Structural lipid and preparation method thereof, structural lipid composition for protecting liver and eyes comprising structural lipid, and product for protecting liver and eyes

Abstract

The invention relates to the field of synthetic biology, in particular to a structural lipid, a preparation method thereof, a structural lipid composition containing the structural lipid and used for protecting liver and eyes, and a product for protecting liver and eyes. The preparation method comprises the following steps: carrying out reaction on mixed oil and lipase, and carrying out enzyme deactivation to prepare structural lipid; wherein the weight ratio of omega3 fatty acid to omega6 fatty acid to omega9 fatty acid in the oil material is (0.8-1.2): (0.8-1.2): (0.8-1.2). The lipase is adopted to carry out hydrolysis reaction on appropriate oil, sn-2 site fatty acid with the liver and eye protection effect can be prepared, and the sn-2 site fatty acid is small in molecular weight and beneficial to absorption.

Description

Technical Field

[0001] This application relates to the field of synthetic biology, specifically to structural lipids and their preparation methods, including structural lipid compositions that protect the liver and eyes, and liver- and eye-protecting products. Background Technology

[0002] With rapid economic development in modern society and significant changes in dietary structure, the number of patients with fatty liver disease has grown rapidly. Fatty liver disease, also known as fatty liver syndrome, is a disease characterized by the breakdown of liver cells into fatty tissue, caused by various factors such as genetic predisposition, environmental factors, and metabolic stress.

[0003] The frequent use of modern electronic devices has led to the absorption of more short-wavelength blue light than ever before, which is extremely harmful to the macula of the eye. Therefore, there is a strong desire to improve symptoms such as dry eye, average visual acuity, glare tolerance, and to enhance visual acuity and relieve dizziness.

[0004] Therefore, there is an urgent need in this field to develop products with liver-protecting and eye-protecting effects. Summary of the Invention

[0005] This application provides one or more embodiments of a liver-protecting and eye-protecting structural lipid, its preparation method, and its application. The technical solutions include the following:

[0006] One or more embodiments of this application provide a method for preparing structural lipids, the preparation method comprising:

[0007] Mixed oils and lipases are reacted to inactivate the enzyme and prepare structured lipids.

[0008] The weight ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the oil is (0.8-1.2):(0.8-1.2):(0.8-1.2).

[0009] Optionally, the oilseeds include a variety of milk thistle seed oil, borage seed oil, conjugated linoleic acid glycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, *Sapindus mukorossi* oil, low-erucic acid rapeseed oil, linseed oil, palm oil, and coconut oil.

[0010] In some specific embodiments of this application, the oilseeds include milk thistle seed oil, borage seed oil, conjugated linoleic acid glycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, and *Sapindus mukorossi* oil.

[0011] Optionally, the oil comprises, by weight:

[0012]

[0013] In some specific embodiments of this application, the oilseeds include low-erucic acid rapeseed oil, borage seed oil, medium-chain triglycerides, perilla seed oil, algal oil DHA, *Sapindus mukorossi* oil, and flaxseed oil;

[0014] Optionally, the oil comprises, by weight:

[0015]

[0016] In some specific embodiments of this application, the oilseeds include milk thistle seed oil, borage seed oil, conjugated linoleic acid triglycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, and *Sapindus mukorossi* oil.

[0017] Optionally, the oil comprises, by weight, parts of oil.

[0018]

[0019]

[0020] In some specific embodiments of this application, the oilseeds include medium-chain triglycerides, perilla seed oil, DHA algal oil, *Sapindus mukorossi* oil, palm oil, and coconut oil;

[0021] Optionally, the oil comprises, by weight:

[0022]

[0023] In some specific embodiments of this application, the preparation method satisfies one or more of the following conditions:

[0024] (1) The weight of the lipase is 3-6 wt% of the weight of the oil; optionally, the lipase includes an immobilized lipase.

[0025] (2) The reaction conditions include: sealed environment, 10-50 Pa, 60-75℃, 24-28 hours; and,

[0026] (3) Enzyme inactivation is performed by thermal inactivation or organic solvent inactivation.

[0027] One or more embodiments of this application provide a structural lipid prepared using the preparation method described above.

