An acer truncatum seed oil-PS complex composition for promoting neural development

Abstract

The present invention discloses an Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development, relating to the field of food nutrition. The composition comprises, in parts by weight, 8 – 12 parts Acer truncatum seed oil, 3 – 6 parts phosphatidylserine, 1 – 2 parts cephalin, 2 – 4 parts linseed oil, 0.5 – 2 parts arachidonic acid phosphatidylinositol, 1 – 3 parts docosahexaenoic acid phospholipid, 0.3 – 1.5 parts sialic acid, 0.2 – 1 part phytosterol, 0.1 – 0.8 part d-alpha-tocopherol, 0.5 – 2 parts co-emulsifier, and 0.05 – 0.3 part antioxidant synergist. The composition forms a synergistic nutritional system meeting neural development requirements and providing comprehensive nutritional support, significantly enhancing efficacy. A brain-targeting lipid-phospholipid hybrid nanocarrier is constructed, combined with a refined process to improve targeting ability and bioavailability while maintaining bioactivity, storage stability, and safety, facilitating application and promotion.

Description

[0001] DESCRIPTION

[0002] An Acer truncatum seed oil-PS complex composition for promoting neural development TECHNICAL FIELD

[0003] The present invention relates to the technical field of food nutrition, and in particular to an Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development.

[0004] BACKGROUND ART

[0005] Neural development is an important physiological stage in human growth, and its developmental status is closely related to many aspects such as brain function and cognitive ability. Accordingly, research on nutritional support for neural development has become an important research direction in the fields of food nutrition and biomedicine. Acer truncatum seed oil, phosphatidylserine, various phospholipids, and functional active ingredients have each been successively confirmed to exert positive effects on neural development by virtue of their respective physiological characteristics related to neural tissue construction and regulation of neuronal activity. Applications and studies of the related single ingredients have become increasingly mature. With the continuous increase in people's demand for health and nutrition, market demand for nutritional supplements for neural development among different populations has continued to grow. The development of composite nutritional compositions having multi-component synergy, high activity, and ready absorption has therefore become an important research direction suited to current demands for nutritional support for neural development and has also promoted technological upgrading and product innovation of related nutritional preparations.

[0006] Current nutritional supplement products for neural development in the market generally have a number of technical shortcomings and cannot adequately satisfy actual nutritional-support needs. Some products employ only a single nutrient component in formulation design and therefore cannot DESCRIPTION provide comprehensive nutritional support for neural development from multiple dimensions and stages, nor can they fully exert synergistic enhancement among components. Some other composite formulation products lack a scientific basis for component compatibility, and unreasonable proportions among components readily lead to mutual restriction of efficacy. Meanwhile, traditional raw-material treatment processes are relatively rough, with insufficient purification of raw materials, easy residue of impurities, and difficulty in ensuring the bioactivity of effective ingredients. Some dosage forms lack targeted design, so effective ingredients are readily lost in vivo and intestinal absorption efficiency is low. In addition, the related preparation processes provide insufficient protection for the components, readily causing oxidation and deterioration of the ingredients and further reducing the practical application effect of the product.

[0007] SUMMARY OF THE INVENTION

[0008] An object of the present invention is to overcome the deficiencies of the prior art by providing an Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development. The composition is obtained by scientifically proportioning multiple active components such as Acer truncatum seed oil and phosphatidylserine, with each raw material having been purified through a specific process so as to ensure component purity and bioactivity. The composition is in the form of a brain-targeting lipid-phospholipid hybrid nanocarrier and is matched with a refined preparation process, in which each stage from raw-material pretreatment to final filling is specifically optimized, thereby realizing synergistic enhancement among the components, accurately providing comprehensive nutritional support for neural development, improving brain-targeting performance and bioavailability of the components, and simultaneously ensuring product stability and safety in use.

[0009] To solve the above technical problems, the present invention provides the DESCRIPTION following technical solutions. In one aspect, the present invention provides an Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development, wherein, in parts by weight, the complex composition comprises 8-12 parts of Acer truncatum seed oil, 3-6 parts of phosphatidylserine, 1-2 parts of cephalin, 2-4 parts of linseed oil, 0.5-2 parts of arachidonic acid phosphatidylinositol, 1-3 parts of docosahexaenoic acid phospholipid, 0.3 -1.5 parts of sialic acid, 0.2-1 part of phytosterol, 0.1-0.8 part of d-alpha-tocopherol, 0.5-2 parts of a co-emulsifier, and 0.05-0.3 part of an antioxidant synergist; wherein the composition is in the form of a brain-targeting lipid-phospholipid hybrid nanocarrier, with linseed oil serving as a base membrane material, phosphatidylserine, cephalin, and arachidonic acid phosphatidylinositol serving as functional membrane auxiliary materials, Acer truncatum seed oil and docosahexaenoic acid phospholipid serving as core materials, and phosphatidylserine serving as a brain-targeting modification group on the carrier surface.

[0010] Further, the Acer truncatum seed oil is low-temperature cold-pressed Acer truncatum kernel oil, with a pressing temperature of <=45 °C, wherein the mass content of nervonic acid is >=6%, the total mass content of unsaturated fatty acids is >=88%, the mass content of linoleic acid is >=30%, the mass content of oleic acid is >=25%, the peroxide value is <=3 mmol / kg, the acid value is <=1.0 mg KOH / g, and the solvent residue is <=0.001%.

[0011] Furthermore, the phosphatidylserine is derived from soybean or sunflower seed and is prepared by a column chromatography separation and purification process, with an effective purity of >=65%, a free fatty acid content of <=1.0%, a moisture content of <=0.5%, an ash content of <=0.3%, and a total heavy metal content of <=0.1 mg / kg; and the cephalin is obtained by molecular distillation purification of soybean concentrated phospholipid, with an effective purity of >=55%, phosphatidyl ethanolamine accounting for >=90% by mass of the cephalin, a free cholesterol content of <=0.5%, a DESCRIPTION moisture content of <=0.5%, and an acid value of <=2.0 mg KOH / g.

