CN119431175A - Preparation method of 3-hydroxybutyric acid betaine salt - Google Patents

Preparation method of 3-hydroxybutyric acid betaine salt Download PDF

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CN119431175A
CN119431175A CN202310975344.3A CN202310975344A CN119431175A CN 119431175 A CN119431175 A CN 119431175A CN 202310975344 A CN202310975344 A CN 202310975344A CN 119431175 A CN119431175 A CN 119431175A
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salt
hydroxybutyric acid
betaine
hydroxybutyrate
acid betaine
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刘剑宏
陈舒明
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Shanghai Kangxin Chemical Co ltd
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Abstract

The invention discloses a 3-hydroxybutyric acid betaine salt, which has the following molecular structure: Wherein M n+ is a metal ion selected from Na +、K+、Mg2+ and Ca 2+;n+ is a charge number representing 1-2 positive charges. The 3-hydroxybutyric acid betaine salt is used as food additive and pharmaceutical active ingredient for reducing or preventing fatty liver. The invention also discloses a preparation method of the 3-hydroxybutyric acid betaine salt.

Description

Preparation method of 3-hydroxybutyric acid betaine salt
Technical Field
The invention belongs to the field of nutritional additives, and particularly relates to a 3-hydroxybutyric acid betaine salt, a preparation method and application thereof.
Background
3-Hydroxybutyric acid is one of three ketone bodies generated by fat metabolism in a human body, has multiple physiological functions, and can help the human body to efficiently generate energy under the condition of no glucose. 3-hydroxybutyric acid is produced in the liver by fat metabolism, and by blood circulation, transports corresponding tissues and organs, and oxidizes in cell mitochondria to generate energy, i.e., it can increase the consumption of fat in the body. K-Clark discloses in patent document CN 110869012A that 3-hydroxybutyric acid and salts/esters thereof have therapeutic effects on liver fat-related diseases, and can be used for treating healthy subjects and subjects suffering from liver fat-related diseases. The patent document CN 109734575A discloses that 3-hydroxybutyric acid and salts thereof can be used as nutritional additives or medicines, and have great health care and medicinal values.
It was found that when running on ketone bodies, the mechanical efficiency of the heart increased by 30%, blood flow increased by 75%, burning fatty acids increased free radicals, and 3-hydroxybutyric acid destroyed them. During ketosis exercise, the burning of intramuscular fat increases 20-fold, even with muscle glycogen available. This means that the cells ignore the normally preferred glucose reserves, but burn more fat.
Betaine is a nutritional additive, can be used in pharmaceutical industry, has effects of resisting fatty liver and protecting kidney, and can also be used in health food and skin caring food. Abdelmalek et al (2009) conducted a randomized control study on 55 non-alcoholic fatty liver disease patients, of which 34 patients were treated and a liver biopsy was performed, and found that the degree of liver steatosis was improved in the oral betaine group compared to the control group, saunderson (1990) test demonstrated that betaine was added to the diet of growing chickens, the body fat of chickens was decreased, the protein content was increased, xu Zirong et al (1998) reported that betaine increased the free carnitine content in the liver of chickens, and that carnitine contributed to the beta-oxidation of long chain fatty acids, xu Zhongna et al (2006) studied on rats fully confirmed the therapeutic effect of betaine on alcoholic fatty liver.
Disclosure of Invention
As manufacturers of 3-hydroxybutyric acid and betaine related fine chemicals, based on their correlation and the same properties in terms of physiological activity as food additives and pharmaceutical active ingredients, a feasible exploration route for compounding the two into salts was conceived, and experiments verified that composite salt crystals with stable properties were obtained. Specifically, the invention comprises the following technical scheme.
In a first aspect, the present invention provides a betaine salt of 3-hydroxybutyric acid having the following molecular structure:
Wherein n is a natural number 1 or 2;M is an alkali metal or alkaline earth metal selected from the group consisting of Na, K, mg and Ca, i.e., M n+ is a metal ion selected from the group consisting of Na +、K+、Mg2+ and Ca 2+;n+ is a charge number representing 1-2 positive charges,
The 3-hydroxybutyric acid is selected from D-3-hydroxybutyric acid, L-3-hydroxybutyric acid and DL-3-hydroxybutyric acid, namely racemic 3-hydroxybutyric acid.
