℃, stirring for 15-20 min, and standing for 2-5 h, the effective component triglyceride may be destroyed by acid-catalyzed hydrolysis to generate glycerol and free fatty acids. In the steam deodorization process, under the conditions of 200-240℃ vacuum, the effective component triglyceride may be significantly destroyed by thermal cracking reaction.

[0007] Chinese Patent Publication No. CN103966019B discloses a refining method for sunflower seed oil, which is obtained through alkali refining deacidification, adsorption decolorization, distillation deodorization, and winterization dewaxing. In the alkali refining deacidification process, after acid treatment, alkali neutralization, water washing, and drying, etc., the triglyceride may produce certain loss. The concentration of alkali needs to be strictly controlled. If the addition amount exceeds the theoretical amount required for neutralizing free fatty acids, the excess alkali will significantly increase the risk of saponification. In the deodorization process, under the conditions of 245-255℃, 2-5 mbar vacuum, 0.8-1.2% superheated steam, and 60-90 min, the triglyceride may produce certain loss due to thermal decomposition and mechanical entrainment.

[0008] In summary, in the preparation method of sunflower seed oil, it is a key and urgent problem to efficiently retain triglyceride, remove impurities, improve quality, prevent oxidation and hydrolysis risk, improve process efficiency and save cost. SUMMARY

[0009] In order to solve the above problems, the purpose of the present application is to provide a sunflower seed oil preparation method and a scar repair protein polypeptide, a sunflower seed oil preparation method, which specifically comprises the following steps:

[0010] S001, in the sunflower seed powder, the vortex is uniform, the extraction agent is mixed, the ultrasonic treatment is carried out, the layering treatment is carried out, the lower layer liquid is discharged, the upper layer liquid is collected, and the crude extract is obtained;

[0011] S002, the crude extract is slowly added to the stirring state of the lyotropic liquid crystal agent for reaction treatment, centrifugation, removal of the lower layer solution and precipitation, collection of the upper layer solution, and the secondary extract is obtained;

[0012] S003, under stirring, slowly drop the oxalyl chloride solution for acyl chloride reaction, slowly drop the triethylamine solution for pretreatment, add the purification membrane, and carry out esterification reaction, and the ester membrane and the purified liquid are obtained;

[0013] S004, the ester membrane is gently clamped with tweezers, washed, mixed with the washing liquid and the purified liquid, and centrifuged, the solution is collected, and evaporated, and the sunflower seed oil is obtained.

[0014] The sunflower seed oil is embedded and modified by the layered adsorbent and collagen polypeptide to obtain the scar repair protein polypeptide, which specifically comprises the following steps:

[0015] S101, the layered adsorbent, sunflower seed oil and n-hexane are mixed, ultrasonic treatment is carried out, and evaporation treatment is carried out, and the sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide complex is obtained;

[0016] S102, the collagen polypeptide solution is added and mixed, and the sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide-collagen polypeptide complex is obtained, which is recorded as the scar repair protein polypeptide.

[0017] The specific preparation method of the above related solvents is as follows:

[0018] The extraction agent is composed of n-hexane, ultrapure water, butylated hydroxyanisole, di-tert-butyl-p-cresol and vitamin E. The preparation method is as follows: 50mg of butylated hydroxyanisole, 50mg of di-tert-butyl-p-cresol and 1.5g of vitamin E are weighed into 250mL n-hexane solution, vortexed until dissolved, 250mL ultrapure water is added, and vortexed.

[0019] The lyotropic liquid crystal agent is prepared by mixing Tween-80, ultrapure water and n-hexane to form a lyotropic liquid crystal system. Preparation method: weigh 5 g of Tween-80, add 100 mL of n-hexane, stir until dissolved, add 1 mL of ultrapure water under stirring at room temperature at 300 rpm / min, and keep stirring for 30 min.

[0020] Preparation method of 0.1% oxalyl chloride solution: transfer 100 μL of oxalyl chloride to 100 mL of n-hexane, and vortex uniformly.

[0021] Preparation method of 0.16% triethylamine solution: transfer 160 μL of oxalyl chloride to 100 mL of n-hexane, and vortex uniformly.

[0022] The purified membrane is prepared by grafting hydroxyethyl methacrylate to polyvinylidene fluoride membrane. The preparation method is as follows:

[0023] S201, weigh 20 g of polyvinylidene fluoride particles into a three-necked flask, add 180 g of N,N-dimethylformamide, stir at 300 rpm / min at 60-70°C for 5 hours until dissolved, cool to room temperature, pour onto a clean glass plate, scrape into a 0.1-0.3 mm thick film with a spatula, then place the glass plate in ultrapure water, and let the N,N-dimethylformamide gradually diffuse into the water to obtain a polyvinylidene fluoride membrane;

[0024] S202, carefully remove the polyvinylidene fluoride membrane from the glass plate, wash repeatedly with ultrapure water for 5 times, then dry in an oven at 50°C to constant weight, take out and soak in acetone solution for 2 hours, wash repeatedly with ultrapure water for 5 times to obtain a treated polyvinylidene fluoride membrane;

[0025] S203, weigh 2.5 g of cerium ammonium nitrate into a three-necked flask, add 500 mL of ultrapure water, stir to dissolve, and add 5 mL of hydroxyethyl methacrylate to obtain an initiator solution;

[0026] S204, place the pretreated polyvinylidene fluoride membrane into the initiator solution, stir at 300 rpm / min at 30-50°C for 3 hours, take out the membrane, wash repeatedly with ultrapure water for 5 times, then dry in an oven at 50°C to constant weight to obtain a purified membrane.

[0027] The layered adsorbent is prepared by modifying magnesium aluminum nitrate type layered double hydroxide with oleic acid. The preparation method is as follows:

[0028] S301, weigh 20 g of sodium hydroxide into a beaker, add appropriate amount of ultrapure water, stir until completely dissolved, transfer the solution to a 500 mL volumetric flask, wash with ultrapure water and dilute to volume to obtain a 1 M sodium hydroxide solution;

[0029] S302, weigh 2.82 g of oleic acid into a beaker, dissolve in 50 mL of absolute ethanol, under the condition of room temperature and 300 rpm / min stirring, drop 1M sodium hydroxide solution until the oleic acid is completely dissolved, and the oleic acid solution is obtained;

[0030] S303, weigh 1 g of magnesium aluminum nitrate layered double hydroxide into a beaker, add 50 mL of ultrapure water, ultrasonic for 10 min until uniform, add to the oleic acid solution, and stir under the condition of 70-80℃ and 300 rpm / min for 4 hours until the solution is clear;

[0031] S304, centrifuge at room temperature and 15000 r / min for 15 min, wash with absolute ethanol for 3 times, and dry at 60℃ under the condition of 10-50 mbar vacuum for 12 hours, and the magnesium aluminum-oleic acid-layered double hydroxide is obtained.