[0028] One or more embodiments of this application provide a structural lipid composition that protects the liver and eyes, the structural lipid composition comprising:

[0029] 1) the structural lipids described above; and,

[0030] 2) Lutein esters and / or Haematococcus pluvialis;

[0031] Optionally, the composition comprises, by weight:

[0032] The structural lipids comprised 99.4-99.7 parts.

[0033] Lutein ester 0.2-0.3 parts, and,

[0034] Haematococcus pluvialis 0.15-0.25 parts.

[0035] One or more embodiments of this application provide a method for preparing the liver-protecting and eye-protecting structural lipid composition described above, the preparation method comprising:

[0036] Structural lipids were prepared using the method described above; and

[0037] The structural lipids are mixed with the lutein esters and / or Haematococcus pluvialis to prepare a liver-protecting and eye-protecting structural lipid composition.

[0038] One or more embodiments of this application provide a liver-protecting and eye-protecting product, the liver-protecting and eye-protecting product comprising the structural lipids or the structural lipid composition described above;

[0039] Optionally, the liver and eye protection products include capsules.

[0040] Compared to traditional technologies, the beneficial effects of this application include: This application utilizes lipase to hydrolyze suitable oils, enabling the preparation of sn-2 position fatty acids with liver-protective and eye-protective effects. These fatty acids have a small molecular weight, facilitating absorption. In particular, the structural lipid mixture prepared by combining sn-2 position fatty acids with lutein esters and Haematococcus pluvialis exhibits significant liver-protective and eye-protective effects. Attached Figure Description

[0041] To more clearly illustrate the technical solutions in the embodiments of this application and to more completely understand this application and its beneficial effects, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0042] Figure 1 To analyze the sample efficacy test results in Test Report (I).

[0043] Figure 2 To analyze the experimental phenotype diagram (40x magnification) in Test Report (I).

[0044] Figure 3 To analyze the sample efficacy test results in Test Report (II).

[0045] Figure 4For analysis of the experimental phenotype diagram (40x magnification) in Test Report (II). Detailed Implementation

[0046] The present application will be further described in detail below with reference to the accompanying drawings, embodiments, and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0047] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for descriptive purposes only and is not intended to be limiting of the application.

[0048] the term

[0049] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0050] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0051] In this application, the terms "multiple", "various", "multiple times", "multi-dimensional", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0052] The terms “combinations of,” “any combination of,” and “any combination of” used in this article include all suitable combinations of any two or more of the listed items.

[0053] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0054] In this document, terms such as “preferred,” “better,” “more suitable,” and “ideal” are merely used to describe implementation methods or examples that achieve better results, and should be understood not to limit the scope of protection of this application.

[0055] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0056] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0057] In this application, the terms "first aspect," "second aspect," "third aspect," "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0058] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0059] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0060] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0061] In this application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass-volume percentage.

[0062] All references to this application are incorporated herein by reference as if each document were individually incorporated herein by reference. Unless they conflict with the purpose and / or technical solution of this application, all cited references are incorporated herein by reference in their entirety and for all purposes. When references are cited in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. Examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0063] One or more embodiments of this application provide a method for preparing structural lipids, the preparation method comprising:

[0064] Mixed oils and lipases are reacted to inactivate the enzyme and prepare structured lipids.

[0065] The weight ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the oil is (0.8-1.2):(0.8-1.2):(0.8-1.2) (for example, 0.8:0.8:0.8, 0.8:0.8:1, 0.8:0.8:1.2, 0.8:1:0.8, 0.8:1:1, 0.8:1:1.2, 0.8:1.2:0.8, 0.8:1.2:1, 0.8:1.2:1.2, 1:1:0.8, 1:1:1.2, 1.2:1.2:0.8, 1.2:1.2:1).

[0066] This application utilizes lipase to hydrolyze suitable oils, which can produce sn-2 fatty acids with liver-protecting and eye-protecting effects. These fatty acids have a small molecular weight and are easily absorbed.

[0067] In some examples of this application, the oilseeds may optionally include a variety of milk thistle seed oil, borage seed oil, conjugated linoleic acid glycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, *Sapindus mukorossi* oil, low-erucic acid rapeseed oil, flaxseed oil, palm oil, and coconut oil.

