Nutritionally fortified edible vegetable oil and its manufacturing process

Abstract

In the field of edible oil processing, we provide nutritionally fortified edible vegetable oils and their manufacturing processes. [Solution] By optimizing the oil processing process, adjusting the refining process parameters, and retaining an appropriate amount of phospholipids and maximizing the retention of intrinsic nutrients such as vitamin E, we developed a nutritionally fortified oil product by adding appropriate amounts of vitamin D and vitamin A. Animal studies have shown that this product exhibits remarkable effects in reducing inflammatory cytokine levels in the brain and improving the brain's resistance to oxidative stress, thus improving cognitive function and providing scientific evidence for the future functional edible oil market.

Description

Technical Field

[0001] The present invention relates to a nutrient - fortified edible vegetable oil and its manufacturing process, belonging to the field of edible oil processing.

Background Art

[0002] Edible vegetable oils are attracting attention because they contain a large amount of unsaturated fatty acids and rich nutritional components. Taking sunflower seed oil as an example, it is manufactured from sunflower seeds through processes such as pressing and refining. In particular, after undergoing refining processes such as degumming, deacidification, dewaxing, decolorization, and deodorization, the contents of beneficial nutritional concomitants such as vitamin E and phospholipids in sunflower seed oil are significantly reduced. Vitamin E is a natural antioxidant and immunomodulator, and also has physiological functions such as preventing infertility, inhibiting the growth of tumor cells, improving arteriosclerosis, and preventing cardiovascular diseases. In daily life, the main source of vitamin E is edible vegetable oils. Phospholipids are important components of cell membranes and play an effective role in activating brain function and improving intelligence, such as enhancing brain function, improving memory, and protecting neurons. The contents of beneficial components such as vitamin E, phytosterols, and squalene in commercially available vegetable oils are low (Shen, M., Zhao, S., Zhang, F. et al. Characterization and authentication of olive, camellia and other vegetable oils by combination of chromatographic and chemometric techniques: role of fatty acids, tocopherols, sterols and squalene. Eur Food Res Technol 247, 411―426 (2021)). Therefore, the development of a manufacturing method for refined vegetable oils that can maintain a high content of nutritional concomitants in vegetable oils is extremely important.

[0003] Conventional oil and fat manufacturing processes primarily involve preparing crude oil by pressing or leaching, followed by a refining process to obtain the final commercial product oil. However, in actual applications, particularly in the refining process, several significant problems exist, such as the loss of nutrients (vitamin E, plant sterols, squalene, etc.) due to high-temperature operations. Insufficient removal of unstable components during the process reduces oxidative stability, posing challenges to preserving the nutritional content and improving the quality of oils and fats. Commonly used improvement methods include cold pressing, physical refining, and enzymatic hydrolysis. However, these methods also have certain limitations. For example, cold pressing results in low oil yield and high costs, physical refining has stringent equipment requirements, and enzymatic hydrolysis is difficult to implement due to its complex and lengthy process.

[0004] Vitamins A and D are essential fat-soluble vitamins for the human body and play a crucial role in maintaining normal physiological functions. Vitamin A, also known as retinol, contributes to maintaining dim light vision and the health of skin and mucous membranes. Vitamin D is a sterol derivative that promotes calcium and phosphorus metabolism and regulates immune function. However, vitamins A and D are only found in animal products, and vegetable oils contain very little of them. Therefore, developing vegetable oils fortified with vitamins A and D is of great importance. Adding vitamins A and D to vegetable oils can significantly improve their functionality, and they have potential effects, particularly in antioxidant activity and improving nervous system function.

[0005] However, due to the refining process, the amount of retained vitamin E and phospholipids and other nutritional supplements is relatively low. Furthermore, vegetable oils fortified with vitamins A and D are still in the early stages of development, and the effects of the specific amounts of retained and added nutritional functional factors on the prevention and intervention of cognitive impairment have not been sufficiently investigated.

[0006] Therefore, developing methods for producing nutrient-fortified vegetable oils with high retention of nutrient supplements is a crucial research topic in the current field of oleotechnology. Through continuous exploration and innovation, it is hoped that safer, more efficient, and environmentally friendly edible vegetable oil production technologies will be developed to meet people's diverse needs regarding health, nutrition, and taste. [Overview of the Initiative]

[0007] To solve the above technical challenges, the present invention optimizes the oil processing process, adjusts the refining process parameters, and develops a nutritionally fortified oil product by appropriately retaining phospholipids and maximizing the retention of unique nutrients such as vitamin E, while also adding appropriate amounts of vitamin D and vitamin A. Animal studies have demonstrated that this product exhibits remarkable effects in reducing inflammatory factor levels in the brain and improving the brain's resistance to oxidative stress, contributing to improved cognitive function and providing a scientific basis for the future functional edible oil market.

[0008] The first object of the present invention is to provide a method for producing refined vegetable oil in which nutrient supplements are highly retained. This method includes a neutralization step, a dewaxing step, a decolorization step, and a deodorization step. Here, the deodorization process employs a two-tower, two-temperature combined deodorization process, specifically as follows: The decolorized oil is pumped into the plate tower, and the first stage of deodorization is performed at 180-230°C for 60-130 minutes. The oil is then pumped into the packed tower, and the second stage of deodorization is performed at 200-250°C for 5-15 minutes.

[0009] In one embodiment, the method specifically includes (1) a neutralization step, (2) a dewaxing step, (3) a decolorization step, and (4) a deodorizing step. (1) Neutralization process (degumming + deacidification): Crude oil is preheated to 40-80°C, and a phosphoric acid solution is added by pump at a flow rate of 1-50 L / h to carry out the hydration degumming reaction to obtain the product after hydration degumming. The product after hydration and degumming is mixed with an alkaline solution and soft water, stirred, and a deoxidation reaction is carried out. After the reaction is complete, the mixture is centrifuged, washed with soft water, centrifuged again, and dried to obtain neutralized oil. (2) Dewaxing process: A crystallization accelerator is added to the neutralized oil, cooled to 3-13°C, stirred, and filtered to obtain dewaxed oil. (3) Decolorization process: The dewaxed oil is heated to 90-120°C, a decolorizing agent is added and adsorbed, and then filtered to obtain decolorized oil. (4) Deodorization process: The decolorized oil is pumped into a plate column and subjected to a first stage of deodorization at 180-230°C for 60-130 minutes. The oil is then pumped into a packed column and subjected to a second stage of deodorization at 200-250°C for 5-15 minutes to obtain refined vegetable oil.

[0010] In one embodiment, the method for producing crude vegetable oil in step (1) is as follows: The pressing process involves removing impurities, crushing, conditioning, compressing, steaming, pressing, and filtering to obtain crude pressed oil. Key control parameters include a steaming temperature of 90-140°C, a steaming time of 20-30 minutes, and a material temperature of 80-120°C during pressing.

[0011] In one embodiment, the flow rate of crude vegetable oil before preheating in step (1) is 18 to 24 T / h, preferably 21 T / h.

[0012] In one embodiment, the preheating temperature of the crude oil in step (1) is 55°C.

[0013] In one embodiment, the concentration of the phosphoric acid solution in step (1) is 75% to 86% (w / w), preferably 80% (w / w).

[0014] In one embodiment, the flow rate of the phosphoric acid solution pumped in step (1) is 0.5 L / h.

[0015] In one embodiment, the hydration degumming reaction in step (1) is carried out at 40-80°C for 20-60 minutes.

[0016] In one embodiment, the flow rate of the alkaline solution in step (1) is 3 to 200 L / h, preferably 104 L / h.

[0017] In one embodiment, the alkaline solution in step (1) is a sodium hydroxide solution with a concentration of 4% to 20% (w / w), preferably 9% (w / w).

[0018] In one embodiment, the stirring speed in step (1) is 20 to 60 Hz, preferably 40 Hz.

[0019] In one embodiment, the deoxidation reaction in step (1) is carried out at 65-90°C for 20-35 minutes.

[0020] In one embodiment, a combination of an electromagnetic pump, an electromagnetic proportional valve, and an electromagnetic flow meter is used for the phosphoric acid solution and alkaline solution in step (1). These components feed back signals to a PLC (Programmable Logic Controller) according to the flow rate of crude oil and the amount of free fatty acids, calculate the theoretical amount to be added, adjust the opening of the proportional control valve, measure and compare the flow rate with the electromagnetic flow meter, and then feed back the results to adjust the opening of the proportional valve in real time, thereby optimizing the precise amount of the medium to be added, such as the phosphoric acid solution and alkaline solution.

[0021] In one embodiment, the crystallization accelerator in step (2) is one or two of diatomaceous earth and perlite.

