Composition for strengthening brain and improving eyesight as well as preparation method, product and application thereof

Abstract

The invention belongs to the technical field of functional food, and particularly relates to a brain-strengthening and eyesight-improving composition as well as a preparation method, a product and application thereof. The composition is prepared from algal oil, linseed oil, arachidonic acid, marigold extract, zeaxanthin suspension oil, NA nervonic acid and PS phosphatidylserine according to a specific weight part ratio. Wherein the marigold extract is prepared by adopting freeze-grinding pretreatment, combining enzymolysis and an ultrasonic-microwave synergistic extraction technology, and performing nanofiltration membrane concentration and vacuum freeze-drying. According to the composition, through the synergistic effect of all the components and an innovative extraction process, the effects of efficiently strengthening the brain and improving eyesight are achieved, and compared with a traditional formula and an extraction method, the component activity retention and the effective component extraction rate are remarkably improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of functional foods, and specifically relates to a composition for strengthening the brain and improving eyesight, and a preparation method, product and application thereof. Background Art

[0002] With the rapid development of technology and significant increases in productivity, electronic products have become deeply integrated into people's work and daily lives. Whether it's long hours spent working in front of a screen or immersive entertainment experiences, the amount of time people spend using their eyes has increased significantly. At the same time, social competition is intensifying, and people are facing increasing pressure from study and life. Chronic mental stress and fatigue are overtaxing the brain. These factors have combined to lead to explosive growth in demand for eye and brain health care.

[0003] Currently, there are numerous products on the market claiming to improve memory and relieve visual fatigue. Among memory-enhancing products, those based on ingredients such as DHA, EPA, vitamins, and zinc are common. Traditional Chinese medicine products often use ingredients such as Alpinia oxyphylla, walnuts, and Gastrodia elata. Lutein esters, zeaxanthin, carotene, and blueberry extracts are particularly popular in products designed to alleviate visual fatigue. Traditional Chinese medicine products often contain ingredients such as goji berries, chrysanthemums, cassia seeds, and bilberries. However, existing technologies present numerous challenges that need to be addressed. First, eye care products offer limited functionality, focusing solely on improving eye problems and failing to meet consumers' diverse health needs for enhancing brain power and memory. Second, existing brain-boosting products often rely on single ingredients or simple formulas. Even with long-term use, some products offer limited results and fail to achieve comprehensive improvements in brain function. Third, the preparation of raw material extracts often relies on single mechanical methods or solvent immersion. These methods not only result in low extraction efficiency but also in limited absorption and utilization by the human body, significantly compromising product efficacy.

[0004] To address these issues, researchers have conducted extensive research, resulting in the emergence of several related invention patents. For example, CN119235021A discloses a composition and its applications for both brain-boosting and vision-enhancing benefits. This composition, comprised of DHA algae oil powder, walnut peptides, and lutein esters, can be formulated into a variety of dosage forms. It effectively improves memory and visual fatigue, and enhances visual persistence. The formulation is scientific, safe, and reliable. Another example is the eye-protecting and brain-enhancing composition disclosed in CN119453474A. By combining extracts of aronia berry, astaxanthin, and DHA, it achieves multi-pathway improvements in vision and cognitive performance, playing a positive role in protecting the retina, promoting blood circulation in the eye, and protecting nerve cells.

[0005] Although these patents have met some of the market demand for eye and brain health products to a certain extent, there is still room for improvement. For example, in terms of raw material selection, the raw material combinations of existing patents have not yet fully explored ingredients with higher eye and brain health benefits; in terms of extraction technology, there is still potential for improvement in the extraction efficiency and activity retention of the active ingredients in the raw materials; and in terms of formula optimization, the synergistic effect between the various ingredients needs to be further strengthened. Therefore, developing a product with better raw material selection, efficient extraction technology, and scientific ingredient ratios that can more effectively achieve multiple benefits such as alleviating visual fatigue, preventing myopia, and enhancing memory has become a technical challenge that needs to be overcome in the current field of eye and brain health products. Summary of the Invention

[0006] In view of the above-mentioned shortcomings, the present invention provides a brain-boosting and eyesight-improving composition with a scientific combination of multiple components, which has multiple functions such as relieving visual fatigue, preventing myopia, and enhancing memory.