[0068] In some examples of this application, the oilseeds include milk thistle seed oil, borage seed oil, conjugated linoleic acid glycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, and *Sapindus mukorossi* oil;

[0069] Optionally, the oil comprises, by weight:

[0070]

[0071] In some examples of this application, the oils include low-erucic acid rapeseed oil, borage seed oil, medium-chain triglycerides, perilla seed oil, algal oil DHA, *Sapindus mukorossi* oil, and flaxseed oil;

[0072] Optionally, the oil comprises, by weight:

[0073]

[0074] In some examples of this application, the oilseeds include milk thistle seed oil, borage seed oil, conjugated linoleic acid triglycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, and *Sapindus mukorossi* oil;

[0075] Optionally, the oil comprises, by weight, parts of oil.

[0076]

[0077] In some examples of this application, the oils include medium-chain triglycerides, perilla seed oil, DHA algal oil, *Sapindus mukorossi* oil, palm oil, and coconut oil;

[0078] Optionally, the oil comprises, by weight:

[0079]

[0080] In some examples of this application, the preparation method satisfies one or more of the following conditions:

[0081] (1) The weight of the lipase is 3-6 wt% of the weight of the oil (e.g., 3 wt%, 3.2 wt%, 3.4 wt%, 3.6 wt%, 3.8 wt%, 4 wt%, 4.2 wt%, 4.4 wt%, 4.6 wt%, 4.8 wt%, 5 wt%, 5.2 wt%, 5.4 wt%, 5.6 wt%, 5.8 wt%, 6 wt%); optionally, the lipase includes immobilized lipase. This application does not particularly limit the type of immobilized lipase, including but not limited to: 435 immobilized lipase, IMNE-100 immobilized enzyme;

[0082] (2) The reaction conditions include: sealed environment, 10-50 Pa (e.g., 10, 15, 20, 25, 30, 35, 40, 45, 50 Pa), 60-75℃ (e.g., 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75℃), 24-28 hours (e.g., 24, 25, 26, 27, 28 hours); and,

[0083] (3) Enzyme inactivation is performed by thermal inactivation or organic solvent inactivation. For example, the product obtained from the reaction is heated at 105-115℃ (e.g., 10, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115℃) for 1-3 hours (e.g., 1, 1.5, 2, 2.5, 3 hours). For example, enzyme inactivation using ethanol includes: mixing the reaction product with edible ethanol at a volume ratio of 0.8-1.2:0.8-1.2 (e.g., 1:1, 0.8:1, 1.2:0.8), reacting at 65-75℃ (e.g., 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75℃) for 3-4 hours (e.g., 3, 3.2, 3.4, 3.6, 3.8, 4 hours), holding at this temperature for 4-6 hours (e.g., 4, 4.2, 4.4, 4.6, 4.8, 5, 5.2, 5.4, 5.6, 5.8, 6 hours), and then removing the edible ethanol.

[0084] One or more embodiments of this application provide a structural lipid prepared using the preparation method described above.

[0085] One or more embodiments of this application provide a structural lipid composition that protects the liver and eyes, the structural lipid composition comprising:

[0086] 1) the structural lipids described above; and,

[0087] 2) Lutein esters and / or Haematococcus pluvialis;

[0088] Optionally, the composition comprises, by weight:

[0089] The structural lipids comprised 99.4-99.7 parts.

[0090] Lutein ester 0.2-0.3 parts, and,

[0091] Haematococcus pluvialis 0.15-0.25 parts.

[0092] One or more embodiments of this application provide a method for preparing the liver-protecting and eye-protecting structural lipid composition described above, the preparation method comprising:

[0093] Structural lipids were prepared using the method described above; and

[0094] The structural lipids are mixed with the lutein esters and / or Haematococcus pluvialis to prepare a liver-protecting and eye-protecting structural lipid composition.

[0095] One or more embodiments of this application provide a liver-protecting and eye-protecting product, the liver-protecting and eye-protecting product comprising the structural lipids or the structural lipid composition described above;

[0096] This application does not impose any specific restrictions on the dosage form of liver-protecting and eye-protecting products, including but not limited to capsules.

[0097] The liver and eye protection products described in this application can be used externally or taken internally.