[0022] In one embodiment, the amount of crystallization accelerator added in step (2) is 0.1% to 2.0% (w / w), preferably 0.62% (w / w). The amount added is relative to the mass of the neutralized oil.

[0023] In one embodiment, cooling in step (2) is performed down to 6.8°C.

[0024] In one embodiment, the stirring time in step (2) is 20 to 30 hours, preferably, the stirring time is 23.8 hours.

[0025] In one embodiment, the heating temperature in step (3) is 105°C.

[0026] In one embodiment, the decolorizing agent in step (3) is one or more of attapulgite clay, activated clay, and activated carbon.

[0027] In one embodiment, the addition amount of the decolorizing agent in step (3) is 0.05% to 4% (w / w), preferably, the addition amount of the decolorizing agent is 1.0% (w / w). The addition amount is relative to the dewaxed oil.

[0028] In one embodiment, the adsorption time in step (3) is 20 to 60 minutes, preferably, the adsorption time is 30 minutes.

[0029] In one embodiment, the deodorization process in step (4) is as follows. Pump the decolorized oil into a plate column at 200°C, and use stripping steam at 0.9 bar to perform the first-stage deodorization for 90 minutes. Then pump it into a packed column at 230°C and use stripping steam at 0.9 bar to perform the second-stage deodorization for 10 minutes to obtain refined vegetable oil.

[0030] In one embodiment, the nutrient accompaniments are one or more of vitamin E, phospholipids, phytosterols, and squalene.

[0031] In one embodiment, the highly retained nutrient accompaniments are as follows. The retention amount of vitamin E in the refined vegetable oil is 200 to 800 mg / kg, the retention amount of phospholipids is 0 to 200 mg / kg (but not 0), the retention amount of phytosterols is 3000 to 10000 mg / kg, and the retention amount of squalene is 50 to 400 mg / kg.

[0032] In one embodiment, the vegetable oil includes, but is not limited to, sunflower seed oil, corn oil, and rapeseed oil. Preferably, the vegetable oil is sunflower seed oil.

[0033] A second object of the present invention is to provide a refined vegetable oil with high retention of nutrient supplements, produced by the method of the present invention.

[0034] A third object of the present invention is to provide a method for improving the retention rate of phospholipids in refined vegetable oil. This method comprises the following steps: (1) Neutralization process (degumming + deacidification): Crude oil is preheated to 40-80°C, and a phosphoric acid solution is added by pump at a flow rate of 1-50 L / h to carry out the hydration degumming reaction to obtain the product after hydration degumming. The product after hydration and degumming is mixed with an alkaline solution and soft water, stirred, and a deoxidation reaction is carried out. After the reaction is complete, the mixture is centrifuged, washed with soft water, centrifuged again, and dried to obtain neutralized oil. (2) Dewaxing process: A crystallization accelerator is added to the neutralized oil, cooled to 3-13°C, stirred, and filtered to obtain dewaxed oil. (3) Decolorization process: The dewaxed oil is heated to 90-120°C, a decolorizing agent is added and adsorbed, and then filtered to obtain decolorized oil. (4) Deodorization process: The decolorized oil is pumped into a plate column and subjected to a first stage of deodorization at 180-230°C for 60-130 minutes. The oil is then pumped into a packed column and subjected to a second stage of deodorization at 200-250°C for 5-15 minutes to obtain refined vegetable oil.

[0035] In one embodiment, improving the retention rate of phospholipids in refined vegetable oil is mainly based on adjusting the parameters of the neutralization step in step (1).

[0036] A fourth object of the present invention is to provide a method for improving the retention rate of vitamin E, plant sterols, and squalene in refined vegetable oil. This method comprises the following steps: (1) Neutralization process (degumming + deacidification): Crude oil is preheated to 40-80°C, and a phosphoric acid solution is added by pump at a flow rate of 1-50 L / h to carry out the hydration degumming reaction to obtain the product after hydration degumming. The product after hydration and degumming is mixed with an alkaline solution and soft water, stirred, and a deoxidation reaction is carried out. After the reaction is complete, the mixture is centrifuged, washed with soft water, centrifuged again, and dried to obtain neutralized oil. (2) Dewaxing process: A crystallization accelerator is added to the neutralized oil, cooled to 3-13°C, stirred, and filtered to obtain dewaxed oil. (3) Decolorization process: The dewaxed oil is heated to 90-120°C, a decolorizing agent is added and adsorbed, and then filtered to obtain decolorized oil. (4) Deodorization process: The decolorized oil is pumped into a plate column and subjected to a first stage of deodorization at 180-230°C for 60-130 minutes. The oil is then pumped into a packed column and subjected to a second stage of deodorization at 200-250°C for 5-15 minutes to obtain refined vegetable oil.

[0037] In one embodiment, improving the retention rate of vitamin E, plant sterols, and squalene in refined vegetable oil is mainly based on adjusting the parameters of the deodorization step (4).

[0038] A fifth object of the present invention is to provide a method for producing nutrient-fortified refined vegetable oil. This method comprises the following steps: Based on refined vegetable oil, nutritional substances are added for nutritional enhancement. These nutrients are one or two of the following: vitamin A and vitamin D.

[0039] In one embodiment, the method specifically includes: The process includes adding vitamin A and vitamin D to refined vegetable oil and stirring to obtain a nutritionally fortified refined vegetable oil, where the amount of vitamin A added is 4000-8000 μg / kg and the amount of vitamin D added is 50-100 μg / kg.

[0040] In one embodiment, the amount of vitamin A added is 6200 mg / kg, and the amount of vitamin D added is 75 μg / kg.

[0041] In one embodiment, the vitamin E content in the fortified refined vegetable oil is 200-800 mg / kg, the phospholipid content is 0-200 mg / kg, the plant sterol retention is 3000-10000 mg / kg, the squalene retention is 50-400 mg / kg, the vitamin A content is 4000-8000 μg / kg, and the vitamin D content is 50-100 μg / kg.

[0042] A sixth object of the present invention is to provide a nutritionally fortified refined vegetable oil produced by the method of the present invention.

[0043] A seventh object of the present invention is to provide applications of the above-mentioned fortified refined vegetable oil in the manufacture of cognitive function improving, antioxidant, or anti-inflammatory products.

[0044] In one embodiment, the product includes, but is not limited to, pharmaceuticals, foods, health supplements, animal feed, feed additives, and food additives.

[0045] Beneficial effects: (1) While phospholipids are highly nutritious, their unique hydrophilic and lipophilic properties mean that excessive amounts in vegetable oils can lead to darkening of the oil's color and deterioration of its flavor, negatively impacting the shelf life and stability of oil products. Currently, the industry commonly uses triple metering pumps as the drive system for acid and alkali addition. Although linked to the crude oil flow rate and free fatty acids, in many cases, it is still necessary to manually adjust the valve stroke and adjust the flow rate using frequency conversion. When the crude oil flow rate fluctuates, the amount of acid and alkali added is often excessive or insufficient, and analysis of the triple pump's flow curve reveals an error of approximately 57%. This error is a cause of the need for excessive acid and alkali addition, hindering subsequent yield improvements.

[0046] Based on the principles of precise and appropriate processing, this invention employs an electromagnetic pump with an engineering plastic pump body, combines an electromagnetic flow meter and a proportional control valve, and designs the additive piping and high-flow return piping according to the flow rate ratio (a schematic diagram of the process control is shown in Figure 2). By feeding back signals to a PLC (Programmable Logic Controller) according to the crude oil flow rate and the amount of free fatty acids, the theoretical amount to be added is calculated, the opening degree of the proportional control valve is adjusted, the flow rate is measured and compared with that using an electromagnetic flow meter, and the opening degree of the proportional valve is controlled in real time by feeding back the results again, thereby optimizing the design of the additive piping for acid solution, alkaline solution, dilution water, and rinse water, as well as the intelligent additive ratio control system, and reducing the error in the amount of added to 10%.

[0047] In summary, by employing a more precise control device in the neutralization process and precisely controlling the phosphate addition ratio in the degumming process according to the phospholipid content of the crude oil, the phospholipid content can be better maintained and controlled.

[0048] (2) The structure of a plate-type deodorizing tower consists of multiple layers of plates installed at regular intervals within a cylindrical shell. Liquid flows from top to bottom through each layer of plates due to gravity and is then discharged from the bottom of the tower. A fluidized liquid layer of a constant thickness is maintained on each layer of plates. Gas rises upward from the bottom of the tower through the liquid layer on each plate due to the pressure difference and is discharged from the top of the tower. Advantages of plate-type deodorizing towers: Plate-type deodorizing towers have a simple structure and are easy to operate. Because of the long residence time within the deodorizing tower, heat-sensitive pigments in oils and fats can be completely decomposed, resulting in an excellent decolorization effect. However, the long residence time generates harmful substances such as trans fatty acids, and nutrients such as vitamin E, plant sterols, and squalene are distilled out with the vapor.