[0007] The technical solution of the present invention is: In one aspect, the present invention provides a composition for strengthening the brain and improving eyesight, comprising the following ingredients in parts by weight: 200-240 parts of algae oil, 60-80 parts of linseed oil, 8-12 parts of arachidonic acid, 4-8 parts of marigold extract, 0.6-2 parts of zeaxanthin suspension oil, 0.02-0.04 parts of NA nervonic acid, and 0.4-0.6 parts of PS phosphatidylserine; The preparation method of the marigold extract comprises the following steps: S1. Pretreatment: Freeze marigold flowers, crush them, and vacuum dry them; S2, primary extraction: mixing the pretreated material with the extraction solvent and performing ultrasound-assisted extraction in the presence of a complex enzyme; S3, secondary extraction: the filter residue after the primary extraction is mixed with the extraction solvent again and subjected to variable amplitude ultrasonic extraction; S4, post-processing: combining the extracts, sequentially performing microfiltration, nanofiltration concentration and drying to obtain the marigold extract; The complex enzyme in step S2 includes cellulase, β-glucanase and hemicellulase; The ultrasound-assisted extraction in step S2 uses dual-frequency ultrasound, with a low frequency of 20-30 kHz and a power of 260-350 W, and a high frequency of 100-120 kHz and a power of 150-220 W; In step S3, the frequency of the variable amplitude ultrasound is periodically changed between 20kHz and 40kHz, the change period is 4-6 minutes, and the ultrasound power is 350-450W.

[0008] Preferably, in some embodiments, the low frequency in step S2 is 30kHz, the power is 350W, and the high frequency is 120kHz, the power is 220W; In other embodiments, the low frequency is 20 kHz, the power is 260 W, and the high frequency is 100 kHz, the power is 150 W.

[0009] Preferably, in some embodiments, the frequency of the variable amplitude ultrasound in step S3 is periodically changed between 20kHz and 40kHz, with a change period of 4 minutes, and the ultrasound power is 450W; In other embodiments, the frequency of the variable amplitude ultrasound changes periodically between 20 kHz and 40 kHz, the change period is 6 minutes, and the ultrasound power is 350W.

[0010] Specifically, the mass percentages of the cellulase, β-glucanase and hemicellulase in step S2 are 2-4:1-3:0.5-1.5.

[0011] Preferably, the mass percentage of the cellulase, β-glucanase and hemicellulase in step S2 is 3:2:1.

[0012] Specifically, the extraction time of the ultrasound-assisted extraction in step S2 is 1-2 hours, and the extraction temperature is 35-40°C.

[0013] Specifically, the extraction solvent in step S2 is a 30-50% ethanol aqueous solution; the pH value of the extraction solvent is 4.5-5.5.

[0014] Preferably, the extraction solvent in step S2 is a 40% ethanol aqueous solution; the pH value of the extraction solvent is 5.0.

[0015] Specifically, during the ultrasound-assisted extraction process in step S2, ultrasound is paused for 1-5 minutes every 10-30 minutes, and the mixed solution is stirred at a rotation speed of 150-250 r / min.

[0016] Preferably, in some embodiments, during the ultrasound-assisted extraction process in step S2, ultrasound is paused for 4 minutes every 25 minutes, and the mixed solution is stirred at a speed of 220 r / min.

[0017] Preferably, in other embodiments, during the ultrasound-assisted extraction process in step S2, ultrasound is paused for 2 minutes every 15 minutes, and the mixed solution is stirred at a speed of 180 r / min.

[0018] Specifically, the extraction time of the variable amplitude ultrasonic method in step S3 is 0.8-1.2 hours, and the extraction temperature is 32-38°C.

[0019] Specifically, the freezing treatment in step S1 is performed at a temperature of -30 to -50°C for 8 to 16 hours; The average particle size of the crushed particles is 0.5-1.0 mm; The vacuum degree of the vacuum drying is 5-15 Pa, the temperature is 20-30° C., and the time is 3-5 hours.

[0020] Specifically, the pore size of the filter membrane used in the microfiltration in step S4 is 0.2-0.8 μm; The membrane used in the nanofiltration has a molecular weight cut-off of 500-1000Da; The drying is vacuum freeze drying, and the moisture content after drying is ≤3.0%.

[0021] Preferably, in some embodiments, the composition comprises the following ingredients in parts by weight: 240 parts of algae oil, 60 parts of linseed oil, 8 parts of arachidonic acid, 8 parts of marigold extract, 0.6 parts of zeaxanthin suspension oil, 0.02 parts of NA nervonic acid, and 0.6 parts of PS phosphatidylserine; In other embodiments, the following ingredients are included in parts by weight: 200 parts of algae oil, 80 parts of linseed oil, 12 parts of arachidonic acid, 4 parts of marigold extract, 2 parts of zeaxanthin suspension oil, 0.04 parts of NA nervonic acid and 0.4 parts of PS phosphatidylserine.