[0098] This application describes the preparation and application of a liver-protecting and eye-protecting structural lipid composition. The composition comprises C8-10 fatty acids, C16-24 fatty acids, conjugated linoleic acid glycerides, and Haematococcus pluvialis oil (astaxanthin oil) as an antioxidant. The reaction is carried out under immobilized enzyme conditions, with 99% sn-2 monoacylglycerols. The product is then homogenized with the addition of Haematococcus pluvialis oil and lutein esters. The outer shell is composed of gelatin, glycerol, water, and sorbitol solution, and the mixture is prepared into a soft capsule gel candy. The structural lipid composition obtained in this application has a good effect on improving liver lipid metabolism, preventing and reducing the progression of fatty liver, eliminating inflammation, inhibiting ocular cell apoptosis, and curbing the increase of myopia. The preparation method of this invention is simple and easy to operate, which is conducive to industrial production.

[0099] Specifically as follows:

[0100] Milk thistle seeds are sourced from Hebei Silk Road Morning Light Oils Co., Ltd.

[0101] Borage seed oil is from: Guyuan Biotechnology (Tianjin) Co., Ltd.

[0102] Conjugated linoleic acid glycerides are from Qingdao Aohai Biotechnology Co., Ltd.

[0103] Medium-chain triglycerides are sourced from Guangzhou Yongyi Food Ingredients Co., Ltd.

[0104] Perilla seed oil: Guyuan Biotechnology (Tianjin) Co., Ltd.

[0105] Algal oil DHA is from Anhui Tiankai Biotechnology Co., Ltd.

[0106] The source of the *Sapindus mukorossi* oil is Hebei Runsheng Biotechnology Co., Ltd.

[0107] Haematococcus pluvialis oil (astaxanthin oil) is from: Chengshan Industrial (Shanghai) Co., Ltd.

[0108] Low-erucic acid rapeseed oil is from Hubei Aoxing Grain and Oil Industry Co., Ltd.

[0109] Flaxseed oil is from Hebei Silk Road Morning Light Oil Co., Ltd.

[0110] Coconut oil is from Kerry Oils & Grains (Tianjin) Co., Ltd.

[0111] Palm oil sourced from: Kerry Oils & Grains (Tianjin) Co., Ltd.

[0112] In the various embodiments and comparative examples:

[0113] Medium and long chain fatty acids: Enterprise standard Q / KRD-0002S-2025.

[0114] Sn-2 fatty acid: GB24894-2010.

[0115] Medium- and long-chain fatty acid triglycerides: Enterprise standard Q / KRD-0002S-2025.

[0116] Medium-chain fatty acids: Enterprise standard Q / KRD-0002S-2025.

[0117] Example 1:

[0118] 1. Take 30 parts of milk thistle seed oil, 25 parts of borage seed oil, 20 parts of conjugated linoleic acid glycerides, 16 parts of medium-chain triglycerides, 5 parts of perilla seed oil, 1.5 parts of algal oil DHA, and 2.5 parts of *Sapindus mukorossi* oil to ensure that the mass ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the obtained oil is 1:1:1. Add the oil to a closed reactor and add 3.5 wt% of 435 immobilized lipase by weight of the oil. The initial reaction system is reacted at 69℃ and under vacuum of 45 Pa for 25 hours to prepare a first-structure lipid with a medium- and long-chain fatty acid triglyceride content of 25 wt%, a medium-chain fatty acid content of 13 wt%, and a sn-2 position fatty acid content of 99 wt%.

[0119] 2. Add anhydrous ethanol (food-grade anhydrous ethanol to inactivate enzymes) to the first structural lipid prepared in step 1 at a volume ratio of 1:1. Stir the reaction at 70°C for 3 hours and keep warm for 6 hours to inactivate the enzyme. Remove the ethanol under vacuum and collect the first structural lipid.

[0120] 3. Add 0.24 parts of lutein ester and 0.2 parts of Haematococcus pluvialis oil (astaxanthin oil) to 99.56 parts of the first structure lipid collected in step 2, mix and homogenize to obtain the second structure lipid, wherein the content of medium- and long-chain fatty acid triglycerides is 30 wt%, the content of medium-chain fatty acids is 14 wt%, and the content of fatty acids at the sn-2 position is 99 wt%.

[0121] 4. Prepare soft capsules (gel candies) by mixing 0.85g of second-structure lipid and 0.35g of gelatin material. The gelatin material includes the following raw materials in the following weight percentages: gelatin 35%, purified water 35%, glycerin 20%, and sorbitol solution (sorbitol content greater than 90%) 10%.