In one embodiment, the equivalent ratio of betaine to 3-hydroxybutyrate in the 3-hydroxybutyrate betaine salt is 1:0.3-3, such as 1:0.5-2 or 1:0.8-1.5.
Preferably, the equivalent ratio of betaine to 3-hydroxybutyrate in the 3-hydroxybutyrate betaine salt is about 1:1.
It is to be understood that the term "about" or "about" when used herein in describing a numerical feature means that the number represented may have an error range or float range of + -10%, + -9%, + -8%, + -7%, + -6% or + -5%.
Preferably, the crotonic acid content as an impurity in the 3-hydroxybutyric acid betaine salt is not more than 0.2wt%, preferably 0.18wt% or less.
In a second aspect the present invention provides a process for the preparation of the above-mentioned 3-hydroxybutyric acid betaine salt,
When the salt is a sodium, potassium or magnesium salt, comprising the steps of:
(1) Dissolving a predetermined equivalence ratio, e.g., an equivalence ratio (1:1), of betaine and a metal salt of 3-hydroxybutyric acid in water to obtain an aqueous solution;
(2) Heating and stirring the aqueous solution, and continuing stirring and reacting for 1-5 hours after the system is dissolved;
(3) Filtering while the system is hot to remove insoluble impurities in the system;
(4) Concentrating the filtrate from step (3), for example, concentrating by distillation under reduced pressure, substantially removing water to form a complex salt;
(5) Adding an organic solvent into the composite salt obtained in the step (4) until crystals are precipitated (crystallization);
(6) Cooling by standing, for example, cooling to below 10deg.C until crystals are substantially completely precipitated (growing crystals);
(7) Filtering, drying the filter cake to obtain 3-hydroxybutyric acid betaine salt crystal, and
When the salt is a calcium salt, the method comprises the steps of:
(1) Dissolving betaine and calcium 3-hydroxybutyrate in a preset equivalent ratio in water to obtain an aqueous solution;
(2) Heating and stirring the aqueous solution, and continuing stirring and reacting for 1-5 hours after the system is dissolved;
(3) Filtering while the system is hot to remove insoluble impurities in the system;
(4) Concentrating the filtrate in the step (3) until crystals are precipitated (crystallization);
(6) Standing and cooling until the crystal is basically completely precipitated (crystal growth);
(7) Filtering and drying to obtain the 3-hydroxybutyric acid betaine calcium salt.
The metal salt of 3-hydroxybutyric acid used in the above step (1) is generally a base-neutralized reaction product of 3-hydroxybutyric acid.
The organic solvent in the step (5) is generally an organic solvent approved for use by food and drug administration, such as the national food and drug administration or the FDA, and is selected from ethanol, isopropanol, isoamyl alcohol, acetone, ethyl acetate or a mixture of two or more thereof, for ensuring the safety of the product. Preferably, the organic solvent is acetone or ethyl acetate.
Preferably, the stirring reaction temperature in the above step (2) is controlled below 65 ℃, for example in the range of 55 ℃ to 60 ℃, to avoid the production of by-product crotonic acid from 3-hydroxybutyric acid.
Preferably, the above method further comprises a step (8) of recovering the organic solvent by distillation or rectification.
It is to be understood that the term "substantially" as used herein refers to a majority or substantial portion of a particular condition, such as 80% or more, 85% or more, 90% or more, 92% or more, 95% or more, or 98% or more of the total.
In a third aspect, the present invention provides the use of the above-described 3-hydroxybutyric acid betaine salt as a food additive and a pharmaceutical active ingredient in foods and medicines.
In particular, the above food is used for reducing or preventing fatty liver or protecting kidney, and the medicine is used for treating fatty liver or protecting kidney.
The sodium/potassium/magnesium/calcium salt of 3-hydroxybutyrate betaine of the invention has a stable crystal structure and a low impurity content, and thus can be sold and used as a compound as a fine chemical, for example, as a food additive and a pharmaceutically active ingredient for preparing foods and medicines having the function of preventing, alleviating or treating fatty liver, or protecting kidneys.