[0049] The structure of a packed deodorizing tower consists of a cylindrical shell filled with packing material to a certain height. Liquid is uniformly distributed to the top of the packing layer by a spray device at the top of the tower, flows from top to bottom along the surface of the packing layer by gravity, and is discharged from the bottom of the tower. Gas passes through the gaps in the packing layer due to the pressure difference and flows from one end of the tower to the other. Advantages of packed deodorizing towers: Because packed deodorizing towers do not have an oil layer and have a large specific surface area, the oil film that is formed is relatively thin, the mass transfer effect per unit time is good, and it can be applied to physical deoxidation. Disadvantages of packed deodorizing towers: The deodorizing effect is not as pronounced as that of plate deodorizing towers, and because the oil film is very thin, the time that the oil to be deodorized remains in the tower is short.

[0050] This invention employs a method that combines a packed column and a plate column, combining the two columns and modifying conventional process piping to achieve stepped temperature control. This allows for flexible and precise adjustment of the temperatures of the two columns according to the quality of the crude oil and the oil in the refining process. Based on the characteristic of the plate column having a long residence time, the deodorization temperature of the plate column is moderately lowered in the first stage. Based on the characteristic of the packed column having a short residence time and high deoxidation capacity, deodorization of the packed column in the second stage employs rapid, short-duration high-temperature heating. This avoids problems such as high residence temperature and long duration of the deodorized oil in the plate-type packed column, resulting in significant loss of nutrient supplements such as vitamin E, plant sterols, and squalene, and thus preserves nutrient supplements in the deodorized oil well.

[0051] (3) The present invention employs a method that uses both packed towers and plate towers, rationally optimizes the piping route, and innovatively utilizes an efficient heater system to achieve temperature control of the two tower bodies. See Figure 3 for details. This ensures deodorization effect, significantly improves energy utilization efficiency, and achieves energy savings and emission reductions.

[0052] (4) The present invention aims to study the synergistic effects of nutrients such as vitamins A, E, D and phospholipids on antioxidant activity and memory improvement by constructing a scopolamine mouse animal model, for example, by fortifying sunflower seed oil with vitamin A or vitamin D in accordance with regulations such as the "National Dietary Guidelines of China" while appropriately retaining nutritional supplements in oils and fats, and by verifying the effective doses in which the above nutrients exert their effects, thereby providing a scientific basis for the claimed blending ratios. [Brief explanation of the drawing]

[0053] [Figure 1] This is a schematic diagram of a conventional acid / alkali addition metering pump used in a neutralization process. Here, 1 is the pump head, 2 is the mechanical device, 3 is the manual stroke adjustment knob, and 4 is the motor. [Figure 2] This is a process flow diagram after the modification of the precise acid / alkali addition system in the neutralization process. Here, 1 is the crude oil transfer tank, 2 is the phosphoric acid storage tank, 3 is the liquid alkali storage tank, 4 is the acid reaction tank, 5 is the alkali reaction tank, 6 is the centrifuge 1, 7 is the centrifuge 2, 8 is the water washing and stirring tank, 9 is the dryer, 10 is the mixer 1, 11 is the mixer 2, 12 is the mixer 3, 13 is the electromagnetic pump, 14 is the electromagnetic proportional valve, 15 is the electromagnetic flow meter, 16 is the electromagnetic pump, 17 is the electromagnetic proportional valve, and 18 is the electromagnetic flow meter. [Figure 3] This is a flow diagram of the two-tower, two-temperature combined deodorization process in the deodorization stage. [Modes for carrying out the invention]

[0054] The following interpretations and explanations will be made using specific examples to further describe the nutrient-enhanced sunflower oil with cognitive-improving function described in the present invention and its manufacturing process, but these interpretations and explanations will not unduly limit the technical solutions of the present invention.

[0055] Drugs and reagents related to the following examples: Scopolamine hydrobromide (98% purity) was purchased from Shanghai Macklin Biochemical Technology Co., Ltd.

[0056] The IL-6 and malondialdehyde (MDA) kits were purchased from Xiamen Huijia Biotechnology Co., Ltd.

[0057] All other common reagents are domestically produced and of analytical purity grade.

[0058] Rivastigmine was purchased from MedChemExpress and its CAS code is 123441-03-2.

[0059] The diatomaceous earth used as a crystallization accelerator was purchased from Linjiang Dayuan Diatomaceous Earth New Material Ecological Environmental Protection Technology Co., Ltd. The activated clay used as an adsorbent was purchased from Huangshan Baiyae Activated Clay Co., Ltd.

[0060] The experimental animals used in the following examples: Male ICR mice (5 weeks old, 21±2g, specific pathogen-free (SPF)) were purchased from Shanghai SLAC Laboratory Animal Co., Ltd. The animals used and the animal experiments conducted complied with the guidelines of the Declaration of Helsinki and were approved by the Laboratory Animal Management and Animal Welfare Ethics Committee of Jiangnan University (JN.No20230615i1000906

[0304] ).

[0061] Behavioral testing methods for the following examples: (1) Open field (OFT) The open field test is primarily used to evaluate the spontaneous activity and exploratory behavior of mice. The open field apparatus consists of an experimental box measuring 40 cm long, 40 cm wide, and 50 cm high, with an infrared camera at the top to track the mouse's path. The test area at the bottom is divided into 16 squares of 10 cm x 10 cm, with the four central squares designated as the central zone. Each mouse is gently lowered from the center point of the open field, and the experimenter immediately leaves the apparatus, allowing the mouse to move freely for 8 minutes. Video recording software is used to record indicators such as the mouse's speed, distance traveled, and time spent in the central zone. After each experiment, any remaining hair, urine, or feces from the mouse are cleaned up, and the experimental box is wiped down with 75% alcohol to remove any odor. After the alcohol is evaporated using paper towels and a hairdryer, the next mouse is tested.

[0062] (2) Novel object recognition (NORT) Novel Object Recognition (NOR) is a learning and memory assessment method that utilizes the instinctive characteristic of rodents to enjoy exploring new things. The experimental setup is the same as that of an open-field test. The experimental period is mainly divided into an adaptation period, a habituation period, and a test period, with 24-hour intervals between each period. During the adaptation period, mice are placed in an experimental box and allowed to freely explore the environment for 8 minutes to reduce the mice's sense of novelty and fear of the unfamiliar environment. During the habituation period, two identical blocks are placed on the opposite side of the experimental setup. The blocks are placed 5 cm away from the corners of the walls on both sides. Mice are released with their backs to the box from the same center point as the previous day, allowing them to freely explore the blocks and environment inside the box for 8 minutes. After 24 hours, one of the blocks is replaced with a new block of a different shape, and the mouse is allowed to freely explore again for 8 minutes using the same method as the habituation period. After each experiment, the experimental box is wiped down with 75% alcohol to remove any odor.

[0063] During the experiment, an infrared camera will track and record the time (T) that the mouse spends exploring new and known objects. The evaluation metric will be calculated using the Discriminant Index (DI). DI = (T new objects - T known objects) / T total exploration. To prevent the mouse from moving or climbing on the blocks, blocks that are too small should not be selected. The blocks may be secured with double-sided tape.

[0064] (3) Y-Maze Test (YMT) The Y-Maze Test (YMT) is primarily used to assess working or reference memory in rodents. The experimental setup consists of three horizontal arms, each measuring 120°, 70cm long, and 15cm high. The horizontal arms are made of opaque polyethylene plastic. During the experiment, the three arms are arbitrarily designated as a, b, and c. After the experiment begins, a mouse is placed in one of the arms, and the order and number of times the mouse enters each arm are recorded using an infrared camera. Consecutive entries and exits into different arms are defined as one alternation (e.g., abc, bac, cba, but not aca).

[0065] The evaluation metric for the Y-maze test is spontaneous alternation = number of alternations / (total number of times entering the arm - 2), and each test session lasts 5 minutes. After each experiment, the experimental box is wiped down with 75% alcohol to remove any lingering odors.

[0066] (4) Morris Water Maze Test (MWM) The Morris water maze test is a classic method for evaluating the spatial memory of rodents. The apparatus consists of a circular pool with a diameter of 120 cm and a height of 60 cm. The pool is divided into four quadrants, indicating the directions of east, south, west, and north. Adjustable light sources are installed on both sides to control the brightness of the water surface, and a camera is mounted above the pool to record experimental data. During the experiment, 40 cm of water is poured into the pool, colored black with ink, and the water temperature is maintained at 21 ± 1°C.

