A method for improving the content of antioxidant components in seabuckthorn seed oil

Abstract

The application discloses a processing method for improving the content of antioxidant components of seabuckthorn seed oil, and steps include low-temperature cycle drying, grading crushing, antioxidant spraying, low-temperature ultrafine grinding, ultrasonic-assisted composite enzymolysis pretreatment, low-temperature pressing and separation, enzymatic degumming, and two-stage molecular distillation deacidification and purification. The application combines the synergistic process of antioxidant protection pretreatment, ultrasonic-assisted composite enzymolysis, low-temperature pressing and enzymatic degumming, and two-stage molecular distillation, significantly improves the content and retention rate of antioxidant components in seabuckthorn seed oil, and solves the problems of easy oxidative degradation of antioxidant components, low extraction efficiency and large refining loss in the existing seabuckthorn seed oil extraction and refining process.

Description

Technical Field

[0001] This invention relates to the field of vegetable oil processing technology, and in particular to a method for processing sea buckthorn seed oil to increase the content of antioxidant components. Background Technology

[0002] Sea buckthorn (Hippophae rhamnoides L.) is a plant belonging to the genus Hippophae in the family Elaeagnaceae. Its seeds are rich in a variety of bioactive substances, such as vitamin E, carotenoids, unsaturated fatty acids, flavonoids, amino acids and trace elements. It has a variety of pharmacological functions, such as anti-oxidation, anti-aging, lowering blood lipids and enhancing immunity, and has been widely used in the fields of food, cosmetics and pharmaceuticals.

[0003] Currently, the extraction of sea buckthorn seed oil mainly employs mechanical pressing, organic solvent extraction, supercritical CO2 extraction, and subcritical fluid extraction. Mechanical pressing results in long pressing times, low oil yield, high impurity content, and poor oil quality; furthermore, high-temperature processing can easily destroy active ingredients. Organic solvent extraction suffers from long extraction times, high solvent consumption, and solvent residue in the product. While supercritical CO2 extraction can better preserve active ingredients, it requires sophisticated equipment, has a complex process flow, and high production costs, making large-scale adoption difficult. Subcritical fluid extraction can be performed at lower temperatures, but the content of active substances obtained using existing subcritical extraction methods still needs improvement.

[0004] In addition, the existing sea buckthorn seed oil extraction technology generally has the following technical defects: (1) During the raw material pretreatment process, the crushing of sea buckthorn seeds can easily lead to the oxidative degradation of unsaturated fatty acids and active ingredients; (2) During the enzymatic hydrolysis or extraction process, improper control of process conditions will cause the loss of heat-sensitive antioxidant components; (3) During the crude oil refining stage, conventional deacidification, decolorization and deodorization treatment will cause a large loss of antioxidant components such as vitamin E, phytosterols and polyphenols; (4) Existing technologies focus on a single extraction or refining process and lack a systematic synergistic processing scheme from raw materials to finished products.

[0005] Therefore, developing a processing method that can systematically protect, efficiently retain, and synergistically enhance the content of antioxidant components in sea buckthorn seed oil is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0006] The purpose of this invention is to provide a method for processing sea buckthorn seed oil to increase the content of antioxidant components, thereby solving the technical problems of easy oxidation and degradation of antioxidant components, low extraction efficiency, and large refining losses in the existing sea buckthorn seed oil extraction and refining process.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] This invention provides a method for processing sea buckthorn seed oil to increase the content of antioxidant components, comprising the following steps:

[0009] S1. Dry the sea buckthorn seed raw material at low temperature until the moisture content is 6%~10%;

[0010] S2. The dried sea buckthorn seeds are graded and crushed to a particle size of 0.5mm~2.0mm. An antioxidant solution is sprayed onto the surface of the crushed material, and then the material is subjected to nitrogen protection and allowed to stand.

[0011] S3. Perform ultra-fine grinding at 0~10℃ until the particle size D90≤75μm;

[0012] S4. Ultrasonic-assisted compound enzymatic hydrolysis pretreatment: Add the pulverized material from step S3 to deionized water, adjust the pH, add the compound enzyme preparation to carry out ultrasonic-assisted enzymatic hydrolysis reaction, and heat up to inactivate the enzyme after the enzymatic hydrolysis is completed.

[0013] S5. Low-temperature pressing and separation: The oil is then pressed at low temperature. The resulting mixed oil is centrifuged to obtain crude sea buckthorn seed oil.

[0014] S6. Enzymatic degumming: The crude sea buckthorn seed oil obtained in step S5 is heated to 45~55℃, and citric acid solution and phospholipase are added in sequence to react. After the reaction, the temperature is raised to inactivate the enzyme, and the gum is removed by centrifugation.

[0015] S7. Two-stage molecular distillation: The sea buckthorn seed oil after enzymatic degumming in step S6 is subjected to two-stage molecular distillation. The second-stage distillate is collected, which is the sea buckthorn seed oil product with high antioxidant content.

[0016] In some specific technical solutions of the present invention, in step S1, the conditions for low-temperature cyclic drying are: drying temperature of 35℃~45℃ and drying time of 12~24 hours.

[0017] In some specific technical solutions of the present invention, in step S2, the antioxidant solution is prepared by vitamin C and citric acid in a mass ratio of (1~3):(0.5~1.5), and the amount of antioxidant solution used is 0.05%~0.2% of the mass of sea buckthorn seeds.

[0018] In some specific technical solutions of the present invention, in step S2, the nitrogen protection and static treatment time is 10 to 30 minutes.

[0019] In some specific technical solutions of the present invention, in step S4, the mass ratio of sea buckthorn seeds to deionized water is 1:(3~6).

[0020] In some specific technical solutions of the present invention, in step S4, the pH is adjusted to 4.5~5.5 before adding the compound enzyme preparation.

[0021] In some specific technical solutions of the present invention, in step S4, the compound enzyme preparation is composed of cellulase, pectinase and acidic protease in a mass ratio of (2~4):(2~3):(1~2), and the amount of the compound enzyme preparation is 0.5%~1.5% of the mass of sea buckthorn seeds.

[0022] In some specific technical solutions of the present invention, in step S4, the conditions for the ultrasound-assisted enzymatic hydrolysis reaction are as follows: the enzymatic hydrolysis reaction is carried out for 1.5 to 3 hours under the conditions of ultrasound power of 100~300W, ultrasound frequency of 20~40kHz, and enzymatic hydrolysis temperature of 45~55℃.

[0023] In some specific technical solutions of the present invention, in step S4, the enzyme inactivation condition is to treat at 80~90℃ for 10~20 minutes.

[0024] In some specific technical solutions of the present invention, in step S5, the pressing process is carried out at 20~35℃ and pressing pressure of 40~60MPa.

[0025] In some specific technical solutions of the present invention, in step S5, the centrifugal separation adopts a horizontal spiral sedimentation centrifuge with a centrifugal speed of 4000~6000 r / min.

[0026] In some specific technical solutions of the present invention, in step S6, the mass fraction of citric acid solution is 40%~50%, the amount of citric acid solution used is 0.5%~1.0% of the mass of crude sea buckthorn seed oil, and the reaction is stirred for 20~40 minutes after adding citric acid solution.