Drawings
FIG. 1 is a process flow diagram of one embodiment of the present invention for preparing betaine sodium salt of 3-hydroxybutyric acid.
FIG. 2 is an HPLC chart of the sodium salt of betaine 3-hydroxybutyrate prepared from example 1.
Detailed Description
The design idea of the invention is that the multifunctional 3-hydroxybutyric acid and betaine are provided for the organism together, so that better effects are exerted on the aspects of fat consumption and the like of the human body. The adopted technical scheme is that the 3-hydroxybutyric acid and betaine are made into compound salt, and the two components are combined into a whole, so that the compound salt is taken as a nutrition additive, and has important significance for improving the physique of a human body and promoting the health of the human body. No product for preparing the two substances into composite salt is known in the market at present.
Betaine is an alkaloid, the chemical name is N, N, N-trimethylglycine, the chemical structure is similar to that of amino acid, and the betaine is an inner salt, and belongs to quaternary ammonium alkali substances. The complex salts therefore require metal ions that match the 3-hydroxybutyrate. In order to ensure edible safety, alkali metal ions and alkaline earth metal ions Na +、K+、Mg2+ and Ca 2+ are selected as metal ions. Namely, the metal salt of 3-hydroxybutyric acid as a reaction material includes sodium salt, potassium salt, magnesium salt and calcium salt. Correspondingly, the 3-hydroxybutyric acid betaine compound salt can be 3-hydroxybutyric acid betaine sodium salt, 3-hydroxybutyric acid betaine potassium salt, 3-hydroxybutyric acid betaine magnesium salt and 3-hydroxybutyric acid betaine calcium salt.
Herein, the term "3-hydroxybutyric acid betaine salt" and "3-hydroxybutyric acid betaine complex salt" mean the same meaning and may be used interchangeably. For convenience of description, it may be simply referred to as "complex salt".
The compound salt at least maintains the respective physiological activity functions of 3-hydroxybutyric acid and betaine, including fatty liver alleviation, kidney protection and the like. The complex salt can be recovered into free 3-hydroxybutyric acid and betaine after being dissolved in water.
In one embodiment, the equivalent ratio of betaine to 3-hydroxybutyrate in the complex salt may be 1:0.3 to 3, for example, about 1:1. Those skilled in the art will readily appreciate that the ratio of the two components betaine and 3-hydroxybutyric acid may be adjusted depending on the specific application due to their different efficacy.
Experiments have shown that the physical stability of a complex salt that does not contain a crystallization mother liquor, such as crystallization water, is higher than that of a complex salt that does contain a crystallization mother liquor, such as crystallization water. Therefore, in the process for preparing a complex salt, when betaine and 3-hydroxybutyrate are subjected to a synthesis reaction in an aqueous solution, after being bound together by ionic bond, hydrogen bond, van der Waals force, etc., crystallization is not performed in the aqueous solution so as not to contain crystal water, but to remove water, and cooled organic solvents such as acetone are used for crystallization, and the resulting complex salt does not contain organic solvents such as acetone components.
It should be understood that in order to ensure the edible safety of the complex salt, the organic solvent is an organic solvent approved for use by the national food and drug administration and the FDA, and may be selected from ethanol, isopropanol, isoamyl alcohol, acetone, ethyl acetate, or a mixture of two or more thereof in any ratio, for example.
FIG. 1 shows a process flow diagram of one embodiment of the present invention for preparing a betaine salt of 3-hydroxybutyric acid.
In order to ensure the edible safety of the compound salt, the generation of impurities in the preparation process of the compound salt is controlled as much as possible. The impurity in the compound salt is mainly crotonic acid (butenoic acid) which is a byproduct generated by 3-hydroxybutyric acid, so that the synthetic reaction temperature of the compound salt is controlled below 65 ℃, and the crotonic acid content in the finished product of the 3-hydroxybutyric acid betaine salt is effectively controlled below 0.2 wt%.
