Nutritionally fortified edible vegetable oil and preparation process thereof

Abstract

The present invention discloses a nutritionally fortified edible vegetable oil and its preparation process, belonging to the field of edible oil processing. By optimizing the oil processing process, adjusting refining process parameters, appropriately retaining phospholipids and maximizing the retention of inherent nutrients such as vitamin E, and adding appropriate amounts of vitamin D and vitamin A, the present invention develops a nutritionally fortified oil product. Animal experiments have verified that the product significantly reduces the levels of brain inflammatory factors and enhances the brain's ability to resist oxidative stress, improving cognition and providing a scientific basis for the future functional edible oil market.

Description

Technical Field

[0001] The invention relates to a nutritionally enhanced edible vegetable oil and a preparation process thereof, and belongs to the field of edible oil processing. Background Art

[0002] Edible vegetable oils have attracted considerable attention due to their high content of unsaturated fatty acids and diverse nutritional profile. Sunflower oil, for example, is produced from sunflower seeds through a series of processes, including pressing and refining. The degumming, deacidification, dewaxing, decolorization, and deodorization processes significantly reduce the content of beneficial nutrients such as vitamin E and phospholipids. Vitamin E is a natural antioxidant and immunomodulator, and also has physiological functions such as combating infertility, inhibiting tumor cell growth, improving atherosclerosis, and preventing cardiovascular disease. The primary source of vitamin E in daily life is edible vegetable oils. Phospholipids, a crucial component of cell membranes, can enhance brain function, improve memory, and protect neurons, offering benefits that enhance brain function and intelligence. Commercially available vegetable oils contain low levels of beneficial ingredients such as vitamin E, phytosterols, and squalene (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, it is extremely important to develop a method for preparing refined vegetable oils that retains a high content of nutritional concomitants in the vegetable oils.

[0003] Conventional processes for preparing oils and fats mainly include pressing or leaching to prepare crude oil, which is then refined into commercially available finished oil through a refining process. However, in practical applications, there are some prominent problems, especially in the refining process. For example, high-temperature operation leads to the loss of nutrients (such as vitamin E, phytosterols, and squalene). Moreover, the failure to fully remove unstable components in the process will lead to poor oxidative stability. In order to improve the nutrient retention and quality of oils and fats, commonly used improvement methods include low-temperature pressing, physical refining, and enzymatic hydrolysis technology. However, these methods also have certain limitations. For example, low-temperature pressing has a low oil yield and high cost, physical refining has strict equipment requirements, and enzymatic hydrolysis technology is difficult to promote due to its complex process and long cycle.

[0004] Vitamin A and vitamin D are essential fat-soluble vitamins for the human body and play an important role in maintaining normal physiological functions. Vitamin A, also known as retinol, helps maintain dark vision and helps maintain healthy skin and mucous membranes. Vitamin D is a sterol derivative that has the functions of promoting calcium and phosphorus metabolism and regulating immune function. However, vitamin A and vitamin D are only found in animal foods, and vegetable oils contain almost no vitamin A and vitamin D. Therefore, it is of great significance to develop a vegetable oil that is nutritionally fortified with vitamin A and vitamin D. The addition of vitamins A and D to vegetable oils can significantly improve their functionality, especially in terms of anti-oxidation and improving nervous system function.

[0005] However, due to the influence of the refining process, the content of retained nutritional companions such as vitamin E and phospholipids is relatively low; and vitamin A and vitamin D fortified vegetable oils are still in the early development stage, and the specific content of retained and added nutritional functional factors on the prevention and intervention effects of cognitive impairment lacks in-depth verification.

[0006] In summary, the preparation of vegetable oils with high-retention nutritional concomitants and nutritional fortification is currently an important research direction in the field of oil and fat engineering. Through continuous exploration and innovation, it is expected that safer, more efficient, and more environmentally friendly edible vegetable oil preparation technologies will be developed to meet people's diverse needs for health, nutrition, and deliciousness. Summary of the Invention

[0007] To solve the above technical problems, the present invention optimizes the oil processing technology, adjusts the refining process parameters, moderately retains phospholipids and maximizes the retention of inherent nutrients such as vitamin E, and adds appropriate amounts of vitamin D and vitamin A to develop a nutritionally fortified oil product. Animal experiments have verified that the product plays a significant role in reducing the level of brain inflammatory factors and improving the brain's ability to resist oxidative stress, and has the effect of improving cognition, 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 preparing refined vegetable oil with high retained nutritional concomitants, the method comprising a neutralization section, a dewaxing section, a decolorization section, and a deodorization section;

[0009] Among them, the deodorization section adopts a double-tower combined dual-temperature deodorization process, as follows:

[0010] The decolorized oil is pumped into a plate tower at 180-230°C for 60-130 minutes for the first stage of deodorization; and then pumped into a packed tower at 200-250°C for 5-15 minutes for the second stage of deodorization.

[0011] In one embodiment, the method specifically includes:

[0012] (1) Neutralization section (degumming + deacidification):

[0013] Preheating the crude oil to 40-80°C, pumping a phosphoric acid solution into the crude oil at a flow rate of 1-50 L / h to perform a hydration degumming reaction to obtain a hydration degumming product;

[0014] The product after hydration degumming, alkaline solution and soft water are mixed and stirred to carry out deacidification reaction; after the reaction is completed, centrifugation is carried out, soft water is added for washing, centrifugation is carried out, and drying is carried out to obtain neutralized oil;

[0015] (2) Dewaxing section:

[0016] Add a crystallizing agent to the neutralized oil, cool it to 3-13°C, stir it, and filter it to obtain dewaxed oil;

[0017] (3) Decolorization section:

[0018] The dewaxed oil is heated to 90-120°C, a decolorizing agent is added for adsorption, and the oil is filtered to obtain decolorized oil;

[0019] (4) Deodorization section:

[0020] The decolorized oil is pumped into a plate tower at 180-230°C for first-stage deodorization for 60-130 minutes; and then pumped into a packed tower at 200-250°C for second-stage deodorization for 5-15 minutes to obtain refined vegetable oil.

[0021] In one embodiment, the preparation method of crude plant oil in step (1) is:

[0022] The crude oil is obtained by adopting the pressing process, which includes impurity removal, crushing, tempering, flaking, steaming, pressing and filtering. The main control parameters include: steaming temperature of 90-140℃, time of 20-30min, and pressing material temperature controlled at 80-120℃.

[0023] In one embodiment, the flow rate of the crude vegetable oil before preheating in step (1) is 18 to 24 T / h; preferably, the flow rate is 21 T / h.

[0024] In one embodiment, the temperature of the crude oil preheating in step (1) is 55°C.

[0025] In one embodiment, the concentration of the phosphoric acid solution in step (1) is 75% to 86% (w / w); preferably, the concentration is 80% (w / w).

[0026] In one embodiment, the flow rate of pumping the phosphoric acid solution in step (1) is 0.5 L / h.

[0027] In one embodiment, the hydration degumming reaction in step (1) is carried out at 40-80° C. for 20-60 minutes.

[0028] In one embodiment, the flow rate of the alkaline solution in step (1) is 3 to 200 L / h; preferably, the flow rate is 104 L / h.

[0029] In one embodiment, the alkaline solution in step (1) is a sodium hydroxide solution with a concentration of 4% to 20% (w / w); preferably, the concentration is 9% (w / w).

[0030] In one embodiment, the stirring speed in step (1) is 20 to 60 Hz; preferably, the stirring speed is 40 Hz.

[0031] In one embodiment, the deacidification reaction in step (1) is carried out at 65-90° C. for 20-35 minutes.

[0032] In one embodiment, the phosphoric acid solution and the alkaline solution in step (1) are prepared using a combination of an electromagnetic pump, an electromagnetic proportional valve, and an electromagnetic flowmeter. The system uses a signal fed back to a PLC (programmable logic controller) based on the crude oil flow rate and the amount of free fatty acids to calculate the theoretical addition amount. The proportional control valve opening is adjusted and compared with the actual flow rate measured by the electromagnetic flowmeter. The proportional valve opening is then fed back to the controller in real time to optimize the precise addition amount of the phosphoric acid solution, alkaline solution, and other media.

[0033] In one embodiment, the crystallization promoting agent in step (2) is one or both of diatomaceous earth and perlite.

[0034] In one embodiment, the amount of the crystallizing agent added in step (2) is 0.1% to 2.0% (w / w); preferably, the amount of the crystallizing agent added is 0.62% (w / w), and the amount added is relative to the mass of the neutralized oil.

[0035] In one embodiment, the temperature is cooled to 6.8°C in step (2).

[0036] In one embodiment, the stirring time in step (2) is 20 to 30 hours; preferably, the stirring time is 23.8 hours.

[0037] In one embodiment, the heating temperature in step (3) is 105°C.