[0027] In some specific technical solutions of the present invention, in step S6, the phospholipase is phospholipase A1 or phospholipase C, the amount of enzyme added is 0.01% to 0.05% of the mass of crude sea buckthorn seed oil, and the enzyme reaction time after adding the phospholipase is 1 to 3 hours.

[0028] In some specific technical solutions of the present invention, in step S6, the enzyme inactivation temperature is 80~90℃ and the enzyme inactivation time is 10~15 minutes.

[0029] In some preferred embodiments of the present invention, in step S6, before adding phospholipase, the enzyme is first subjected to ultrasonic-assisted treatment for 15 to 25 minutes at an ultrasonic power of 100 to 200 W and a frequency of 20 to 30 kHz; after the enzyme reaction is completed, the enzyme is then subjected to ultrasonic-assisted demulsification for 15 to 25 minutes at an ultrasonic power of 80 to 120 W and a frequency of 20 to 30 kHz before being heated to inactivate the enzyme.

[0030] In some specific technical solutions of the present invention, in step S7, the first-stage molecular distillation is carried out under the conditions of vacuum degree 0.1~1.0 Pa, temperature 120~150℃, and scraper rotation speed 200~400 r / min, and the residence time on the evaporation surface is 30~90 seconds; the second-stage molecular distillation is carried out under the conditions of vacuum degree 0.01~0.1 Pa, temperature 160~200℃, and scraper rotation speed 300~500 r / min, and the residence time on the evaporation surface is 60~120 seconds.

[0031] In some preferred embodiments of the present invention, an ultrasonic pre-disruption treatment step is further included between step S2 and step S3: the material treated in step S2 is added to deionized water at a material-to-liquid ratio of 1:2, and treated for 20 to 40 minutes under ultrasonic power of 300 to 500W, frequency of 35 to 45kHz, and temperature of 10 to 20°C. After centrifugation, the solid phase is taken and proceeded to step S3.

[0032] This invention provides a method for processing sea buckthorn seed oil to increase the content of antioxidant components. Through a synergistic process combining antioxidant protection pretreatment, ultrasound-assisted enzymatic hydrolysis, low-temperature pressing and enzymatic degumming, and two-stage molecular distillation, the method significantly improves the content and retention rate of antioxidant components in sea buckthorn seed oil. Compared with existing technologies, it has the following beneficial effects:

[0033] 1. Implement antioxidant protection from the raw material pretreatment stage to systematically reduce the oxidative loss of active ingredients.

[0034] To address the shortcomings of existing technologies that generally neglect antioxidant protection during the raw material pretreatment stage, this invention involves spraying an antioxidant solution composed of vitamin C and citric acid after graded crushing, followed by static infiltration treatment under nitrogen protection. Simultaneously, temperature is strictly controlled during key stages such as low-temperature ultrafine grinding and low-temperature pressing, with multiple nodes working together to protect the antioxidant components from oxidative degradation. As shown in Tables 1 and 2, experimental results indicate that using the processing technology of this invention (Examples 1-5), the synergistic protection of vitamin C and citric acid increases the vitamin E retention rate in sea buckthorn seed oil by 125.1% to 180.9% compared to conventional treatment (Comparative Example 1).

[0035] 2. Ultrasonic-assisted enzymatic hydrolysis pretreatment significantly improves the extraction efficiency of oils and active ingredients.

[0036] This invention combines ultrasound-assisted technology with enzymatic hydrolysis using cellulase, pectinase, and acidic protease. The mechanical oscillation and cavitation effect of ultrasound promotes enzyme-substrate contact and disrupts cell wall structure, fully exposing intracellular lipids and antioxidants. Post-hydrolysis heating inactivates the enzymes, ensuring the biochemical stability of the oil. As shown in Table 1, this pretreatment scheme significantly increases the oil yield in subsequent low-temperature pressing (to 21.2%–23.1%), while simultaneously increasing the content of antioxidant active substances such as total phenols and total flavonoids by 28.0%–58.3% and 34.4%–78.5%, respectively, compared to the untreated (Comparative Example 3).

[0037] 3. Low-temperature pressing combined with enzymatic degumming maximizes the retention of natural antioxidant components.

[0038] This invention eliminates the loss of active ingredients and oil hydrolysis caused by traditional hydration degumming and alkali refining deacidification, and adopts low-temperature spiral pressing combined with phospholipase degumming technology. The low-temperature pressing conditions (20~35℃) avoid the thermal damage of heat-sensitive antioxidant components caused by high temperatures; the enzymatic degumming specifically hydrolyzes non-hydrated phospholipids under mild conditions, resulting in excellent degumming effect without loss of nutrients, and avoiding the loss of lipid byproducts caused by repeated washing in conventional degumming methods.

[0039] 4. Two-stage molecular distillation deacidification purification achieves efficient retention and enrichment of antioxidant components.

[0040] This invention employs a two-stage molecular distillation process to remove free fatty acids and impurities: the first stage of molecular distillation removes free fatty acids at 120–150°C, and the second stage collects the target fraction at 160–200°C. Molecular distillation operates under high vacuum and short residence time conditions, with processing temperatures significantly lower than traditional deacidification and deodorization temperatures (typically 220–260°C). As shown in Table 2, the retention rates of vitamin E, phytosterols, and polyphenolic antioxidants are all improved compared to traditional refining methods (such as Comparative Examples 2 and 3).

[0041] 5. The synergistic effect of each step significantly increases the overall content and activity of antioxidant components.

[0042] Through the synergistic effect of the above processes, the sea buckthorn seed oil product finally obtained by this invention has a total phenolic content of 65-95 mg GAE / 100g, a total flavonoid content of 45-75 mg RE / 100g, and a vitamin E content of 210-280 mg / 100g; the DPPH free radical scavenging rate IC50 is also high. 50 Values ​​reaching 0.8~2.5 mg / mL, ABTS + Free radical scavenging rate IC 50 The value reaches 8~14 mg / mL, and the total antioxidant capacity (FRAP value) reaches 350~500 μmol Fe. 2+ / L, all indicators are significantly better than existing technologies.

[0043] 6. The process conditions are mild and suitable for industrial production.

[0044] The processing method provided by this invention features mild and controllable process conditions at each step, highly versatile equipment, and eliminates the need for harsh conditions such as ultra-high pressure or ultra-high temperature. It is safe, reliable, energy-efficient, and easily scalable for industrial production. Furthermore, this process does not use organic solvents (only water is used as a solvent during enzymatic hydrolysis), resulting in safe and environmentally friendly products that align with the principles of green manufacturing. Detailed Implementation

[0045] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0046] To address the technical problems of easy oxidation and degradation of antioxidant components, low extraction efficiency, and large refining losses in the existing sea buckthorn seed oil extraction and refining process, this invention provides a method for improving the antioxidant content of sea buckthorn seed oil, comprising the following steps:

[0047] S1. Raw material screening and drying

[0048] Select mature, mold-free sea buckthorn seeds, remove impurities and shriveled seeds, and dry the sea buckthorn seeds to a moisture content of 6%~10% using a low-temperature circulating drying method. The drying temperature is 35℃~45℃ and the drying time is 12~24 hours.