Examples
The invention is further illustrated by the following examples. It is to be understood that these examples are for illustrative purposes only and are not limiting of the invention. Various changes and modifications may be made by one skilled in the art in light of the teachings of this invention, and are intended to fall within the scope of this invention. For example, those skilled in the art use metal salts of chiral 3-hydroxybutyric acid, such as D-3-hydroxybutyric acid, with betaines to prepare complex salts, and the synthesis process is similar to the process conditions described below for the calcium, magnesium, sodium, and potassium salts of racemic 3-hydroxybutyric acid.
The amounts, amounts and concentrations of various substances are referred to herein, wherein the percentages refer to percentages by mass unless otherwise specified.
In the examples herein, if no specific description is made regarding the operating temperature, this temperature is generally referred to as room temperature (15-35 ℃).
HPLC detection method of the components in the betaine salt of 3-hydroxybutyric acid in examples:
instrument liquid chromatograph Agilent1260;
chromatographic column: agilent ZORBAX SB-Aq 4.6mm×250mm×5 μm or equivalent;
Mobile phase: 0.1% aqueous perchloric acid solution: acetonitrile=95:5 (v/v);
The flow rate is 1.0ml/min;
The column temperature is 30 ℃,
A differential refraction detector with a detection wavelength of 214nm;
the sample injection volume is 10 μl, and the component content is calculated by area normalization.
Wherein, the retention time of betaine is 2.599min, the retention time of 3-hydroxybutyric acid is 4.864min, and the retention time of crotonic acid is 15.181min.
EXAMPLE 1 preparation of the sodium salt of betaine 3-hydroxybutyrate
The process for salifying the sodium 3-hydroxybutyrate and the betaine comprises the following steps:
1. 25.00g (0.198 mol) of sodium 3-hydroxybutyrate and 23.23g (0.198 mol) betaine were weighed into a three-necked flask, and 100ml of deionized water was added thereto and stirred.
2. The electric heater was started and slowly warmed to 55 ℃.
3. After the system in the flask is dissolved, the reaction is continued for 3 hours under stirring within the range of 55-60 ℃.
4. Filtering while hot to remove insoluble impurities in the system.
5. Starting a vacuum pump, decompressing and concentrating the reaction system until the reaction system is sticky, and stopping concentration.
6. 150Ml of acetone was slowly added dropwise, at which point white crystals began to precipitate.
7. Cooling to below 10deg.C, and stirring for 1 hr.
8. Suction filtration and baking of the wet product in an oven at 80 ℃ for 4 hours, and obtaining 47.1g of white crystalline 3-hydroxybutyric acid betaine sodium salt with the yield of 97.8%.
9. The purity of the product and crotonate was determined by HPLC, see Table 1, and the chromatogram shown in FIG. 2.
Table 1 purity of each component in 3-hydroxybutyric acid betaine sodium salt measured by HPLC
3-Hydroxybutyric acid 42.123%
Betaine (betaine) 57.828%
Crotonic acid 0.049%
EXAMPLE 2 preparation of the potassium salt of 3-hydroxybutyric acid betaine
The preparation process of the 3-hydroxybutyric acid betaine potassium salt comprises the following steps:
1. 28.17g (0.198 mol) of potassium 3-hydroxybutyrate and 23.23g (0.198 mol) betaine were weighed into a three-necked flask, followed by adding 120ml of deionized water thereto and stirring.
2. The electric heater was started and slowly warmed to 55 ℃.
3. After the system in the flask is dissolved, the reaction is continued for 3 hours under stirring within the range of 55-60 ℃.
4. Filtering while hot to remove insoluble impurities in the system.
5. Starting a vacuum pump, decompressing and concentrating the reaction system until the reaction system is sticky, and stopping concentration.
6. 150Ml of acetone was slowly added dropwise, at which point white crystals began to precipitate.
7. Cooling to below 10deg.C, and stirring for 1 hr.
8. Suction filtration, drying the wet product in an oven at 80 ℃ for 4 hours to obtain 48.8g of white crystalline 3-hydroxybutyric acid betaine potassium salt with the yield of 95 percent.