[0022] In another aspect, the present invention provides a method for preparing the aforementioned composition, comprising the following steps: (1) Mixing the formulated amounts of algae oil, linseed oil, and arachidonic acid to obtain a uniform oil phase mixture; (2) adding the marigold extract to the oil phase mixture obtained in step (1) and mixing well to obtain a mixture A; (3) adding the zeaxanthin suspension oil to the mixture A obtained in step (2), mixing well, and obtaining a mixture B; (4) Add NA neuraminic acid and PS phosphatidylserine to the mixture B obtained in step (3) and mix well to obtain a composition.

[0023] In another aspect, the present invention provides a functional food, characterized in that it comprises the aforementioned composition.

[0024] Specifically, the health care product also includes auxiliary materials.

[0025] The excipients include but are not limited to: one or more of fillers, disintegrants, lubricants, and flavoring agents.

[0026] The filler includes but is not limited to one or more of lactose, microcrystalline cellulose, starch, and mannitol.

[0027] The disintegrant includes, but is not limited to, one or more of sodium carboxymethyl starch, cross-linked polyvinylpyrrolidone, and low-substituted hydroxypropyl cellulose.

[0028] The lubricant includes, but is not limited to, one or more of magnesium stearate, talc, silicon dioxide, and polyethylene glycol.

[0029] The flavoring agent includes but is not limited to one or more of steviol glycosides, aspartame, sucrose, and flavors.

[0030] In another aspect, the present invention provides the use of the aforementioned composition or functional food in the preparation of a brain-enhancing and eye-improving product.

[0031] Specifically, the product is a functional food or medicine.

[0032] The beneficial effects of the present invention are: (1) The present invention achieves an organic combination of multiple functions such as relieving visual fatigue, preventing myopia, and enhancing memory through the scientific combination of multiple functional ingredients, overcoming the shortcomings of existing eye protection products with single functions, and meeting the needs of consumers for improving brain power and enhancing memory at the same time.

[0033] (2) The present invention adopts multiple extraction methods to prepare marigold extract, thereby improving the extraction efficiency. Furthermore, through scientific concentration and drying processes, high-quality active ingredients are obtained, thereby improving the absorption and utilization efficiency of the product in the body.

[0034] (3) The formula design of the present invention is scientific and reasonable, and the components work synergistically to improve memory and visual fatigue symptoms, increase visual persistence, and have a significant brain-boosting and vision-enhancing effect. (4) The preparation method of the present invention is simple and feasible, with a clear process flow, easy for industrial production, and scientific quality control to ensure the stability and consistency of the product. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 The following are typical graphs of the average fluorescence intensity of apoptotic cells in the eyes of zebrafish in each group, where A is the normal control group; B is the model control group; C is coenzyme Q; D is Example 1; E is Example 2; F is Comparative Example 1; G is Comparative Example 2; H is Comparative Example 3; I is Comparative Example 4; the green fluorescent particles pointed by the red arrows are apoptotic cells.

[0036] Figure 2 The figure shows the retinal pigment epithelium diameter / sclera diameter of each group of zebrafish, where A is the normal control group; B is the MO standard control group; C is the model control group; E is Example 1; D is Atropine sulfate monohydrate; F is Example 2; G is Comparative Example 1; H is Comparative Example 2; I is Comparative Example 3; and J is Comparative Example 4.

[0037] Figure 3 The following are typical diagrams of the movement trajectories of zebrafish in each group, where A is the normal control group; B is the Ginkgo Biloba and Cistanche tablets; C is Example 1; D is Example 2; E is Comparative Example 1; F is Comparative Example 2; G is Comparative Example 3; and H is Comparative Example 4. The blue box is the blue area of ​​the cross maze, which is the quantitative area. DETAILED DESCRIPTION

[0038] The present invention will be further clarified and fully described below by way of examples. The following examples are only a portion of the present invention and are not intended to limit the present invention, but are merely for illustration. The experimental methods used in the following examples are all routine experiments unless otherwise specified, and the materials and reagents used in the following examples are all commercially available unless otherwise specified.