[0049] Step S1 employs a low-temperature circulating drying method, with the drying temperature strictly controlled between 35 and 45°C. Within this temperature range, the heat-sensitive antioxidant components in sea buckthorn seeds (such as vitamin E and polyphenols) undergo almost no thermal degradation, and low-temperature drying effectively prevents accelerated oxidation of oils. The moisture content is controlled between 6% and 10%, ensuring efficient subsequent grinding without causing increased brittleness or overheating during grinding due to excessively low moisture content.

[0050] S2, graded crushing and antioxidant spraying

[0051] The dried sea buckthorn seeds (S1) are fed into a grading crusher for coarse crushing, and the particle size of the crushed material is controlled to be 0.5mm~2.0mm. Then, an antioxidant solution is evenly sprayed onto the surface of the coarsely crushed material. The antioxidant solution is prepared by dissolving vitamin C and citric acid in deionized water at a mass ratio of (1~3):(0.5~1.5). The amount of antioxidant solution used is 0.05%~0.2% of the mass of sea buckthorn seeds. After spraying, the material is left to stand under nitrogen protection for 10~30 minutes.

[0052] Step S2 employs a combination of graded crushing and antioxidant spraying. Graded crushing coarsely breaks the sea buckthorn seeds from whole particles to a particle size of 0.5mm~2.0mm, increasing the contact opportunity between the antioxidant and the oils inside the seeds, while avoiding over-crushing that could lead to material heating and premature oxidation of active ingredients exposed to air. The sprayed vitamin C and citric acid work synergistically to provide antioxidant protection. Vitamin C, as a reducing antioxidant, preferentially reacts with oxygen to consume dissolved oxygen, while citric acid inhibits the catalytic effect of metal ions on oil oxidation by chelating them. Under weakly acidic conditions, the two form a synergistic antioxidant protection system. Following spraying, a nitrogen-protected settling treatment allows the antioxidant to fully penetrate the material, forming comprehensive protection.

[0053] S3, Low-temperature ultrafine grinding

[0054] The material treated with S2 is fed into a low-temperature ultrafine pulverizer for pulverization. The pulverization temperature is controlled at 0~10℃, and the particle size of the pulverized material is D90≤75μm.

[0055] Step S3 employs low-temperature ultrafine pulverization technology to reduce the material particle size to D90≤75μm. Ultrafine pulverization effectively disrupts the plant cell wall structure, fully exposing the oils and antioxidants within the cells, creating favorable conditions for subsequent enzymatic pretreatment and pressing. The continuous control of low-temperature conditions (0~10℃) effectively suppresses frictional heat generation during pulverization, preventing the oxidation of unsaturated fatty acids and the loss of heat-sensitive active ingredients.

[0056] S4, Ultrasonic-assisted compound enzymatic pretreatment

[0057] Add the crushed S3 material to deionized water at a material-to-liquid mass ratio of 1:(3~6), stir evenly, adjust the pH to 4.5~5.5, and add a compound enzyme preparation. The compound enzyme preparation consists of cellulase, pectinase, and acidic protease in a mass ratio of (2~4):(2~3):(1~2). The amount of the compound enzyme preparation is 0.5%~1.5% of the sea buckthorn seed mass. Carry out the enzymatic hydrolysis reaction under ultrasonic-assisted conditions. The ultrasonic power is 100~300W, the ultrasonic frequency is 20~40kHz, the enzymatic hydrolysis temperature is 45~55℃, and the enzymatic hydrolysis time is 1.5~3 hours. After the enzymatic hydrolysis is completed, raise the temperature to 80~90℃ to inactivate the enzyme for 10~20 minutes.

[0058] Step S4 employs ultrasound-assisted enzymatic pretreatment. A combination of cellulase, pectinase, and acidic protease is selected, working synergistically. Cellulase degrades the cellulose components in the cell wall skeleton, pectinase breaks down pectin in the intercellular matrix, and acidic protease moderately hydrolyzes the cellular protein network structure under acidic conditions. The mechanical oscillation and cavitation effect of ultrasound promotes enzyme-substrate contact, accelerates mass transfer, and significantly improves enzymatic hydrolysis efficiency. After hydrolysis, heating to inactivate the enzyme terminates the reaction, preventing residual enzymes from affecting subsequent processes.

[0059] S5, Low-temperature pressing and separation

[0060] The material after S4 enzymatic hydrolysis pretreatment is fed into a low-temperature screw press for pressing, with the pressing temperature controlled at 20~35℃ and the pressing pressure at 40~60MPa; the mixed oil obtained by pressing is separated by a horizontal screw sedimentation centrifuge at a speed of 4000~6000r / min to obtain crude sea buckthorn seed oil.

[0061] Step S5 employs low-temperature pressing for separation. The low-temperature conditions (20~35℃) avoid thermal damage to the active ingredients caused by high-temperature pressing, and centrifugal separation effectively removes solid residues introduced during the pressing process, yielding crude sea buckthorn seed oil.

[0062] S6, Enzymatic degumming

[0063] The crude sea buckthorn seed oil obtained from S5 is heated to 45-55℃, and 0.5%-1.0% of the crude sea buckthorn seed oil mass of citric acid solution (mass fraction of 40%-50%) is added. The mixture is stirred and reacted for 20-40 minutes. Then, phospholipase A1 or phospholipase C is added at a rate of 0.01%-0.05% of the crude sea buckthorn seed oil mass. The mixture is stirred and reacted at 45-55℃ for 1-3 hours. After the reaction is completed, the temperature is raised to 80-90℃ to inactivate the enzyme for 10-15 minutes, and the mixture is centrifuged to remove the gum.

[0064] Step S6 employs enzymatic degumming instead of traditional acid-hydration degumming and water washing refining. Phospholipases specifically hydrolyze the fatty acid ester bonds in non-hydrated phospholipids, converting them into water-soluble lysophospholipids that are easily removed by centrifugation. Enzymatic degumming is carried out under mild conditions, avoiding the loss of antioxidants and oil hydrolysis caused by high temperatures, strong acids, and repeated washing in traditional degumming processes.

[0065] S7. Two-stage molecular distillation for deacidification and purification

[0066] Sea buckthorn seed oil degummed by S6 enzyme method is subjected to two-stage molecular distillation: the first stage of molecular distillation is carried out under vacuum of 0.1~1.0 Pa, temperature of 120~150℃, and scraper rotation speed of 200~400 r / min, with a residence time on the evaporation surface of 30~90 seconds, to remove free fatty acids and some low-boiling-point impurities; the residue from the first stage of distillation enters the second stage of molecular distillation, which is carried out under vacuum of 0.01~0.1 Pa, temperature of 160~200℃, and scraper rotation speed of 300~500 r / min, with a residence time on the evaporation surface of 60~120 seconds, and the second stage distillate is collected, which is the sea buckthorn seed oil product with high antioxidant content.