[0067] The experiment is divided into two stages: place navigation and spatial probe. After each swimming test, the mouse is dried with a clean towel or hairdryer and kept warm. In the training stage, a platform is placed 1 cm below the water surface and fixed in a certain quadrant. At the start of each test, the mouse is placed on the side facing the pool wall, introduced into the water from a certain quadrant, and its swimming path is tracked with an infrared camera. The test time limit is 60 seconds. The time it takes for the mouse to find the platform is recorded as escape latency. Once the mouse finds the platform, it can stay on it for 5 seconds. If the mouse does not find the platform within the time limit, the escape latency is recorded as 60 seconds, and the mouse is guided to stay on the platform for 15 seconds. During the guidance process, the mouse should not be placed directly on the platform, but rather guided using a guidance stick to guide its swimming direction, allowing it to learn how to sense and climb the platform and memorize its location.

[0068] Each mouse undergoes four training tests per day, entering the water from a different quadrant each time. A minimum of 30 minutes is allowed between two consecutive tests. Training continues for five days. On the sixth day, a spatial probe test is conducted. The platform is removed from the pool. The mouse enters the water from the pool wall in the quadrant diagonally opposite the platform's original position. The test time is limited to 60 seconds. Parameters such as the mouse's swimming speed, latency, swimming path, and the number of times it passes the platform's original position (number of platform crossings) are recorded. The mouse is not given scopolamine during the spatial probe phase.

[0069] Method for measuring nutrients in the following example: (1) Phospholipids Measurements were performed in accordance with GB / T 5537-2008, "Inspection of Grains and Oils: Measurement of Phospholipid Content."

[0070] (2) Vitamin E Measurements were performed in accordance with GB 5009.82-2016 "National Standard for Food Safety in China: Measurement of Vitamins A, D, and E in Food."

[0071] (3) Plant sterols Measurements were performed in accordance with GB / T 25223-2010, "Measurement of Sterol Composition and Total Sterol Amount of Animal and Vegetable Oils and Fats."

[0072] (4) Squalene Measurements were performed in accordance with LS / T 6120-2017, "Corn and Oil Testing: Measurement of Squalene in Vegetable Oils."

[0073] Statistical methods relating to the following examples: All experimental results were statistically analyzed using SPSS (Statistical Product and Service Solutions) statistical processing software. Differences between groups were evaluated using t-tests, and a statistically significant difference was considered to exist if P < 0.05.

[0074] The t-test is a type of probability and statistics method, primarily used for normally distributed data with small sample sizes and unknown population standard deviations. The p-value represents the probability of obtaining the current sample if the original hypothesis is true (P<0.05 means that the probability of the latter data sequence yielding the same results as the former data sequence is less than 0.05, i.e., the correlation between the two is less than 0.05, which is statistically significant). In the following, "t" and "P" will be considered to have the same meaning.

[0075] Method for producing crude oil in the following examples: (1) Production of crude sunflower seed oil: The production of crude sunflower seed oil employs a pressing process. Sunflower seeds are hulled, washed, crushed, tempered, flattened, steamed, pressed, and filtered to obtain the pressed crude oil. Key control parameters include a steaming temperature of 130°C for 25 minutes and a material temperature of 90°C during pressing.

[0076] (2) Production of crude corn oil: Crude corn oil is produced using a pressing process. Corn germ is washed, crushed, seasoned, flattened, steamed, pressed, and filtered to obtain pressed crude oil. Key control parameters include a steaming temperature of 100°C for 25 minutes and a material temperature of 90°C during pressing.

[0077] (3) Production of crude rapeseed oil: Crude rapeseed oil is produced using a pressing process. The rapeseed is washed, crushed, seasoned, flattened, steamed, pressed, and filtered to obtain the crude pressed oil. Key control parameters include a steaming temperature of 120°C for 25 minutes and a material temperature of 100°C during pressing.

[0078] Apparatus relating to the following example: (1) Metering pump (conventional control piping before modification): Figure 1 shows a schematic diagram of a conventional additive metering pump for acid / alkali in a neutralization process. A conventional additive metering pump consists of a pump head (1), a mechanical device (2), a manual stroke adjustment knob (3), and a motor (4). Driven by the motor (4), the pump head (1) operates in conjunction with the mechanical device (2), transporting the medium so that it flows in and out of the pump head (1). The stroke (3) is adjusted with the manual stroke adjustment knob to achieve flow rate control. Depending on the flow rate requirements, two or three units can be combined in series.

[0079] (2) Intelligent control system (modified control piping used in the present invention): Figure 2 shows the process flowchart after modification of the acid / alkali precision addition system in the neutralization process. 1 is the crude oil transfer tank, 2 is the phosphoric acid storage tank, 3 is the liquid alkali storage tank, 4 is the acid reaction tank, 5 is the alkali reaction tank, 6 is the centrifuge 1, 7 is the centrifuge 2, 8 is the water washing and stirring tank, 9 is the dryer, 10 is the mixer 1, 11 is the mixer 2, 12 is the mixer 3, 13 is the electromagnetic pump, 14 is the electromagnetic proportional valve, 15 is the electromagnetic flow meter, 16 is the electromagnetic pump, 17 is the electromagnetic proportional valve, and 18 is the electromagnetic flow meter.

[0080] After passing through the crude oil transfer tank (1), the crude oil is mixed with a phosphoric acid solution in mixer 1 (10) and enters the acid reaction tank (4) where it undergoes a thorough hydration and degumming reaction. Next, it is mixed with an alkaline solution in mixer 2 (11) and enters the alkaline reaction tank (5), where it reacts thoroughly and flows into centrifuge 1 (6) where it is centrifuged. Soft water is added and mixed in mixer 3 (12), then enters the washing tank (8) where it is washed with water. After that, it enters centrifuge 2 (7), is centrifuged, and is dried in the dryer (9) to obtain neutralized oil.

[0081] When adding phosphoric acid solution or alkaline solution, instead of using a conventional metering pump, a combination of electromagnetic pumps (13, 16), electromagnetic proportional valves (14, 17), and electromagnetic flow meters (15, 18) is used. The signal is fed back to the PLC (Programmable Logic Controller) according to the crude oil flow rate and the amount of free fatty acids, the theoretical amount to be added is calculated, the opening of the proportional control valve is adjusted, the actual flow rate is measured with the electromagnetic flow meter and compared, and the opening of the proportional valve is adjusted again with feedback in real time to optimize the precise amount of the medium to be added, such as phosphoric acid solution or alkaline solution.

[0082] (3) Two-tower, two-temperature combined deodorization device: Figure 3 shows the flowchart for the two-tower, two-temperature combined deodorization process. The decolorized oil is first heated with the deodorized oil in heat exchanger 3, then enters heat exchanger 2 for secondary heating. Afterward, it enters the plate column for the first stage of deodorization, before entering heat exchanger 1 (heater) for further heating. This prepares it for the second stage of deodorization in the packed column. The deodorization process is carried out under negative pressure (dotted lines indicate negative pressure piping), and the deodorized distillate is collected in a collector. Throughout the entire process, the residual heat of the deodorized oil is effectively utilized to provide the first stage deodorization temperature, and heating / heat exchange is performed using a heating furnace to achieve the instantaneous high temperature required for the second stage packed column.

[0083] In the examples, unless otherwise specified, the solvent used in the solution shall be water. Unless otherwise specified, the reaction temperature shall be room temperature (25°C).

[0084] For phosphoric acid and NaOH solutions, a combination of an electromagnetic pump, electromagnetic proportional valve, and electromagnetic flow meter is used. Signals are fed back to a PLC (Programmable Logic Controller) according to the crude oil flow rate and the amount of free fatty acids. The theoretical amount to be added is calculated, the opening of the proportional control valve is adjusted, the actual flow rate is measured with the electromagnetic flow meter and compared, and the opening of the proportional valve is adjusted again in real time using this feedback mechanism to optimize the precise amount of the medium to be added, such as phosphoric acid or NaOH solution.

[0085] The packing material in the packed column is specifically low-pressure-loss stainless steel structural packing from Sulzer GmbH of Switzerland.