[0039] The purchase sources of the raw materials of the present invention are shown in Table 1: Table 1 Sources of raw materials

[0040] Example 1 1.1 Formulation A composition for strengthening the brain and improving eyesight, the formula of which is shown in Table 2: Table 2 Formulation of Example 1

[0041] 1.2 Preparation method 1.2.1 Preparation method of marigold extract (1) Pretreatment: Fresh and pest-free marigold flowers were selected. After removing the stems and sepals, they were first placed in a freezer at -40°C for 12 hours to allow the water in the cells to fully crystallize. Subsequently, an ultra-low temperature grinder was used to crush the frozen flowers into particles with an average particle size of approximately 0.8 mm. These particles were then placed in a vacuum drying oven and dried at a vacuum degree of 10 Pa and a temperature of 25°C for 4 hours to remove some of the bound water, destroy the cell structure, and improve the efficiency of subsequent extraction. (2) Single ultrasonic extraction: The pretreated marigold granules were added to a mixed liquid consisting of 45% ethanol aqueous solution, acetic acid-sodium acetate buffer solution (pH 5.0) containing 0.3% (m / m) cellulase, 0.2% (m / m) β-glucanase and 0.1% (m / m) hemicellulase at a material-liquid ratio of 1:12 (mass of marigold granules: total volume of mixed liquid). The mixed system was transferred to an ultrasonic extraction device and dual-frequency ultrasonic synergistic extraction was used. The low-frequency ultrasonic frequency was set at 30 kHz and the power range was 350 W; the high-frequency ultrasonic frequency was set at 120 kHz and the power range was 220 W. The extraction was carried out at a temperature of 30 °C for 2 hours. During the extraction, the ultrasonic wave was paused for 4 minutes every 25 minutes, and the mixed liquid was stirred at a speed of 220 r / min to ensure full contact between the materials and promote the release of the active ingredients in the cells.

[0042] (3) Solid-liquid separation and secondary ultrasonic extraction: After the first ultrasonic extraction, the mixed liquid was subjected to solid-liquid separation by a plate and frame filter press to obtain a filter residue and a primary extract. The filter residue was added to a 45% ethanol aqueous solution at a solid-liquid ratio of 1:8 and transferred to the ultrasonic extraction device again. This time, variable amplitude ultrasonic extraction was used. The ultrasonic frequency was periodically changed between 20kHz and 40kHz with a change cycle of 4 minutes. The ultrasonic power was constant at 450W, and the extraction was carried out at 38°C for 0.8 hours. Within this parameter range, low-frequency ultrasound can deeply break up the stubborn cell structure remaining after the first extraction, and high-frequency ultrasound accelerates the diffusion of residual effective ingredients.

[0043] (4) Merging and concentration: The secondary extract was combined with the primary extract, first passed through a microfiltration membrane (pore size 0.45 μm) to remove larger particle impurities, and then concentrated using a nanofiltration membrane with a molecular weight cutoff of 800 Da. The extract was concentrated to 1 / 4 of its original volume at a pressure of 0.4 MPa to enrich the target component. (5) Drying: The concentrate was transferred to a vacuum freeze dryer and pre-frozen at a pre-freezing temperature of -50°C for 3 hours to completely freeze the concentrate. The concentrate was then sublimated and dried at a vacuum degree of 2Pa and a temperature of 32°C until the moisture content dropped below 2.5%, thereby obtaining high-quality marigold extract powder.

[0044] 1.2.2 Preparation method of the composition for strengthening the brain and improving eyesight Combine the formulated amounts of algae oil, linseed oil, and arachidonic acid and stir at 50°C to form a uniform oil phase mixture. Slowly add the marigold extract powder to this oil phase mixture and continue stirring for 30 minutes to fully disperse it. Next, add the zeaxanthin suspension oil and continue stirring for 20 minutes. Finally, add NA nervonic acid and PS phosphatidylserine and stir at 30°C for 15 minutes to obtain a uniform and stable brain-boosting and eye-improving composition.

[0045] Example 2 2.1 Formulation A composition for strengthening the brain and improving eyesight, the formula of which is shown in Table 3: Table 3 Formulation of Example 2

[0046] 2.1 Preparation method The difference from Example 1 lies in steps (2) and (3) in the preparation method of marigold extract, and the rest is the same as Example 1.