[0122] This embodiment also prepared a comparative soft capsule, specifically:

[0123] Compared to soft capsule 1, the preparation method of soft capsule 1 differs from the above-mentioned soft capsule preparation method only in the oil composition in step 1. Specifically, the oil consists of: 20 parts milk thistle seed oil, 20 parts borage seed oil, 20 parts conjugated linoleic acid glycerides, 20 parts medium-chain triglycerides, 10 parts algal oil DHA, and 10 parts *Sapindus mukorossi* oil. The weight ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the oil is 1:4:2. As a result, the content of the fatty acid at the n-2 position is 75 wt%.

[0124] Example 2:

[0125] 1. Take 35 parts of low-erucic acid rapeseed oil, 20 parts of borage seed oil, 20 parts of flaxseed oil, 15 parts of medium-chain triglycerides, 5 parts of perilla seed oil, 2.5 parts of *Sapindus mukorossi* oil, and 2.5 parts of algal oil DHA, so that the mass ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the resulting oil is 1:1:1. Add the oil to a closed reactor, add 3 wt% of IMNE-100 immobilized enzyme by weight of the oil, react for 26 hours at a reaction temperature of 68℃ and a vacuum of 50 Pa to prepare the first structure lipid, wherein the content of medium- and long-chain fatty acid triglycerides is 28%, the content of medium-chain fatty acids is 13.2 wt%, and the content of fatty acids at the sn-2 position is 99.5%.

[0126] 2. Add anhydrous ethanol to the first structural lipid in step 1 at a volume ratio of 1:1 (ethanol to oil), react at 70°C for 4 hours, keep warm for 6 hours, remove the anhydrous ethanol under vacuum, and collect the first structural lipid.

[0127] 3. Mix 99.65 parts of the first structure lipid with 0.20 parts of lutein ester and 0.15 parts of Haematococcus pluvialis (astaxanthin oil), and homogenize to obtain the second structure lipid, wherein the content of medium and long chain fatty acid triglycerides is 28%, the content of medium chain fatty acids is 12%, and the content of fatty acids at the sn-2 position is 99.8 wt%.

[0128] 4. Following step 4 of Example 1, the second structural lipid and the gelatin material are used to prepare a soft capsule.

[0129] Example 3:

[0130] 1. Take 25 parts of medium-chain triglycerides, 34 parts of milk thistle seed oil, 15 parts of borage seed oil, 15 parts of conjugated linoleic acid triglycerides, 5 parts of perilla seed oil, 3 parts of *Sapindus mukorossi* oil, and 2.7 parts of algal oil DHA to make the mass ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the resulting oils 1:1:1; add the oils to a closed reactor, add 4 wt% of the weight of the immobilized enzyme IMNE-100, and react in the reactor for 24 hours at a temperature of 68℃ and a vacuum of 40 Pa to prepare the first structured lipid, wherein the content of medium- and long-chain fatty acid triglycerides is 45 wt%, the content of medium-chain fatty acids is 32 wt%, and the content of fatty acids at the sn-2 position is 99 wt%.

[0131] 2. The first structure lipid was vaporized to inactivate the enzyme for 2 hours at 110°C, and the first structure lipid was collected.

[0132] 3. Mix 99.6 parts of the first structural lipid collected in step 2 with 0.2 parts of lutein ester and 0.2 parts of Haematococcus pluvialis oil (astaxanthin oil) and homogenize to prepare the second structural lipid. The second structural lipid contains 28 wt% medium- and long-chain fatty acid triglycerides, 14 wt% medium-chain fatty acids, and 99 wt% fatty acids at the sn-2 position.

[0133] 4. Following step 4 of Example 1, the second structural lipid and the gelatin material are used to prepare a soft capsule.

[0134] Example 4:

[0135] 1. Take 30 parts palm oil, 20 parts perilla seed oil, 20 parts DHA algal oil, 15 parts medium-chain triglycerides, 10 parts coconut oil, and 5 parts *Sapindus mukorossi* oil to make the mass ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the resulting oils 1:1:1. Add the oils to a closed reactor, add 3.5 wt% of 435 immobilized lipase by weight of the oils, and react for 24 hours at 70℃ and 50 Pa vacuum to prepare the first structured lipid, wherein the content of medium- and long-chain fatty acid triglycerides is 28 wt%, the content of medium-chain fatty acids is 13 wt%, and the content of fatty acids at the sn-2 position is 99.5 wt%.