[0086] <Example 1> A method for producing refined sunflower seed oil with high retention of nutrient supplements, comprising: (1) a neutralization step; (2) a dewaxing step; (3) a decolorization step; and (4) a deodorization step. (1) Neutralization process (degumming + deoxidation): Modified control piping was used. For the crude sunflower seed oil in the crude oil transfer tank, the flow rate of the crude sunflower seed oil was maintained at 21 T / h, the temperature was preheated to 55°C, and an 80% (w / w) phosphoric acid solution was accurately pumped in at a flow rate of 0.5 L / h. After mixing in mixer 1, the mixture was introduced into a reaction tank at 55°C, and the hydration degumming reaction was carried out for 30 minutes to obtain the product after hydration degumming. After the reaction was complete, a 9% (w / w) NaOH solution was pumped into the product after hydration and degumming at a flow rate of 104 L / h, and mixed with soft water at a flow rate of 750 L / h in mixer 2. After mixing, the mixture was introduced into an alkaline reaction tank, and a deoxidation reaction was carried out for 30 minutes at a stirring speed of 42 Hz and 70°C. After the reaction was complete, the mixture was centrifuged with a back pressure of 3.0 bar using centrifuge 1 for soap removal and placed in mixer 3. Soft water was added to mixer 3 at a flow rate of 772 L / h and mixed, then introduced into a washing tank for washing, centrifuged with a back pressure of 3.0 bar using centrifuge 2, and then vacuum dried in a drying oven at 105°C for 20 minutes to obtain neutralized oil. (2) Dewaxing process: 0.62% (w / w) of diatomaceous earth was added to the neutralized oil, cooled to 6.8°C, stirred for 23.8 hours, and filtered to obtain dewaxed oil. (3) Decolorization process: The dewaxed oil was heated to 105°C under vacuum, and 0.5% (w / w) of decolorized activated clay was added using 0.9 bar of stripping steam. Adsorption was carried out for approximately 30 minutes, and the oil was filtered to obtain the decolorized oil. (4) Deodorization process (two-tower, two-temperature combined deodorization process): The decolorized oil was heated to 190°C in heat exchanger 3, introduced into heat exchanger 2, and subjected to secondary heating to 210°C. Subsequently, the decolorized oil was pumped into the plate tower, and the first stage of deodorization was performed for 90 minutes using 0.9 bar of stripping steam. Subsequently, the oil was introduced into heat exchanger 1 (heater) and heated to 230°C. The oil was then pumped into the packed column, and the second stage of deodorization was performed for 10 minutes using 0.9 bar of stripping steam. The vacuum system was maintained at 2 mbar throughout the entire deodorization process to obtain refined sunflower seed oil.

[0087] The refined sunflower seed oil obtained contained aflatoxin B1 ≤ 10 μg / kg and benzo[a]pyrene ≤ 10 μg / kg, and all other major physicochemical and food safety indicators met the requirements of the Chinese national standard GB / T 10464 Sunflower Seed Oil.

[0088] <Comparative Example 1> A conventional method for producing refined sunflower seed oil, comprising (1) a neutralization step, (2) a dewaxing step, (3) a decolorization step, and (4) a deodorization step. (1) Neutralization process (degumming + deoxidation): The conventional control piping from before the modification was used. Under motor drive, the flow rate of crude sunflower seed oil in the crude oil transfer tank was set to 21 T / h, and the temperature was preheated to 55°C. The pump head and mechanical device were used to adjust the stroke using a manual stroke adjustment knob, maintaining a flow rate of 0.5 L / h for pumping in an 80% (w / w) phosphoric acid solution. The mixture was then mixed in mixer 1 and introduced into the acid reaction tank, where a hydration degumming reaction was carried out at 55°C for 30 minutes. After the reaction was complete, a 9% (w / w) NaOH solution was pumped into mixer 2 at a flow rate of 104 L / h and mixed with soft water at a flow rate of 750 L / h in mixer 2. The flow rate control method was the same as for the phosphoric acid solution in (1). After mixing, the mixture was introduced into an alkaline reaction tank and a deoxidation reaction was carried out for 30 minutes at a stirring speed of 42 Hz and 70°C. After the reaction was complete, the mixture was centrifuged with a back pressure of 3.0 bar using desommé centrifuge 1 and placed in mixer 3. Soft water was added to mixer 3 at a flow rate of 772 L / h and mixed, then introduced into a washing tank and washed with water. After centrifuging with a back pressure of 3.0 bar using centrifuge 2, the mixture was vacuum dried in a dryer at 105°C for 20 minutes to obtain neutralized oil. (2) Dewaxing process: The procedure was carried out in the same manner as in step (2) of Example 1. (3) Decolorization process: The procedure was carried out in the same manner as in step (2) of Example 1. (4) Deodorization process: The decolorized oil was introduced into the deodorization process. In the deodorization process, the same temperature was used for both the packed column and the plate column, controlled to approximately 230°C, with a deodorization time of 92 minutes and a stripping steam of 0.9 bar. The vacuum system was maintained at 2 mbar throughout the entire deodorization process to obtain conventionally refined sunflower seed oil.

[0089] Table 1 shows the test results for the major nutrients (phospholipids, vitamin E, plant sterols, and squalene) in the refined oils prepared in Example 1 and Comparative Example 1.

[0090] JPEG2026123773000002.jpg65170

[0091] Comparative Example 2 The specific embodiment is the same as in Example 1, the only difference being that step (5) was prepared as follows. (5) Deodorization process: The decolorized oil was heated to 200°C in heat exchanger 3, introduced into heat exchanger 2, and subjected to secondary heating to 210°C. Then, the decolorized oil was pumped into a plate tower, and the first stage of deodorization was performed for 90 minutes using 0.9 bar of stripping steam. Next, it was introduced into heat exchanger 1 (heater) and heated to 260°C. The oil was pumped into a packed tower, and the second stage of deodorization was performed for 10 minutes using 0.9 bar of stripping steam. The vacuum system was maintained at 2 mbar throughout the entire deodorization process to obtain refined sunflower seed oil.

[0092] Comparative Example 3 The specific embodiment is the same as in Example 1, the only difference being that step (5) was prepared as follows. (5) Deodorization process: The decolorized oil was heated to 210°C in heat exchanger 3, introduced into heat exchanger 2, and subjected to secondary heating to 220°C. Subsequently, the decolorized oil was pumped into the plate tower, and the first stage of deodorization was performed for 90 minutes using 0.9 bar of stripping steam. Next, the oil was introduced into heat exchanger 1 (heater) and heated to 260°C. The oil was then pumped into the packed column, and the second stage of deodorization was performed for 10 minutes using 0.9 bar of stripping steam. The vacuum system was maintained at 2 mbar throughout the entire deodorization process to obtain refined sunflower seed oil.

[0093] Comparative Example 4 The specific embodiment is the same as in Example 1, the only difference being that step (5) was prepared as follows. (5) Deodorization process: The decolorized oil was heated to 220°C in heat exchanger 3, introduced into heat exchanger 2, and subjected to secondary heating to 230°C. Subsequently, the decolorized oil was pumped into the plate tower, and the first stage of deodorization was performed for 100 minutes using 0.9 bar of stripping steam. Next, the oil was introduced into heat exchanger 1 (heater) and heated to 260°C. The oil was pumped into the packed column, and a second stage of deodorization was performed for 10 minutes using 0.9 bar of stripping steam. The vacuum system was maintained at 2 mbar throughout the entire deodorization process to obtain refined sunflower seed oil.

[0094] Performance tests were conducted on the refined sunflower seed oil obtained, and the test results are as follows. JPEG2026123773000003.jpg45170JPEG2026123773000004.jpg48170

[0095] Example 2 The specific embodiment is the same as in Example 1, the only difference being that crude sunflower seed oil was prepared into crude corn oil.

[0096] The refined corn oil obtained had aflatoxin B1 ≤ 20 μg / kg and benzo[a]pyrene ≤ 10 μg / kg, and other major physicochemical indicators and food safety requirements met the requirements of the Chinese national standard "GB / T 19111 Corn Oil".

[0097] Comparative Example 5 The specific embodiment is the same as in Comparative Example 1, the only difference being that crude sunflower seed oil was prepared into crude corn oil.

[0098] Table 4 shows the test results for the main nutrients (phospholipids, vitamin E, plant sterols, and squalene) in the refined oils prepared in Example 2 and Comparative Example 5.

[0099] JPEG2026123773000005.jpg48170

[0100] Example 3 The specific embodiment is the same as in Example 1, the only difference being that crude sunflower seed oil was prepared into crude rapeseed oil.

[0101] The refined rapeseed oil obtained had aflatoxin B1 ≤ 10 μg / kg and benzo[a]pyrene ≤ 10 μg / kg, and other major physicochemical indicators and food safety requirements met the requirements of the Chinese national standard "GB / T 1536 Rapeseed Oil".

[0102] Comparative Example 6 The specific embodiment is the same as in Comparative Example 1, the only difference being that crude sunflower seed oil was prepared into crude rapeseed oil.

[0103] Table 5 shows the test results for the main nutrients (phospholipids, vitamin E, plant sterols, and squalene) in the refined oils prepared in Example 3 and Comparative Example 6.