[0038] In one embodiment, the decolorizing agent in step (3) is one or more of attapulgite clay, activated clay, and activated carbon.

[0039] In one embodiment, the amount of the decolorizing agent added in step (3) is 0.05% to 4% (w / w); preferably, the amount of the decolorizing agent added is 1.0% (w / w), and the amount added is relative to the dewaxed oil.

[0040] In one embodiment, the adsorption time in step (3) is 20 to 60 minutes; preferably, the adsorption time is 30 minutes.

[0041] In one embodiment, the deodorization section in step (4) is:

[0042] The decolorized oil was pumped into a plate tower at 200°C and stripping steam at 0.9 bar for a first-stage deodorization of 90 minutes; and then pumped into a packed tower at 230°C and stripping steam at 0.9 bar for a second-stage deodorization of 10 minutes to obtain refined vegetable oil.

[0043] In one embodiment, the nutritional companion is one or more of vitamin E, phospholipids, phytosterols, and squalene.

[0044] In one embodiment, the high retention nutritional companion is:

[0045] The retained amount of vitamin E in refined vegetable oil is 200-800 mg / kg, the retained amount of phospholipids is 0-200 mg / kg (but not 0), the retained amount of phytosterols is 3000-10000 mg / kg, and the retained amount of squalene is 50-400 mg / kg.

[0046] In one embodiment, the vegetable oil includes but is not limited to sunflower oil, corn oil, and rapeseed oil; preferably, the vegetable oil is sunflower oil.

[0047] A second object of the present invention is a refined vegetable oil with high retention of nutritional concomitants, obtained by the process according to the invention.

[0048] A third object of the present invention is to provide a method for improving the phospholipid retention rate in refined vegetable oil, comprising the following steps:

[0049] (1) Neutralization section (degumming + deacidification):

[0050] Preheating the crude oil to 40-80°C, pumping a phosphoric acid solution into the crude oil at a flow rate of 1-50 L / h to perform a hydration degumming reaction to obtain a hydration degumming product;

[0051] The product after hydration degumming, alkaline solution and soft water are mixed and stirred to carry out deacidification reaction; after the reaction is completed, centrifugation is carried out, soft water is added for washing, centrifugation is carried out, and drying is carried out to obtain neutralized oil;

[0052] (2) Dewaxing section:

[0053] Add a crystallizing agent to the neutralized oil, cool it to 3-13°C, stir it, and filter it to obtain dewaxed oil;

[0054] (3) Decolorization section:

[0055] The dewaxed oil is heated to 90-120°C, a decolorizing agent is added for adsorption, and the oil is filtered to obtain decolorized oil;

[0056] (4) Deodorization section:

[0057] The decolorized oil is pumped into a plate tower at 180-230°C for first-stage deodorization for 60-130 minutes; and then pumped into a packed tower at 200-250°C for second-stage deodorization for 5-15 minutes to obtain refined vegetable oil.

[0058] In one embodiment, improving the phospholipid retention rate in refined vegetable oil is mainly based on the parameter adjustment of the neutralization section of step (1).

[0059] A fourth object of the present invention is to provide a method for improving the retention rate of vitamin E, phytosterols, and squalene in refined vegetable oils, comprising the following steps:

[0060] (1) Neutralization section (degumming + deacidification):

[0061] Preheating the crude oil to 40-80°C, pumping a phosphoric acid solution into the crude oil at a flow rate of 1-50 L / h to perform a hydration degumming reaction to obtain a hydration degumming product;

[0062] The product after hydration degumming, alkaline solution and soft water are mixed and stirred to carry out deacidification reaction; after the reaction is completed, centrifugation is carried out, soft water is added for washing, centrifugation is carried out, and drying is carried out to obtain neutralized oil;

[0063] (2) Dewaxing section:

[0064] Add a crystallizing agent to the neutralized oil, cool it to 3-13°C, stir it, and filter it to obtain dewaxed oil;

[0065] (3) Decolorization section:

[0066] The dewaxed oil is heated to 90-120°C, a decolorizing agent is added for adsorption, and the oil is filtered to obtain decolorized oil;

[0067] (4) Deodorization section:

[0068] The decolorized oil is pumped into a plate tower at 180-230°C for first-stage deodorization for 60-130 minutes; and then pumped into a packed tower at 200-250°C for second-stage deodorization for 5-15 minutes to obtain refined vegetable oil.

[0069] In one embodiment, the method for improving the retention rate of vitamin E, phytosterols, and squalene in refined vegetable oil is mainly based on the parameter adjustment of the deodorization section in step (4).

[0070] A fifth object of the present invention is to provide a method for preparing a nutritionally enhanced refined vegetable oil, comprising the steps of:

[0071] Nutrients are added to refined vegetable oil for nutritional fortification, wherein the nutrients are one or both of vitamin A and vitamin D.

[0072] In one embodiment, the method specifically includes:

[0073] Vitamin A and vitamin D are added to refined vegetable oil and stirred to obtain nutritionally fortified refined vegetable oil; wherein the added amount of vitamin A is 4000-8000 μg / kg, and the added amount of vitamin D is 50-100 μg / kg.

[0074] In one embodiment, the amount of vitamin A added is 6200 mg / kg, and the amount of vitamin D added is 75 μg / kg.

[0075] In one embodiment, the nutritionally fortified refined vegetable oil has a vitamin E content of 200-800 mg / kg, a phospholipid content of 0-200 mg / kg, a phytosterol retention amount of 3000-10000 mg / kg, a squalene retention amount of 50-400 mg / kg, a vitamin A content of 4000-8000 μg / kg, and a vitamin D content of 50-100 μg / kg.

[0076] The sixth object of the present invention is the nutritionally enhanced refined vegetable oil prepared by the method of the present invention.

[0077] The seventh object of the present invention is to provide the use of the above-mentioned nutritionally fortified refined vegetable oil in the preparation of products for improving cognitive function or anti-oxidation or anti-inflammatory.

[0078] In one embodiment, the product includes but is not limited to medicines, foods, health products, feeds, feed additives, and food additives.

[0079] Beneficial effects:

[0080] (1) Although lecithin has a high nutritional value, due to its special hydrophilic and lipophilic properties, excessive content in vegetable oils will accelerate the darkening of the oil color and aftertaste, thereby affecting the safety and stability of oil products during storage. Currently, most of the industry uses triple metering pumps as the driving system for acid and alkali addition. Although it is linked to the flow rate of crude oil and free fatty acids, most still require manual adjustment of the valve stem stroke and the use of frequency conversion to adjust the flow rate. Once the crude oil flow rate fluctuates, the addition of acid and alkali is often excessive or insufficient. Analysis of the flow curve of the triple pump shows that there is an error of about 57%. This error is also the reason for the need to add excessive acid and alkali and the subsequent inability to improve the yield.

[0081] The present invention adopts the concept of precise and appropriate processing, adopts an electromagnetic pump with an engineering plastic pump body, and combines an electromagnetic flow meter and a proportional control valve, and designs an addition pipeline and a large return pipeline according to the flow ratio (the process control diagram is shown in FIG). Figure 2 ), combined with the crude oil flow rate and the amount of free fatty acids, the signal is fed back to the PLC (Programmable Logic Controller) to calculate the theoretical addition amount. The proportional control valve opening is adjusted and compared with the actual flow rate measured by the electromagnetic flowmeter. The proportional valve opening is then adjusted in real time based on the feedback. This optimizes the design of the acid, alkali, dilution water, and wash water addition pipelines and the intelligent addition proportion control system, reducing the addition amount error to 10%.

[0082] In summary, more precise control equipment is used in the neutralization section, and the proportion of phosphoric acid added in the degumming section is accurately adjusted according to the phospholipid content of the crude oil, so as to better retain and control the phospholipid content.

[0083] (2) The structure of the plate deodorization tower is to arrange several layers of plates horizontally at a certain interval within a cylindrical shell. The liquid flows from top to bottom through each layer of plates by gravity and is discharged from the bottom of the tower. A certain thickness of flowing liquid layer is maintained on each layer of plates. The gas, driven by the pressure difference, rises from the bottom of the tower through the liquid layer on each plate and is discharged from the top of the tower. Advantages of the plate deodorization tower: The plate deodorization tower has a simple structure and strong operability. The residence time in the deodorization tower is long, which allows the heat-sensitive pigments in the oil to be completely decomposed, thereby achieving a good decolorization effect. However, the long residence time also causes harmful substances such as trans fatty acids to be produced. At the same time, nutrients such as vitamin E, phytosterols, and squalene are distilled out with the steam.