[0047] (2) Single ultrasonic extraction: The pretreated marigold granules were added to a mixed liquid consisting of 45% ethanol aqueous solution, acetic acid-sodium acetate buffer solution (pH 5.0) containing 0.3% (m / m) cellulase, 0.2% (m / m) β-glucanase and 0.1% (m / m) hemicellulase at a material-liquid ratio of 1:12 (mass of marigold granules: total volume of mixed liquid). The mixed system was transferred to an ultrasonic extraction device and dual-frequency ultrasonic synergistic extraction was used. The low-frequency ultrasonic frequency was set at 20 kHz and the power range was 260 W; the high-frequency ultrasonic frequency was set at 100 kHz and the power range was 150 W. The extraction was carried out at a temperature of 35°C for 1 hour. During the extraction, the ultrasonic wave was paused for 2 minutes every 15 minutes, and the mixed liquid was stirred at a speed of 180 r / min to ensure full contact between the materials and promote the release of the active ingredients in the cells.

[0048] (3) Solid-liquid separation and secondary ultrasonic extraction: After the first ultrasonic extraction, the mixed liquid was subjected to solid-liquid separation by a plate and frame filter press to obtain a filter residue and a primary extract. The filter residue was added to a 45% ethanol aqueous solution at a solid-liquid ratio of 1:8 and transferred to the ultrasonic extraction device again. This time, variable amplitude ultrasonic extraction was used. The ultrasonic frequency was periodically changed between 20kHz and 40kHz with a change cycle of 6 minutes. The ultrasonic power was constant at 350W, and the extraction was carried out at 32°C for 1.2 hours. Within this parameter range, low-frequency ultrasound can deeply break up the stubborn cell structure remaining after the first extraction, and high-frequency ultrasound accelerates the diffusion of residual effective ingredients.

[0049] Comparative Example A comparative example was set with reference to Example 1. The differences between the comparative example and Example 1 are shown in Table 4: Table 4

[0050] Effect Example 1 Evaluation of the efficacy of anti-blue light eye damage 1.1 Testing materials 1.1.1 Sample Information The brain-boosting and eye-improving compositions prepared in Examples 1-2 and Comparative Examples 1-4 all used DMSO as the solvent.

[0051] Positive control: Coenzyme Q10, yellow powder, batch number K2211223, Shanghai Aladdin Biochemical Technology Co., Ltd., solvent is DMSO.

[0052] 1.1.2 Experimental animals Zebrafish were maintained in aquaculture water at 28°C (water quality: 200 mg of instant sea salt added per liter of reverse osmosis water, conductivity 450–550 μS / cm; pH 6.5–8.5; hardness 50–100 mg / L CaCO3). Zebrafish were bred and provided by the Fish Farming Center of Hangzhou Huante Biotechnology Co., Ltd. under the experimental animal use license number SYXK (Zhejiang) 2022-0004. Their husbandry and management complied with the requirements of AAALAC accreditation (certification number: 001458) and the IACUC ethics review number: IACUC-2025-12427-01.

[0053] 1.1.3 Instruments, consumables, and reagents Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China); blue light analyzer (50w450nm, Huiheng Light Source Technology Mall, China); motorized focus continuous zoom fluorescence microscope (AZ100, Nikon, Japan); ultrasonic cleaner (JP-010T, Shenzhen Jiemeng Cleaning Equipment Co., Ltd., China).

[0054] Methylcellulose (Lot No. C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China); dimethyl sulfoxide (DMSO, Lot No. BCCD8942, Sigma, Switzerland); acridine orange (AO, Lot No. C15109250, Shanghai MacLean Biochemical Technology Co., Ltd., China); pronase E (Lot No. G12511Y118034, Shanghai Yuanye Biotechnology Co., Ltd., China).

[0055] 1.2 Detection method A zebrafish model of blue light-induced eye damage was established by randomly selecting wild-type AB zebrafish at 1 dpf and irradiating them with blue light. At 3 dpf, well-developed model zebrafish were randomly assigned to 6-well plates, with 30 zebrafish treated per well (experimental group). The corresponding samples (concentrations shown in Table 5) and a positive control, Coenzyme Q10 at a concentration of 62.5 μg / mL, were administered in water. A normal control group (not exposed to blue light but given water) and a model control group (exposed to blue light but given water) were also established. Each well contained 3 mL of water. After one day of treatment at 28°C, zebrafish in each experimental group were stained with AO in the dark for 30 minutes. After three washes with standard dilution water, 10 randomly selected zebrafish from each experimental group were photographed under a fluorescence microscope. Data were analyzed and collected using Image J software. The average fluorescence intensity of apoptotic cells in the zebrafish eyes was calculated and used to evaluate the anti-blue light eye damage efficacy of the samples. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software, and p < 0.05 indicated that the difference was statistically significant.