[0104] JPEG2026123773000006.jpg48170

[0105] Example 4 A conventional method for producing refined sunflower seed oil, comprising (1) a neutralization step, (2) a dewaxing step, (3) a decolorization step, and (4) a deodorization step. (1) Neutralization process (degumming + deoxidation): The conventional control piping from before the modification was used. Under motor drive, the flow rate of crude sunflower seed oil in the crude oil transfer tank was set to 21 T / h, and the temperature was preheated to 55°C. Using the pump head and mechanical device, the stroke was adjusted using a manual stroke adjustment knob to maintain a flow rate of 0.5 L / h for pumping in an 80% (w / w) phosphoric acid solution. This solution was mixed in mixer 1 and introduced into the acid reaction tank, where a hydration degumming reaction was carried out at 55°C for 30 minutes. After the reaction was complete, a 9% (w / w) NaOH solution was pumped into mixer 2 at a flow rate of 104 L / h and mixed with soft water at a flow rate of 750 L / h in mixer 2. The flow rate control method was the same as for the phosphoric acid solution in (1). After mixing, the solution was introduced into the alkaline reaction tank, and a deoxidation reaction was carried out at a stirring speed of 42 Hz and 70°C for 30 minutes. After the reaction was complete, the mixture was centrifuged at a back pressure of 3.0 bar using a de-soaping centrifuge 1 and placed in a mixer 3. Soft water was added to the mixer 3 at a flow rate of 772 L / h and mixed. Next, it was introduced into a washing tank and washed with water. After centrifuging at a back pressure of 3.0 bar using a centrifuge 2, it was vacuum-dried in a dryer at 105°C for 20 minutes to obtain neutralized oil. (2) Dewaxing process: The procedure was carried out in the same manner as in step (2) of Example 1. (3) Decolorization process: The procedure was carried out in the same manner as in step (2) of Example 1. (4) Deodorization process: The decolorized oil was heated to 190°C in heat exchanger 3, introduced into heat exchanger 2, and subjected to secondary heating to 210°C. Subsequently, the decolorized oil was pumped into the plate tower, and the first stage of deodorization was performed for 90 minutes using 0.9 bar of stripping steam. Subsequently, the oil was introduced into heat exchanger 1 (heater) and heated to 230°C. The oil was pumped into the packed column, and a second stage of deodorization was performed for 10 minutes using 0.9 bar of stripping steam. The vacuum system was maintained at 2 mbar throughout the entire deodorization process to obtain refined sunflower seed oil.

[0106] Performance tests were conducted on the refined sunflower seed oil obtained, and the test results are as follows: The phospholipid content in refined sunflower seed oil was 12.9 mg / kg, with a yield of 0.25%. The vitamin E content was 620 mg / kg, with a yield of 88.6%. The plant sterol content was 4328 mg / kg, with a yield of 86.6%. The squalene content was 78.0 mg / kg, with a yield of 78.0%.

[0107] Example 5 The refined sunflower seed oil obtained in Example 1 was fortified with nutrients. The specific steps are as follows: (1) Preparation of vitamin A premix oil and vitamin D premix oil Vitamin A premix oil: Using a balance, 0.893 kg of vitamin A crystals were weighed and dissolved in 1.607 kg of refined sunflower seed oil prepared in Example 1 to prepare 2.5 kg of vitamin A 1.0 MIU raw oil. 0.0936 kg of vitamin A 1.0 MIU raw oil was weighed and mixed with 1.9064 kg of refined sunflower seed oil prepared in Example 1 to prepare vitamin A premix oil.

[0108] Vitamin D premix oil: 0.15 g of vitamin D crystals were weighed and dissolved in 1 kg of refined sunflower seed oil prepared in Example 1 to prepare a 0.15 g / kg vitamin D premix oil.

[0109] (2) Vitamin A premix oil and vitamin D premix oil were mixed with the refined sunflower seed oil obtained in Example 1 in proportions of 0.625% and 0.050% (w / w), respectively, and stirred for a further 15 minutes to obtain nutrient-fortified sunflower seed oil. The added concentration of vitamin A was 6200 μg / kg and the added concentration of vitamin D was 75 μg / kg.

[0110] The nutrient-enhanced sunflower seed oil obtained as described above was filled into yellow, diamond-shaped PET bottles capable of blocking light. Vitamins A and D were measured, and the product was deemed acceptable if the error from the theoretical design was controlled to within 10%.

[0111] Comparative Example 7 The refined sunflower seed oil obtained in Example 1 was fortified with nutrients. The specific steps are as follows: A vitamin A premix oil was prepared according to step 1 of Example 5, and this premix oil was mixed with the refined sunflower seed oil obtained in Example 1 at a ratio of 0.625% (w / w). The mixture was then stirred for a further 15 minutes to obtain a nutritionally fortified sunflower seed oil.

[0112] Comparative Example 8 The refined sunflower seed oil obtained in Example 1 was fortified with nutrients. The specific steps are as follows: A vitamin D premix oil was prepared according to step 1 of Example 5. This premix oil was mixed with the refined sunflower seed oil obtained in Example 1 at a ratio of 0.050% (w / w), and the mixture was stirred for a further 15 minutes to obtain a nutrient-fortified sunflower seed oil.

[0113] Comparative Example 9 The refined sunflower seed oil obtained in Example 4 was fortified with nutrients. The specific steps are as follows: A vitamin A premix oil was prepared according to step 1 of Example 5, and this premix oil was mixed with the refined sunflower seed oil obtained in Example 4 at a ratio of 0.625% (w / w). The mixture was then stirred for a further 15 minutes to obtain a nutritionally fortified sunflower seed oil.

[0114] Comparative Example 10 The refined sunflower seed oil obtained in Example 4 was fortified with nutrients. The specific steps are as follows: A vitamin D premix oil was prepared according to Step 1 of Example 5, and mixed with the refined sunflower seed oil obtained in Example 4 at a ratio of 0.050% (w / w). The mixture was then stirred for a further 15 minutes to obtain a nutrient-fortified sunflower seed oil.

[0115] Example 6 The refined sunflower seed oil obtained in Example 4 was fortified with nutrients. The specific steps are as follows: Vitamin A and vitamin D premix oils were prepared according to Step 1 of Example 5. These were mixed with the refined sunflower seed oil obtained in Example 1 at a ratio of 0.625% (w / w) and 0.050% (w / w), respectively, and stirred for a further 15 minutes to obtain fortified sunflower seed oil. The added vitamin A concentration was 6200 μg / kg, and the added vitamin D concentration was 75 μg / kg.

[0116] Example 7 Mice were housed in a light-dark cycle consisting of 12 hours of light and 12 hours of dark, provided with ample water and a specially formulated diet, and the experiment was started one week after pre- rearing. The experimental design was optimized to minimize the number of animals used and the pain inflicted on the animals.

[0117] Animal studies using healthy 5-week-old ICR male mice demonstrated that the prepared, fortified sunflower seed oil has a protective effect on cognitive function in the brain.

[0118] The specific experimental method is as follows: The animal experiments were validated using the above-mentioned 5-week-old healthy ICR male mice. All experimental procedures were performed in accordance with the regulations of the Jiangnan University Laboratory Animal Management and Animal Welfare Ethics Committee, under controlled conditions of a 12 / 12 hour lighting cycle (8:00-20:00), constant temperature (23±2℃), and constant humidity (60±5%).

[0119] The above 5-week-old healthy ICR male mice were randomly divided into six groups of 10 mice each after a one-week acclimatization period: a control group (Con), a scopolamine model group (Sop), a rivastigmine-positive control group (Riv), a vitamin D group (D), a vitamin D + vitamin A group (DA), a phospholipid group (P), a phospholipid + vitamin A group (PA), and a phospholipid + vitamin A + vitamin D group (PAD).

[0120] 1. Feed production Feed A: Casein 140 g / kg, L-Cystine 1.8 g / kg, Corn starch 495.7 g / kg, Maltodextrin 10 125 g / kg, Sucrose 100 g / kg, Cellulose 50 g / kg, Mineral Mix S10022M (purchased from Jiangsu Cooperative Biotechnology Co., Ltd.) 35 g / kg, the remainder being corn starch. Sunflower seed oil 40 g / kg obtained in Example 4 was added to Feed A. In the other groups of this experiment, different sunflower seed oils were used as needed. Feed B: Based on Feed A, 40 g / kg of sunflower seed oil, fortified with vitamin D alone as obtained in Comparative Example 10, was added. Feed C: Based on Feed A, 40 g / kg of sunflower seed oil fortified with vitamin D and vitamin A, as obtained in Example 6, was added. Feed D: Based on Feed A, 40 g / kg of sunflower seed oil obtained in Example 1 was added. This sunflower seed oil retains an appropriate amount of phospholipids through the process of the present invention. Feed E: Based on Feed A, 40 g / kg of sunflower seed oil, which retains phospholipids and is fortified with vitamin A, as obtained in Comparative Example 7, was added.