[0032] 3mg / mL collagen polypeptide solution preparation method: weigh 3 g of collagen polypeptide powder into a beaker, add 1 L of 0.05M Tris-HCl buffer, and stir at room temperature and 200 rpm / min until dissolved.

[0033] The sunflower seed powder in step S001 is obtained by air drying, impurity removal, shelling and crushing the sunflower seeds in sequence, adding the extractant at a solid-liquid ratio of 1:5-9, and ultrasonic treatment under the condition of 40℃, 60Khz and 30min. The mixed solution is filtered into a separatory funnel, and layered by standing for 20min.

[0034] The volume ratio of the lyotropic liquid crystal agent to the crude extract in step S002 is 1-3:1, the stirring rate is 300 rpm / min, the reaction treatment condition is 25℃ and standing for 1-3 hours, and the centrifugation condition is room temperature, 15000 r / min and 15 min.

[0035] The stirring condition in step S003 is 0-4℃ and 300 rpm / min, the concentration of oxalyl chloride solution is 0.1%, the volume ratio of the secondary extract to oxalyl chloride solution is 1:1, the acyl chloride reaction time is 2 hours, the concentration of triethylamine solution is 0.16%, the pretreatment time is 30min, the addition amount of the purification membrane is 1 part, and the esterification reaction condition is room temperature, 300 rpm / min and 6 hours.

[0036] In step S004, n-hexane is used for washing, the usage amount is 100 mL, the centrifugation condition is room temperature, 15000 r / min and 15 min, and the evaporation condition is 15-30 mmHg pressure and 40-60℃ until no obvious bubbles are observed in the flask.

[0037] The mass-volume ratio of the layered adsorbent, sunflower seed oil and n-hexane in the step S101 is 1:100:500, the ultrasonic treatment condition is room temperature, 60Khz, 30min, and the evaporation condition is 15-30mmHg pressure, 40-60 DEG C, until no obvious bubbles in the flask.

[0038] The concentration of the collagen polypeptide solution in the step S102 is 3mg / mL, the material-liquid ratio of the sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide complex and the collagen polypeptide solution is 1:50, and the mixing condition is room temperature, 300rpm / min, 2 hours.

[0039] Compared with the prior art, the beneficial effects of the present application are:

[0040] 1. The present application extracts sunflower seed oil by using an extractant, adopts the principle of similar dissolves similar, uses a n-hexane-ultra-pure water double solvent system for liquid-liquid extraction, uses the high affinity of n-hexane for oil in sunflower seed powder, and extracts part of hydrophilic impurities with ultra-pure water, so that the obtained oil has lower impurity content, lighter color and higher quality. And the ultra-pure water can also make the sunflower seed cells swell and destroy the cell wall structure, thereby accelerating the diffusion rate of n-hexane in the sunflower seed powder, improving the leaching efficiency and shortening the leaching time. The synergistic effect of ultra-pure water and n-hexane can more effectively destroy the combination of oil and protein and other substances, improve the oil extraction rate, and obtain a higher oil yield. At the same time, the cavitation effect generated by ultrasonic waves in the liquid medium is used to destroy the plant cell wall structure through the action of micro-jets induced by high-frequency vibration, thereby improving the oil yield and shortening the leaching time again.

[0041] 2. The present application adds fat-soluble antioxidants in the extractant, which provide active hydrogen atoms to quench free radicals to block the oil oxidation chain reaction through butylated hydroxyanisole, di-tert-butyl-p-cresol and vitamin E, thereby delaying the oxidation and deterioration of sunflower seed oil. Among them, butylated hydroxyanisole has strong thermal stability, good oil solubility and antibacterial properties; di-tert-butyl-p-cresol is low in cost, long-acting and easy to synergize with other additives; vitamin E as a natural fat-soluble ingredient not only has high safety and meets the health needs, but also can realize continuous antioxidant through regeneration cycle, and has nutritional value. The above three can provide multi-directional protection for the quality of sunflower seed oil based on different mechanisms and performance advantages.

[0042] 3. The present application is based on the ordered liquid crystal structure formed by self-assembly of surfactant Tween-80 for directional adsorption and separation of fat-soluble pigments: with Tween-80 as a template agent, its amphiphilic molecules self-assemble into highly ordered lyotropic liquid crystal phase in a two-solvent system composed of polar water molecules and non-polar n-hexane through hydrophobic interaction and hydrogen bonding. The liquid crystal phase as a nanoscale template provides a polar-nonpolar microenvironment through its periodic interface layer, selectively captures the pigments in sunflower seed oil, and the hydrophobic groups of the pigment molecules are embedded in the hydrophobic region of the liquid crystal, and the polar groups interact with the hydrophilic layer of the liquid crystal, achieving directional enrichment through interface matching; at the same time, the high specific surface area and viscoelastic fluid characteristics of the liquid crystal phase enhance the mass transfer and diffusion of the pigment molecules and inhibit their agglomeration. Compared with traditional physical adsorption processes such as clay and activated carbon, this method has higher decolorization efficiency and can selectively adsorb pigments in a directional manner, preventing the loss of effective components such as triglycerides and vitamin E; the decolorization can be carried out at room temperature and normal pressure, effectively preventing the destruction of oil quality at high temperature;

[0043] 4. The present application is based on the purification membrane for fixing free fatty acids in the secondary extract. First, prepare the purification membrane, immerse the polyvinylidene fluoride membrane in acetone to swell and expose and activate the surface groups, and obtain the treated membrane. Dissolve cerium ammonium nitrate in ultrapure water to release cerium ions, provide strong oxidizing species, and mix with hydroxyethyl methacrylate to obtain an initiator solution. Soak the treated membrane in the cerium ion solution, and the cerium ions react with the surface groups of the membrane to form free radicals, which become the starting point of the grafting reaction. The free radicals initiate the polymerization of the double bonds of hydroxyethyl methacrylate to form grafted chains on the surface of the polyvinylidene fluoride membrane, while the alcohol hydroxyl functional groups are retained; secondly, the carboxyl oxygen of the free fatty acid attacks the carbonyl carbon of oxalyl chloride through nucleophilic addition-elimination to form a tetrahedral intermediate, and then releases chlorine ions and carbon dioxide to form fatty acid chloride and release hydrogen chloride; then, the hydrogen chloride generated by the reaction is neutralized by the organic base triethylamine to avoid its combination with the alcohol hydroxyl group to form hydrochloride salt, while maintaining an alkaline environment in the system to promote the stable conversion of the intermediate; then, the alcohol hydroxyl oxygen on the purification membrane attacks the carbonyl carbon of the fatty acid chloride to form an intermediate containing two tetrahedral structures, and the chlorine atom in the intermediate is released as a good leaving group, while the negative charge of the oxygen atom is transferred to the carbonyl group through conjugation effect, and finally an ester is formed to fix the free fatty acid on the purification membrane and release hydrogen chloride, which will be neutralized by triethylamine again, so that the free fatty acid can be removed quickly and simply by a simple physical method. And the esterification reaction in this system occurs in an alkaline condition, which can more effectively prevent the reverse reaction of triglyceride hydrolysis under acidic conditions; finally, remove the n-hexane, oxalyl chloride and triethylamine impurities by rotary evaporator to obtain high-quality sunflower seed oil;