[0012] Furthermore, the arachidonic acid phosphatidylinositol is prepared by a microbial fermentation method or an egg-yolk phospholipid separation and purification method, with a purity of >=95%, a retention rate of the phosphatidylinositol head-group structure of >=98%, a binding rate of arachidonic acid in the arachidonic acid phosphatidylinositol of >=92%, a moisture content of <=0.3%, no protein residue, and a heavy metal content of <=0.05 mg / kg; and the docosahexaenoic acid phospholipid is of a phospholipid-bound type and is derived from Schizochy trium fermentation phospholipid or egg-yolk phospholipid, with a mass content of DHA of >=45%, a purity of phosphatidylcholine of >=80%, no free DHA residue, and an oxidation value of <=2 mmol / kg.

[0013] Furthermore, the sialic acid is N-acetylneuraminic acid derived from milk or bird's nest, with a purity of >=98%, an effective content of >=98%, and a moisture content of <=0.5%; the phytosterol is a mixture of beta-sitosterol, stigmasterol, and campesterol in a mass ratio of 5:3:2, with a total purity of >=95%, a proportion of beta-sitosterol of >=55%, no phytosterol ester impurity, and a melting point of 130-140 °C; and the d-alpha-tocopherol is natural vitamin E extracted from soybean oil or sunflower-seed oil, with a purity of >=96%, d-alpha-tocopherol accounting for >=92% of total vitamin E, and no synthetic vitamin E added.

[0014] Furthermore, the co-emulsifier is a combination of sucrose fatty acid ester and polyglycerol fatty acid ester in a mass ratio of 2:1, wherein the HLB value of the sucrose fatty acid ester is 8-10 and the HLB value of the poly glycerol fatty acid ester is 10-12; and the antioxidant synergist is a combination of ascorbyl palmitate and rosemary extract in a mass ratio of 3:1, wherein the carnosic acid content in the rosemary extract is >=5%.

[0015] Furthermore, the brain-targeting lipid-phospholipid hybrid nanocarrier has an average particle size of 140-160 nm, a poly dispersity index (PDI) of DESCRIPTION

[0016] <=0.15, and a zeta potential of -34 to -36 mV; the encapsulation efficiencies of the core effective components are all >=90%; the particle-size change rate in simulated gastric fluid in vitro for 2 h is <=5%; and the particle-size change rate in simulated intestinal fluid in vitro for 4 h is <=7%.

[0017] Furthermore, the preparation method of the complex composition specifically comprises the following steps:

[0018] 51. Raw-material pretreatment and weighing: weighing all components according to the parts-by-weight ratio; subjecting the Acer truncatum seed oil and linseed oil to vacuum filtration through an organic filter membrane; subjecting phosphatidylserine, cephalin, arachidonic acid phosphatidylinositol, and docosahexaenoic acid phospholipid to vacuum drying; separately sieving the sialic acid, phytosterol, d-alpha-tocopherol, co-emulsifier, and antioxidant synergist; and placing all pretreated raw materials in dry, light-protected sealed containers for subsequent use;

[0019] 52. Preparation of an oil phase: adding the pretreated and weighed Acer truncatum seed oil and linseed oil into a constant-temperature stirring vessel, starting the stirring device and controlling the water-bath temperature at 40-50 °C, then successively adding arachidonic acid phosphatidylinositol, docosahexaenoic acid phospholipid, phytosterol, and d-alpha-tocopherol after stirring, adjusting the stirring speed and continuing stirring until a uniform, transparent, particle-free, and non-layered lipid-soluble oil phase is formed, and keeping the system under stirring and constant temperature for subsequent use;

[0020] 53. Preparation of an aqueous phase: preparing a phosphate buffer having a pH of 7.0-7.8 and adding the buffer into a constant-temperature stirring vessel, starting water-bath heating and adjusting the stirring speed, wherein the water-bath temperature is 40-50 °C and is kept consistent with that of the oil phase, then successively adding the pretreated phosphatidylserine, cephalin, co-emulsifier, antioxidant synergist, and sialic DESCRIPTION acid, adjusting the stirring speed and continuing stirring until a uniform, clear, and precipitate-free aqueous phase is formed, and, after standing, keeping the system under constant temperature and stirring for subsequent use;

[0021] 54. Emulsification and forming of the nanocarrier: keeping the water-bath temperatures of the two stirring vessels consistent, slowly adding the oil phase into the aqueous phase through a constant-flow pump while maintaining stirring during the addition, increasing the stirring speed after completion of the addition to perform high-speed shearing so as to form a primary emulsion, and then introducing the primary emulsion into a high-pressure homogenizer for cyclic homogenization, with sampling performed to detect particle size until the average particle size reaches 140-160 nm and the poly dispersity index PDI is <=0.15;

[0022] 55. Post-treatment and filling of the final product: transferring the composite nano-composition to pasteurization equipment for sterilization, cooling to room temperature after sterilization, then transferring to a sterile filling workshop for sealed filling in a light-protected sterile environment, immediately sealing after filling, placing the finished product in a low-temperature light-protected environment for storage, and simultaneously conducting sampling inspection of various indicators, whereby a qualified product is the final product.

[0023] Furthermore, the weighing process is carried out in a light-protected, dry, and dust-free environment; the temperature for vacuum drying is 40 °C, the drying time is 2 h, and nitrogen protection is used during drying; the pore size of the organic filter membrane is 0.22 um, the filtration pressure is 0.1-0.15 MPa, the filtration time is 10-15 min; and the sieve is an 80-mesh stainless-steel sieve.

[0024] Furthermore, during dropwise addition of the oil phase, the stirring speed is maintained at 400 r / min; the high-speed shearing speed is 10,000-12,000 r / min, and the shearing time is 3-5 min; the high-pressure homogenizer is a DESCRIPTION plunger high-pressure homogenizer, the homogenization pressure is 600-800 bar, cyclic homogenization is performed 3-5 times, the homogenization time for each cycle is 2-3 min, and the interval between two homogenization cycles is 1 min; particle-size detection is performed with a laser particle-size analyzer, the detection range is 0.1-1000 nm, and the detection temperature is 25 + / - 2 °C.