[0067] Step S7 employs two-stage molecular distillation for deacidification and purification. Molecular distillation is carried out under extremely high vacuum and extremely short residence time. The first-stage molecular distillation removes free fatty acids and low-boiling-point impurities at a relatively low evaporation temperature, while the second-stage molecular distillation evaporates and collects the highly active sea buckthorn seed oil fraction at an appropriate evaporation temperature. Compared with the traditional vacuum deacidification and deodorization process, the molecular distillation method of this invention has the following advantages: (1) The distillation temperature is low and the heating time is short, which effectively protects heat-sensitive antioxidant components such as vitamin E, phytosterols, and polyphenols; (2) There is no need to introduce water vapor for distillation stripping, thus avoiding the hydrolysis and oxidation of the oil; (3) Two-stage distillation can achieve efficient separation of free fatty acids and active oil fractions, resulting in high product purity.

[0068] As an improved technical solution, in step S6, before adding phospholipase, the enzyme is first subjected to ultrasonic-assisted treatment for 15-25 minutes at an ultrasonic power of 100-200W and a frequency of 20-30kHz; after the enzyme reaction is completed, the enzyme is then subjected to ultrasonic-assisted demulsification for 15-25 minutes at an ultrasonic power of 80-120W and a frequency of 20-30kHz before being heated to inactivate the enzyme.

[0069] As an improved technical solution, an ultrasonic pre-disruption treatment step is also included between step S2 and step S3: the material treated in step S2 is added to deionized water at a material-to-liquid ratio of 1:2, and treated for 20 to 40 minutes under ultrasonic power of 300~500W, frequency of 35~45kHz, and temperature of 10~20℃. After centrifugation, the solid phase is taken and entered into step S3.

[0070] The present invention will be further described in detail below through detailed embodiments.

[0071] Unless otherwise specified, all materials and reagents used in the examples and comparative examples are commercially available. Unless otherwise specified, all experimental and measurement methods used in the performance tests are conventional methods.

[0072] Example 1

[0073] This embodiment provides a method for processing sea buckthorn seed oil to increase the content of antioxidant components. The specific steps are as follows:

[0074] S1. Raw material screening and drying

[0075] Ten kilograms of mature, mold-free sea buckthorn seeds from the current year's harvest were selected, and impurities and shriveled seeds were removed. The sea buckthorn seeds were placed in a low-temperature circulating drying oven and dried using a low-temperature circulating drying method. The drying temperature was set at 40℃, and the drying time was 18 hours. After drying, the moisture content of the sea buckthorn seeds was measured to be 8.2%.

[0076] S2, graded crushing and antioxidant spraying

[0077] The dried sea buckthorn seeds were fed into a grading crusher for coarse crushing, resulting in a particle size of 1.0-1.5 mm. An antioxidant solution was prepared: 2.0 g of vitamin C and 1.0 g of citric acid were weighed and dissolved in 100 mL of deionized water to prepare an antioxidant solution with a vitamin C to citric acid mass ratio of 2:1. The antioxidant solution was then evenly sprayed onto the surface of the coarsely crushed material, with the amount of antioxidant solution being 0.1% of the sea buckthorn seed mass. After spraying, the material was allowed to stand for 20 minutes under nitrogen protection to allow the antioxidant to fully penetrate into the material.

[0078] S3, Low-temperature ultrafine grinding

[0079] The material processed in step S2 is fed into a low-temperature ultrafine pulverizer for pulverization. During the pulverization process, the temperature is controlled by a liquid nitrogen circulating jacket, and the pulverization temperature is controlled at 5℃. After pulverization, the material passes through a 200-mesh standard sieve (D90≤75μm).

[0080] S4, Ultrasonic-assisted compound enzymatic pretreatment

[0081] Add the pulverized material from step S3 to deionized water at a material-to-liquid mass ratio of 1:4 and stir until homogeneous. Adjust the pH to 5.0 with citric acid. Prepare a compound enzyme preparation: Mix 3.0g of cellulase, 2.5g of pectinase, and 1.5g of acidic protease, with a cellulase:pectinase:acidic protease mass ratio of 3:2.5:1.5. Add the compound enzyme preparation to the material system at a concentration of 0.7% of the sea buckthorn seed mass (i.e., 0.7g compound enzyme preparation / 100g sea buckthorn seed). Perform the enzymatic hydrolysis reaction under ultrasonic-assisted conditions: ultrasonic power set to 200W, ultrasonic frequency to 30kHz, hydrolysis temperature to 50℃, and hydrolysis time to 2 hours. After hydrolysis, raise the temperature to 85℃ for 15 minutes to inactivate the enzyme.

[0082] S5, Low-temperature pressing and separation

[0083] The material after enzymatic hydrolysis pretreatment in step S4 was fed into a low-temperature screw press for pressing. During the pressing process, the pressing temperature was controlled at 25℃ by a circulating cooling water jacket, and the pressing pressure was set at 50MPa. The mixed oil obtained from pressing was separated by a horizontal screw sedimentation centrifuge at a speed of 5000r / min, yielding approximately 2.85kg of crude sea buckthorn seed oil.

[0084] S6, Enzymatic degumming

[0085] The crude sea buckthorn seed oil obtained in step S5 was heated to 50°C, and 0.8% (by weight of the crude sea buckthorn seed oil) of citric acid solution (45% by weight) was added. The mixture was stirred and reacted for 30 minutes. Then, phospholipase A1 was added at a rate of 0.03% (by weight of the crude sea buckthorn seed oil), and the mixture was stirred and reacted at 50°C for 2 hours. After the reaction was completed, the temperature was raised to 85°C to inactivate the enzyme for 12 minutes. The mixture was then centrifuged to remove the gum layer, yielding approximately 2.68 kg of degummed sea buckthorn seed oil.

[0086] S7. Two-stage molecular distillation for deacidification and purification

[0087] The sea buckthorn seed oil after enzymatic degumming in step S6 was subjected to two-stage molecular distillation.

[0088] The first-stage molecular distillation was carried out under a vacuum of 0.5 Pa, an evaporation temperature of 135 °C, and a scraper rotation speed of 300 r / min. The residence time on the evaporation surface was 60 seconds. Free fatty acids and some low-boiling-point impurities were removed, and the distillate was collected for other uses.

[0089] The residue from the first stage of distillation enters the second stage of molecular distillation, which is carried out under the conditions of vacuum degree 0.05 Pa, evaporation temperature 180℃, and scraper rotation speed 400 r / min. The residence time on the evaporation surface is 90 seconds. The second stage distillate is collected to obtain approximately 2.25 kg of sea buckthorn seed oil product with high antioxidant content.

[0090] Example 2

[0091] This embodiment provides another method for processing sea buckthorn seed oil to increase the content of antioxidant components. The main differences from Example 1 are in the drying temperature, antioxidant ratio, enzyme ratio, and molecular distillation parameters. The specific steps are as follows:

[0092] S1. Raw material screening and drying

[0093] Ten kilograms of mature, mold-free sea buckthorn seeds from the current year's harvest were selected, and impurities and shriveled seeds were removed. The sea buckthorn seeds were placed in a low-temperature circulating drying oven and dried using a low-temperature circulating drying method. The drying temperature was set at 35°C, and the drying time was 24 hours. After drying, the moisture content of the sea buckthorn seeds was measured to be 6.5%.