[0056] 1.3 Test results Under the experimental conditions, Examples 1-2 and Comparative Examples 1-4 all have the effect of preventing blue light eye damage, which is specifically manifested by reducing the average fluorescence intensity of apoptotic cells in the eyes. Moreover, the anti-blue light eye damage effect of Examples 1-2 is better than that of Comparative Examples 1-4. See Table 5 for details. Figure 1 .

[0057] Table 5 Results of the experimental evaluation of the efficacy of samples against blue light eye damage (n = 10)

[0058] Note: Compared with the model control group, *** indicates p < 0.001, * indicates p < 0.05; compared with the normal control group, no 井 The number indicates p>0.05, ## Indicates p<0.01, ### Indicates p<0.001.

[0059] As can be seen from the results in Table 5, Examples 1-2 can all significantly reduce the fluorescence intensity of blue-light-induced apoptotic cells in the zebrafish eye. The efficacy of Example 2 is better than that of the positive control coenzyme Q10. Although Example 1 is slightly inferior to coenzyme Q10, both are better than Comparative Examples 1-4, and the effect of Example 2 is closest to the normal state. The improvement effects of Comparative Examples 1-4 are weak, and Comparative Example 4 is even not statistically significant, further highlighting the significant advantages of the examples in resisting blue light eye damage. The overall data show that Examples 1-2 exhibit excellent anti-blue light eye damage potential by inhibiting ocular cell apoptosis, and can be used as a preferred solution for anti-blue light eye protection products.

[0060] Effect Example 2 Evaluation of the efficacy of preventing myopia 2.1 Test materials 2.1.1 Sample information The brain-boosting and eye-improving compositions prepared in Examples 1-2 and Comparative Examples 1-4 use standard dilution water as the solvent.

[0061] Positive control: Atropine sulfate monohydrate, white crystals, batch number I2111034, Shanghai Aladdin Biochemical Technology Co., Ltd., solvent is standard dilution water.

[0062] 2.1.2 Experimental animals Zebrafish were maintained in aquaculture water at 28°C (water quality: 200 mg of instant sea salt added per liter of reverse osmosis water, conductivity 450–550 μS / cm; pH 6.5–8.5; hardness 50–100 mg / L CaCO3). Zebrafish were bred and provided by our company's fish farming center under the experimental animal use license number SYXK (Zhejiang) 2022-0004. Their husbandry and management complied with the requirements of AAALAC accreditation (certification number: 001458) and the IACUC ethics review number: IACUC-2025-12427-01.

[0063] 2.1.3 Instruments, consumables and reagents Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); microinjector (IM300, Narishige, Japan); needle puller (PC-10, Narishige, Japan); precision electronic balance (CP214, OHAUS, USA); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China).

[0064] Methylcellulose (Batch No. C2004046, Shanghai Aladdin Biochemical Technology Co., Ltd., China).

[0065] 2.2 Evaluation of the efficacy of preventing myopia One-cell-stage wild-type AB strain zebrafish were randomly selected and plated in 6-well plates, with 30 zebrafish treated per well. All experimental groups, except the normal control group and the MO standard control group, were injected with lumican-MO to establish a zebrafish myopia model. After two days of treatment at 28°C, the samples (concentrations shown in Table 6) and a positive control, atropine sulfate monohydrate, were administered in water (3 mL per well). A normal control group, a MO standard control group, and a model control group were also established. After three days of treatment at 28°C, 10 zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The ratio of retinal pigment epithelium diameter to scleral diameter was analyzed, and the myopia prevention efficacy of the samples was evaluated using this metric. Statistical analysis results are expressed as mean ± SE. Statistical analysis was performed using SPSS software. P < 0.05 indicated statistical significance.

[0066] 2.3 Test results Under the experimental conditions, Examples 1-2 and Comparative Examples 1-4 all have the effect of preventing myopia, which is specifically manifested by increasing the ratio of retinal pigment epithelium diameter / sclera diameter. The myopia prevention effect of Examples 1-2 is better than that of Comparative Examples 1-4. See Table 6 for details. Figure 2 .

[0067] Table 6 Results of the experimental evaluation of the effectiveness of samples in preventing myopia (n = 10)

[0068] Note: Compared with the model control group, *** indicates p < 0.001, * indicates p < 0.05; compared with the normal control group, ### Indicates p<0.001.