[0136] 2. The first structural lipid was mixed with anhydrous ethanol at a volume ratio of 1:1, and reacted at 70°C for 4 hours. After incubation for 3 hours, the ethanol was removed under vacuum to inactivate the enzyme.

[0137] 3. The second structure lipid was collected by homogenizing 99.6 parts of the first structure lipid with 0.25 parts of lutein ester and 0.15 parts of Haematococcus pluvialis (astaxanthin oil). The content of medium- and long-chain fatty acid triglycerides was 27 wt%, the content of medium-chain fatty acids was 13 wt%, and the content of fatty acid at the sn-2 position was 99.5 wt%.

[0138] 4. Referring to step 4 of Example 1, the second structural lipids collected in step 3 are combined with the gelatin material to prepare soft capsules.

[0139] Example 5:

[0140] 1. Take 35 parts of milk thistle seed oil, 31 parts of borage seed oil, 14 parts of conjugated linoleic acid glycerides, 8 parts of medium-chain triglycerides, 5 parts of perilla seed oil, 3 parts of algal oil DHA, and 4 parts of *Sapindus mukorossi* oil to make the mass ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the resulting oilseeds 1:1:1; add the oilseeds to a closed reactor, add 6% of the weight of IMNE-100 immobilized lipase, and react for 25 hours at 72℃ and 35Pa vacuum to obtain a first-structure lipid with a medium- and long-chain fatty acid triglyceride content of 25%, a medium-chain fatty acid content of 15wt%, and a sn-2 position fatty acid content of 99wt%.

[0141] 2. Add anhydrous ethanol to the first structural lipid at a volume ratio of 1:1, stir and react at 70°C for 3 hours, keep warm, vacuum for 6 hours, remove ethanol, and collect the first structural lipid, in which medium-chain fatty acid triglycerides account for 35% and medium-chain fatty acids account for 15%.

[0142] 3. Mix 0.28 parts of lutein ester, 0.25 parts of Haematococcus pluvialis oil (astaxanthin oil), and 99.47 parts of the first structure lipid and stir until homogenized to obtain the second structure lipid.

[0143] 4. Following step 4 of Example 1, the second structural lipid and the gelatin material are used to prepare a soft capsule.

[0144] Performance testing

[0145] Analysis and Testing Report (I)

[0146] Table 1

[0147]

[0148] I. Overview of Laboratory Methods

[0149] 1. Detection Principle

[0150] Zebrafish embryos develop vision rapidly; juveniles, developing for only four days after fertilization, already rely on visual cues for hunting and avoidance behaviors. Zebrafish not only share significant similarities with human eyes in structures such as the cornea, lens, choroid, retina, vascularization, and nerve innervation, but also exhibit conserved gene expression, cellular composition, and tissue structure, thus serving as a viable model for vision protection. Damage to DBP (dibutyl phthalate) in zebrafish eyes results in an increased number of dead cells, thus establishing a zebrafish eye injury model.

[0151] 2. Materials and Methods

[0152] 2.1 Detection system and sample size

[0153] Experimental system: wild AB strain zebrafish

[0154] Zebrafish age: five days after fertilization

[0155] Sample size per group: 30 samples per well, 1 well per group

[0156] Adult fish rearing and breeding methods: Follow our company's standard rearing and breeding methods.

[0157] 2.2 Experimental Instruments and Reagents

[0158] Stereoscopic fluorescence microscope (SZX7, Olympus, Japan), dbp (CAS:2508-19-2), ginsenoside Rg1.

[0159] 3. Experimental Procedure

[0160] The zebrafish under test were divided into four groups: blank control group, model control group, positive control group, and sample treatment group. The test substance stock solution (i.e., the contents of the soft capsules) was diluted with buffer water in a proportional manner to prepare a series of appropriate concentration gradient solutions, with a dilution factor of not less than 10 times, and kept for later use.

[0161] The blank control group received no treatment, the model control group received dbp, the positive control group received dbp and ginsenoside Rg1 (1wt%), and the sample treatment group received dbp and sample. After a period of culture, the number of apoptotic zebrafish eye cells was detected.