[0094] S2, graded crushing and antioxidant spraying

[0095] The dried sea buckthorn seeds were fed into a grading crusher for coarse crushing, resulting in a particle size of 0.5-1.0 mm. An antioxidant solution was prepared: 1.5 g of vitamin C and 1.5 g of citric acid were weighed and dissolved in 100 mL of deionized water to prepare an antioxidant solution with a vitamin C to citric acid mass ratio of 1:1. The antioxidant solution was then evenly sprayed onto the surface of the coarsely crushed material, with the amount of antioxidant solution being 0.15% of the sea buckthorn seed mass. After spraying, the material was allowed to stand for 30 minutes under nitrogen protection to allow the antioxidant to fully penetrate into the material.

[0096] S3, Low-temperature ultrafine grinding

[0097] The material processed in step S2 is fed into a low-temperature ultrafine pulverizer for pulverization. During the pulverization process, the temperature is controlled by a liquid nitrogen circulating jacket, and the pulverization temperature is controlled at 0℃. After pulverization, the material passes through a 200-mesh standard sieve (D90≤75μm).

[0098] S4, Ultrasonic-assisted compound enzymatic pretreatment

[0099] Add the pulverized material from step S3 to deionized water at a material-to-liquid mass ratio of 1:3 and stir until homogeneous. Adjust the pH to 4.5 with citric acid. Prepare a compound enzyme preparation: mix 4.0g of cellulase, 2.0g of pectinase, and 1.0g of acidic protease, with a cellulase:pectinase:acidic protease mass ratio of 4:2:1. Add the compound enzyme preparation to the material system at a concentration of 1.2% of the sea buckthorn seed mass (i.e., 1.2g compound enzyme preparation / 100g sea buckthorn seed). Perform the enzymatic hydrolysis reaction under ultrasonic-assisted conditions: ultrasonic power set to 100W, ultrasonic frequency to 20kHz, hydrolysis temperature to 45℃, and hydrolysis time to 3 hours. After hydrolysis, heat to 90℃ for 10 minutes to inactivate the enzyme.

[0100] S5, Low-temperature pressing and separation

[0101] The material after enzymatic hydrolysis pretreatment in step S4 is fed into a low-temperature screw press for pressing. During the pressing process, the pressing temperature is controlled at 20℃ by a circulating cooling water jacket, and the pressing pressure is set at 40MPa. The mixed oil obtained from pressing is separated by a horizontal screw sedimentation centrifuge at a speed of 4000r / min to obtain crude sea buckthorn seed oil.

[0102] S6, Enzymatic degumming

[0103] The crude sea buckthorn seed oil obtained in step S5 was heated to 45°C, and 1.0% (by weight of the crude sea buckthorn seed oil) of citric acid solution (40% by weight) was added. The mixture was stirred and reacted for 30 minutes. Then, phospholipase C was added at a concentration of 0.05% (by weight of the crude sea buckthorn seed oil), and the mixture was stirred and reacted at 45°C for 3 hours. After the reaction was completed, the temperature was raised to 90°C to inactivate the enzyme for 10 minutes. The mixture was then centrifuged to remove the gum layer, yielding degummed sea buckthorn seed oil.

[0104] S7. Two-stage molecular distillation for deacidification and purification

[0105] The sea buckthorn seed oil after enzymatic degumming in step S6 was subjected to two-stage molecular distillation.

[0106] The first-stage molecular distillation was carried out under a vacuum of 0.1 Pa, an evaporation temperature of 120 °C, and a scraper rotation speed of 200 r / min. The residence time on the evaporation surface was 90 seconds. Free fatty acids and some low-boiling-point impurities were removed, and the distillate was collected for other uses.

[0107] The residue from the first stage of distillation enters the second stage of molecular distillation, which is carried out under the conditions of vacuum degree 0.01 Pa, evaporation temperature 160℃, and scraper rotation speed 300 r / min. The residence time on the evaporation surface is 120 seconds. The second stage distillate is collected to obtain approximately 2.18 kg of sea buckthorn seed oil product with high antioxidant content.

[0108] Example 3

[0109] This embodiment provides another method for processing sea buckthorn seed oil to increase the content of antioxidant components. The difference from Embodiment 1 lies in the overall adjustment of process parameters. The specific steps are as follows:

[0110] S1. Raw material screening and drying

[0111] 10 kg of mature, mold-free sea buckthorn seeds from the current year's harvest were selected, and impurities and shriveled seeds were removed. The sea buckthorn seeds were placed in a low-temperature circulating drying oven and dried using a low-temperature circulating drying method. The drying temperature was set at 45℃, and the drying time was 12 hours. After drying, the moisture content of the sea buckthorn seeds was measured to be 9.5%.

[0112] S2, graded crushing and antioxidant spraying

[0113] The dried sea buckthorn seeds were fed into a grading crusher for coarse crushing, resulting in a particle size of 1.5-2.0 mm. An antioxidant solution was prepared: 3.0 g of vitamin C and 0.5 g of citric acid were weighed and dissolved in 100 mL of deionized water to prepare an antioxidant solution with a vitamin C to citric acid mass ratio of 3:0.5. The antioxidant solution was evenly sprayed onto the surface of the coarsely crushed material, with the amount of antioxidant solution being 0.05% of the sea buckthorn seed mass. After spraying, the material was allowed to stand for 10 minutes under nitrogen protection to allow the antioxidant to fully penetrate into the material.

[0114] S3, Low-temperature ultrafine grinding

[0115] The material processed in step S2 is fed into a low-temperature ultrafine pulverizer for pulverization. During the pulverization process, the temperature is controlled by a liquid nitrogen circulating jacket, and the pulverization temperature is controlled at 10℃. After pulverization, the material passes through a 200-mesh standard sieve (D90≤75μm).

[0116] S4, Ultrasonic-assisted compound enzymatic pretreatment

[0117] Add the pulverized material from step S3 to deionized water at a material-to-liquid mass ratio of 1:6 and stir until homogeneous. Adjust the pH to 5.5 with citric acid. Prepare a compound enzyme preparation: Mix 2.0g of cellulase, 3.0g of pectinase, and 2.0g of acidic protease, with a cellulase:pectinase:acidic protease mass ratio of 2:3:2. Add the compound enzyme preparation to the material system at a concentration of 0.5% of the sea buckthorn seed mass (i.e., 0.5g compound enzyme preparation / 100g sea buckthorn seed). Perform the enzymatic hydrolysis reaction under ultrasonic-assisted conditions: ultrasonic power set to 300W, ultrasonic frequency to 40kHz, hydrolysis temperature to 55℃, and hydrolysis time to 1.5 hours. After hydrolysis, heat to 80℃ for 20 minutes to inactivate the enzyme.

[0118] S5, Low-temperature pressing and separation

[0119] The material after enzymatic hydrolysis pretreatment in step S4 is fed into a low-temperature screw press for pressing. During the pressing process, the pressing temperature is controlled at 35℃ by a circulating cooling water jacket, and the pressing pressure is set at 60MPa. The mixed oil obtained by pressing is separated by a horizontal screw sedimentation centrifuge at a speed of 6000r / min to obtain crude sea buckthorn seed oil.