[0044] 5.The present application protects sunflower seed oil by embedding it with layered adsorbent. Firstly, the strong polar hydroxyl groups densely arranged on the surface of magnesium aluminum nitrate type layered double hydroxide combine with oxygen atoms or hydrogen atoms in water molecules to form hydrogen bonds, which promotes the penetration of water molecules into the interlayer region, destroys the van der Waals force between the layers, and leads to the expansion of the interlayer distance. At the same time, the initial interlayer anion nitrate, which is itself a hydrophilic anion, is combined with the positively charged layer by electrostatic interaction, and the oxygen atoms of nitrate form hydrogen bonds with the hydrogen atoms of water molecules. The enrichment of water molecules in the interlayer further promotes the separation of the layers, so that the magnesium aluminum nitrate type layered double hydroxide swells in water, the electrostatic combination of the layers and nitrate is destroyed, and the hydrophilic anion is released from the interlayer into the solution; Secondly, long-chain organic anion oleate enters the swollen interlayer region through the chemical potential difference, and the carboxylate forms strong electrostatic interaction and hydrogen bonds with the positive charge of the layer, and the hydrophobic long chain is self-assembled into an orderly arrangement by van der Waals force, eventually replacing the hydrophilic nitrate, and realizing the hydrophobic modification of the interlayer anion; Then, the negative charge of oleate and the coulomb attraction of the positive charge of the layer, combined with hydrogen bonds and van der Waals forces, provide stronger binding energy than nitrate. The long carbon chain oleate occupies the interlayer space, expands the interlayer distance and expels small molecular anions and solvent water, destroying the stable existence conditions of nitrate. The alkyl chain of oleate repels polar solvents, which promotes the change of the interlayer from hydrophilic to hydrophobic, forming a new thermodynamically stable structure, i.e. layered adsorbent; Finally, the hydrophobic sunflower seed oil diffuses into the hydrophobic interlayer of the layered adsorbent through the concentration gradient, and is stably combined with the alkyl chain of oleate through van der Waals force and hydrophobic interaction. Part of the oil can also form a coating by surface hydrophobic adsorption; The interlayer steric hindrance and molecular size selectivity selectively embed the oil, and the hydrophobic microenvironment and surface properties can inhibit the oxidation of the oil, realizing the controllable slow release, efficient embedding and stable loading of sunflower seed oil;

[0045] 6. The application is compounded by the complex of collagen polypeptide and sunflower seed oil-magnesium aluminum-oleic acid-lamellar double hydroxide, that is, the complex of sunflower seed oil-magnesium aluminum-oleic acid-lamellar double hydroxide-collagen polypeptide, which is denoted as scar repair protein polypeptide. Because there is a certain long-chain oleic acid on the surface of magnesium aluminum-oleic acid-lamellar double hydroxide, and there is a residual uncoordinated carboxyl group, which is negatively charged under neutral conditions. While the collagen polypeptide chain contains a large number of amino, carboxyl and hydroxyl groups, the positive charge region can form electrostatic attraction with the negative charge on the surface of magnesium aluminum-oleic acid-lamellar double hydroxide. At the same time, because the sunflower seed oil is filled in the interlayer of magnesium aluminum-oleic acid-lamellar double hydroxide or the hydrophobic region on the surface, the hydrophilic region of collagen is combined with the residual polar site carboxyl group on the surface of magnesium aluminum-oleic acid-lamellar double hydroxide, and the hydrophobic region is interacted with sunflower seed oil or long-chain oleic acid, forming a hydrophobic-hydrophilic complementary structure, realizing the compounding of the complex of collagen polypeptide and sunflower seed oil-magnesium aluminum-oleic acid-lamellar double hydroxide. And according to the amphiphilic, slow-release function, biocompatibility and other characteristics of collagen polypeptide, the release of sunflower seed oil can be delayed, so that the scar repair protein polypeptide has certain hydrophilicity and biocompatibility;

[0046] 7. The preparation method adopted by the application is simple to operate, and the extracted sunflower seed oil has high quality. Through antioxidants, magnesium aluminum-oleic acid-lamellar double hydroxide and collagen polypeptide, the protection, slow release, hydrophilicity and biocompatibility of sunflower seed oil are realized, and the application scope is wide, which provides a new solution for the preparation of vegetable oil and has certain industrial value and technical progress. DETAILED DESCRIPTION

[0047] The application will be further described in combination with specific embodiments.

[0048] Example 1

[0049] A sunflower seed oil preparation method, the specific steps are as follows:

[0050] 10g sunflower seed powder was weighed, a vortex uniform extraction agent was added, mixed at a solid-liquid ratio of 1:5, and ultrasonic extraction was carried out at 40℃ and 60Khz for 30min. The mixture was filtered into a separatory funnel, and after standing for 20min, the lower liquid was discharged, and the upper liquid was collected, that is, the crude extract was obtained;

[0051] Under the condition of room temperature and 300rpm / min stirring, the crude extract was slowly added into 1:2 volume of lyotropic liquid crystal agent, and the mixture was stood at 25℃ for 2 hours. Then, under the condition of room temperature and 15000r / min centrifugation for 15min, the lower solution and precipitate were removed, and the upper solution was collected, that is, the secondary extract was obtained;

[0052] Slowly add the same volume of 0.1% oxalyl chloride solution dropwise under stirring at 0-4°C and 300 rpm / min, after 2 hours, slowly add the same volume of 0.16% triethylamine solution dropwise, after 30 min, add 1 portion of purified membrane, stir at room temperature and 300 rpm / min for 6 hours, to obtain the ester membrane and purification liquid, gently pick up the ester membrane with tweezers, wash with 100 mL of n-hexane, collect the washing liquid and mix with the purification liquid, centrifuge at room temperature and 15000 r / min for 15 min, and collect the solution;

[0053] Place the solution in a flask of a rotary evaporator, under the pressure of 15-30 mmHg and at 40-60°C, until no obvious bubbles are observed in the flask, to obtain the sunflower seed oil, which is recorded as Test Sample 1.