[0025] Compared with the prior art, the Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development provided by the present invention has the following beneficial effects:

[0026] I. By scientifically and rationally combining a plurality of nutritional components and forming a synergistic system in light of the physiological activity characteristics of each component, the present invention accurately meets nutritional requirements in the process of neural development. Various lipid and phospholipid components can supplement structural substances required for neural-tissue construction, while functional ingredients such as sialic acid and phytosterol assist the growth and differentiation of nerve cells from multiple dimensions. Raw materials from different sources that have been purified through specific processes effectively ensure the activity and purity of the respective components and avoid interference of impurities with the efficacy of the composition. At the same time, the limitations of any single raw material in promoting neural development are overcome, so that the functions of the respective ingredients complement one another and provide more comprehensive nutritional support for neural development, thereby greatly improving the promoting effect.

[0027] II. By constructing the dosage form of a brain-targeting lipid-phospholipid DESCRIPTION hybrid nanocarrier and combining the dosage form with a refined stepwise preparation process, the present invention simultaneously improves targeting performance and bioavailability of the composition. A carrier structure using lipids as membrane materials and functional phospholipids as auxiliary materials endows the composition with brain-targeting properties, enabling nutritional ingredients to act accurately on brain nerve tissue and reducing loss of effective components in vivo. The nanoscale carrier form further improves intestinal absorption efficiency. Throughout the refined raw-material pretreatment and preparation operations, the bioactivity of each component is effectively retained, and, with antioxidant-related ingredients and process control suppressing oxidation and deterioration of the raw materials, the composition exhibits good storage stability and safety in use, thereby facilitating product promotion and application.

[0028] Other advantages, objectives, and features of the present invention will be set forth in part in the following description, and in part will become apparent to those skilled in the art upon examination of the following disclosure, or may be learned from practice of the invention.

[0029] BRIEF DESCRIPTION OF THE DRAWINGS

[0030] To describe the technical solutions in the embodiments of the present invention or in the prior art more clearly, the drawings required for describing the embodiments or the prior art are briefly introduced below. Obviously, the drawings in the following description merely illustrate certain embodiments of the present invention, and those of ordinary skill in the art can derive other drawings therefrom without inventive effort.

[0031] Figure 1 is a process flow chart of the preparation process of the Acer DESCRIPTION truncatum seed oil-PS complex composition for promoting neural development;

[0032] Figure 2 is a process flow chart of raw-material pretreatment for the Acer truncatum seed oil-PS complex composition for promoting neural development;

[0033] Figure 3 is a process flow chart of the preparation steps of the lipid-phospholipid hybrid nanocarrier.

[0034] DETAILED DESCRIPTION

[0035] In order to further illustrate the technical means and effects adopted by the present invention to achieve the intended objectives, the specific implementation, structure, features, and effects of the present invention are described in detail below in conjunction with the accompanying drawings and preferred embodiments.

[0036] Embodiment 1

[0037] Preparation of the Acer truncatum seed oil-PS complex composition for promoting neural development.

[0038] In this embodiment, an Acer truncatum seed oil-PS complex composition for promoting neural development is prepared. In parts by weight, the selected raw materials are: 10 parts of Acer truncatum seed oil, 4.5 parts of phosphatidylserine, 1.5 parts of cephalin, 3 parts of linseed oil, 1.2 parts of arachidonic acid phosphatidylinositol, 2 parts of docosahexaenoic acid phospholipid, 0.9 part of sialic acid, 0.6 part of phytosterol, 0.45 part of d-alpha-tocopherol, 1.2 parts of a co-emulsifier, and 0.17 part of an antioxidant synergist. The co-emulsifier is a combination of sucrose fatty acid ester and polyglycerol fatty acid ester in a mass ratio of 2: 1, and the antioxidant synergist is a combination of ascorbyl palmitate and rosemary extract in a mass ratio of 3 : 1. DESCRIPTION

[0039] 51. Raw-material pretreatment and weighing: weighing is carried out in a light-protected, dry, and dust-free environment, and all raw materials are accurately weighed according to the above parts-by-weight ratio, with raw-material identification and records being prepared. The Acer truncatum seed oil and linseed oil are subjected to vacuum filtration through an organic filter membrane having a pore size of 0.22 um, with the filtration pressure controlled at 0.12 MPa and the filtration time at 12 min. During filtration, the filtrate-collection container is kept dry and sealed, and, after filtration, the filtrates are placed in light-protected containers. Phosphatidylserine, cephalin, arachidonic acid phosphatidylinositol, and docosahexaenoic acid phospholipid are subjected to vacuum drying at 40 °C for 2 h under nitrogen protection and then transferred to dry, light-protected containers. Sialic acid, phytosterol, d-alpha-tocopherol, the co-emulsifier, and the antioxidant synergist are separately passed through an 80-mesh stainless-steel sieve, and all pretreated raw materials are placed in dry, light-protected sealed containers for subsequent use.

[0040] 52. Preparation of an oil phase: the pretreated Acer truncatum seed oil and linseed oil are added together into a constant-temperature stirring vessel. After the opening of the vessel is closed, the stirring device is started, the water-bath temperature is adjusted to 45 °C and maintained at a constant temperature, and the mixture is continuously stirred at 300 rpm for 5 min so that the two oils are fully and uniformly mixed. After completion of stirring, arachidonic acid phosphatidylinositol, docosahexaenoic acid phospholipid, phytosterol, and d-alpha-tocopherol are successively added into the vessel. After each raw material is added, stirring is maintained at 300 rpm for 3 min to ensure preliminary DESCRIPTION dispersion of the raw material before the next raw material is added. After all lipid-soluble raw materials have been added, the stirring speed is increased to 500 rpm, and stirring is continued for 10 min until a uniform, transparent, particle-free, and non-layered lipid-soluble oil phase is formed, which is then kept under stirring and constant temperature for subsequent use.