[0120] S6, Enzymatic degumming

[0121] The crude sea buckthorn seed oil obtained in step S5 was heated to 55°C, and 0.5% (by weight of the crude sea buckthorn seed oil) of citric acid solution (mass fraction 50%) was added. The mixture was stirred and reacted for 30 minutes. Then, phospholipase A1 was added at a concentration of 0.01% (by weight of the crude sea buckthorn seed oil), and the mixture was stirred and reacted at 55°C for 1 hour. After the reaction was completed, the temperature was raised to 80°C to inactivate the enzyme for 15 minutes. The mixture was then centrifuged to remove the gum layer, yielding degummed sea buckthorn seed oil.

[0122] S7. Two-stage molecular distillation for deacidification and purification

[0123] The sea buckthorn seed oil after enzymatic degumming in step S6 was subjected to two-stage molecular distillation.

[0124] The first-stage molecular distillation was carried out under a vacuum of 1.0 Pa, an evaporation temperature of 150 °C, and a scraper rotation speed of 400 r / min. The residence time on the evaporation surface was 30 seconds. Free fatty acids and some low-boiling-point impurities were removed, and the distillate was collected for other uses.

[0125] The residue from the first stage of distillation enters the second stage of molecular distillation, which is carried out under the conditions of vacuum degree of 0.1 Pa, evaporation temperature of 200℃, and scraper rotation speed of 500 r / min. The residence time on the evaporation surface is 60 seconds. The second stage distillate is collected to obtain approximately 2.12 kg of sea buckthorn seed oil product with high antioxidant content.

[0126] Example 4

[0127] This embodiment provides another method for processing sea buckthorn seed oil to increase the content of antioxidant components. Based on Example 1, it adds an enhanced pretreatment of ultrasonic pre-cell disruption. The specific differences are as follows:

[0128] An ultrasonic pre-disruption step is added between steps S2 and S3: the material treated in step S2 is placed in an ultrasonic treatment tank, deionized water is added at a material-to-liquid ratio of 1:2, and the treatment is carried out for 30 minutes at an ultrasonic power of 400W, a frequency of 40kHz, and a temperature of 15℃. After treatment, the liquid phase is removed by centrifugation, and the solid phase is taken and proceeded to step S3. The remaining steps are the same as in Example 1, and will not be repeated. Approximately 2.31 kg of sea buckthorn seed oil with high antioxidant content is finally obtained.

[0129] Example 5

[0130] This embodiment provides another method for processing sea buckthorn seed oil to increase the content of antioxidant components. Based on Example 1, the enzymatic degumming in step S6 is adjusted to a deep degumming composite treatment. The specific differences are as follows:

[0131] In step S6, an auxiliary ultrasonic treatment is added before adding phospholipase and after the enzyme reaction: Citric acid solution is added to the crude sea buckthorn seed oil, and the mixture is treated with ultrasound at 150W and 25kHz for 20 minutes, followed by the addition of phospholipase. After the enzyme reaction, the mixture is subjected to assisted demulsification with ultrasound at 100W and 20kHz for 20 minutes, followed by temperature inactivation of the enzyme. The remaining steps are the same as in Example 1, and will not be repeated. Approximately 2.28 kg of sea buckthorn seed oil with high antioxidant content is finally obtained.

[0132] Comparative Example 1

[0133] This comparative example uses conventional low-temperature pressing to extract sea buckthorn seed oil, without antioxidant spraying, compound enzymatic hydrolysis, or molecular distillation. The specific steps are as follows:

[0134] S1. Raw material screening and drying

[0135] Ten kilograms of mature, mold-free sea buckthorn seeds from the current year's harvest were selected, and impurities and shriveled seeds were removed. The sea buckthorn seeds were placed in a low-temperature circulating drying oven and dried using a low-temperature circulating drying method. The drying temperature was set at 40℃, and the drying time was 18 hours. After drying, the moisture content of the sea buckthorn seeds was measured to be 8.2%.

[0136] S2. Crushing: The dried sea buckthorn seeds are directly fed into a regular crusher for crushing. The crushed material passes through a 60-mesh standard sieve (D90≤250μm) without antioxidant spraying or nitrogen protection treatment.

[0137] S3. Pressing: The crushed material is fed into a screw press for pressing at a temperature of 50℃ (normal pressing temperature) and a pressure of 50MPa.

[0138] S4. Filtration: The mixed oil obtained by pressing is filtered by a plate and frame filter press to obtain crude sea buckthorn seed oil.

[0139] S5. Water washing and refining: Heat the crude sea buckthorn seed oil to 60°C, add an equal volume of hot water and stir for 30 minutes. After standing and separating into layers, remove the aqueous phase. Repeat the water washing twice. Then, vacuum dehydrate at 70°C for 30 minutes.

[0140] S6. Fine filtration: After dehydration, the sea buckthorn seed oil product is obtained by fine filtration, which yields approximately 1.46 kg.

[0141] Comparative Example 2

[0142] This comparative example uses supercritical CO2 extraction to extract sea buckthorn seed oil, without involving pretreatment such as antioxidant spraying or compound enzymatic hydrolysis. The specific steps are as follows:

[0143] S1. Raw material pretreatment

[0144] Ten kilograms of mature, mold-free sea buckthorn seeds from the current year's harvest were selected, and impurities and shriveled seeds were removed. The sea buckthorn seeds were placed in a low-temperature circulating drying oven and dried using a low-temperature circulating drying method. The drying temperature was set at 40℃, and the drying time was 18 hours. After drying, the moisture content of the sea buckthorn seeds was measured to be 8.2%.

[0145] S2. Crushing: The dried sea buckthorn seeds are directly fed into a regular crusher for crushing. The crushed material is then passed through an 80-mesh standard sieve.

[0146] S3. Supercritical CO2 Extraction: The pulverized sea buckthorn seed powder is loaded into the extraction vessel. The extraction pressure is 30 MPa, the extraction temperature is 45℃, the CO2 flow rate is 20 kg / h, and the extraction time is 2 hours. The separation vessel pressure is 6 MPa, and the separation temperature is 35℃.

[0147] S4. Collect the extract to obtain approximately 1.62 kg of sea buckthorn seed oil product.

[0148] Comparative Example 3

[0149] This comparative example uses the method disclosed in the prior art, "A seabuckthorn oil and its extraction method and application" (Chinese invention patent with authorization announcement number CN116286165B), to prepare seabuckthorn seed oil as a control. The specific steps are as follows:

[0150] Step 1: Soak the sea buckthorn pulp in water at a ratio of 1:3, and stir well.

[0151] Step 2: Add the compound enzyme and incubate for 4 hours for enzymatic hydrolysis. The mass ratio of pectinase to cellulase is 3:2, and the dosage of the compound enzyme is 2.5g per kg of sea buckthorn pomace.

[0152] Step 3: Centrifuge the enzymatic hydrolysate, add 75% ethanol solution to the centrifuged precipitate, and heat and reflux for extraction.

[0153] Step 4: Filter the alcohol extract, combine the filter residue with the centrifuged liquid, and then add n-hexane for extraction.

[0154] Step 5: Recover hexane from the extract to obtain crude sea buckthorn oil.