[0121] Feed F: Based on Feed A, 40 g / kg of sunflower seed oil, which retains phospholipids and is fortified with vitamins A and D, as obtained in Example 5, was added.

[0122] 2. Mouse experiments The control group (Con) received daily intraperitoneal injections of physiological saline, while the mice in the other groups received daily intraperitoneal injections of 3 mg / kg·bw of scopolamine (scopolamine hydrobromide was dissolved in sterile physiological saline to prepare a scopolamine solution for later use), with a total intraperitoneal injection volume of 10 mL / kg·bw. bw represents the body weight of the mouse.

[0123] Simultaneously with the modeling process, mice were fed daily as part of the intervention. They were fed once a day, with 4g per mouse each time. Each group of mice was given free access to water.

[0124] Specifically, it is as follows: Control group (Con): Received one intraperitoneal injection of physiological saline daily (10 mL / kg·bw), and fed daily with feed A. Scopolamine model group (Sop): 3 mg / kg·bw of scopolamine was administered intraperitoneally daily (modeling), with an injection volume of 10 mL / kg·bw, and feed was diet A. Rivastigmine-positive control group (Riv): Daily intraperitoneal injection of 3 mg / kg·bw of scopolamine (modeling), with an injection volume of 10 mL / kg·bw. Daily, 30 minutes after modeling, rivastigmine was again injected intraperitoneally at a dose of 2 mg / kg·bw, and feed A was administered. Vitamin D group (D): 3 mg / kg·bw of scopolamine was administered intraperitoneally daily, with an injection volume of 10 mL / kg·bw, and feed B was given. Vitamin D + Vitamin A complex (DA): 3 mg / kg·bw of scopolamine was administered intraperitoneally daily, with a total injection volume of 10 mL / kg·bw, and feed C was provided. Phospholipid group (P): 3 mg / kg·bw of scopolamine was administered intraperitoneally daily (10 mL / kg·bw), and feed was diet D. Phospholipids + Vitamin A group (PA): 3 mg / kg·bw of scopolamine was administered intraperitoneally daily, with a total injection volume of 10 mL / kg·bw, and feed E was provided. Phospholipids + Vitamin A group + Vitamin D group (PAD): 3 mg / kg·bw of scopolamine was administered intraperitoneally daily, with a total injection volume of 10 mL / kg·bw, and feed F was provided.

[0125] Following four weeks of continuous modeling and intervention, the mice underwent behavioral tests including the Open Field (OFT), Novel Object Recognition (NORT), Y-Maze Test (YMT), and Morris Water Maze Test (MWM).

[0126] After fasting for 12 hours, the mice were anesthetized by intraperitoneal injection of 10% chloral hydrate, and then euthanized by cervical dislocation. Blood and cerebral cortex samples were collected from the mice for further study.

[0127] The results are as follows: 1. Results of mouse behavioral experiments The Open Field Test (OFT) was used to investigate the intervention effect of the edible oils obtained in the above examples and comparative examples on the exploratory behavior of scopolamine-treated mice. In a new environment, the exploratory behavior of mice can be evaluated by the amount of time spent in the central area. The longer the time spent in the central area, the greater the animals' motivation to explore the open space, which reflects high exploratory motivation and low anxiety.

[0128] JPEG2026123773000007.jpg81170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0129] As shown in Table 6, the Sop model group showed a significant decrease in both time spent in the central region and percentage compared to the blank group. The added functional factors, phospholipids and vitamin A, showed some improvement effects. Among these, the DA, P, PA, and PAD groups were able to significantly increase both time and percentage in the central region (p<0.01). The VD group did not show a significant effect in the open-field trial, but the combined effect of vitamin A and vitamin D was statistically significant, and experimental data showed it to be superior to single-component vitamin D. Among the multi-component intervention groups, the tripartite combination of phospholipids, vitamin A, and vitamin D showed the best effect, and was superior to the combination of two components.

[0130] The Y-maze test was used to examine the intervention effect of edible oils obtained in the above examples and comparative examples on the working memory of mice. Spontaneous alternation reflects the working memory capacity of mice. Working memory is the ability to retain and manipulate information within a short period of time. In the Y-maze test, mice tended to explore new arms rather than return to recently visited arms. This indicates that they need to remember the places they have recently been. Successful spontaneous alternation of mice indicates that they have good working memory.

[0131] As shown in Table 7, the model group showed a significant difference compared to the blank group (Con) (P<0.05), indicating successful modeling. With the exception of group P, all other intervention groups were able to significantly improve the number of spontaneous rotations in mice, and the combination of phospholipids, vitamin A, and vitamin D was superior to the combination of two components.

[0132] JPEG2026123773000008.jpg81170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0133] A novel object recognition test was employed to investigate the interventional effect of the edible oils obtained in the above examples and comparative examples on the novel object recognition impairment in mice. In the NOR test, mice are exposed to two objects: one known and one novel. Healthy mice typically spend more time exploring the novel object because they have a greater interest and curiosity towards it. The discrimination index can effectively reflect the memory function of mice, as their ability to recognize the novel object and spend more time with it indicates that their memory function is normal.

[0134] As shown in Table 8, the Sop model group showed a significant decrease in the discrimination index (DI) compared to the blank group, indicating successful modeling. Vitamin D and vitamin A intake showed a significant improvement in the DI index (p<0.01), indicating that scopolamine impairs learning and cognitive memory in mice. From the experimental data, all four intervention groups were able to significantly improve the discrimination index of the model mice (P<0.01). Numerically, the combined effect of phospholipids, vitamin A, and vitamin D was superior to the two-component intervention group, followed by phospholipids + vitamin A.

[0135] JPEG2026123773000009.jpg81170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0136] The Morris Water Maze (MWM) was used to evaluate the improvement effect of the edible oils obtained in the above examples and comparative examples on spatial memory in mice. By having mice search for hidden platforms in a pool, their ability to remember and utilize environmental cues can be evaluated. This is particularly suitable for studying spatial memory disorders such as those caused by scopolamine.

[0137] During the place navigation period, the escape latency of mice gradually decreased with increasing training days, demonstrating the effectiveness of the training. Statistical analysis of escape latency on day 5 revealed a significant difference between the model group and the blank group. Vitamin D alone (P<0.01), phospholipids (P<0.01), and the combination of vitamin D and vitamin A (P<0.01) significantly shortened the escape latency of mice. Furthermore, the combination of phospholipids, vitamin A, and vitamin D showed the best effect, with a significant difference compared to the other intervention groups (P<0.01).

[0138] JPEG2026123773000010.jpg81170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0139] Platform crossing count refers to the number of times a mouse crosses the location of a hidden platform and reflects the mouse's ability to accurately remember the target platform location. This is primarily used to assess whether a mouse can accurately remember the platform location after repeated training. A higher crossing count indicates stronger spatial orientation memory in the mouse. Platform area crossing count refers to the number of times a mouse crosses a region containing the target platform (usually defined as a region slightly larger than the platform itself) and investigates the mouse's memory of the platform's approximate location. This metric reflects the mouse's overall memory ability for spatial cues and is somewhat broader than platform crossing count. Platform area cumulative time refers to the amount of time a mouse spends within the target platform area and reflects the degree of the mouse's preference for that area and whether it recognizes the platform's approximate location. This metric is used to assess whether a mouse has a high exploratory motivation and cognitive association with the platform area. A longer cumulative time indicates a clearer memory of the area in the mouse.

[0140] As shown in Table 10, at the spatial probe stage, the number of platform crossings and cumulative platform area time in the model group were significantly lower than in the blank group (P<0.01). Regarding the number of platform crossings, the improvement effect of phospholipid alone was not significant, but the combination of phospholipid and vitamin A significantly increased the number of platform crossings in the model mice (P<0.01). Regarding cumulative platform area time, all intervention groups were able to significantly increase the cumulative platform area time in mice. Similarly, the improvement effect of the combination of phospholipid and vitamin A was numerically superior to that of single-component phospholipids.

[0141] JPEG2026123773000011.jpg62170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0142] 2. The effects of the above-mentioned vegetable oils on oxidative stress in mice. Numerous studies have revealed that oxidative stress is one of the important mechanisms for the development of neurodegenerative diseases. When faced with oxidative stress, the body can remove excess oxygen radicals by increasing the activity of antioxidant enzymes or by utilizing naturally occurring antioxidants. Currently, there are many indicators that reflect the level of oxidation in the body, such as GSH (glutathione), MDA (malondialdehyde), SOD (total superoxide dismutase), and GPX (glutathione peroxidase). In this study, these indicators were measured in the blood and cerebral cortex, and the effects of vitamin A, vitamin D, phospholipids, and a combination of these on the antioxidant activity of mice were evaluated. As MDA accumulates, biomolecules such as nucleic acids and proteins undergo cross-linking polymerization, causing cytotoxicity. Therefore, the MDA content is an important indicator that reflects the degree of oxidative damage suffered by the body.