[0084] The structure of a packed deodorizing tower is to fill a cylindrical shell with a certain height of packing. The liquid is evenly distributed on the top of the packing layer through the tower top spray device. It flows from top to bottom along the packing surface through the packing layer by gravity and is discharged from the bottom of the tower. The gas passes through the gaps in the packing layer under the pressure difference and flows from one end of the tower to the other. Advantages of a packed deodorizing tower: Because there is no oil level and the specific surface area is large, the oil film formed is relatively thin, the mass transfer effect per unit time is good, and it can be used for physical deacidification. Disadvantages of a packed deodorizing tower: The deodorization effect is not as significant as that of a plate deodorizing tower. Since the liquid film of oil is very thin, the residence time of the deodorized oil in the tower is short.

[0085] The present invention adopts a method of combining a packed tower with a plate tower to form a double-tower combination. By transforming traditional process pipelines, segmented temperature control can be achieved, and the temperature of the double towers can be flexibly and accurately adjusted according to the quality of crude oil and oil products in the refining process. According to the characteristic of the long residence time of the plate tower, the deodorization temperature of the plate tower is appropriately lowered in the first stage. According to the characteristics of the short residence time and strong deacidification ability of the packed tower, rapid, short-term and high-temperature heating is adopted for the deodorization of the packed tower in the second stage, thereby avoiding the problems of high temperature and long residence time of the deodorized oil in the plate-type packed tower, and large loss of nutritional accompanying substances such as vitamin E, phytosterols and squalene, and better retaining the nutritional accompanying substances in the deodorized oil.

[0086] (3) The present invention adopts a method of combining a packed tower with a plate tower, rationally optimizes the direction of the pipeline, and innovatively utilizes a set of efficient heater systems to achieve temperature control of the two tower bodies. Figure 3 , thereby ensuring the deodorization effect while significantly improving energy utilization efficiency and achieving energy conservation and emission reduction.

[0087] (4) The present invention, on the basis of moderately retaining the nutritional accompaniments in oils and fats, carries out nutritional fortification of vitamin A or vitamin D in sunflower oil, according to the "Dietary Guidelines for Chinese Residents" and other regulations. By building a scopolamine mouse animal model, the synergistic effects of vitamins A, E, D and phospholipids and other nutrients in anti-oxidation and memory improvement are studied, and the effective doses of the above nutrients to exert their effects are verified, providing scientific support for the claimed formula ratio. BRIEF DESCRIPTION OF THE DRAWINGS

[0088] Figure 1 This is a schematic diagram of a traditional acid / alkali metering pump in the neutralization section; 1 is the pump head, 2 is the mechanical device, 3 is the manual stroke adjustment knob, and 4 is the motor.

[0089] Figure 2 This is the process flow chart after the transformation of the acid / alkali precision addition system in the neutralization section; among them, 1 is the crude oil relay 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 1# centrifuge, 7 is the 2# centrifuge, 8 is the water washing and stirring tank, 9 is the dryer, 10 is the 1# mixer, 11 is the 2# mixer, 12 is the 3# mixer, 13 is the electromagnetic pump, 14 is the electromagnetic proportional valve, 15 is the electromagnetic flowmeter, 16 is the electromagnetic pump, 17 is the electromagnetic proportional valve, and 18 is the electromagnetic flowmeter.

[0090] Figure 3 This is the process flow chart of double-tower combined dual-temperature deodorization in the deodorization section. DETAILED DESCRIPTION

[0091] The nutritionally fortified sunflower oil with improved cognitive function and the manufacturing process of the present invention will be further explained and illustrated below with reference to specific embodiments. However, such explanation and illustration do not constitute an undue limitation to the technical solution of the present invention.

[0092] The drugs and reagents involved in the following examples:

[0093] Scopolamine hydrobromide (98% purity) was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0094] IL-6 and malondialdehyde (MDA) kits were purchased from Xiamen Huijia Biotechnology Co., Ltd.

[0095] The other commonly used reagents were all domestically produced analytical grade.

[0096] Rivastigmine was purchased from MedChemExpress, CAS: 123441-03-2.

[0097] The crystallization promoting agent diatomite was purchased from Linjiang Dayuan Diatomite New Material Ecological Environmental Protection Technology Co., Ltd.

[0098] The adsorbent activated clay was purchased from Huangshan Baiyue Activated Clay Co., Ltd.

[0099] The experimental animals used in the following examples are:

[0100] Male ICR mice (5 weeks old, 21 ± 2 g, specific pathogen-free (SPF)) were purchased from Shanghai Slake Laboratory Animal Co., Ltd. The animals used and the animal experiments performed adhered to the guidelines of the Declaration of Helsinki and were approved by the Experimental Animal Care and Animal Welfare Ethics Committee of Jiangnan University (JN.No20230615i1000906

[304] ).

[0101] The behavioral testing methods involved in the following embodiments are:

[0102] (1) Open field (OFT)

[0103] The open field test is primarily used to evaluate the spontaneous activity and exploratory behavior of mice. The open field apparatus consists of a 40 cm long, 40 cm wide, and 50 cm high experimental box, with an infrared camera above to track the mouse's path. The bottom test area is divided into 16 10×10 cm squares, with the four central squares defined as the center area. Each mouse is gently lowered from the center of the open field. The person immediately leaves the experimental apparatus and allows the mouse to move freely for 8 minutes. Video recording software records the mouse's speed, distance traveled, time spent in the center area, and other indicators. After each experiment, the mouse's hair, urine, and feces are cleaned, and the experimental box is wiped with 75% alcohol to eliminate the effects of odor. The alcohol is evaporated with a paper towel and a hair dryer before the next mouse is tested.

[0104] (2) Novel Object Recognition (NORT)

[0105] The novel object recognition (NOR) test is a learning and memory assessment method developed by leveraging rodents' natural tendency to explore novel objects. The experimental setup is similar to the open-field test, with the experimental period consisting of an acclimation period, a familiarization period, and a test period, each separated by 24 hours. During the acclimation period, mice were placed in an experimental chamber and allowed to freely explore the environment for 8 minutes to eliminate the novelty and fear of unfamiliar surroundings. During the familiarization period, two identical blocks were placed on the opposite side of the experimental chamber, 5 cm from either corner. The mice were placed in the center of the previous side, facing away from the blocks, and allowed to freely explore the blocks and the surroundings for 8 minutes. After 24 hours, one of the blocks was replaced with a new block of a different shape, and the mice were allowed to freely explore the blocks and the surroundings for another 8 minutes, following the familiarization protocol. After each experiment, the experimental chamber was wiped clean with 75% alcohol to eliminate odors.

[0106] During the experiment, an infrared camera tracked the time (T) the mice spent exploring the new and old objects. The discrimination index (DI) was calculated as (T new object - T old object) / T total exploration. The blocks should not be too small to prevent the mice from moving or climbing on them. Double-sided tape can be used to secure the blocks.

[0107] (3) Y-maze (YMT)

[0108] The Y-Maze Test (YMT) is primarily used to assess working or reference memory in rodents. The experimental apparatus consists of three horizontal arms, each 70 cm long and 15 cm high, angled 120° apart. The arms are made of opaque polyethylene plastic. During the experiment, the three arms are arbitrarily designated as abc. At the start of the experiment, mice are placed into any arm, and an infrared camera records the order and number of times they enter each arm. Successive entries into different arms are defined as a single alternation (e.g., abc, bac, cba, but not aca).

[0109] The Y-maze performance was evaluated using the following criteria: spontaneous alternation = number of alternations / (total number of arm entries - 2). Each test lasted 5 minutes. After each experiment, the experimental chamber was wiped clean with 75% alcohol to eliminate the effects of odor.

[0110] (4) Morris water maze test (MWM)

[0111] The Morris water maze test is one of the classic methods for assessing spatial memory in 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, marked as east, south, west, and north. Adjustable light sources are set on both sides to adjust the brightness of the water surface, and a camera is installed on the top of the pool to record experimental data. During the experiment, the pool is filled with 40 cm of water, dyed black with ink, and the water temperature is maintained at 21±1°C.

[0112] The experiment is divided into two phases: place navigation and spatial probe. After each swimming test, the mice will be dried and kept warm with a clean towel or hair dryer. During the training phase, a platform is placed 1 cm below the water surface and fixed in a certain quadrant. At the beginning of each trial, the mouse is placed on the side facing the wall of the pool and enters the water from a certain quadrant. An infrared camera is used to track the swimming path of the mouse. The test time limit is 60 seconds, and the time required for the mouse to find the platform is recorded as the escape latency. When the mouse finds the platform, it can stay on the platform for 5 seconds. If the platform is not found 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, it is necessary to avoid placing the mouse directly on the platform. Instead, use a guide stick to guide the mouse in the swimming direction to help it learn how to perceive and climb the platform, so as to remember its location.