[0041] 53. Preparation of an aqueous phase: a phosphate buffer is prepared as a base solution for the aqueous phase. The prepared phosphate buffer is added into another constant-temperature stirring vessel, the water-bath heating device is started, the temperature is adjusted to 45 °C so as to be consistent with that used for preparation of the oil phase, and the stirring speed is adjusted to 400 rpm. The pretreated phosphatidylserine, cephalin, co-emulsifier, antioxidant synergist, and sialic acid are successively added into the phosphate buffer. After each raw material is added, stirring is maintained at 400 rpm for 5 min, so that the raw material is preliminarily dispersed in the buffer solution. After all water-soluble raw materials have been added, stirring is continued for 15 min until a uniform, clear, and precipitate-free aqueous phase is formed, after which the mixture is allowed to stand and is then kept under constant temperature and stirring for subsequent use.

[0042] 54. Emulsification and forming of the nanocarrier: the water-bath temperatures of the constant-temperature stirring vessels containing the oil phase and the aqueous phase are both maintained at 45 °C. The oil phase is slowly added into the aqueous phase through a constant-flow pump at a uniform dropping rate so as to ensure that the oil phase is dispersed into the aqueous phase in the form of droplets. During the addition, the stirring speed of the aqueous DESCRIPTION phase is maintained at 400 rpm to prevent droplet coalescence of the oil phase. After the addition of the oil phase is completed, the stirring speed of the aqueous phase is increased to 11,000 rpm for high-speed shearing for 4 min, so that the oil phase is fully emulsified in the aqueous phase to form a primary emulsion. The primary emulsion is then introduced into a high-pressure homogenizer for cyclic homogenization, with the homogenization pressure controlled at 700 bar and the cyclic homogenization performed 4 times, each time for 2 min with an interval of 1 min between cycles. After homogenization, sampling is performed to detect particle size, and, after confirmation that the average particle size of the composition is 149 nm, PDI is 0.13, and zeta potential is -36 mV, the qualified composite nanocarrier is obtained.

[0043] S5. Post-treatment and filling of the final product: the prepared composite nano-composition is transferred into pasteurization equipment for sterilization. After sterilization, the composition is naturally cooled to room temperature in a clean environment. The cooled composition is then transferred into a sterile filling workshop, where sealed filling is performed in a light-protected sterile environment. The filling amount is controlled according to a predetermined standard, and sealing is carried out immediately after filling so as to prevent entry of external impurities and oxidation of the composition components. The sealed finished product is stored in a designated light-protected dry environment. At the same time, samples are randomly taken from the finished product for detection of average particle size, polydispersity index PDI, zeta potential, and encapsulation efficiencies of the effective components, and, after completion of the inspection, qualified finished-product quality records are established. DESCRIPTION

[0044] Embodiment 2

[0045] Specification characterization tests of the raw materials of the complex composition.

[0046] All raw materials used for preparation of the complex composition are subjected to specification characterization tests. Standard industry methods are used for all test methods, the testing process strictly follows operating procedures, each test index is tested in triplicate, and the average value is taken as the final test result. The specific testing procedures and results for the respective raw materials are as follows.

[0047] Acer truncatum seed oil:

[0048] The Acer truncatum seed oil is low-temperature cold-pressed Acer truncatum kernel oil. Gas chromatography is used to determine the fatty-acid composition and the mass contents of the respective fatty acids, potentiometric titration is used to determine the peroxide value and acid value, and gas chromatography-mass spectrometry is used to determine solvent residue. The test results show that the pressing temperature of the Acer truncatum seed oil is 43 °C, the mass content of nervonic acid is 6.1%, the total mass content of unsaturated fatty acids is 89%, the mass content of linoleic acid is 34%, the mass content of oleic acid is 27%, the peroxide value is 2.3 mmol / kg, the acid value is 0.6 mg KOH / g, and the solvent residue is 0.0008%.

[0049] Phosphatidylserine:

[0050] The phosphatidylserine is soybean-derived and is prepared by a column chromatography separation and purification process. High-performance liquid chromatography is used to determine effective purity and free fatty acid content, DESCRIPTION the Karl Fischer method is used to determine moisture content, gravimetry is used to determine ash content, and atomic absorption spectrophotometry is used to determine total heavy metal content. The test results show that the effective purity of the phosphatidylserine is 68%, the free fatty acid content is 0.7%, the moisture content is 0.3%, the ash content is 0.2%, and the total heavy metal content is 0.07 mg / kg.

[0051] Cephalin:

[0052] The cephalin is obtained by molecular distillation purification of soybean concentrated phospholipid. High-performance liquid chromatography is used to determine effective purity and the mass percentage of phosphatidylethanolamine in the cephalin, an enzymatic method is used to determine free cholesterol content, and the detection methods for moisture content and ash content are the same as those used for phosphatidylserine. The test results show that the effective purity of the cephalin is 58%, phosphatidylethanolamine accounts for 92% by mass of the cephalin, the free cholesterol content is 0.3%, the moisture content is 0.4%, and the acid value is 1.7 mg KOH / g.

[0053] Arachidonic acid phosphatidylinositol:

[0054] The arachidonic acid phosphatidylinositol is prepared by a microbial fermentation method. High-performance liquid chromatography is used to determine purity and the binding rate of arachidonic acid in the arachidonic acid phosphatidylinositol, gel permeation chromatography is used to determine protein residues, atomic absorption spectrophotometry is used to determine heavy metal content, and the Karl Fischer method is used to determine moisture content. The test results show that the purity of the arachidonic acid phosphatidylinositol is DESCRIPTION

[0055] 97%, the retention rate of the phosphatidylinositol head-group structure is 98.8%, the binding rate of arachidonic acid is 94%, the moisture content is 0.2%, no protein residue is detected, and the heavy metal content is 0.03 mg / kg.

[0056] Docosahexaenoic acid phospholipid:

[0057] The docosahexaenoic acid phospholipid is of a phospholipid-bound type and is derived from Schizochytrium fermentation phospholipid. High-performance liquid chromatography is used to determine the mass content of DHA, the purity of phosphatidylcholine, and the presence of free DHA residue, and potentiometric titration is used to determine the oxidation value. The test results show that the mass content of DHA in the docosahexaenoic acid phospholipid is 46%, the purity of phosphatidylcholine is 87%, no free DHA residue is detected, and the oxidation value is 1.7 mmol / kg.