[0069] As can be seen from the results in Table 6, both Example 1 and Example 2 can significantly improve the ratio of retinal pigment epithelium diameter / sclera diameter, among which Example 1 has better efficacy than the positive control Atropine sulfate monohydrate, and Example 2 is slightly inferior to the positive control, but both are significantly better than Comparative Examples 1 to Comparative Examples 4; the improvement effects of Comparative Examples 1 to Comparative Examples 4 are weaker, further highlighting the significant advantages of the examples in preventing myopia.

[0070] Effect Example 3 Evaluation of the effect of promoting brain development 3.1 Test materials 3.1.1 Sample information The brain-boosting and eye-improving compositions prepared in Examples 1-2 and Comparative Examples 1-4 use standard dilution water as the solvent.

[0071] Positive control: Ginkgo biloba and Cistanche tablets, brown tablets, batch number 3313N903, Amway (China) Daily Products Co., Ltd., solvent is standard dilution water.

[0072] 3.1.2 Experimental animals Zebrafish were maintained in aquaculture water at 28°C (water quality: 200 mg of instant sea salt added per liter of reverse osmosis water, conductivity 450–550 μS / cm; pH 6.5–8.5; hardness 50–100 mg / L CaCO3). Zebrafish were bred and provided by our company's fish farming center under the experimental animal use license number SYXK (Zhejiang) 2022-0004. Their husbandry and management complied with the requirements of AAALAC accreditation (certification number: 001458) and the IACUC ethics review number: IACUC-2025-12427-01.

[0073] 3.1.3 Instruments, consumables and reagents Dissecting microscope (SZX7, OLYMPUS, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); precision electronic balance (CP214, OHAUS, USA); zebrafish behavior analysis system (Zebra Lab3.22.3.31, Viewpoint, France); 6-well plate (Zhejiang Bellanbo Biotechnology Co., Ltd., China).

[0074] 3.2 Detection method 5-day-old wild-type AB strain zebrafish were randomly selected and placed in beakers. Thirty zebrafish were treated in each beaker (experimental group). Water-soluble samples (concentrations shown in Table 7) were administered, along with a positive control, Ginkgo biloba and Cistanche deserticola tablets, at a concentration of 125 μg / mL. A normal control group was also established. Each beaker held 20 mL of water. After one day of treatment at 28°C, five zebrafish were randomly selected from each experimental group and placed in a cross-shaped module. The module was divided into four zones: yellow, blue, red, and green. Each group had six modules. Data were collected using a behavioral analyzer. The percentage of the total distance traveled within the blue zone relative to the total distance traveled within the entire zone within 10 minutes was analyzed. This metric was used to evaluate the efficacy of the samples in promoting brain development. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software. P < 0.05 indicated statistical significance.

[0075] 3.3 Test results Under the experimental conditions, Examples 1-2 and Comparative Examples 1-4 all have the effect of promoting brain development. And the effect of Examples 1-2 on promoting brain development is better than that of Comparative Examples 1-4. See Table 7 for details. Figure 3 .

[0076] Table 7 Results of the experimental evaluation of the samples’ efficacy in promoting brain development (n = 6)

[0077] Note: Compared with the normal control group, *p<0.05, ***p<0.001.

[0078] From the results in Table 7, it can be seen that Examples 1 and 2 can significantly increase the proportion of zebrafish movement in the blue area, among which Example 1 is more effective than the positive control Ginkgo Biloba and Cistanche Tablets, and Example 2 is equivalent to the positive control, and both are significantly better than Comparative Examples 1 to 4; although Comparative Examples 1-4 have a certain promoting effect, the effect is relatively weak, which further highlights the significant advantages of the examples in promoting brain development.

[0079] The above detailed description is a specific description of one feasible embodiment of the present invention and is not intended to limit the scope of the present invention. It should be noted that any equivalent implementation or modification that does not depart from the present invention should be included within the scope of the technical solution of the present invention. Therefore, the scope of protection of the patent of this invention should be based on the attached requirements.