[0113] Each mouse underwent four training trials daily, entering the water from a different quadrant each time, with at least 30 minutes between trials. Training lasted for five days; on the sixth day, a spatial exploration test was conducted. The platform was removed from the pool, and the mouse entered the water from the wall of the quadrant diagonally opposite the platform's original position. The test lasted 60 seconds. Parameters such as swimming speed, latency, swimming path, and the number of platform crossings were recorded. Scopolamine was not administered to the mice during the spatial exploration phase.

[0114] The nutrient detection methods involved in the following embodiments are:

[0115] (1) Phospholipids

[0116] Refer to GB / T 5537-2008 Grain and oil inspection-Determination of phospholipid content.

[0117] (2) Vitamin E

[0118] Refer to GB 5009.82-2016 National Food Safety Standard Determination of Vitamins A, D and E in Foods.

[0119] (3) Phytosterols

[0120] Refer to GB / T 25223-2010 Determination of sterol composition and total sterol content in animal and vegetable oils and fats.

[0121] (4) Squalene

[0122] Refer to LS / T 6120-2017 Inspection of grains and oils—Determination of squalene in vegetable oils.

[0123] The statistical methods involved in the following embodiments are:

[0124] All experimental results were statistically analyzed using SPSS (Statistical Product and Service Solutions) statistical processing software, and the differences between the groups were analyzed using t-test, with P < 0.05 indicating statistical significance.

[0125] It's important to note that the t-test is a probabilistic statistical method primarily used for small sample sizes and normal distributions with unknown population standard deviations. The P value represents the probability of obtaining the current sample if the null hypothesis is correct (P < 0.05 means the probability of obtaining the former with the latter is less than 0.05, meaning the correlation between the two is less than 0.05, indicating a statistically significant difference). In the following text, "t" and "P" have the same meaning.

[0126] The preparation method of crude oil in the following embodiment:

[0127] (1) Preparation of sunflower seed crude oil:

[0128] Crude sunflower seed oil is prepared by a pressing process. The sunflower seeds are shelled, cleaned, crushed, tempered, rolled, steamed, pressed, and filtered to obtain the crude oil. The main control parameters include: steaming temperature of 130°C, time of 25 minutes, and pressing material temperature controlled at 90°C.

[0129] (2) Preparation of crude corn oil:

[0130] Corn crude oil is prepared by a pressing process. The corn germ is cleaned, crushed, tempered, rolled, steamed, pressed, and filtered to obtain pressed crude oil. The main control parameters include: steaming temperature 100℃, time 25min, and pressing material temperature controlled at 90℃.

[0131] (3) Preparation of crude rapeseed oil:

[0132] Rapeseed crude oil is prepared by a pressing process. The rapeseed is cleaned, crushed, tempered, rolled, steamed, pressed, and filtered to obtain the pressed crude oil. The main control parameters include: steaming temperature of 120°C, time of 25 minutes, and pressing temperature of 100°C. The apparatus involved in the following embodiments:

[0133] (1) Metering pump (traditional control pipeline before transformation):

[0134] Schematic diagram of traditional acid / alkali dosing pump in neutralization process Figure 1 As shown, a conventional 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) works in conjunction with the mechanical device (2), allowing the conveying medium to flow in and out of the pump head (1). The stroke (3) is adjusted by the manual stroke adjustment knob to achieve the purpose of regulating the flow rate. Depending on the flow rate requirements, two or three pumps can be connected in series.

[0135] (2) Intelligent control system (the modified control pipeline used in the present invention):

[0136] The process flow chart after the transformation of the acid / alkali precise addition system in the neutralization section is as follows Figure 2 As shown, 1 is a crude oil relay tank, 2 is a phosphoric acid storage tank, 3 is a liquid alkali storage tank, 4 is an acid reaction tank, 5 is an alkali reaction tank, 6 is a 1# centrifuge, 7 is a 2# centrifuge, 8 is a water washing mixing tank, 9 is a dryer, 10 is a 1# mixer, 11 is a 2# mixer, 12 is a 3# mixer, 13 is an electromagnetic pump, 14 is an electromagnetic proportional valve, 15 is an electromagnetic flowmeter, 16 is an electromagnetic pump, 17 is an electromagnetic proportional valve, and 18 is an electromagnetic flowmeter.

[0137] The crude oil passes through the crude oil relay tank (1) and is mixed with phosphoric acid solution in the 1# mixer (10) and enters the acid reaction tank (4) for full hydration and degumming reaction. The crude oil is then mixed with alkaline solution in the 2# mixer (11) and enters the alkaline reaction tank (5) for full reaction. The crude oil flows into the 1# centrifuge (6) for centrifugation. Soft water is added to the crude oil and the crude oil is mixed in the 3# mixer (12) and enters the water washing tank (8) for water washing. The crude oil then enters the 2# centrifuge (7) for centrifugation and is dried in the dryer (9) to obtain neutralized oil.

[0138] When adding phosphoric acid solution and alkaline solution, electromagnetic pumps (13, 16), electromagnetic proportional valves (14, 17), and electromagnetic flowmeters (15, 18) are respectively combined to replace the original metering pumps, and the crude oil flow rate and the amount of free fatty acids are combined to feed back signals to a PLC (programmable logic controller) to calculate the theoretical addition amount. The opening of the proportional control valve is adjusted and compared with the actual flow rate measured by the electromagnetic flowmeter, and the opening of the proportional valve is adjusted in real time to optimize the accurate addition amount of the phosphoric acid solution, alkaline solution and other media.

[0139] (3) Double-tower combined dual-temperature deodorization device:

[0140] The deodorization process flow chart of double tower combination and double temperature deodorization is as follows Figure 3 As shown;

[0141] After the bleached oil is heated with deodorized oil in heat exchanger #3 for the first time, it enters heat exchanger #2 for a second heating. It then enters the plate tower for the first stage of deodorization. It then enters heat exchanger #1 (heater) to raise the temperature before entering the packed tower for the second stage of deodorization. The deodorization process is carried out under negative pressure (the dotted line represents the negative pressure pipeline). The deodorized distillate is collected in a collector. The entire process effectively utilizes the waste heat of the deodorized oil to maintain the deodorization temperature in the first stage, and utilizes heating and heat exchange in the furnace to achieve the instantaneous high temperature in the packed tower in the second stage.

[0142] For the solutions involved in the examples, if the solvent is not specified, water is used as the solvent; if the reaction temperature is not specified, room temperature (25° C.) is used.

[0143] Phosphoric acid solution and NaOH solution are added using a solenoid pump, solenoid proportional valve, and electromagnetic flowmeter. The system, combined with crude oil flow and free fatty acid levels, feeds back signals to a PLC (Programmable Logic Controller) to calculate the theoretical addition amount. The proportional control valve opening is then adjusted, compared to the actual flow rate measured by the electromagnetic flowmeter, and feedback is then used to adjust the proportional valve opening in real time to optimize the precise addition amount of the phosphoric acid solution, NaOH solution, and other media.

[0144] The packing in the packed tower is specifically Swiss Sulzer low pressure drop stainless steel structured packing.

[0145] Example 1

[0146] A method for preparing refined sunflower oil with high retention of nutritional concomitants comprises the following steps:

[0147] (1) Neutralization section (degumming + deacidification): the modified control pipeline is used

[0148] The crude sunflower oil in the crude oil relay tank is maintained at a flow rate of 21 T / h, the temperature is preheated to 55°C, and a phosphoric acid solution with a concentration of 80% (w / w) is accurately pumped in at a flow rate of 0.5 L / h. The solution is mixed in the 1# mixer and then enters the acid reaction tank at 55°C for hydration degumming reaction for 30 minutes to obtain a hydration degummed product;

[0149] After the reaction is completed, the hydrated degummed product is pumped with a 9% (w / w) NaOH solution at a flow rate of 104 L / h and soft water at a flow rate of 750 L / h, mixed in a 2# mixer, and then transferred to an alkali reaction tank with a stirring speed of 42 Hz for deacidification reaction at 70°C for 30 minutes. After the reaction is completed, the product is centrifuged in a 1# soap removal centrifuge at a back pressure of 3.0 bar and transferred to a 3# mixer. Soft water is added to the 3# mixer at a flow rate of 772 L / h and then transferred to a water washing tank for water washing. After centrifugation in a 2# centrifuge at a back pressure of 3.0 bar, the product is vacuum dried in a dryer at 105°C for 20 minutes to obtain a neutralized oil.

[0150] (2) Dewaxing section:

[0151] 0.62% (w / w) diatomaceous earth was added to the neutralized oil, cooled to 6.8°C, stirred for 23.8 hours, and filtered to obtain dewaxed oil;

[0152] (3) Decolorization section:

[0153] The dewaxed oil was heated to 105°C under vacuum, stripping steam was 0.9 bar, 0.5% (w / w) decolorizing activated clay was added and adsorbed for about 30 minutes, and filtered to obtain the decolorized oil;

[0154] (4) Deodorization section (double-tower combination dual-temperature deodorization process):

[0155] The decolorized oil is heated to 190°C by the 3# heat exchanger and enters the 2# heat exchanger for a second heating to 210°C. The decolorized oil is then pumped into the plate tower for the first stage of deodorization for 90 minutes with stripping steam at 0.9 bar.