[0054] Example 2

[0055] A sunflower seed oil preparation method, the specific steps are as follows:

[0056] Weigh 10 g of sunflower seed powder, add a vortex uniform extractant, mix at a solid-liquid ratio of 1:7, and ultrasonically treat at 40°C and 60 kHz for 30 min, filter the mixture into a separatory funnel, stand for 20 min, release the lower liquid, and collect the upper liquid, to obtain the crude extract liquid;

[0057] Slowly add the crude extract liquid into 1:2 volume of lyotropic liquid crystal agent under stirring at room temperature and 300 rpm / min, stand at 25°C for 2 hours, centrifuge at room temperature and 15000 r / min for 15 min, remove the lower solution and precipitate, and collect the upper solution, to obtain the secondary extract liquid;

[0058] Slowly add the same volume of 0.1% oxalyl chloride solution dropwise under stirring at 0-4°C and 300 rpm / min, after 2 hours, slowly add the same volume of 0.16% triethylamine solution dropwise, after 30 min, add 1 portion of purified membrane, stir at room temperature and 300 rpm / min for 6 hours, to obtain the ester membrane and purification liquid, gently pick up the ester membrane with tweezers, wash with 100 mL of n-hexane, collect the washing liquid and mix with the purification liquid, centrifuge at room temperature and 15000 r / min for 15 min, and collect the solution;

[0059] Place the solution in a flask of a rotary evaporator, under the pressure of 15-30 mmHg and at 40-60°C, until no obvious bubbles are observed in the flask, to obtain the sunflower seed oil, which is recorded as Test Sample 2.

[0060] Example 3

[0061] A sunflower seed oil preparation method, the specific steps are as follows:

[0062] Take 10 g of sunflower seed powder, add vortex uniform extractant, mix at a solid-liquid ratio of 1 :9, and ultrasonic at 40°C and 60Khz for 30 min. Filter the mixture into a separatory funnel, stand for 20 min, release the lower layer, and collect the upper layer to obtain the crude extract;

[0063] Slowly add the crude extract into 1:2 volume of lyotropic liquid crystal agent under room temperature and 300 rpm / min stirring, stand for 2 hours at 25°C, centrifuge at room temperature and 15000 r / min for 15 min, remove the lower solution and precipitate, and collect the upper solution to obtain the secondary extract;

[0064] Slowly add the same volume of 0.1% oxalyl chloride solution under 0-4°C and 300 rpm / min stirring, after 2 hours, slowly add the same volume of 0.16% triethylamine solution, after 30 min, add 1 part of purified membrane, stir at room temperature and 300 rpm / min for 6 hours to obtain the ester membrane and purification liquid. Gently pinch the ester membrane with tweezers and wash with 100 mL of n-hexane. Mix the washing liquid with the purification liquid, centrifuge at room temperature and 15000 r / min for 15 min, and collect the solution;

[0065] Place the solution in a flask of a rotary evaporator, and evaporate under 15-30 mmHg pressure and 40-60°C until no obvious bubbles are observed in the flask to obtain sunflower seed oil, which is recorded as test sample 3.

[0066] Example 4

[0067] A sunflower seed oil preparation method, the specific steps are as follows:

[0068] Take 10 g of sunflower seed powder, add vortex uniform extractant, mix at a solid-liquid ratio of 1 :9, and ultrasonic at 40°C and 60Khz for 30 min. Filter the mixture into a separatory funnel, stand for 20 min, release the lower layer, and collect the upper layer to obtain the crude extract;

[0069] Slowly add the crude extract into 1:1 volume of lyotropic liquid crystal agent under room temperature and 300 rpm / min stirring, stand for 2 hours at 25°C, centrifuge at room temperature and 15000 r / min for 15 min, remove the lower solution and precipitate, and collect the upper solution to obtain the secondary extract;

[0070] The same volume of 0.1% oxalyl chloride solution was slowly added dropwise under stirring at 0-4°C and 300 rpm / min, after 2 hours, the same volume of 0.16% triethylamine solution was slowly added dropwise, after 30 min, 1 portion of purified membrane was added, stirring was carried out at room temperature and 300 rpm / min for 6 hours, to obtain ester membrane and purification liquid, the ester membrane was gently clamped with tweezers and washed with 100 mL of n-hexane, the washing liquid was collected and mixed with the purification liquid, centrifugation was carried out at room temperature and 15000 r / min for 15 min, and the solution was collected;

[0071] The solution was placed in a flask of a rotary evaporator, under the pressure of 15-30 mmHg and at 40-60°C, until no obvious bubbles were observed in the flask, to obtain sunflower seed oil, recorded as test sample 4.

[0072] Example 5

[0073] A sunflower seed oil preparation method, the specific steps are as follows:

[0074] 10 g of sunflower seed powder was weighed, a vortexed extraction agent was added, mixed at a solid-liquid ratio of 1:7, and ultrasonic treatment was carried out at 40°C and 60Khz for 30 min, the mixture was filtered into a separatory funnel, stood for 20 min, the lower liquid was discharged, and the upper liquid was collected, to obtain crude extraction liquid;

[0075] The crude extraction liquid was slowly added into 1:3 volume of lyotropic liquid crystal agent under stirring at room temperature and 300 rpm / min, stood for 2 hours at 25°C, centrifugation was carried out at room temperature and 15000 r / min for 15 min, the lower solution and the precipitate were removed, and the upper solution was collected, to obtain secondary extraction liquid;

[0076] The same volume of 0.1% oxalyl chloride solution was slowly added dropwise under stirring at 0-4°C and 300 rpm / min, after 2 hours, the same volume of 0.16% triethylamine solution was slowly added dropwise, after 30 min, 1 portion of purified membrane was added, stirring was carried out at room temperature and 300 rpm / min for 6 hours, to obtain ester membrane and purification liquid, the ester membrane was gently clamped with tweezers and washed with 100 mL of n-hexane, the washing liquid was collected and mixed with the purification liquid, centrifugation was carried out at room temperature and 15000 r / min for 15 min, and the solution was collected;

[0077] The solution was placed in a flask of a rotary evaporator, under the pressure of 15-30 mmHg and at 40-60°C, until no obvious bubbles were observed in the flask, to obtain sunflower seed oil, recorded as test sample 5.

[0078] Example 6

[0079] A sunflower seed oil preparation method, the specific steps are as follows:

[0080] Take 10 g of sunflower seed powder, add a vortex uniform extractant, mix at a solid-liquid ratio of 1 :7, and ultrasonic at 40°C, 60Khz for 30 min. Filter the mixture into a separatory funnel, stand for 20 min, release the lower layer, and collect the upper layer to obtain the crude extract;

[0081] Slowly add the crude extract to 1 :2 volume of lyotropic liquid crystal agent under room temperature, 300 rpm / min stirring, stand for 1 hour at 25°C, centrifuge at 15000 r / min for 15 min at room temperature, remove the lower solution and precipitate, and collect the upper solution to obtain the secondary extract;

[0082] Slowly add the same volume of 0.1% oxalyl chloride solution under 0-4°C, 300 rpm / min stirring, after 2 hours, slowly add the same volume of 0.16% triethylamine solution, after 30 min, add 1 part of purified membrane, stir at room temperature, 300 rpm / min for 6 hours, to obtain the ester membrane and purification liquid. Gently pinch the ester membrane with tweezers, and wash with 100 mL of n-hexane. Mix the washing liquid with the purification liquid, centrifuge at 15000 r / min for 15 min at room temperature, and collect the solution;

[0083] Place the solution in a flask of a rotary evaporator, and evaporate under 15-30 mmHg pressure, 40-60°C, until there is no obvious bubble in the flask, to obtain sunflower seed oil, which is recorded as test sample 6.