[0058] Sialic acid:

[0059] The sialic acid is milk-derived N-acetylneuraminic acid. High-performance liquid chromatography is used to determine purity and effective content, liquid chromatography-mass spectrometry is used to determine whether other neuraminic-acid isomer impurities are present, and the Karl Fischer method is used to determine moisture content. The test results show that the purity of the sialic acid is 98.9%, the effective content is 98.7%, the moisture content is 0.4%, and no other neuraminic-acid isomer impurity is detected.

[0060] Phytosterol:

[0061] The phytosterol is a mixture of beta-sitosterol, stigmasterol, and campesterol in a mass ratio of 5:3:2. High-performance liquid chromatography is used to determine total purity and the proportion of beta-sitosterol, gas chromatography is DESCRIPTION used to determine whether phytosterol ester impurities are present, and differential scanning calorimetry is used to determine the melting point. The test results show that the total purity of the phytosterol is 97%, the proportion of beta-sitosterol is 56%, no phytosterol ester impurity is detected, and the melting point is 136 °C. d-alpha-Tocopherol:

[0062] The d-alpha-tocopherol is natural vitamin E extracted from soybean oil. High-performance liquid chromatography is used to determine purity and the proportion of d-alpha-tocopherol in total vitamin E, and liquid chromatography-mass spectrometry is used to determine whether synthetic vitamin E has been added. The test results show that the purity of d-alpha-tocopherol is 98%, d-alpha-tocopherol accounts for 94% of total vitamin E, and no synthetic vitamin E addition is detected.

[0063] Co-emulsifier:

[0064] The co-emulsifier is a combination of sucrose fatty acid ester and polyglycerol fatty acid ester. The hydrophilic-lipophilic balance method is used to determine the HLB values of the two components, respectively. The test results show that the HLB value of the sucrose fatty acid ester is 9 and the HLB value of the polyglycerol fatty acid ester is 11.

[0065] Antioxidant synergist:

[0066] The antioxidant synergist is a combination of ascorbyl palmitate and rosemary extract. High-performance liquid chromatography is used to determine the carnosic acid content in the rosemary extract. The test results show that the camosic acid content in the rosemary extract is 7%.

[0067] Embodiment 3 DESCRIPTION

[0068] Optimization of key preparation conditions of the complex composition.

[0069] Using the average particle size, polydispersity index PDI, and zeta potential of the complex composition as core evaluation indices, single-factor optimization experiments are carried out on four key conditions in the preparation process, namely the oil-phase water-bath temperature, the pH value of the phosphate buffer in the aqueous phase, the high-speed shearing time, and the number of high-pressure homogenization cycles. Three gradient parameters are set for each condition while other preparation conditions remain unchanged. By detecting the core indices of the composition prepared under different gradient parameters, the influence of each condition on the preparation effect of the composition is analyzed. The parameter gradients in the optimization experiments are all controlled within a reasonable range. The specific optimization process is as follows.

[0070] Optimization of oil-phase water-bath temperature:

[0071] Three gradients of 40 °C, 45 °C, and 50 °C are set for the oil-phase water-bath temperature, while other preparation conditions remain unchanged. After preparation of the complex composition is completed, the average particle size, polydispersity index PDI, and zeta potential of the composition under each gradient are determined with a laser particle-size analyzer. At 40 °C, the fluidity of the oily raw materials is slightly poor, the raw-material dispersion rate is relatively slow, and the uniformity of the oil phase is insufficient, so the particle size of the prepared composition is relatively large; at 50 °C, the water-bath temperature is relatively high, and slight thermal dispersion of some raw materials occurs, thereby affecting the subsequent emulsification effect; at 45 °C, the oily DESCRIPTION raw materials have moderate fluidity and good compatibility with the other lipid-soluble raw materials, the oil phase is uniform and stable, and the prepared composition shows the best core indices overall.

[0072] Optimization of pH value of the phosphate buffer in the aqueous phase:

[0073] Three gradients of pH 7.0, 7.4, and 7.8 are set for the phosphate buffer in the aqueous phase, while other preparation conditions remain unchanged. After preparation is completed, the core indices are detected. At pH 7.0, the acid-base environment of the buffer is somewhat acidic, and the dissolving and dispersing effects of some water-soluble raw materials are unsatisfactory, resulting in slight precipitation in the aqueous phase; at pH 7.8, the acid-base environment of the buffer is somewhat alkaline, and slight structural changes occur in some phospholipid raw materials; at pH 7.4, the acid-base environment of the buffer is moderate, all water-soluble raw materials can be fully dissolved and dispersed, the aqueous phase is clear and free of precipitation, and the prepared composition reaches a relatively favorable level in the core indices.

[0074] Optimization of high-speed shearing time:

[0075] Three gradients of 3 min, 4 min, and 5 min are set for the high-speed shearing time, while other preparation conditions remain unchanged. After preparation is completed, the core indices are detected. At 3 min, the shearing time is short and the shearing intensity is insufficient, so the uniformity of the primary emulsion is poor and the particle-size distribution of the composition is broad; at 5 min, the shearing time is excessively long, and heat accumulation generated by shearing causes signs of emulsion breaking in part of the primary emulsion, thereby affecting the structure of the nanocarrier; at 4 min, the shearing DESCRIPTION time is moderate, the uniformity of the primary emulsion is good, the oil phase is sufficiently dispersed in the aqueous phase, the particle-size distribution of the prepared composition is concentrated, and the PDI value is relatively low.