[0156] After that, it enters the 1# heat exchanger (heater) to heat up to 230℃, and the oil is pumped into the packed tower for the second stage of deodorization for 10 minutes, with stripping steam at 0.9 bar;

[0157] The vacuum system is maintained at 2 mbar throughout the deodorization process to obtain refined sunflower oil.

[0158] The refined sunflower oil contains aflatoxin B1 ≤ 10 μg / kg and benzo[a]pyrene ≤ 10 μg / kg, and other major physical, chemical and food safety indicators meet the requirements of the national standard "GB / T 10464 Sunflower Oil".

[0159] Comparative Example 1

[0160] A method for preparing traditional refined sunflower oil comprises the following steps:

[0161] (1) Neutralization section (degumming + deacidification): using the traditional control pipeline before transformation

[0162] Driven by a motor, the crude sunflower seed oil in the crude oil relay tank has a flow rate of 21 T / h and is preheated to 55° C. 80% (w / w) phosphoric acid solution is pumped in at a flow rate of 0.5 L / h through a pump head and a mechanical device using a manual stroke adjustment knob to adjust the stroke. The solution is mixed in a #1 mixer and then enters an acid reaction tank for a hydration degumming reaction at 55° C. for 30 minutes.

[0163] After the reaction is completed, a 9% (w / w) NaOH solution is pumped into the 2# mixer at a flow rate of 104 L / h, and soft water is mixed in the 2# mixer at a flow rate of 750 L / h. The flow rate control method is the same as the phosphoric acid solution in (1); after mixing, the mixture enters the alkali reaction tank, the stirring speed is 42 Hz, and the deacidification reaction is carried out at 70°C for 30 minutes; after the reaction is completed, the mixture is centrifuged in the 1# soap removal centrifuge at a back pressure of 3.0 bar and placed in the 3# mixer, and soft water is added to the 3# mixer at a flow rate of 772 L / h for mixing, and then enters the water washing tank for water washing, and after centrifugation in the 2# centrifuge at a back pressure of 3.0 bar, it is vacuum dried in a dryer at 105°C for 20 minutes to obtain a neutralized oil;

[0164] (2) Dewaxing section:

[0165] Same as step (2) in Example 1;

[0166] (3) Decolorization section:

[0167] Same as step (2) in Example 1;

[0168] (4) Deodorization section:

[0169] The decolorized oil enters the deodorization section. The deodorization section uses a packed tower and a plate tower with the same temperature, controlled at about 230°C, a deodorization time of 92 minutes, and a stripping steam of 0.9 bar. The vacuum system is maintained at 2 mbar throughout the deodorization section to obtain the original traditional refined sunflower oil.

[0170] The test results of the main nutrients (phospholipids, vitamin E, phytosterols, squalene) of the refined oils prepared in Example 1 and Comparative Example 1 are shown in Table 1.

[0171] Table 1

[0172]

[0173] Comparative Example 2

[0174] The specific implementation is the same as that of Example 1, except that the adjustment step (5) is:

[0175] (5) Deodorization section:

[0176] The decolorized oil is heated to 200°C by the 3# heat exchanger, and then enters the 2# heat exchanger for a second heating to 210°C. The decolorized oil is then pumped into the plate tower for the first stage of deodorization for 90 minutes with 0.9 bar stripping steam. After that, the oil enters the 1# heat exchanger (heater) to heat up to 260°C, and then is pumped into the packed tower for the second stage of deodorization for 10 minutes with 0.9 bar stripping steam.

[0177] The vacuum system is maintained at 2 mbar throughout the deodorization process to obtain refined sunflower oil.

[0178] Comparative Example 3

[0179] The specific implementation is the same as that of Example 1, except that the adjustment step (5) is:

[0180] (5) Deodorization section:

[0181] The decolorized oil is heated to 210°C by the 3# heat exchanger and then enters the 2# heat exchanger for a second heating to 220°C. The decolorized oil is then pumped into the plate tower for the first stage of deodorization for 90 minutes with stripping steam at 0.9 bar.

[0182] After that, it enters the 1# heat exchanger (heater) to heat up to 260℃, and the oil is pumped into the packed tower for the second stage of deodorization for 10 minutes, with stripping steam at 0.9 bar;

[0183] The vacuum system is maintained at 2 mbar throughout the deodorization process to obtain refined sunflower oil.

[0184] Comparative Example 4

[0185] The specific implementation is the same as that of Example 1, except that the adjustment step (5) is:

[0186] (5) Deodorization section:

[0187] The decolorized oil is heated to 220°C by the 3# heat exchanger and then enters the 2# heat exchanger for a second heating to 230°C. The decolorized oil is then pumped into the plate tower for the first stage of deodorization for 100 minutes with stripping steam at 0.9 bar.

[0188] After that, it enters the 1# heat exchanger (heater) to heat up to 260℃, and the oil is pumped into the packed tower for the second stage of deodorization for 10 minutes, with 0.9 bar of stripping steam; the vacuum system is maintained at 2 mbar throughout the deodorization process to obtain refined sunflower oil.

[0189] The obtained refined sunflower oil was subjected to performance testing, and the test results are as follows:

[0190] Table 2

[0191]

[0192] Table 3

[0193] Nutrient categories Example 1 Yield (%) Comparative Example 2 Yield (%) Comparative Example 3 Yield (%) Comparative Example 4 Yield (%) phospholipids 1.60 1.53 1.51 1.50 Vitamin E 94.3 80.9 71.6 67.3 Phytosterols 90.0 80.2 77.3 74.7 Squalene 80.0 76.0 70.0 66.0

[0194] Example 2

[0195] The specific implementation is the same as that of Example 1, except that the crude sunflower seed oil is replaced with crude corn oil.

[0196] The refined corn oil contains aflatoxin B1 ≤ 20 μg / kg and benzo[a]pyrene ≤ 10 μg / kg, and its other main physical and chemical indicators and food safety meet the requirements of the national standard "GB / T 19111 Corn Oil".

[0197] Comparative Example 5

[0198] Specific implementation method is the same as comparative example 1, except that the sunflower seed crude oil is changed to corn crude oil.

[0199] The test results of the main nutrients (phospholipids, vitamin E, phytosterols, squalene) of the refined oils prepared in Example 2 and Comparative Example 5 are shown in Table 4.

[0200] Table 4

[0201]

[0202] Example 3

[0203] The specific implementation is the same as that of Example 1, except that the crude sunflower seed oil is replaced with crude rapeseed oil.

[0204] The refined rapeseed oil contains aflatoxin B1 ≤ 10 μg / kg and benzo[a]pyrene ≤ 10 μg / kg, and other major physical and chemical indicators and food safety meet the requirements of the national standard "GB / T 1536 Rapeseed Oil".

[0205] Comparative Example 6

[0206] The specific implementation method is the same as that of Comparative Example 1, except that the crude sunflower seed oil is replaced with crude rapeseed oil.

[0207] The test results of the main nutrients (phospholipids, vitamin E, phytosterols, squalene) of the refined oils prepared in Example 3 and Comparative Example 6 are shown in Table 5.

[0208] Table 5

[0209]

[0210] Example 4

[0211] A method for preparing traditional refined sunflower oil comprises the following steps:

[0212] (1) Neutralization section (degumming + deacidification): using the traditional control pipeline before transformation

[0213] Driven by a motor, the crude sunflower seed oil in the crude oil relay tank has a flow rate of 21T / h and is preheated to 55°C. The stroke is adjusted manually by a stroke adjustment knob through the pump head and a mechanical device to maintain an 80% (w / w) phosphoric acid solution pumped in at a flow rate of 0.5L / h, mixed in the 1# mixer, and then enters the acid reaction tank for a hydration degumming reaction at 55°C for 30 minutes. After the reaction is completed, a 9% (w / w) NaOH solution and a 2# mixer are pumped in at a flow rate of 104L / h and a 750L / h. Soft water is mixed in the 2# mixer, and the flow control method is the same as that of the phosphoric acid solution in (1). After mixing, the mixture enters the alkaline reaction tank, the stirring speed is 42 Hz, and the deacidification reaction is carried out at 70°C for 30 minutes. After the reaction, it is centrifuged in the 1# soap removal centrifuge at a back pressure of 3.0 bar and placed in the 3# mixer. Soft water is added to the 3# mixer at a flow rate of 772 L / h for mixing, and then enters the water washing tank for water washing. After centrifugation in the 2# centrifuge at a back pressure of 3.0 bar, it is vacuum dried in a dryer at 105°C for 20 minutes to obtain neutralized oil.