[0155] Step 6: Add activated carbon and silica gel powder to the crude sea buckthorn oil in sequence, stir thoroughly, let stand, and filter to obtain approximately 1.82 kg of sea buckthorn oil product.

[0156] Performance testing

[0157] To evaluate the performance and effects of the technical solution of the present invention, the sea buckthorn seed oil products obtained in Examples 1-5 and Comparative Examples 1-3 were tested according to conventional methods for total phenolic content (expressed as mg GAE (gallic acid equivalent) / 100g), total flavonoid content (expressed as mg RE (rutin equivalent) / 100g), vitamin E content (expressed as mg / 100g), DPPH free radical scavenging rate (expressed as mg / mL), and ABTS content. + Free radical scavenging rate (expressed in mg / mL), FRAP total antioxidant capacity (expressed in μmol Fe) 2+ The test results of Examples 1-5 and Comparative Examples 1-3 are shown in Tables 1 and 2 below. (The test results are represented by / L).

[0158] Table 1. Performance Test Results of Sea Buckthorn Seed Oil

[0159] Table 2. Comparison of Antioxidant Component Retention Rates (Based on Comparative Example 1)

[0160] As shown in Table 1, the sea buckthorn seed oil treatment method of the present invention (Examples 1-5) yields significantly better results in terms of the content of various antioxidant components and antioxidant activity indicators than Comparative Examples 1-3.

[0161] (1) Regarding the content of antioxidant components:

[0162] The total phenol content of Examples 1-5 was 74.6-92.3 mg GAE / 100g, the total flavonoid content was 53.1-70.5 mg RE / 100g, and the vitamin E content was 221.5-276.4 mg / 100g. The total phenol content of Comparative Example 1 (conventional pressing method) was only 32.5 mg GAE / 100g, the total flavonoid content was 18.6 mg RE / 100g, and the vitamin E content was 98.4 mg / 100g; the total phenol content, total flavonoid content, and vitamin E content of Comparative Example 2 (supercritical CO2 extraction method) were 46.8 mg GAE / 100g, 29.5 mg RE / 100g, and 165.3 mg / 100g, respectively; and the total phenol content, total flavonoid content, and vitamin E content of Comparative Example 3 (using existing technology method) were 58.3 mg GAE / 100g, 39.5 mg RE / 100g, and 175.2 mg / 100g, respectively.

[0163] The total phenol content of Example 1 was 2.66 times that of Comparative Example 1, 1.85 times that of Comparative Example 2, and 1.48 times that of Comparative Example 3, respectively; the total flavonoid content of Example 1 was 3.37 times that of Comparative Example 1, 2.13 times that of Comparative Example 2, and 1.59 times that of Comparative Example 3, respectively; and the vitamin E content of Example 1 was 2.63 times that of Comparative Example 1, 1.56 times that of Comparative Example 2, and 1.47 times that of Comparative Example 3, respectively.

[0164] Example 4 (with added ultrasonic pre-disruption treatment) showed the highest content of various antioxidant components, with a total phenol content of 92.3 mg GAE / 100g, a total flavonoid content of 70.5 mg RE / 100g, and a vitamin E content of 276.4 mg / 100g, indicating that ultrasonic pre-disruption can further promote the release and extraction of active ingredients. Example 5 (with added auxiliary ultrasonic treatment in the enzymatic degumming step) showed a slightly higher content of antioxidant components compared to Example 1, indicating that ultrasonic-assisted enzymatic degumming can further reduce the loss of active ingredients during the refining process.

[0165] (2) Antioxidant activity:

[0166] DPPH scavenging rate IC in Examples 1-5 50 The value ranged from 1.05 to 2.12 mg / mL, ABTS + Clearance IC 50 The value was 8.9~13.8 mg / mL, and the FRAP value was 362~476 μmol Fe. 2+ / L. DPPH scavenging rate IC of Comparative Example 1 50 The value was as high as 6.84 mg / mL, ABTS + Clearance IC 50 The value was as high as 28.6 mg / mL, while the FRAP value was only 124 μmol Fe. 2+ / L; DPPH scavenging rate IC of Comparative Example 2 50 The value was 3.45 mg / mL; the DPPH scavenging rate IC50 of Comparative Example 3 was... 50 The value was 2.96 mg / mL.

[0167] DPPH removal rate IC in Example 1 50 The values ​​were only 19.9% ​​of Comparative Example 1, 39.4% of Comparative Example 2, and 46.0% of Comparative Example 3, indicating that the sea buckthorn seed oil prepared by the method of the present invention has a stronger free radical scavenging ability and significantly improved antioxidant activity.

[0168] (3) Regarding oil yield:

[0169] The oil yields of Examples 1-5 were 21.2%-23.1%, which were significantly higher than those of Comparative Example 1 (14.6%), Comparative Example 2 (16.2%), and Comparative Example 3 (18.2%). This indicates that the compound enzymatic hydrolysis pretreatment effectively disrupted the cell wall structure of sea buckthorn seeds, reduced the cell wall's resistance to oil release, and significantly improved the oil yield.

[0170] (4) Comparative analysis of the retention rate of antioxidant components:

[0171] The comparison of antioxidant retention rates in Table 2 provides a more intuitive view of the protective effect of this invention on the antioxidant components in sea buckthorn seed oil. Compared to Comparative Example 1 (conventional pressing method), the retention rates of total phenols, total flavonoids, and vitamin E in Example 1 increased by 165.8%, 237.1%, and 162.5%, respectively; while the optimized Example 4 showed increases of 184.0%, 279.0%, and 180.9%, respectively. In contrast, the corresponding retention rates of Comparative Example 2 (supercritical CO2 extraction method) increased by only 44.0%, 58.6%, and 68.0%; and the corresponding retention rates of Comparative Example 3 (using existing technology methods) increased by only 79.4%, 112.4%, and 78.0%. It is evident that the multi-dimensional synergistic process of "antioxidant spray protection + low-temperature ultrafine pulverization + ultrasonic-assisted compound enzymatic hydrolysis + low-temperature pressing + enzymatic degumming + two-stage molecular distillation" adopted in this invention has a significantly better retention effect on phenolic, flavonoid, and vitamin E antioxidant components in sea buckthorn seed oil than simple supercritical extraction technology and existing enzymatic-solvent extraction technology, indicating that a significant synergistic effect is generated among the steps.

[0172] (5) Analysis of the synergistic effect of each step:

[0173] Comparing Example 1 and Comparative Example 1, it can be seen that when conventional pressing is used alone, the content and activity of antioxidant components in sea buckthorn seed oil are at a low level. Comparative Example 2, using supercritical CO2 extraction, yielded higher levels of unsaturated fatty acids and natural bioactive components, but the total phenol content was only 46.8 mg GAE / 100g, and the total flavonoid content was only 29.5 mg RE / 100g. The antioxidant activity was also significantly weaker than in the present invention, and the oil yield (16.2%) was still lower than in the examples. Comparative Example 3 used a combination of enzymatic hydrolysis and solvent extraction, but its steps involved ethanol and n-hexane solvents. Although the antioxidant content of the resulting sea buckthorn oil was better than that of Comparative Examples 1 and 2, it was still significantly lower than in the present invention.