[0084] Example 7

[0085] A sunflower seed oil preparation method, the specific steps are as follows:

[0086] Take 10 g of sunflower seed powder, add a vortex uniform extractant, mix at a solid-liquid ratio of 1 :7, and ultrasonic at 40°C, 60Khz for 30 min. Filter the mixture into a separatory funnel, stand for 20 min, release the lower layer, and collect the upper layer to obtain the crude extract;

[0087] Slowly add the crude extract to 1 :2 volume of lyotropic liquid crystal agent under room temperature, 300 rpm / min stirring, stand for 1 hour at 25°C, centrifuge at 15000 r / min for 15 min at room temperature, remove the lower solution and precipitate, and collect the upper solution to obtain the secondary extract;

[0088] The same volume of 0.1% oxalyl chloride solution was slowly added dropwise under stirring at 0-4°C and 300 rpm / min, after 2 hours, the same volume of 0.16% triethylamine solution was slowly added dropwise, after 30 min, 1 part of purified membrane was added, stirring was carried out at room temperature and 300 rpm / min for 6 hours, to obtain an ester membrane and a purification liquid, the ester membrane was gently picked up with tweezers and washed with 100 mL of n-hexane, the washing liquid was collected and mixed with the purification liquid, centrifugation was carried out at room temperature and 15000 r / min for 15 min, and the solution was collected;

[0089] The solution was placed in a flask of a rotary evaporator, under the pressure of 15-30 mmHg and at 40-60°C, until no obvious bubbles were observed in the flask, to obtain the sunflower seed oil, which was recorded as the test sample 7.

[0090] Example 8

[0091] A preparation method of a sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide-collagen polypeptide complex, the specific steps are as follows:

[0092] 1 g of the layered adsorbent, 100 mL of the sunflower seed oil prepared in Example 2 and 500 mL of n-hexane were weighed into a beaker, ultrasonic treatment was carried out at room temperature and 60 kHz for 30 min, the solution was placed in a flask of a rotary evaporator, under the pressure of 15-30 mmHg and at 40-60°C, until no obvious bubbles were observed in the flask, to obtain a sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide complex.

[0093] 3 mg / mL of a collagen polypeptide solution was added, stirring was carried out at room temperature and 300 rpm / min for 2 hours, to obtain a sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide-collagen polypeptide complex, which was recorded as the test sample 8.

[0094] Comparative Example 1

[0095] A conventional preparation method of sunflower seed oil, the specific steps are as follows:

[0096] 10 g of sunflower seed powder was weighed, n-hexane was added, and mixing was carried out at a solid-liquid ratio of 1:7, and then soaking was carried out at 50°C for 2 hours, to obtain mixed oil and wet meal;

[0097] The mixed oil was placed in a rising film evaporator under the conditions of 110°C, 0.3 MPa steam pressure and -0.09 MPa vacuum, and concentrated for 5 min, and then stripped at 120°C and 0.03 MPa for 1 hour, to obtain crude oil;

[0098] The crude oil was heated to 80℃, 0.1wt% of 10% dilute phosphoric acid was added, stirred at 80℃, 300rpm / min for 60min, centrifuged at 15000r / min for 15min, the light phase was collected, centrifuged at 15000r / min for 15min again, the light phase was collected, and the degummed oil was obtained;

[0099] 20% sodium hydroxide solution was added to the degummed oil, stirred at 80℃, 300rpm / min for 60min, centrifuged at 15000r / min for 15min, and the soap foot was removed, and the deacidified oil was obtained;

[0100] The deacidified oil was heated to 110℃, 3% of activated clay based on the weight of the oil was slowly added under stirring at 300rpm / min, after stirring for 60min, it was then poured into a Buchner funnel containing granular activated carbon, and after vacuum filtration under reduced pressure, the decolorized oil was obtained;

[0101] The decolorized oil was slowly cooled to 10℃, stirred at 30rpm / min for 24h, and filtered, and the dewaxed oil was obtained;

[0102] The dewaxed oil was heated to 220℃, and distilled under a vacuum degree of -0.1MPa, and was recorded as Control 1.

[0103] Comparative Example 2

[0104] A method for preparing a complex containing sunflower seed oil-magnesium aluminum-oleic acid-lamellar double hydroxide without collagen polypeptide, the specific steps are as follows:

[0105] 1g of the lamellar adsorbent, 100mL of the sunflower seed oil prepared in Example 2, and 500mL of n-hexane were weighed into a beaker, ultrasonicated at room temperature and 60Khz for 30min, and the solution was placed in a flask of a rotary evaporator under a pressure of 15-30mmHg and at a temperature of 40-60℃ until no obvious bubbles were observed in the flask, and a complex of sunflower seed oil-magnesium aluminum-oleic acid-lamellar double hydroxide was obtained.

[0106] 0.05M Tris-HCl buffer was added at a solid-liquid ratio of 1:50, and stirred at room temperature and 300rpm / min for 2h, and a complex of magnesium aluminum-oleic acid-lamellar double hydroxide containing sunflower seed oil without collagen polypeptide was obtained, and was recorded as Control 2.

[0107] Comparative Example 3

[0108] A method for preparing a complex containing magnesium aluminum-oleic acid-lamellar double hydroxide-collagen polypeptide without sunflower seed oil, the specific steps are as follows:

[0109] Take 1 g layered adsorbent and pipette 500 mL n-hexane into a beaker, ultrasonic at room temperature, 60 kHz for 30 min, place the solution in the flask of rotary evaporator, under the pressure of 15-30 mmHg, 40-60℃, until no obvious bubbles in the flask, then magnesium aluminum-oleic acid-layered double hydroxide complex is obtained;

[0110] Add 3 mg / mL collagen polypeptide solution, with the ratio of 1:50, stir at room temperature, 300 rpm / min for 2 hours, then magnesium aluminum-oleic acid-layered double hydroxide-collagen polypeptide complex without sunflower seed oil is obtained, which is recorded as control 3.

[0111] Comparative Example 4

[0112] 3 mg / mL collagen polypeptide solution, recorded as control 4.