[0214] (2) Dewaxing section:

[0215] Same as step (2) in Example 1;

[0216] (3) Decolorization section:

[0217] Same as step (2) in Example 1;

[0218] (4) Deodorization section:

[0219] The decolorized oil is heated to 190°C by the 3# heat exchanger and enters the 2# heat exchanger for a second heating to 210°C. The decolorized oil is then pumped into the plate tower for the first stage of deodorization for 90 minutes with stripping steam at 0.9 bar.

[0220] After that, it enters the 1# heat exchanger (heater) to heat and raise the temperature to 230℃, and the oil is pumped into the packed tower for the second stage of deodorization for 10 minutes, with steam stripping at 0.9 bar; the vacuum system is maintained at 2 mbar throughout the deodorization process to obtain refined sunflower oil.

[0221] The obtained refined sunflower oil was subjected to performance testing, and the test results are as follows:

[0222] The content of phospholipids in the refined sunflower seed oil is 12.9 mg / kg, with a yield of 0.25%; the content of vitamin E is 620 mg / kg, with a yield of 88.6%; the content of phytosterol is 4328 mg / kg, with a yield of 86.6%; and the content of squalene is 78.0 mg / kg, with a yield of 78.0%.

[0223] Example 5

[0224] The refined sunflower oil obtained in Example 1 is subjected to nutritional fortification, and the specific steps are as follows:

[0225] (1) Preparation of vitamin A and vitamin D premixed oil

[0226] Vitamin A premixed oil:

[0227] 0.893 kg of vitamin A crystals were weighed using a balance and dissolved in 1.607 kg of the refined sunflower oil prepared in Example 1 to obtain 2.5 kg of vitamin A 1.0 MIU raw material oil; 0.0936 kg of vitamin A 1.0 MIU raw material oil was weighed and mixed into 1.9064 kg of the refined sunflower oil prepared in Example 1 to prepare the vitamin A premixed oil.

[0228] Vitamin D premixed oil:

[0229] 0.15 g of vitamin D crystals were weighed and dissolved in 1 kg of the refined sunflower oil prepared in Example 1 to obtain 0.15 g / kg of vitamin D premixed oil.

[0230] (2) Vitamin A premixed oil and vitamin D premixed oil were mixed into the refined sunflower oil obtained in Example 1 at a ratio of 0.625% and 0.050% (w / w), respectively, and stirring was continued for 15 minutes to obtain nutritionally fortified sunflower oil; wherein the added concentration of vitamin A was 6200 μg / kg, and the added concentration of vitamin D was 75 μg / kg.

[0231] The sunflower oil fortified with nutrients obtained above was filled into yellow diamond PET bottles that could be protected from light; and the vitamins A and D were determined. The error between the control and theoretical design was within 10%, indicating that the product was qualified.

[0232] Comparative Example 7

[0233] The refined sunflower oil obtained in Example 1 is subjected to nutritional fortification, and the specific steps are as follows:

[0234] Vitamin A premix oil was prepared according to step 1 of Example 5, and was mixed into the refined sunflower oil obtained in Example 1 at a ratio of 0.625% (w / w). The mixture was stirred for 15 minutes to obtain nutritionally fortified sunflower oil.

[0235] Comparative Example 8

[0236] The refined sunflower oil obtained in Example 1 is subjected to nutritional fortification, and the specific steps are as follows:

[0237] Vitamin D premix oil was prepared according to step 1 of Example 5, and was mixed into the refined sunflower oil obtained in Example 1 at a ratio of 0.050% (w / w). The mixture was stirred for 15 minutes to obtain nutritionally fortified sunflower oil.

[0238] Comparative Example 9

[0239] The refined sunflower oil obtained in Example 4 was subjected to nutritional enhancement, and the specific steps were as follows:

[0240] Vitamin A premix oil was prepared according to step 1 of Example 5, and was mixed into the refined sunflower oil obtained in Example 4 at a ratio of 0.625% (w / w). The mixture was stirred for 15 minutes to obtain nutritionally fortified sunflower oil.

[0241] Comparative Example 10

[0242] The refined sunflower oil obtained in Example 4 was subjected to nutritional enhancement, and the specific steps were as follows:

[0243] Vitamin D premix oil was prepared according to step 1 of Example 5, and was mixed into the refined sunflower oil obtained in Example 4 at a ratio of 0.050% (w / w). The mixture was stirred for 15 minutes to obtain nutritionally fortified sunflower oil.

[0244] Example 6

[0245] The refined sunflower oil obtained in Example 4 was subjected to nutritional enhancement, and the specific steps were as follows:

[0246] Vitamin A and vitamin D premixed oils were prepared according to step 1 of Example 5 and mixed into the refined sunflower oil obtained in Example 1 at a ratio of 0.625% (w / w) and 0.050% (w / w), respectively. Stirring was continued for 15 minutes to obtain nutritionally fortified sunflower oil; wherein the added concentration of vitamin A was 6200 μg / kg, and the added concentration of vitamin D was 75 μg / kg.

[0247] Example 7

[0248] Mice were housed under a 12-hour light-dark cycle and provided with ample water and custom-made chow. Experiments were conducted after a 1-week pre-feeding period. The experimental design was optimized to minimize the number of animals used and the pain caused to the animals.

[0249] Animal experiments were conducted using 5-week-old healthy ICR male mice to demonstrate that the prepared nutritionally fortified sunflower oil has the effect of protecting brain cognition.

[0250] The specific experimental methods are:

[0251] The above-mentioned 5-week-old healthy ICR male mice were used for animal experiments, with a 12 / 12-h light cycle (8:00-20:00), constant temperature (23±2°C), and constant humidity (60±5%). All experimental operations were carried out in accordance with the requirements of the Experimental Animal Management and Animal Welfare Ethics Committee of Jiangnan University.

[0252] The 5-week-old healthy ICR male mice were adaptively fed for 1 week and then randomly divided into 6 groups: control group (Con), scopolamine model group (Sop), rivastigmine positive control group (Riv), vitamin D group (D), vitamin D + vitamin A group (DA), phospholipid group (P), phospholipid + vitamin A group (PA), and phospholipid + vitamin A + vitamin D group (PAD), with 10 mice in each group.

[0253] 1. Feed preparation

[0254] Feed A: 140 g / kg casein, 1.8 g / kg L-cysteine, 495.7 g / kg corn starch, 10125 g / kg maltodextrin, 100 g / kg sucrose, 50 g / kg cellulose, 35 g / kg Mineral Mix S10022M (mixed minerals S10022M, purchased from Jiangsu Collaborative Biotechnology Co., Ltd.), and the remainder corn starch; 40 g / kg sunflower oil obtained in Example 4 was added to feed A. Different sunflower oils were used in other groups of this experiment according to experimental needs.

[0255] Feed B: Feed A was supplemented with 40 g / kg of sunflower oil fortified with vitamin D obtained in Comparative Example 10;

[0256] Feed C: Feed A supplemented with 40 g / kg of sunflower oil fortified with vitamin D and vitamin A obtained in Example 6;

[0257] Feed D: 40 g / kg of sunflower oil obtained in Example 1 was added to Feed A. This sunflower oil retained an appropriate amount of phospholipids through the process of the present invention.

[0258] Feed E: 40 g / kg of sunflower oil fortified with phospholipids and vitamin A obtained in Comparative Example 7 was added to Feed A.

[0259] Feed F: Feed A was supplemented with the retained phospholipids obtained in Example 5 and 40 g / kg of sunflower oil fortified with vitamin A and vitamin D.

[0260] 2. Mouse experiments

[0261] The Con group received daily intraperitoneal injections of normal saline, while the other groups received daily intraperitoneal injections of 3 mg / kg·bw scopolamine (scopolamine hydrobromide dissolved in sterile normal saline, referred to as scopolamine solution) at a volume of 10 mL / kg·bw (where bw represents the body weight of the mouse).

[0262] During the modeling, mice were given feed every day for intervention, feeding once a day, 4 g per mouse each time; mice in each group had free access to water.

[0263] The details are as follows:

[0264] Control group (Con): The rats were intraperitoneally injected with normal saline once a day at a volume of 10 mL / kg·bw and fed with feed A daily;

[0265] Scopolamine model group (Sop): 3 mg / kg·bw scopolamine (modeling) was intraperitoneally injected daily, with an injection volume of 10 mL / kg·bw, and fed with feed A;

[0266] Rivastigmine positive control group (Riv): scopolamine 3 mg / kg·bw was intraperitoneally injected daily (model establishment), with an injection volume of 10 mL / kg·bw. Rivastigmine was injected intraperitoneally again 30 minutes after model establishment every day, with an injection dose of 2 mg / kg·bw. The rats were fed diet A.