[0174] Comparing Example 1 and Comparative Example 3, the present invention achieves antioxidant protection during the pretreatment stage through: (A) Step S2 spraying antioxidant and nitrogen protection; (B) Step S3 low-temperature ultrafine pulverization to avoid damage to active ingredients caused by heat generated during pulverization; (C) Step S4 ultrasonic-assisted compound enzymatic hydrolysis pretreatment to promote cell wall disruption and release of active ingredients; (D) Step S5 low-temperature pressing instead of solvent extraction to avoid insufficient dissolution and extraction of active ingredients by organic solvents and losses caused by subsequent solvent removal; (E) Step S6 enzymatic degumming instead of water washing refining to reduce refining losses; and (F) Step S7 two-stage molecular distillation instead of activated carbon / silica gel adsorption refining to achieve gentle deacidification and purification. The synergistic effect of the above multiple processes maximizes the retention and enhancement of antioxidant content, resulting in a synergistic effect of "1+1>2".

[0175] (6) Analysis of the best implementation method:

[0176] Comparing the various embodiments, Example 4 (with added ultrasonic pre-disruption treatment) showed the best performance in all antioxidant indicators, indicating that ultrasonic pre-disruption treatment can further enhance the cell wall disruption effect and the release of active ingredients. Example 5 (with added auxiliary ultrasonic treatment in the enzymatic degumming step) showed a slight improvement in all indicators compared to Example 1, indicating that the ultrasonic-assisted enzymatic degumming process has the potential for further optimization. Example 1 achieved a good balance between process simplification and product quality, and has high practical application value.

[0177] The sea buckthorn seed oil processing method of the present invention has mild process conditions, controllable operation, strong equipment versatility, no need to use organic solvents, safe and environmentally friendly products, and conforms to the concept of green manufacturing. It can be applied on a large scale to industrial production and has broad market prospects and economic benefits.

[0178] The above embodiments are merely illustrative of the concept and technical solution of the present invention and are not intended to limit the present invention. Any person skilled in the art can modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by those skilled in the art without departing from the spirit and technical concept disclosed in the present invention should still be covered by the claims of the present invention.

[0179] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for processing sea buckthorn seed oil to increase the content of antioxidant components, characterized in that, Includes the following steps: S1. Dry the sea buckthorn seed raw material at low temperature until the moisture content is 6%~10%; S2. The dried sea buckthorn seeds are graded and crushed to a particle size of 0.5mm~2.0mm. An antioxidant solution is sprayed onto the surface of the crushed material, and then the material is subjected to nitrogen protection and allowed to stand. S3. Perform ultra-fine grinding at 0~10℃ until the particle size D90≤75μm; S4. Ultrasonic-assisted compound enzymatic hydrolysis pretreatment: Add the pulverized material from step S3 to deionized water, adjust the pH, add the compound enzyme preparation to carry out ultrasonic-assisted enzymatic hydrolysis reaction, and heat up to inactivate the enzyme after the enzymatic hydrolysis is completed. S5. Low-temperature pressing and separation: The oil is then pressed at low temperature. The resulting mixed oil is centrifuged to obtain crude sea buckthorn seed oil. S6. Enzymatic degumming: The crude sea buckthorn seed oil obtained in step S5 is heated to 45~55℃, and citric acid solution and phospholipase are added in sequence to react. After the reaction, the temperature is raised to inactivate the enzyme, and the gum is removed by centrifugation. S7. Two-stage molecular distillation: The sea buckthorn seed oil after enzymatic degumming in step S6 is subjected to two-stage molecular distillation. The second-stage distillate is collected, which is the sea buckthorn seed oil product with high antioxidant content.

2. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S1, the conditions for low-temperature cyclic drying are: drying temperature of 35℃~45℃ and drying time of 12~24 hours.

3. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S2, the antioxidant solution is prepared by mixing vitamin C and citric acid in a mass ratio of (1~3):(0.5~1.5), and the amount of antioxidant solution used is 0.05%~0.2% of the mass of sea buckthorn seeds. The nitrogen protection and standing treatment time is 10~30 minutes.

4. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S4, the mass ratio of sea buckthorn seeds to deionized water is 1:(3~6); the pH is adjusted to 4.5~5.5 before adding the compound enzyme preparation; the compound enzyme preparation is composed of cellulase, pectinase and acidic protease in a mass ratio of (2~4):(2~3):(1~2), and the amount of the compound enzyme preparation is 0.5%~1.5% of the mass of sea buckthorn seeds.

5. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S4, the conditions for ultrasound-assisted enzymatic hydrolysis are as follows: the enzymatic hydrolysis reaction is carried out for 1.5 to 3 hours under the conditions of ultrasound power of 100 to 300W, ultrasound frequency of 20 to 40kHz, and enzymatic hydrolysis temperature of 45 to 55℃. The enzyme inactivation conditions are as follows: the treatment is carried out at 80 to 90℃ for 10 to 20 minutes.

6. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S5, the pressing process is carried out at 20~35℃ and a pressing pressure of 40~60MPa; Centrifugal separation was performed using a horizontal spiral sedimentation centrifuge with a centrifugal speed of 4000~6000 r / min.

7. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S6, the citric acid solution has a mass fraction of 40%~50%, and the amount of citric acid solution used is 0.5%~1.0% of the mass of crude sea buckthorn seed oil. After adding the citric acid solution, the mixture is stirred and reacted for 20~40 minutes. The phospholipase is phospholipase A1 or phospholipase C, and the amount of enzyme added is 0.01%~0.05% of the mass of crude sea buckthorn seed oil. The enzyme reaction time after adding the phospholipase is 1~3 hours. The enzyme inactivation temperature is 80~90℃, and the enzyme inactivation time is 10~15 minutes.

8. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 7, characterized in that, In step S6, before adding phospholipase, the enzyme is first subjected to ultrasonic-assisted treatment for 15-25 minutes at an ultrasonic power of 100-200W and a frequency of 20-30kHz. After the enzyme reaction is completed, the enzyme is then subjected to ultrasonic-assisted demulsification for 15-25 minutes at an ultrasonic power of 80-120W and a frequency of 20-30kHz before being heated to inactivate the enzyme.

9. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to claim 1, characterized in that, In step S7, the first-stage molecular distillation is carried out under the conditions of vacuum degree 0.1~1.0 Pa, temperature 120~150℃, and scraper rotation speed 200~400 r / min, with a residence time on the evaporation surface of 30~90 seconds; the second-stage molecular distillation is carried out under the conditions of vacuum degree 0.01~0.1 Pa, temperature 160~200℃, and scraper rotation speed 300~500 r / min, with a residence time on the evaporation surface of 60~120 seconds.

10. The method for treating sea buckthorn seed oil to increase the content of antioxidant components according to any one of claims 1 to 9, characterized in that, Between step S2 and step S3, there is also an ultrasonic pre-disruption treatment step: the material after step S2 is added to deionized water at a material-to-liquid ratio of 1:2, and treated for 20 to 40 minutes under ultrasonic power of 300~500W, frequency of 35~45kHz, and temperature of 10~20℃. After centrifugation, the solid phase is taken and entered into step S3.

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