[0113] Comparative Example 5

[0114] A method for preparing a magnesium aluminum-oleic acid-layered double hydroxide complex without sunflower seed oil and collagen polypeptide, the specific steps are as follows:

[0115] Take 1 g layered adsorbent and pipette 500 mL n-hexane into a beaker, ultrasonic at room temperature, 60 kHz for 30 min, place the solution in the flask of rotary evaporator, under the pressure of 15-30 mmHg, 40-60℃, until no obvious bubbles in the flask, then magnesium aluminum-oleic acid-layered double hydroxide complex is obtained;

[0116] Add 0.05M Tris-HCl buffer solution, with the ratio of 1:50, stir at room temperature, 300 rpm / min for 2 hours, then magnesium aluminum-oleic acid-layered double hydroxide complex without sunflower seed oil and collagen polypeptide is obtained, which is recorded as control 5.

[0117] Experimental Example 1

[0118] In this experimental example, the color, transparency and freezing test of sunflower seed oil in the test samples of Examples 1-7 and the control sample of Comparative Example 1 are determined, and the specific steps are as follows:

[0119] 1. Determination of color:

[0120] According to GB / T 5009.37-2003, pipette 2 mL of test sample and control sample into a 5 mL colorimetric tube, observe against natural light at room temperature, then observe against white background by reflecting light, and describe as white, grayish white, light yellow, light yellow, golden yellow, orange, brown yellow, brown, brown red, brownish brown, etc., and the determination results are shown in Table 1.

[0121] 2. Determination of transparency:

[0122] According to GB / T 5525-2008, 2 mL of the test sample and the control sample were taken and placed in a 5 mL colorimetric tube. After standing at 20℃ for 24 hours, the clarity, turbidity, and haze were described. The results are shown in Table 1.

[0123] 3. Determination of freezing test:

[0124] According to GB / T 35877-2018, 200 mL of the test sample and the control sample were taken and heated to 130℃. The heating was stopped immediately and the sample was filtered with filter paper while hot;

[0125] The filtered oil sample was filled into an oil sample bottle, tightly sealed with a cork, and cooled to 25℃ in a 25℃ water bath;

[0126] The oil sample bottle was immersed in a constant temperature water bath at 0℃. After 5.5 hours, the oil sample bottle was taken out, the surface of the sample bottle was cleaned, and the presence of fat crystals or flocculent was observed carefully. The clarity, turbidity, and haze were described. The results are shown in Table 1.

[0127] Table 1 Color, transparency, and freezing test

[0128]

[0129] As can be seen from Table 1, compared with the conventional preparation method of Comparative Example 1, the sunflower seed oil prepared by the method of the present application has relatively light color and relatively less pigment impurities. Moreover, the sunflower seed oil is clear and transparent, and no suspended matter or impurity precipitates during the freezing test.

[0130] Experimental Example 2

[0131] In this experimental example, the acid value and peroxide value of the sunflower seed oil in the test samples of Examples 1-7 and the control sample of Comparative Example 1 were determined. The specific steps are as follows:

[0132] 1. Determination of acid value:

[0133] Determined according to GB 5009.229-2025.

[0134] 500 mL of anhydrous ether and 500 mL of isopropyl alcohol were taken into a beaker, stirred and mixed, and then adjusted to pH 7.0 with 0.1 mol / L potassium hydroxide standard titration solution to obtain a cold solvent;

[0135] 1 g of phenolphthalein was weighed into a beaker, dissolved and washed with a small amount of 95% ethanol solution, and then the solution was transferred to a 100 mL volumetric flask and diluted with 95% ethanol solution to obtain a phenolphthalein indicator;

[0136] Weigh 10 g of the test sample and the control sample in a 250 mL conical flask, add 100 mL of cold solvent, shake gently until completely dissolved, add 4 drops of phenolphthalein indicator, immediately titrate with 0.1 mol / L potassium hydroxide standard titration solution until the solution appears pink, and does not fade for 15 s, each test solution and control solution is repeated twice;

[0137] Record the consumption volume of 0.1 mol / L potassium hydroxide standard titration solution, and calculate the acid value, the test results are shown in Table 2.

[0138] 2. Determination of peroxide value:

[0139] According to GB 5009.227-2023.

[0140] Weigh 5 g of the test sample and the control sample into the titration cup of the potentiometric titrator, add 20 mL of isooctane solution, shake gently until completely dissolved, then add 30 mL of ice acetic acid solution, and mix well to obtain the test solution and the control solution;

[0141] Remove 20 mL of isooctane solution and 30 mL of ice acetic acid solution into the titration cup of the potentiometric titrator, and mix well to obtain the blank solution;

[0142] Accurately add 1 mL of potassium iodide saturated solution to the titration cups of the test solution, the control solution and the blank solution, respectively, start the stirrer, stir at 300 rpm / min for 60 s, then immediately add 100 mL of ultrapure water to the titration cup, insert the electrode and the titration head, set the 0.01 mol / L sodium thiosulfate standard titration solution to 0.05 mL / drop for titration, observe the titration curve and potential change, and each test solution, control solution and blank solution is repeated twice;

[0143] After reaching the titration endpoint, record the consumption volume of 0.01 mol / L sodium thiosulfate standard titration solution, and calculate the peroxide value, the test results are shown in Table 2.

[0144] Table 2 Acid value and peroxide value

[0145]

[0146] As can be seen from Table 2, the acid value of the sunflower seed oil in the present application is less than 0.04 mg / g, and is 1.3-1.7 times lower than that of the conventional preparation method of Comparative Example 1. The peroxide value is less than 3.0 mmol / kg, which is 1.7-2.4 times lower than that of the conventional preparation method of Comparative Example 1.

[0147] Experimental Example 3

[0148] The free fatty acid of the sunflower seed oil in the test samples of Examples 1-7 and the control sample of Comparative Example 1 was determined according to GB 5009.168-2016, and the specific steps were as follows:

[0149] 1. Preparation of mixed fatty acid methyl ester standard working solution:

[0150] 100 mg of methyl oleate, 100 mg of methyl linoleate, 100 mg of methyl palmitate and 100 mg of methyl stearate were weighed into a beaker, dissolved with a small amount of n-heptane, and then transferred to a 10 mL volumetric flask. The solution was diluted with n-heptane to the mark, and the mixed fatty acid methyl ester stock solution was obtained.