[0267] Vitamin D group (D): scopolamine 3 mg / kg·bw was intraperitoneally injected daily, the injection volume was 10 mL / kg·bw, and feed B was fed;

[0268] Vitamin D + vitamin A group (DA): scopolamine 3 mg / kg·bw was intraperitoneally injected daily, the injection volume was 10 mL / kg·bw, and feed C was fed;

[0269] Phospholipid group (P): scopolamine 3 mg / kg·bw was intraperitoneally injected daily, the injection volume was 10 mL / kg·bw, and feed was fed with feed D;

[0270] Phospholipid + vitamin A group (PA): scopolamine 3 mg / kg·bw was intraperitoneally injected daily, the injection volume was 10 mL / kg·bw, and feed E was fed;

[0271] Phospholipid + vitamin A + vitamin D group (PAD): 3 mg / kg·bw scopolamine was intraperitoneally injected daily, with an injection volume of 10 mL / kg·bw, and feed F was fed.

[0272] After 4 weeks of continuous modeling and intervention, the mice were subjected to behavioral tests, including open field (OFT), novel object recognition (NORT), Y-maze (YMT) and Morris water maze (MWM) tests.

[0273] Subsequently, after fasting for 12 hours, the mice were anesthetized by intraperitoneal injection of 10% chloral hydrate and then killed by cervical dislocation. The blood and cerebral cortex of the mice were collected for subsequent studies.

[0274] Here are the results:

[0275] 1. Results of mouse behavioral experiments

[0276] The open field test (OFT) was used to investigate the effects of the edible oils obtained in the above examples and comparative examples on the exploratory behavior of scopolamine-treated mice. In a novel environment, the exploratory behavior of mice can be assessed by the amount of time they spend in the central area. Longer stays in the central area indicate that the animals are more willing to explore the open space, reflecting higher exploratory motivation and lower anxiety.

[0277] Table 6 Nutritionally fortified sunflower oil improves the length of time in the central area of scopolamine model mice

[0278] Grouping Center area time (seconds) Con 72.95±3.52 Sop <![CDATA[42.34±7.99 ## ]]> Riv <![CDATA[118.75±9.11 ** ]]> VD 50.81±8.74 DA <![CDATA[78.29±8.45 ** ]]> P <![CDATA[73.31±4.35 ** <!-- 15 -->]]> PA <![CDATA[74.18±8.89 ** ]]> PAD <![CDATA[94.96±9.38 ** ]]>

[0279] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0280] As shown in Table 6, the Sop model group significantly reduced the time and percentage spent in the central area compared to the blank group. The added functional factors phospholipids and vitamin A had a certain improvement effect. Among them, the DA, P, PA, and PAD groups significantly increased the time and percentage spent in the central area (p < 0.01). The effect of the VD group in the open field test was not obvious, but the combined use of vitamin A and vitamin D made the improvement statistically significant, and the experimental data showed that it was better than single-component vitamin D. Among the multi-component intervention groups, the combination of phospholipids, vitamin A, and vitamin D had the best effect, better than the two-component combination.

[0281] A Y-maze was used to investigate the effect of the edible oils obtained in the above examples and comparative examples on the working memory of mice. Spontaneous alternation reflects the working memory ability of mice. Working memory involves the ability to retain and manipulate information in the short term. In the Y-maze experiment, mice tend to explore new arms and avoid returning to the arms they recently explored, which requires them to remember where they have recently been. If mice can successfully perform spontaneous alternation, it shows that they have good working memory.

[0282] The results are shown in Table 7. There was a significant difference between the model group and the blank group (Con) (P < 0.05), indicating that the model was successfully established. Except for the P group, the other intervention groups were able to significantly increase the number of spontaneous alternations in mice, and the combined use of phospholipids, vitamin A, and vitamin D was better than that of the two components.

[0283] Table 7 Nutritionally fortified sunflower oil improves the number of spontaneous alternations in scopolamine model mice

[0284] Grouping Spontaneous alternation (times) Con 0.408±0.02 Sop <![CDATA[0.327±0.03 # ]]> Riv <![CDATA[0.466±0.02 ** ]]> VD <![CDATA[0.469±0.02 ** ]]> DA <![CDATA[0.429±0.01 ** ]]> P 0.337±0.01 PA <![CDATA[0.388±0.03 * ]]> PAD <![CDATA[0.458±0.04 ** ]]>

[0285] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0286] A novel object recognition experiment was used to explore the intervention effect of the edible oils obtained in the above examples and comparative examples on mice's difficulty in recognizing novel objects. In the NOR experiment, mice were exposed to two objects: a familiar object and a new object. Healthy mice typically spend more time exploring new objects because they have more interest and curiosity in new objects. If mice can recognize new objects and spend more time on them, this indicates that their memory function is normal, so the discrimination index can effectively reflect the memory function of mice.

[0287] The results, as shown in Table 8, show that the discrimination index (DI) in the Sop model group was significantly lower than in the blank group, indicating successful model establishment. The intake of vitamin D and vitamin A significantly improved the DI (p < 0.01), suggesting that scopolamine impairs learning and cognitive memory in mice. The experimental data show that all four intervention groups significantly improved the discrimination index of the model mice (p < 0.01). Numerically, the combined use of phospholipids, vitamin A, and vitamin D was superior to the two-component intervention group, followed by phospholipids + vitamin A.

[0288] Table 8 Nutritionally fortified sunflower oil improves the discrimination index of scopolamine model mice

[0289] Grouping Discrimination Index Con 0.546±0.03 Sop <![CDATA[0.126±0.01 ## ]]> Riv <![CDATA[0.457±0.02 ** ]]> VD <![CDATA[0.577±0.03 ** ]]> DA <![CDATA[0.400±0.03 ** ]]> P <![CDATA[0.316±0.01 ** ]]> PA <![CDATA[0.407±0.04 ** ]]> PAD <![CDATA[0.420±0.01 ** ]]>

[0290] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0291] The Morris Water Maze (MWM) was used to assess the effects of the edible oils obtained in the above examples and comparative examples on improving spatial memory in mice. By having mice search for a hidden platform in a pool of water, their ability to remember and utilize environmental cues can be assessed. This is particularly useful for studying spatial memory impairments, such as those caused by scopolamine.

[0292] During the positioning cruise, the mice's escape latency gradually shortened with the increase in training days, indicating that the training was effective. Statistical analysis of the escape latency on the fifth day revealed significant differences between the model group and the blank group. Vitamin D alone (P < 0.01), phospholipids (P < 0.01), and vitamin D combined with vitamin A (P < 0.01) significantly shortened the mice's escape latency. The combined intervention of phospholipids, vitamin A, and vitamin D had the best effect, with significant differences compared to the other intervention groups (P < 0.01).

[0293] Table 9 Nutritionally fortified sunflower oil shortens the escape latency of scopolamine model mice

[0294]

[0295]

[0296] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0297] The number of platform crossings refers to the number of times the mouse crosses the location of the hidden platform, reflecting the mouse's ability to accurately remember the location of the target platform. It is mainly used to evaluate whether the mouse can accurately remember the location of the platform after repeated training. The more crossings, the stronger the mouse's spatial positioning memory. The number of platform area crossings refers to the number of times the mouse crosses the area where the target platform is located (usually set to an area slightly larger than the platform), which examines the mouse's memory of the approximate location of the platform. This indicator reflects the mouse's overall memory ability for spatial cues and is slightly broader than the number of platform crossings. The cumulative time in the platform area refers to the time the mouse stays in the target platform area, reflecting the mouse's preference for the area and its cognition of the approximate location of the platform. This indicator is used to evaluate whether the mouse has a high willingness to explore and cognitive association with the platform area. The longer the cumulative time, the clearer the mouse's memory of the area.

[0298] As shown in Table 10, during the spatial exploration phase, the model group had significantly lower platform crossing times and cumulative platform time than the blank group (P < 0.01). Regarding platform crossing times, lecithin alone had no significant improvement effect, but lecithin combined with vitamin A significantly increased the number of platform crossings in the model mice (P < 0.01). Regarding cumulative platform time, all intervention groups significantly increased the cumulative platform time in the mice, and the improvement effect of lecithin combined with vitamin A was numerically superior to that of lecithin alone.