[0143] Blood and cerebral cortex collection and processing: Blood was collected from the retroorbital venous sinus and placed in an EP tube containing heparin sodium. The sample was then centrifuged at 3,000 rpm for 15 minutes, and the supernatant was collected for measurement of IL-6 and MDA (malondialdehyde) levels.

[0144] After blood collection, the mouse cerebral cortex was carefully extracted, and a 10% tissue homogenate was rapidly prepared in pre-cooled sterile saline. This homogenate was then centrifuged at 4°C at 12,000 revolutions / minute for 10 minutes, and the supernatant was collected for further analysis. Relevant biochemical indicators were measured using an enzyme-linked immunosorbent assay (ELISA) kit according to the instructions provided by the manufacturer (Xiamen Huijia Biotechnology Co., Ltd.). The cerebral cortex supernatant was used to detect IL-6 and MDA (malondialdehyde). The remaining samples were stored at -80°C for use in subsequent experiments.

[0145] As shown in Table 11, vitamin D and phospholipid intake tended to lower serum malondialdehyde levels, and the combined use of phospholipids and vitamin A significantly reduced serum MDA levels (P<0.01). Regarding malondialdehyde levels in the cerebral cortex, the model group showed a significant difference compared to the blank group (P<0.01), and the combined intake of vitamin D or the three functional factors significantly reduced malondialdehyde levels in the cerebral cortex (P<0.01). Furthermore, experimental data showed that the combined effects of vitamin D and vitamin A, and phospholipids and vitamin A, were superior to the use of vitamin D or phospholipids alone (P<0.01). Overall, the combination of phospholipids and vitamin A played a greater role in reducing MDA levels in model mice compared to other intervention groups.

[0146] JPEG2026123773000012.jpg92170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0147] 3. Effects of the above-mentioned vegetable oils on the inflammatory response in mice. Research indicates that cognitive impairment and memory loss are influenced by oxidative stress in the brain, and that inflammatory responses also damage neuronal cells and synaptic structures. Among the physiological responses of the body, oxidative stress often induces inflammation. When an inflammatory response occurs, it itself triggers oxidative stress, and the two mutually exacerbate each other. IL-6 is a major factor in causing abnormal secretion of inflammatory cytokines in cells and the development of inflammatory responses.

[0148] As shown in Table 12, serum and cerebral cortical inflammatory cytokine IL-6 levels in the model group were higher than in the blank group (P<0.01). In terms of blood indicators, all five intervention groups significantly reduced IL-6 levels in the model mice, and the combination of vitamin D + vitamin A and phospholipid + vitamin A was superior to vitamin D or phospholipid alone, based on experimental data. In the cerebral cortex, all intervention groups except the phospholipid group significantly reduced IL-6 levels, and the combination of vitamin D + vitamin A and phospholipid + vitamin A was superior to vitamin D or phospholipid alone, based on experimental data.

[0149] JPEG2026123773000013.jpg92170 All values ​​are expressed as mean ± standard deviation (mean ± SD). *p<0.05, **p<0.01 (comparison with the Sop group). #p<0.05, ##p<0.01 (comparison with the Con group), n=10.

[0150] Comparative Example 11 This method was based on Example 2, with the only difference being that the heat exchanger 3 in step (4) was omitted and the temperature was directly heated to 210°C. Otherwise, it is identical to Example 2.

[0151] The results are as follows: JPEG2026123773000014.jpg54170

[0152] Comparative Example 12 A method for producing corn oil, comprising the following steps: In the alkaline refining process, crude corn oil was first heated to 40°C, then food-grade phosphoric acid with a concentration of 75% was added at a rate of 0.15% of the oil volume, and the mixture was acid-treated for 20 minutes. The acid-treated mixture was then neutralized with an alkaline solution and an alkaline solution with a concentration of 15Be for 15 minutes. After that, it was centrifuged.

[0153] A continuous dewaxing method using tubes was employed. Alkali-refined corn oil was crystallized and dewaxed through 12 crystal tubes. Specifically, the alkali-refined corn oil entered the first crystal tube from a pump, and then flowed sequentially into the remaining crystal tubes by overflow. Condensing coils were wound around the outside of the crystal tubes to gradually lower the oil temperature to 5°C. After that, the oil was kept warm for 4 hours to promote crystallization and crystal growth, and then the oil temperature was raised to 20°C for filtration. The dewaxed corn oil was pumped into a decolorization tower for decolorization at a decolorization temperature of 118°C, with an activated clay addition amount of 2.5% of the oil weight, a decolorization time of 25 mins, and a pressure of 0.010 MPa. The clay was filtered to obtain decolorized oil.

[0154] The deodorization stage employed a two-temperature, two-column deodorization process. Decolorized oil was pumped into a packed column, and volatile substances such as fatty acids, hydrocarbons, and aldehydes were removed at a packed column temperature of 220°C for 15 minutes. The oil then flowed into a plate column by gravity, where it underwent thermal decolorization at a plate column temperature of 210°C for 30 minutes of steam stripping. After that, it entered the packed column and remained there for 13 minutes, where secondary fatty acids were removed at a temperature of 230°C to obtain deodorized oil.

[0155] The deodorized oil was cooled in an exchanger to room temperature before being filled. Before filling, nitrogen gas was purged into the oil tank, and then the oil was poured into the tank. Various indicator tests were conducted on the finished oil product. The results are as follows: JPEG2026123773000015.jpg47170

[0156] While the present invention is disclosed by the above-described preferred embodiments, these do not limit the invention. Those skilled in the art can make various modifications and alterations without departing from the spirit and scope of the invention. Therefore, the scope of protection of the present invention shall be defined by the claims.

Claims

1. A method for simultaneously improving the retention rate of phospholipids, vitamin E, plant sterols, and squalene in refined vegetable oil, characterized by comprising the following steps: (1) neutralization step, (2) dewaxing step, (3) decolorization step, and (4) deodorization step. (1) Neutralization process (degumming + deacidification): Crude oil is preheated to 55°C, and a phosphoric acid solution is added by pump at a flow rate of 1 to 50 L / h to carry out a hydration degumming reaction to obtain the product after hydration degumming. The product after hydration and degumming is mixed with an alkaline solution and soft water, stirred, and a deoxidation reaction is carried out. After the reaction is complete, the mixture is centrifuged, washed with soft water, centrifuged again, and dried to obtain neutralized oil. (2) Dewaxing process: A crystallization accelerator is added to the neutralized oil, cooled to 6.8°C, stirred, and filtered to obtain dewaxed oil. (3) Decolorization process: The dewaxed oil is heated to 105°C, a decolorizing agent is added and adsorbed, and then filtered to obtain decolorized oil. (4) Deodorization process: The decolorized oil is pumped into a plate column and first heated to 190°C, then subjected to secondary heating to 210°C for the first stage of deodorization for 90 minutes. The oil is then pumped into a packed column and subjected to the second stage of deodorization at 230°C for 10 minutes to obtain refined vegetable oil. Here, for the phosphoric acid solution and alkaline solution, a combination of an electromagnetic pump, electromagnetic proportional valve, and electromagnetic flow meter is used. The signal is fed back to the PLC according to the crude oil flow rate and the amount of free fatty acids, the theoretical amount to be added is calculated, the opening of the proportional control valve is adjusted, the flow rate is measured and compared with the electromagnetic flow meter, and the opening of the proportional valve is adjusted in real time by feeding back the results, thereby optimizing the precise amount of phosphoric acid solution and alkaline solution to be added.

2. A refined vegetable oil produced by the method described in claim 1.

3. A method for producing nutrient-fortified refined vegetable oil, A method characterized by comprising the step of adding a nutritional substance to a refined vegetable oil according to claim 2 for nutritional enhancement, wherein the nutritional substance is one or two of vitamin A and vitamin D.

4. The aforementioned method, The method according to claim 3, characterized in that it includes the step of adding vitamin A and vitamin D to refined vegetable oil and stirring to obtain a nutritionally fortified refined vegetable oil, wherein the amount of vitamin A added is 4000 to 8000 μg / kg and the amount of vitamin D added is 50 to 100 μg / kg.

5. Nutrient-fortified refined vegetable oil produced by the method of claim 3 or 4.

6. Use of the nutritionally fortified refined vegetable oil according to claim 5 in the manufacture of a product for improving cognitive function or for antioxidant or anti-inflammatory purposes.

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