Claims

1. A composition for strengthening the brain and improving eyesight, characterized in that: The invention comprises the following ingredients in parts by weight: 200-240 parts of algae oil, 60-80 parts of linseed oil, 8-12 parts of arachidonic acid, 4-8 parts of marigold extract, 0.6-2 parts of zeaxanthin suspension oil, 0.02-0.04 parts of NA nervonic acid and 0.4-0.6 parts of PS phosphatidylserine; The preparation method of the marigold extract comprises the following steps: S1. Pretreatment: Freeze marigold flowers, crush them, and vacuum dry them; S2, primary extraction: mixing the pretreated material with the extraction solvent and performing ultrasound-assisted extraction in the presence of a complex enzyme; S3, secondary extraction: the filter residue after the primary extraction is mixed with the extraction solvent again and subjected to variable amplitude ultrasonic extraction; S4, post-processing: combining the extracts, sequentially performing microfiltration, nanofiltration concentration and drying to obtain the marigold extract; The complex enzyme in step S2 includes cellulase, β-glucanase and hemicellulase; The ultrasound-assisted extraction in step S2 uses dual-frequency ultrasound, with a low frequency of 20-30 kHz and a power of 260-350 W, and a high frequency of 100-120 kHz and a power of 150-220 W; In step S3, the frequency of the variable amplitude ultrasound is periodically changed between 20kHz and 40kHz, the change period is 4-6 minutes, and the ultrasound power is 350-450W.

2. The composition according to claim 1, characterized in that In step S2, the low frequency is 30kHz, the power is 350W, the high frequency is 120kHz, and the power is 220W; Or the low frequency is 20kHz, the power is 260W, the high frequency is 100kHz, and the power is 150W; In step S3, the frequency of the variable amplitude ultrasound is periodically varied between 20 kHz and 40 kHz, with a variation period of 4 minutes and an ultrasound power of 450 W; Or the frequency of the variable amplitude ultrasound changes periodically between 20kHz and 40kHz, the change period is 6 minutes, and the ultrasound power is 350W.

3. The composition according to claim 1, characterized in that The mass percentages of the cellulase, β-glucanase and hemicellulase in step S2 are 2-4:1-3:0.5-1.

5.

4. The composition according to claim 3, characterized in that The mass percentages of the cellulase, β-glucanase and hemicellulase in step S2 are 3:2:

1.

5. The composition according to claim 1, characterized in that The freezing treatment in step S1 is performed at a temperature of -30 to -50°C for 8 to 16 hours; The average particle size of the crushed particles is 0.5-1.0 mm; The vacuum drying is performed at a vacuum degree of 5-15 Pa, a temperature of 20-30° C., and a time of 3-5 hours; The extraction time of the ultrasonic-assisted extraction in step S2 is 1-2 hours, and the extraction temperature is 35-40° C.; During the ultrasound-assisted extraction process in step S2, the ultrasound is paused for 1-5 minutes every 10-30 minutes, and the mixed solution is stirred at a speed of 150-250 r / min; The extraction solvent in step S2 is a 30-50% ethanol aqueous solution; the pH value of the extraction solvent is 4.5-5.5; The extraction time of the variable amplitude ultrasonic method in step S3 is 0.8-1.2 hours, and the extraction temperature is 32-38°C; The pore size of the filter membrane used in the microfiltration in step S4 is 0.2-0.8 μm; The membrane used in the nanofiltration has a molecular weight cut-off of 500-1000Da; The drying is vacuum freeze drying, and the moisture content after drying is ≤3.0%.

6. The composition according to any one of claims 1 to 5, characterized in that The invention comprises the following ingredients in parts by weight: 240 parts of algae oil, 60 parts of linseed oil, 8 parts of arachidonic acid, 8 parts of marigold extract, 0.6 parts of zeaxanthin suspension oil, 0.02 parts of NA nervonic acid and 0.6 parts of PS phosphatidylserine; Or it includes the following ingredients in parts by weight: 200 parts of algae oil, 80 parts of linseed oil, 12 parts of arachidonic acid, 4 parts of marigold extract, 2 parts of zeaxanthin suspension oil, 0.04 parts of NA nervonic acid and 0.4 parts of PS phosphatidylserine.

7. A method for preparing the composition according to any one of claims 1 to 6, comprising the steps of: (1) Mixing the formulated amounts of algae oil, linseed oil, and arachidonic acid to obtain a uniform oil phase mixture; (2) adding the marigold extract to the oil phase mixture obtained in step (1) and mixing well to obtain a mixture A; (3) adding the zeaxanthin suspension oil to the mixture A obtained in step (2), mixing well, and obtaining a mixture B; (4) Add NA neuraminic acid and PS phosphatidylserine to the mixture B obtained in step (3) and mix well to obtain a composition.

8. A functional food, characterized in that The invention comprises the composition according to any one of claims 1 to 6.

9. Use of the composition according to any one of claims 1 to 6 or the functional food according to claim 8 in the preparation of a product for strengthening the brain and improving eyesight.

10. The use according to claim 9, characterized in that The product is a functional food or medicine.

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