[0076] Optimization of number of high-pressure homogenization cycles:

[0077] Three gradients of 3 cycles, 4 cycles, and 5 cycles are set for the number of high-pressure homogenization cycles, while other preparation conditions remain unchanged. After preparation is completed, the core indices are detected. At 3 cycles, the number of homogenization cycles is insufficient and the homogenization effect is inadequate, so the particle size of the composition is relatively large and the PDI value is slightly high; at 5 cycles, the number of homogenization cycles is excessive, the nanocarrier of the composition is subjected to repeated pressure impact, and the zeta potential shows slight fluctuation; at 4 cycles, the homogenization effect is sufficient, the nanocarrier structure of the composition is stable, and the average particle size, PDI, and zeta potential all exhibit relatively favorable values. DESCRIPTION

[0078] Comparison of optimization indices for key preparation conditions:

[0079] The test results of the core indices under different gradient parameters for the four key preparation conditions in this embodiment are shown in the following table.

[0080] The table data show that changes in the gradient parameters of the four key preparation conditions all have significant influences on the average particle size, DESCRIPTION polydispersity index PDI, and zeta potential of the complex composition, and the indices show regular changing trends with the parameter gradients. When the oil-phase water-bath temperature and the pH value of the aqueous phase deviate from the intermediate gradients, the average particle sizes of the compositions increase to different extents and the PDI values rise, indicating that the dispersion and fusion effects of the raw materials are reduced; when the high-speed shearing time and the number of high-pressure homogenization cycles are insufficient or excessive, the particle-size distribution of the composition broadens and the stability of the zeta potential decreases. The intermediate gradient parameters for the four conditions, namely 45 °C, pH 7.4, 4 min, and 4 cycles, provide the best overall preparation effect.

[0081] Embodiment 4

[0082] Performance tests of the brain-targeting lipid-phospholipid hybrid nanocarrier of the complex composition.

[0083] Comprehensive performance tests are carried out on the brain-targeting lipid-phospholipid hybrid nanocarrier of the complex composition prepared in Embodiment 1. Average particle size, polydispersity index PDI, zeta potential, encapsulation efficiency of the core effective components, stability in simulated gastric fluid in vitro, and stability in simulated intestinal fluid in vitro are selected as test indices. Standard industry methods are used for all test methods, each test index is tested six times, and the average value is taken as the final test result. The specific test procedures are as follows.

[0084] Determination of average particle size, polydispersity index PDI, and zeta potential: DESCRIPTION

[0085] A laser particle-size analyzer is used to determine the average particle size, polydispersity index PDI, and zeta potential of the nanocarrier. Before testing, the sample is equilibrated in a constant-temperature environment at 25 °C for 30 min, the detection temperature is controlled at 25 °C, the detection range is 0.1-1000 nm, and, during the testing process, the sample is kept in a uniformly dispersed state to avoid particle aggregation. After completion of the testing, the data from the six measurements are statistically processed and the average values are calculated to obtain the specific values of the average particle size, polydispersity index PDI, and zeta potential.

[0086] Determination of encapsulation efficiency of the core effective components:

[0087] Acer truncatum seed oil, phosphatidylserine, and docosahexaenoic acid phospholipid are selected as the core effective components of the complex composition, and high-performance liquid chromatography is used to determine the encapsulation efficiencies of the three components, respectively. Before testing, the sample is pretreated, the nanocarrier and free effective components are separated by high-speed centrifugation, the supernatant is collected to determine the contents of the free components, and the encapsulation efficiencies are calculated in combination with the total contents of the respective components in the sample. Each component is tested six times, and the average value is taken as the final encapsulation efficiency.

[0088] Determination of stability in simulated gastric fluid in vitro:

[0089] The acid-base environment and temperature conditions of human gastric fluid are simulated to prepare simulated gastric fluid. The nanocarrier of the complex composition is placed into the simulated gastric fluid and incubated at a DESCRIPTION constant temperature of 37 °C for 2 h. After incubation is completed, the sample is taken out and the average particle size is determined with a laser particle-size analyzer, and the change rate relative to the particle size before incubation is calculated. This test is repeated six times, and the average value is taken as the test result for stability in simulated gastric fluid in vitro.

[0090] Determination of stability in simulated intestinal fluid in vitro:

[0091] The acid-base environment and temperature conditions of human intestinal fluid are simulated to prepare simulated intestinal fluid. The nanocarrier of the complex composition is placed into the simulated intestinal fluid and incubated at a constant temperature of 37 °C for 4 h. After incubation is completed, the sample is taken out and the average particle size is determined with a laser particle-size analyzer, and the change rate relative to the particle size before incubation is calculated. This test is repeated six times, and the average value is taken as the test result for stability in simulated intestinal fluid in vitro.

[0092] Comparison of nanocarrier performance test indices:

[0093] The test results of the respective performance indices of the brain-targeting lipid-phospholipid hybrid nanocarrier of the complex composition in this embodiment are shown in the following table. DESCRIPTION

[0094] The table data show that the values of the respective performance indices of the nanocarrier of the complex composition exhibit good overall performance, and the consistency of the six repeated tests is relatively high, indicating that the prepared nanocarrier has stable performance. The average particle size is 148 nm, which falls within the reasonable range of 100-200 nm and is beneficial to transport and brain-targeted delivery of the nanocarrier in vivo; the PDI value is 0.14, which is relatively low, indicating that the particle-size distribution of the nanocarrier is highly concentrated, the particles are uniform, and there is no obvious particle-size difference; the zeta potential is -35 mV, which falls within the range of -30 to -40 mV, and this potential value can render the nanocarrier system sufficiently stable and effectively reduce aggregation among particles. The encapsulation efficiencies of Acer truncatum seed oil, phosphatidylserine, and docosahexaenoic acid phospholipid are 93%, 91%, and 92%, respectively, indicating that the nanocarrier has a good encapsulation effect on the core effective components. The particle-size change rates in simulated gastric fluid in vitro for 2 h and in simulated intestinal fluid in vitro for 4 h are 5% and 7%, respectively, indicating that the nanocarrier still maintains relatively good structural stability in digestive-fluid environments.

[0095] Embodiment 5

[0096] Tests of antioxidant performance and storage stability of the complex DESCRIPTION composition.