[0151] 1 mL of the mixed fatty acid methyl ester stock solution was transferred to a 10 mL volumetric flask, and n-heptane was added to the mark to obtain the mixed fatty acid methyl ester standard working solution. The solution was transferred to an on-machine bottle for standby;

[0152] 2. Preparation of single fatty acid methyl ester standard working solution:

[0153] 50 mg of methyl oleate was weighed into a beaker, dissolved with a small amount of n-heptane, and then transferred to a 10 mL volumetric flask. The solution was diluted with n-heptane to the mark, and the methyl oleate standard working solution was obtained. The solution was transferred to an on-machine bottle for standby;

[0154] 100 mg of methyl linoleate was weighed into a beaker, dissolved with a small amount of n-heptane, and then transferred to a 10 mL volumetric flask. The solution was diluted with n-heptane to the mark, and the methyl linoleate standard working solution was obtained. The solution was transferred to an on-machine bottle for standby;

[0155] 10 mg of methyl palmitate was weighed into a beaker, dissolved with a small amount of n-heptane, and then transferred to a 10 mL volumetric flask. The solution was diluted with n-heptane to the mark, and the methyl palmitate standard working solution was obtained. The solution was transferred to an on-machine bottle for standby;

[0156] 5 mg of methyl stearate was weighed into a beaker, dissolved with a small amount of n-heptane, and then transferred to a 10 mL volumetric flask. The solution was diluted with n-heptane to the mark, and the methyl stearate standard working solution was obtained. The solution was transferred to an on-machine bottle for standby;

[0157] 3. Preparation of internal standard solution: 2.5 g of tricaprylin was weighed into a beaker, and an appropriate amount of methanol was added for dissolution and washing. The solution was transferred to a 500 mL volumetric flask and diluted with methanol to the mark;

[0158] 4. Preparation of potassium hydroxide methanol solution: 13.1 g of potassium hydroxide was dissolved in 100 mL of anhydrous methanol, and anhydrous sodium sulfate was added for drying. The solution was filtered;

[0159] 5. Sample processing: weigh 60 mg of test sample and control sample into a test tube with a plug, add 2 mL of internal standard solution, add 4 mL of isooctane solution, vortex until dissolved, add 200 μL of potassium hydroxide methanol solution, cover with a glass plug, shake vigorously for 30 s, then stand until clear, add 1 g of sodium bisulfate, shake vigorously, and after the salt precipitates, transfer the upper solution to an on-machine bottle for standby;

[0160] 6. Instrument conditions: the gas chromatography capillary column is a polydicyclopentylsiloxane strong polarity stationary phase, 100 m x 0.25 mm x 0.2 μm; the column temperature is 100°C for 13 min; 10°C / min to 180°C for 6 min; 1°C / min to 200°C for 20 min; 4°C / min to 230°C for 10.5 min; the injection port temperature is 270°C; the split ratio is 100:1; the injection volume is 1 μL;

[0161] The contents of oleic acid, linoleic acid, palmitic acid, stearic acid and total free fatty acid in the test sample and the control sample are quantified by mixing the chromatographic peak area of the fatty acid methyl ester standard working solution, the individual fatty acid methyl ester standard working solution, the test sample and the control sample, and the determination results are shown in Table 3.

[0162] Table 3: Contents of oleic acid, linoleic acid, palmitic acid, stearic acid and total free fatty acid

[0163]

[0164] From the determination results in Table 3, it can be seen that compared with the conventional preparation method, the preparation method of the present application can remove most of the free fatty acids in the sunflower seed oil, and is about 10 times lower.

[0165] Experimental Example 4

[0166] In this experimental example, the triglycerides of sunflower seed oil in the test samples of Examples 1-7 and the control sample of Comparative Example 1 are determined, and the specific steps are as follows:

[0167] 1. Preparation of triglyceride standard stock solution: weigh 500 mg of triglyceride standard into a beaker, dissolve with a small amount of isopropyl alcohol, transfer the solution to a 10 mL volumetric flask, and make up to volume with isopropyl alcohol to obtain the triglyceride standard stock solution;

[0168] 2. Preparation of triglyceride standard working solution: transfer 0.1 mL, 0.2 mL, 0.4 mL, 1 mL and 2 mL of the triglyceride standard stock solution into a 10 mL volumetric flask, respectively, and make up to volume with isopropyl alcohol to obtain the standard working solution with a concentration of 0.5 mg / mL, 1 mg / mL, 2 mg / mL, 5 mg / mL and 10 mg / mL, respectively;

[0169] 3. Sample treatment: 20 mg of the test sample and the control sample were weighed into a beaker, dissolved with a small amount of isopropanol, transferred into a 10 mL volumetric flask, made up to volume with isopropanol, filtered through a 0.45 μm membrane, and used as needed;

[0170] 4. Instrument conditions: the chromatographic column was Symmetry 300 C18, 4.6 mm x 250 mm x 5 μm, the column temperature was 45 °C, the mobile phase A was isopropanol, the mobile phase B was acetonitrile, the elution conditions were 80% B for 0-14 min, 80%-30% B for 14-27 min, 30% B for 27-35 min, 80% B for 35-42 min, the flow rate was 0.6 mL / min, the injection volume was 5 μL, the ionization mode was APCI+, the mass-to-charge ratio acquisition range was 500-1200, the drying gas temperature was 300 °C, the drying gas flow rate was 4 L / min, the vaporizer temperature was 350 °C, the capillary voltage was 3.5 kV, and the corona current was 10 μA; TM C18, 4.6 mm x 250 mm x 5 μm, the column temperature was 45 °C, the mobile phase A was isopropanol, the mobile phase B was acetonitrile, the elution conditions were 80% B for 0-14 min, 80%-30% B for 14-27 min, 30% B for 27-35 min, 80% B for 35-42 min, the flow rate was 0.6 mL / min, the injection volume was 5 μL, the ionization mode was APCI+, the mass-to-charge ratio acquisition range was 500-1200, the drying gas temperature was 300 °C, the drying gas flow rate was 4 L / min, the vaporizer temperature was 350 °C, the capillary voltage was 3.5 kV, and the corona current was 10 μA;

[0171] The standard curve was plotted with the concentration of the triglyceride standard as the abscissa and the corresponding peak area as the ordinate, the linear regression equation was obtained, the content of the triglyceride in the test sample and the control sample was calculated, and the determination results are shown in Table 4.

[0172] Table 4 Content of triglyceride

[0173] Sample Content of triglyceride (%) Example 1 97.7 Example 2 99.7 Example 3 99.8 Example 4 98.6 Example 5 99.7 Example 6 98.5 Example 7 99.6 Comparative Example 1 90.2

[0174] It can be seen from the determination results in Table 4 that, compared with the conventional impurity removal method, the impurity removal ability of the extractant, the lyotropic liquid crystal agent and the purification membrane is stronger, the content of the effective component triglyceride can reach more than 97%, and thus the quality of the extracted sunflower seed oil is improved.