[0162] II. Applicability and Limitations

[0163] For testing the eye protection efficacy of health products and their raw materials, the sample must be soluble in water or prepared into a uniformly dispersed suspension in water.

[0164] III. Data Processing Methods

[0165] SPSS 26 software was used for analysis, and GraphPad Prism 8 was used for plotting. Data statistics are expressed as mean ± standard deviation. Experimental results were tested for normality using SPSS software. If the data conformed to normality, the significance was determined by the T-parameter test or analysis of variance (ANOVA). If the data did not conform to normality, nonparametric tests were used (P-value < 0.05 indicates statistical significance).

[0166] IV. Judgment Criteria

[0167] Compared with the blank control group, the number of apoptotic zebrafish eye cells was significantly increased in the model control group (P<0.05), indicating that the modeling was successful.

[0168] Compared with the model control group, the number of apoptotic zebrafish eye cells in the sample treatment group was significantly less than that in the model control group (P<0.05), indicating that the sample has the effect of protecting eyesight.

[0169] V. Preliminary Experiment Results

[0170] There was no significant difference between the 1% concentration sample treatment group and the blank control group, therefore 1% concentration is the maximum safe concentration.

[0171] VI. Test Results

[0172] Table 2

[0173]

[0174] The bar chart of the sample efficacy test results is shown in the figure: Figure 1Figure caption: *: P < 0.05; **: P < 0.01; ***: P < 0.001; The more *, the smaller the P value, and the more significant the difference. Experimental phenotypic graph (40x magnification): Figure 2 .

[0175] Analysis and Testing Report (II)

[0176] Table 3

[0177]

[0178] I. Brief Description of Laboratory Methods

[0179] 1. Detection Principle

[0180] Zebrafish livers contain many lipid metabolism enzymes homologous to those found in mammals, mirroring mammalian defense mechanisms against exogenous chemicals, including enzyme induction and oxidative stress. Furthermore, the transparent bodies of juvenile zebrafish make it easy to observe toxic phenotypes. Ethanol can induce liver damage in zebrafish, mimicking human liver disease. Yolk sac absorption is closely related to liver function; therefore, assessing yolk sac absorption capacity can evaluate liver function in samples.

[0181] 2. Materials and Methods

[0182] 2.1 Experimental Instruments and Reagents

[0183] Stereoscopic microscope (SZX7, Olympus, Japan), anhydrous ethanol (CAS: 64-17-5), metadoxine (Zhejiang Zhenyuan Pharmaceutical Co., Ltd.)

[0184] 2.2 Detection system and sample size

[0185] Experimental system: wild AB strain zebrafish

[0186] Zebrafish age: 3 days after fertilization

[0187] Sample size per group: 30 samples per well, 1 well per group

[0188] Adult fish rearing and breeding methods: Follow our company's standard rearing and breeding methods.

[0189] 3. Experimental Procedure

[0190] We divided the zebrafish into four groups: a blank control group, a model control group, a positive control group, and a sample treatment group. The blank control group received no treatment. The model control group, positive control group, and sample treatment group all received anhydrous ethanol in the same volume as the water used for fish rearing (ethanol was ingested by the zebrafish through dissolution in the water). The positive control group and sample treatment group received ethanol along with metadoxine (1 wt%) and the test sample solution, respectively. After a period of culture, the liver-protective effect of the samples was evaluated by testing the yolk sac absorption capacity.

[0191] II. Applicability and Limitations

[0192] For testing of liver protection in health products, food and their raw materials, the sample must be soluble in water or prepared into a uniformly dispersed suspension in water.

[0193] III. Data Processing Methods

[0194] SPSS 26 software was used for analysis, and GraphPad Prism 8 was used for plotting. Data statistics are expressed as mean ± standard deviation. The experimental results were tested for normality using SPSS software. If the data conformed to normality, the significance was determined by the T-parameter test or analysis of variance (ANOVA). If the data did not conform to normality, nonparametric tests were used (P-value < 0.05 indicates statistical significance).

[0195] IV. Judgment Criteria

[0196] Based on the fact that the experiment met the validity requirements, the yolk sac area of ​​the model control group was significantly increased compared with that of the blank control group (P<0.05), which means that the modeling was successful.