[0299] Table 10 Nutritionally fortified sunflower oil improves the spatial exploration ability of scopolamine model mice

[0300] Grouping Number of platform crossings (times) Platform area cumulative time (seconds) Con 3.67±0.94 2.47±0.25 Sop <![CDATA[1.50±0.50 ## ]]> <![CDATA[0.53±0.09 ## ]]> Riv <![CDATA[2.00±0.09 ** ]]> <![CDATA[1.13±0.34 * ]]> VD <![CDATA[3.40±0.49 ** ]]> <![CDATA[1.65±0.26 ** ]]> DA <![CDATA[3.67±0.47 ** ]]> <![CDATA[1.48±0.20 ** ]]> P 1.80±0.75 <![CDATA[1.67±0.25 ** ]]> PA <![CDATA[3.50±0.50 ** ]]> <![CDATA[2.00±0.40 ** ]]> PAD <![CDATA[2.80±0.40 ** ]]> <![CDATA[1.57±0.33 ** ]]>

[0301] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0302] 2. Effects of the above vegetable oils on oxidative stress in mice

[0303] Numerous studies have shown that oxidative stress is one of the important pathogenesis of neurodegenerative diseases. When facing oxidative stress, the body can increase the activity of antioxidant enzymes or use its own natural antioxidants to eliminate excess oxygen free radicals. Currently, there are many indicators that reflect the body's oxidation level, such as GSH (glutathione), MDA (malondialdehyde), SOD (total superoxide dismutase), and GPX (glutathione peroxidase). This study evaluated the effects of vitamin A, vitamin D, phospholipids, and compound groups on the antioxidant effects of mice by measuring these indicators in the blood and cerebral cortex. The gradual accumulation of MDA causes cross-linking and polymerization of biological macromolecules such as nucleic acids and proteins, causing cytotoxicity. Therefore, MDA content is an important indicator of the degree of oxidative damage to the body.

[0304] Blood and cerebral cortex collection and processing: Blood was collected from the retro-orbital venous sinus and placed in EP tubes containing sodium heparin. The tubes were then centrifuged at 3,000 rpm for 15 minutes, and the supernatant was collected for the determination of IL-6 and MDA (malondialdehyde) levels.

[0305] After blood collection, the cerebral cortex of the mice was carefully extracted and quickly homogenized to 10% in pre-chilled sterile saline. The homogenate was then centrifuged at 12,000 rpm for 10 minutes at 4°C, and the supernatant was collected for further analysis. Enzyme-linked immunosorbent assay (ELISA) kits were used to measure relevant biochemical markers according to the manufacturer's instructions (Xiamen Huijia Biotechnology Co., Ltd.). The supernatant of the cerebral cortex was used to measure IL-6 and malondialdehyde (MDA). The remaining sample was stored at −80°C for subsequent use.

[0306] As shown in Table 11, the intake of vitamin D and phospholipids tended to reduce serum malondialdehyde levels, and phospholipids combined with vitamin A significantly reduced serum MDA levels (P < 0.01). Regarding cerebral cortical malondialdehyde levels, there was a significant difference between the model group and the blank group (P < 0.01). Vitamin D or the combined intake of the three functional factors significantly reduced cerebral cortical malondialdehyde levels (P < 0.01). Furthermore, experimental data showed that the combined use of vitamin D and vitamin A, and phospholipids and vitamin A, was more effective than either 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 than the other intervention groups.

[0307] Table 11 Nutritionally fortified sunflower oil reduces malondialdehyde levels in the blood and cerebral cortex of scopolamine model mice

[0308] Grouping Serum malondialdehyde (nmol / L) Malondialdehyde in cerebral cortex (nmol / gpro) Con 1.82±0.07 2.09±0.08 Sop 2.08±0.17 <![CDATA[3.27±0.19 ## ]]> Riv 2.20±0.15 <![CDATA[2.37±0.22 ** ]]> VD 1.88±0.13 <![CDATA[2.28±0.28 ** ]]> DA 1.81±0.06 <![CDATA[1.99±0.22 ** ]]> P 1.85±0.13 3.81±0.26 PA <![CDATA[1.60±0.09 ** ]]> <![CDATA[1.70±0.20 ** ]]> PAD 1.89±0.15 <![CDATA[2.46±0.18 ** ]]>

[0309] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0310] 3. Effects of the above vegetable oils on inflammatory response in mice

[0311] Research indicates that cognitive impairment and memory loss are not only influenced by oxidative stress in the brain, but also by inflammatory responses, which can damage neurons and synaptic structures. Among physiological responses, oxidative stress often induces inflammatory responses. Inflammatory responses, in turn, trigger oxidative stress, mutually reinforcing and exacerbating the condition. IL-6 is a key factor in disrupted inflammatory cytokine release and the onset of inflammatory responses.

[0312] As shown in Table 12, the levels of the inflammatory factor IL-6 in the serum and cerebral cortex of the model group were higher than those in the blank group (P < 0.01). Among the blood indicators, all five intervention groups significantly reduced IL-6 levels in the model mice, and the combined use of vitamin D + vitamin A and phospholipids + vitamin A was superior to either vitamin D or phospholipids alone. In the cerebral cortex, except for the phospholipid group, all other intervention groups significantly reduced IL-6 levels, and the combined use of vitamin D + vitamin A and phospholipids + vitamin A was superior to either vitamin D or phospholipids alone.

[0313] Table 12 Nutritionally fortified sunflower oil reduces IL-6 levels in the blood and cerebral cortex of scopolamine model mice

[0314] Grouping Serum IL-6 (pg / mL) Cerebral cortex IL-6 (pg / mgpro) Con 40.84±1.05 274.84±20.79 Sop <![CDATA[48.69±0.88 ## ]]> <![CDATA[448.00±13.72 ## ]]> Riv 49.28±1.45 <![CDATA[251.01±25.25 ** ]]> VD <![CDATA[40.39±1.76 ** ]]> <![CDATA[338.81±6.14 ** ]]> DA <![CDATA[35.02±1.48 ** ]]> <![CDATA[240.76±9.95 ** ]]> P <![CDATA[40.03±1.72 ** ]]> 426.24±18.64 PA <![CDATA[36.44±2.52 ** ]]> <![CDATA[166.32±3.47 ** ]]> PAD <![CDATA[36.13±1.95 ** ]]> <![CDATA[339.92±14.38 ** ]]>

[0315] All values are expressed as mean ± SD. *p < 0.05, **p < 0.01, compared with the Sop group; #p < 0.05, ##p < 0.01, compared with the Con group, n = 10.

[0316] Although the present invention has been disclosed above in terms of preferred embodiments, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A method for simultaneously improving the retention rate of phospholipids, vitamin E, phytosterols and squalene in refined vegetable oils, characterized in that: The steps include: (1) Neutralization section (degumming + deacidification): The crude oil is preheated to 55°C, and a phosphoric acid solution is pumped into the crude oil at a flow rate of 1-50 L / h to perform a hydration degumming reaction to obtain a hydration degumming product; The product after hydration degumming, alkaline solution and soft water are mixed and stirred to carry out deacidification reaction; after the reaction is completed, centrifugation is carried out, soft water is added for washing, centrifugation is carried out, and drying is carried out to obtain neutralized oil; (2) Dewaxing section: Add a crystallizing agent to the neutralized oil, cool it to 6.8°C, stir it, and filter it to obtain dewaxed oil; (3) Decolorization section: Heat the dewaxed oil to 105°C, add a decolorizing agent for adsorption, and filter to obtain decolorized oil; (4) Deodorization section: The decolorized oil was pumped into a plate tower and heated to 190°C, then heated to 210°C for the first stage of deodorization for 90 minutes; then pumped into a packed tower at 230°C for the second stage of deodorization for 10 minutes to obtain refined vegetable oil. Phosphoric acid solution and alkaline solution are produced using a combination of an electromagnetic pump, electromagnetic proportional valve, and electromagnetic flowmeter. The system, in conjunction with crude oil flow and the amount of free fatty acids, feeds back signals to the PLC, which calculates the theoretical addition amount. This is then compared with the actual flow rate measured by the electromagnetic flowmeter by adjusting the opening of the proportional control valve. The feedback then feeds back to adjust the proportional valve opening in real time, optimizing the precise addition amounts of the phosphoric acid solution and alkaline solution.

2. The refined vegetable oil prepared by the method according to claim 1.

3. A method for preparing a nutritionally enhanced refined vegetable oil, characterized in that: The steps include: Nutrients are added to the refined vegetable oil according to claim 2 for nutritional fortification, wherein the nutrients are one or both of vitamin A and vitamin D.

4. The method according to claim 3, characterized in that The method specifically includes: Vitamin A and vitamin D are added to refined vegetable oil and stirred to obtain nutritionally fortified refined vegetable oil; wherein the added amount of vitamin A is 4000-8000 μg / kg, and the added amount of vitamin D is 50-100 μg / kg.

5. The nutritionally enhanced refined vegetable oil prepared by the method according to claim 3 or 4.

6. Use of the nutritionally fortified refined vegetable oil according to claim 5 in the preparation of products for improving cognitive function or anti-oxidation or anti-inflammatory effects.

Images