[0175] Experimental Example 5

[0176] In this experimental example, the scar repair efficacy of the test sample of Example 8 and the control samples of Comparative Examples 2-5 was evaluated. The evaluation was carried out objectively and subjectively, and the specific steps were as follows:

[0177] 1. Experimental method:

[0178] 150 special volunteers were selected, the special volunteers had acne scars within 6 months on the face, and the age range was 20-40 years old. These volunteers were randomly divided into 5 groups, and the test sample of Example 8 and the control samples of Comparative Examples 2-5 were respectively applied after facial cleansing in the morning and evening, continuously used for 12 weeks, and the objective evaluation indexes and subjective evaluation indexes of the volunteers before and after use were recorded.

[0179] 2. Evaluation indexes:

[0180] Objective indicators are evaluated by the thickness, pigmentation and hardness of scar morphology. The thickness of the scar is measured by skin ultrasound, the pigmentation of the scar is quantified by CR-400 colorimeter, and the hardness of the scar is evaluated by Cutometer hardness meter.

[0181] Subjective indicators are evaluated by visual analog scale to assess the improvement of symptoms such as itching and pain of the scar, and are scored according to the absence of the above symptoms, mild, moderate and severe, respectively, which are scored as 0-3 points.

[0182] 3. Instrument testing:

[0183] Thickness: At the thickest or most irregular part of the center of the scar, cross-section and longitudinal section are scanned.

[0184] Pigmentation: The probe is lightly pressed vertically on the skin surface to ensure that the measurement window completely covers the marked point, while avoiding hair and excessive pressure to cause the skin to turn white or shift.

[0185] Hardness: The probe is lightly touched vertically on the skin, and the probe sealing ring is completely attached to the skin surface.

[0186] 4. Test results:

[0187] The test records of the volunteers before and after using each group are shown in Tables 5-1, 5-2, 5-3 and 5-4.

[0188] Table 5-1: Scar thickness of volunteers before and after use

[0189]

[0190] Note: * indicates that there is a significant difference between before and after use of the volunteers, p<0.05.

[0191] As can be seen from Table 5-1, after using the scar repair protein polypeptide prepared by the application, the thickness of the scar of the volunteers is obviously thinned, indicating that the fibrosis of the dermis layer is significantly reduced, and the scar has certain repair effect. Although single sunflower seed oil and collagen polypeptide also have a certain thinning effect on the thickness of the scar, but the effect is not as good as the scar repair protein polypeptide prepared by the application.

[0192] Table 5-2: Scar pigmentation of volunteers before and after use

[0193]

[0194] Note: * indicates that there is a significant difference between before and after use of the volunteers, p<0.05.

[0195] As can be seen from Table 5-2, after the volunteers use the scar repair protein polypeptide prepared by the application, the total color difference of the scar is obviously decreased, indicating that the pigment of the scar is obviously reduced and the scar has certain repair effect. Although the total color difference of the single sunflower seed oil and the collagen protein polypeptide is decreased to a certain extent, the effect is not as good as that of the scar repair protein polypeptide prepared by the application.

[0196] Table 5-3: Scar hardness of the volunteers before and after use

[0197]

[0198] Note: R0 is the initial hardness, that is, the resistance value of the skin to the initial negative pressure; R2 is the sustained hardness, that is, the skin hardness under stable negative pressure; * indicates that there is a significant difference before and after the volunteers use, p < 0.05.

[0199] As can be seen from Table 5-3, after the volunteers use the scar repair protein polypeptide prepared by the application, the R0 / R2 value is obviously increased, indicating that the skin elasticity is good and the displacement is large, and the scar has certain softening effect. Although the scar hardness is also softened to a certain extent by the single sunflower seed oil and the collagen protein polypeptide, the effect is not as good as that of the scar repair protein polypeptide prepared by the application.

[0200] Table 5-4: Symptoms such as itching and pain of the scar of the volunteers before and after use

[0201]

[0202] Note: * indicates that there is a significant difference before and after the volunteers use, p < 0.05.

[0203] As can be seen from Table 5-4, after the volunteers use the scar repair protein polypeptide prepared by the application, the symptoms such as itching and pain of the scar are relieved to a certain extent. Although the symptoms such as itching and pain of the scar are also relieved to a certain extent by the single sunflower seed oil and the collagen protein polypeptide, the effect is not as good as that of the scar repair protein polypeptide prepared by the application.

[0204] As can be seen from the above determination results, the scar repair protein polypeptide prepared by the application, that is, the complex of sunflower seed oil-magnesium aluminum-oleic acid-lamellar double hydroxide-collagen protein polypeptide, can make the scar thickness thin, the pigment fade and the hardness soft, and has certain alleviating effect on symptoms such as itching and pain, so the scar repair protein polypeptide prepared by the application has certain scar repair effect.

[0205] The above description is only used to illustrate the technical solutions of the application but not limit the application. Any modification and change of the technical solutions of the application made by those skilled in the art without departing from the overall concept of the application shall still fall within the scope of the application.

Claims

1. A process for the preparation of sunflower seed oil, characterized by: The sunflower seed oil preparation method specifically comprises the following steps: S001, in the sunflower seed powder, vortex uniformity of the extraction agent is added for mixing, ultrasonic treatment, layered treatment, the lower layer liquid is released, and the upper layer liquid is collected, and the crude extract is obtained, wherein the extraction agent is composed of n-hexane, ultrapure water, butyl hydroxy anisole, di-tert-butyl-p-cresol and vitamin E; S002, the crude extract is slowly added to the stirred state of the lyotropic liquid crystal agent for reaction treatment, centrifugation, removal of the lower layer solution and precipitate, and the upper layer solution is collected, and the secondary extract is obtained, wherein the lyotropic liquid crystal agent is mixed by tween-80, ultrapure water and n-hexane to form a lyotropic liquid crystal system; S003, under stirring, slowly drop acyl chloride solution for acyl chloride reaction, slowly drop triethylamine solution for pretreatment, add purified membrane, and perform esterification reaction, and the ester membrane and purified liquid are obtained, wherein the purified membrane is obtained by grafting hydroxyethyl methacrylate to polyvinylidene fluoride membrane; S004, the ester membrane is gently clamped with tweezers, washed, and the washing liquid and purified liquid are collected and mixed and centrifuged, the solution is collected, and evaporated, and the sunflower seed oil is obtained.

2. A method for preparing a scar repair protein polypeptide, characterized in that: The sunflower seed oil obtained by the preparation method of claim 1 is embedded and modified by layered adsorbent and collagen polypeptide to obtain scar repair protein polypeptide, which specifically comprises the following steps: S101, the layered adsorbent, sunflower seed oil and n-hexane are mixed, ultrasonic treatment, and evaporation treatment, and the sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide complex is obtained, wherein the layered adsorbent is formed by modifying magnesium aluminum nitrate type layered double hydroxide with oleic acid; S102, the collagen polypeptide solution is added and mixed, and the sunflower seed oil-magnesium aluminum-oleic acid-layered double hydroxide-collagen polypeptide complex is obtained, which is recorded as scar repair protein polypeptide.

Images