[0197] Compared with the model control group, the yolk sac area in the sample treatment group was significantly reduced (P<0.05), indicating that the sample has a liver-protective effect.

[0198] V. Preliminary Experimental Test Results

[0199] There was no significant difference between the 1% concentration sample group and the blank control group, therefore 1% concentration is the maximum safe concentration.

[0200] VI. Test Results (See attached table on the next page)

[0201] Table 4

[0202]

[0203] The bar chart of the efficacy test results for this sample is shown below. Figure 3 As shown in the figure: *: P < 0.05; **: P < 0.01; ***: P < 0.001; The more *, the smaller the P value, indicating a more significant difference.

[0204] Phenotypic diagram of the efficacy test results of this sample (magnification 40x): Figure 4 .

[0205] Table 5. Summary Table of Clinical Application Cases

[0206]

[0207] Table 6 Summary of Clinical Application Cases (Part Two)

[0208]

[0209] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0210] The embodiments described above are merely illustrative of several implementation methods of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings of this application, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A method for preparing a structural lipid, characterized in that, The preparation method includes: Mixed oils and lipases are reacted to inactivate the enzyme and prepare structured lipids. The weight ratio of ω3 fatty acids, ω6 fatty acids, and ω9 fatty acids in the oil is (0.8-1.2):(0.8-1.2):(0.8-1.2). Optionally, the oilseeds include a variety of milk thistle seed oil, borage seed oil, conjugated linoleic acid glycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, *Sapindus mukorossi* oil, low-erucic acid rapeseed oil, linseed oil, palm oil, and coconut oil.

2. The method for preparing structural lipids according to claim 1, characterized in that, The oilseeds include milk thistle seed oil, borage seed oil, conjugated linoleic acid glycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, and *Sapindus mukorossi* oil. Optionally, the oil comprises, by weight:

3. The method for preparing structural lipids according to claim 1, characterized in that, The oils include low-erucic acid rapeseed oil, borage seed oil, medium-chain triglycerides, perilla seed oil, algal oil DHA, *Sapindus mukorossi* oil, and flaxseed oil; Optionally, the oil comprises, by weight:

4. The method for preparing structural lipids according to claim 1, characterized in that, The oilseeds include milk thistle seed oil, borage seed oil, conjugated linoleic acid triglycerides, medium-chain triglycerides, perilla seed oil, algal oil DHA, and *Sapindus mukorossi* oil. Optionally, the oil comprises, by weight, parts of oil.

5. The method for preparing structural lipids according to claim 1, characterized in that, The oils include medium-chain triglycerides, perilla seed oil, DHA algal oil, *Sapindus mukorossi* oil, palm oil, and coconut oil; Optionally, the oil comprises, by weight:

6. The method for preparing the structured lipid according to any one of claims 1 to 5, characterized in that, The preparation method satisfies one or more of the following conditions: (1) The weight of the lipase is 3-6 wt% of the weight of the oil; optionally, the lipase includes an immobilized lipase. (2) The reaction conditions include: sealed environment, 10-50 Pa, 60-75℃, 24-28 hours; and, (3) Enzyme inactivation is performed by thermal inactivation or organic solvent inactivation.

7. A structural lipid, characterized in that, Prepared using the preparation method described in any one of claims 1 to 6.

8. A structural lipid composition for liver and eye protection, characterized in that, The structured lipid composition comprises: 1) The structural lipid of claim 7; and, 2) Lutein esters and / or Haematococcus pluvialis; Optionally, the composition comprises, by weight: The structural lipids comprised 99.4-99.7 parts. Lutein ester 0.2-0.3 parts, and, Haematococcus pluvialis 0.15-0.25 parts.

9. The method for preparing the liver-protecting and eye-protecting structural lipid composition according to claim 8, characterized in that, The preparation method includes: Structural lipids are prepared using the preparation method described in any one of claims 1 to 6; and The structural lipids are mixed with the lutein esters and / or Haematococcus pluvialis to prepare a liver-protecting and eye-protecting structural lipid composition.

10. A liver-protecting and eye-protecting product, characterized in that, The liver-protecting and eye-protecting product includes the structural lipids of claim 7 or the structural lipid composition of claim 8; Optionally, the liver and eye protection products include capsules.

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