[0097] With peroxide value and acid value as core antioxidant evaluation indices, basic tests of the antioxidant performance of the complex composition are carried out using standard industry methods, and three different storage conditions, namely room temperature protected from light, room temperature under light exposure, and low temperature protected from light, are set so as to investigate changes in peroxide value and acid value of the composition at five time points of 0 day, 7 days, 14 days, 21 days, and 28 days during storage and to analyze storage stability. All tests are performed under light-protected conditions, each time point is tested in triplicate, and the average value is taken as the final test result. The specific testing procedures are as follows.

[0098] Basic antioxidant performance test:

[0099] Potentiometric titration is used to determine the peroxide value and acid value of the freshly prepared complex composition. Before testing, the sample is equilibrated at room temperature, titration is carried out according to the standard operating procedure, titration data are recorded, and the peroxide value and acid value are calculated. This test is repeated three times, and the average value is taken as the result of the basic antioxidant performance test.

[0100] Determination under room-temperature light-protected storage conditions:

[0101] The complex composition is stored in an environment at room temperature protected from light. Samples are taken at five time points of 0 day, 7 days, 14 days, 21 days, and 28 days of storage. Potentiometric titration is used to determine the peroxide value and acid value of the samples. Each test is repeated three times, the average value is taken, and changes in the indices at the different time points DESCRIPTION are recorded.

[0102] Determination under room-temperature light-exposure storage conditions:

[0103] The complex composition is stored in an environment at room temperature under light exposure, with light intensity kept constant. Samples are taken at five time points of 0 day, 7 days, 14 days, 21 days, and 28 days of storage. Potentiometric titration is used to determine the peroxide value and acid value of the samples. Each test is repeated three times, the average value is taken, and changes in the indices at the different time points are recorded.

[0104] Determination under low-temperature light-protected storage conditions:

[0105] The complex composition is stored in a low-temperature light-protected environment, with the temperature stably maintained at 4 °C. Samples are taken at five time points of 0 day, 7 days, 14 days, 21 days, and 28 days of storage. Potentiometric titration is used to determine the peroxide value and acid value of the samples. Each test is repeated three times, the average value is taken, and changes in the indices at the different time points are recorded.

[0106] Comparison of antioxidant and storage-stability indices of the complex composition:

[0107] The test results of the core antioxidant indices of the complex composition at different time points under the three storage conditions in this embodiment are shown in the following table. DESCRIPTION DESCRIPTION

[0108] The table data show that the peroxide value and acid value of the complex composition both increase with extension of storage time, and the storage conditions have a significant influence on the rate of increase of the indices, with obvious differences in the extent of changes under different storage conditions. Under the three storage conditions, the peroxide value and acid value on day 0 are identical, being 1.8 mmol / kg and 0.5 mg KOH / g, respectively, indicating that the freshly prepared composition has good antioxidant performance and no initial oxidation phenomenon. Under room-temperature light-exposure conditions, the peroxide value and acid value of the composition increase most rapidly, reaching 2.8 mmol / kg and 0.9 mg KOH / g, respectively, on day 28; under room-temperature light-protected conditions, the indices increase more slowly; and under low-temperature light-protected conditions, the increases are the smallest. These results show that the composition has relatively good storage stability, particularly under low-temperature light-protected conditions. DESCRIPTION

[0109] The above description only sets forth preferred embodiments of the present invention and is not intended to limit the present invention in any form. Although the present invention has been disclosed above with reference to the preferred embodiments, the present invention is not limited thereto. Any person skilled in the art may make certain changes or modifications to the disclosed technical content without departing from the scope of the technical solution of the present invention, and all simple modifications, equivalent changes, and modifications made to the above embodiments according to the technical spirit of the present invention shall still fall within the scope of the technical solution of the present invention.

Claims

1. CLAIMS1. An Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development, characterized in that, in parts by weight, the complex composition comprises 8-12 parts of Acer truncatum seed oil, 3-6 parts of phosphatidylserine, 1-2 parts of cephalin, 2-4 parts of linseed oil, 0.5-2 parts of arachidonic acid phosphatidylinositol, 1-3 parts of docosahexaenoic acid phospholipid, 0.3-1.5 parts of sialic acid, 0.2-1 part of phytosterol, 0.1-0.8 part of d-alpha-tocopherol, 0.5-2 parts of a co-emulsifier, and 0.05-0.3 part of an antioxidant synergist; wherein the composition is in the form of a brain-targeting lipid-phospholipid hybrid nanocarrier, with linseed oil serving as a base membrane material, phosphatidylserine, cephalin, and arachidonic acid phosphatidylinositol serving as functional membrane auxiliary materials, Acer truncatum seed oil and docosahexaenoic acid phospholipid serving as core materials, and phosphatidylserine serving as a brain-targeting modification group on the carrier surface.

2. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 1, characterized in that the Acer truncatum seed oil is low-temperature cold-pressed Acer truncatum kernel oil, with a pressing temperature of <=45 °C, wherein the mass content of nervonic acid is >=6%, the total mass content of unsaturated fatty acids is >=88%, the mass content of linoleic acid is >=30%, the mass content of oleic acid is >=25%, the peroxide value is <=3 mmol / kg, the acid value is <=1.0 mg KOH / g, and the solvent residue is <=0.001%.

3. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 1, characterized in that the phosphatidylserine is derived from soybean or sunflower seed and is prepared by a column chromatography separation and purification process, with an effective purity of >=65%, a free fatty acid content of <=1.0%, a moisture content of <=0.5%, an ash content of <=0.3%, and a total heavy metal content of <=0.1 mg / kg; and the cephalin is obtained byCLAIMS molecular distillation purification of soybean concentrated phospholipid, with an effective purity of >=55%, phosphatidylethanolamine accounting for >=90% by mass of the cephalin, a free cholesterol content of <=0.5%, a moisture content of <=0.5%, and an acid value of <=2.0 mg KOH / g.

4. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 1, characterized in that the arachidonic acid phosphatidylinositol is prepared by a microbial fermentation method or an egg-yolk phospholipid separation and purification method, with a purity of >=95%, a retention rate of the phosphatidylinositol head-group structure of >=98%, a binding rate of arachidonic acid in the arachidonic acid phosphatidylinositol of >=92%, a moisture content of <=0.3%, no protein residue, and a heavy metal content of <=0.05 mg / kg; and the docosahexaenoic acid phospholipid is of a phospholipid-bound type and is derived from Schizochytrium fermentation phospholipid or egg-yolk phospholipid, with a mass content of DHA of >=45%, a purity of phosphatidylcholine of >=80%, no free DHA residue, and an oxidation value of <=2 mmol / kg.

5. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 1, characterized in that the sialic acid is N-acetylneuraminic acid derived from milk or bird's nest, with a purity of >=98%, an effective content of >=98%, and a moisture content of <=0.5%; the phytosterol is a mixture of beta-sitosterol, stigmasterol, and campesterol in a mass ratio of 5:3:2, with a total purity of >=95%, a proportion of beta-sitosterol of >=55%, no phytosterol ester impurity, and a melting point of 130-140 °C; and the d-alpha-tocopherol is natural vitamin E extracted from soybean oil or sunflower- seed oil, with a purity of >=96%, d-alpha-tocopherol accounting for >=92% of total vitamin E, and no synthetic vitamin E added.

6. The Acer truncatum seed oil-phosphatidylserine (PS) complexCLAIMS composition for promoting neural development according to claim 1, characterized in that the co-emulsifier is a combination of sucrose fatty acid ester and polyglycerol fatty acid ester in a mass ratio of 2:1, wherein the HLB value of the sucrose fatty acid ester is 8-10 and the HLB value of the poly glycerol fatty acid ester is 10-12; and the antioxidant synergist is a combination of ascorbyl palmitate and rosemary extract in a mass ratio of 3:1, wherein the carnosic acid content in the rosemary extract is >=5%.

7. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 1, characterized in that the brain-targeting lipid-phospholipid hybrid nanocarrier has an average particle size of 140-160 nm, a polydispersity index (PDI) of <=0.15, and a zeta potential of -34 to -36 mV; the encapsulation efficiencies of the core effective components are all >=90%; the particle-size change rate in simulated gastric fluid in vitro for 2 h is <=5%; and the particle-size change rate in simulated intestinal fluid in vitro for 4 h is <=7%.

8. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 1, characterized in that the preparation method of the complex composition comprises the following steps:

51. Raw-material pretreatment and weighing: weighing all components according to the parts-by -weight ratio; subjecting the Acer truncatum seed oil and linseed oil to vacuum filtration through an organic filter membrane; subjecting phosphatidylserine, cephalin, arachidonic acid phosphatidylinositol, and docosahexaenoic acid phospholipid to vacuum drying; separately sieving the sialic acid, phytosterol, d-alpha-tocopherol, co-emulsifier, and antioxidant synergist; and placing all pretreated raw materials in dry, light-protected sealed containers for subsequent use;52. Preparation of an oil phase: adding the pretreated and weighed Acer truncatum seed oil and linseed oil into a constant-temperature stirring vessel,CLAIMS starting the stirring device and controlling the water-bath temperature at 40-50 °C, then successively adding arachidonic acid phosphatidylinositol, docosahexaenoic acid phospholipid, phytosterol, and d-alpha-tocopherol after stirring, adjusting the stirring speed and continuing stirring until a uniform, transparent, particle-free, and non-layered lipid-soluble oil phase is formed, and keeping the system under stirring and constant temperature for subsequent use;53. Preparation of an aqueous phase: preparing a phosphate buffer having a pH of 7.0-7.8 and adding the buffer into a constant-temperature stirring vessel, starting water-bath heating and adjusting the stirring speed, wherein the water-bath temperature is 40-50 °C and is kept consistent with that of the oil phase, then successively adding the pretreated phosphatidylserine, cephalin, co-emulsifier, antioxidant synergist, and sialic acid, adjusting the stirring speed and continuing stirring until a uniform, clear, and precipitate-free aqueous phase is formed, and, after standing, keeping the system under constant temperature and stirring for subsequent use;54. Emulsification and forming of the nanocarrier: keeping the water-bath temperatures of the two stirring vessels consistent, slowly adding the oil phase into the aqueous phase through a constant-flow pump while maintaining stirring during the addition, increasing the stirring speed after completion of the addition to perform high-speed shearing so as to form a primary emulsion, and then introducing the primary emulsion into a high-pressure homogenizer for cyclic homogenization, with sampling performed to detect particle size until the average particle size reaches 140-160 nm and the polydispersity index PDI is <=0.15;55. Post-treatment and filling of the final product: transferring the composite nano-composition to pasteurization equipment for sterilization, cooling to room temperature after sterilization, then transferring to a sterile filling workshop for sealed filling in a light-protected sterile environment, immediately sealing after filling, placing the finished product in aCLAIMS low-temperature light-protected environment for storage, and simultaneously conducting sampling inspection of various indicators, whereby a qualified product is the final product.

9. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 8, characterized in that, in step SI, the weighing process is carried out in a light-protected, dry, and dust-free environment; the temperature for vacuum drying is 40 °C, the drying time is 2 h, and nitrogen protection is used during drying; the pore size of the organic filter membrane is 0.22 um, the filtration pressure is 0.1-0.15 MPa, the filtration time is 10-15 min; and the sieve is an 80-mesh stainless-steel sieve.

10. The Acer truncatum seed oil-phosphatidylserine (PS) complex composition for promoting neural development according to claim 8, characterized in that, in step S4, the stirring speed is maintained at 400 r / min during dropwise addition of the oil phase; the high-speed shearing speed is 10,000-12,000 r / min, and the shearing time is 3-5 min; the high-pressure homogenizer is a plunger high-pressure homogenizer, the homogenization pressure is 600-800 bar, cyclic homogenization is performed 3-5 times, the homogenization time for each cycle is 2-3 min, and the interval between two homogenization cycles is 1 min; particle-size detection is performed with a laser particle-size analyzer, the detection range is 0.1-1000 nm, and the detection temperature is 25 + / - 2 °C.

Images