9. The purity of the product and crotonate was determined by HPLC and is shown in table 2.
Table 2 purity of each component in the potassium salt of 3-hydroxybutyric acid betaine measured by HPLC
EXAMPLE 3 preparation of magnesium salt of betaine 3-hydroxybutyrate
The preparation process of the 3-hydroxybutyric acid betaine magnesium salt comprises the following steps:
1. 22.9g (0.099 mol) of magnesium 3-hydroxybutyrate and 23.23g (0.198 mol) of betaine were weighed into a three-necked flask, followed by 250ml of deionized water and stirred.
2. The electric heater was started and slowly warmed to 55 ℃.
3. After the system in the flask is dissolved, the reaction is continued for 3 hours under stirring within the range of 55-60 ℃.
4. Filtering while hot to remove insoluble impurities in the system.
5. The vacuum pump was started, the reaction system was concentrated under reduced pressure until turbidity appeared, and the concentration was stopped.
6. 100Ml of ethyl acetate was slowly added dropwise, at which time a large amount of white crystals precipitated.
7. Cooling to below 10deg.C, and stirring for 1 hr.
8. Suction filtration, drying the wet product in an oven at 80 ℃ for 4 hours to obtain 44.2g of white crystalline 3-hydroxybutyric acid betaine magnesium salt with the yield of 96 percent.
9. The purity of the product and crotonate was determined by HPLC and is shown in table 3.
Table 3 purity of each component in magnesium salt of 3-hydroxybutyrate betaine measured by HPLC
3-Hydroxybutyric acid 42.432%
Betaine (betaine) 57.385%
Crotonic acid 0.183%
EXAMPLE 4 preparation of the calcium salt of 3-hydroxybutyric acid betaine
The preparation process of the 3-hydroxybutyric acid betaine calcium salt comprises the following steps:
1. 24.4g (0.099 mol) of calcium 3-hydroxybutyrate and 23.23g (0.198 mol) of betaine were weighed into a three-necked flask, and 380ml of deionized water was added thereto and stirred.
2. The electric heater was started and slowly warmed to 55 ℃.
3. After the system in the flask is dissolved, the reaction is continued for 3 hours under stirring within the range of 55-60 ℃.
4. Filtering while hot to remove insoluble impurities in the system.
5. Starting a vacuum pump, decompressing and concentrating the reaction system until a large amount of crystallization occurs, and stopping concentration.
6. Cooling to below 10deg.C, and stirring for 1 hr.
7. Suction filtration, drying the wet product in an oven at 80 ℃ for 4 hours to obtain 43.1g of white crystalline 3-hydroxybutyric acid betaine calcium salt with the yield of 91 percent.
9. The purity of the product and crotonate was determined by HPLC and is shown in table 4.
Table 4 purity of each component in calcium salt of 3-hydroxybutyrate betaine measured by HPLC
3-Hydroxybutyric acid 42.143%
Betaine (betaine) 57.815%
Crotonic acid 0042%
The above embodiments are only for illustrating the technical solution of the present invention, but not for limiting the same, and various modifications and variations of the present invention will be apparent to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1.一种3-羟基丁酸甜菜碱盐,具有如下分子结构:1. A 3-hydroxybutyric acid betaine salt having the following molecular structure: 其中,n是自然数1或2;M是碱金属或者碱土金属,选自Na、K、Mg和Ca;即Mn+是金属离子,选自Na+、K+、Mg2+和Ca2+;n+是电荷数,代表1-2个正电荷,Wherein, n is a natural number 1 or 2; M is an alkali metal or alkaline earth metal selected from Na, K, Mg and Ca; that is, Mn + is a metal ion selected from Na + , K + , Mg2 + and Ca2 + ; n + is the charge number, representing 1-2 positive charges, 所述3-羟基丁酸选自D-3-羟基丁酸、L-3-羟基丁酸和DL-3-羟基丁酸。The 3-hydroxybutyric acid is selected from D-3-hydroxybutyric acid, L-3-hydroxybutyric acid and DL-3-hydroxybutyric acid. 2.如权利要求1所述的3-羟基丁酸甜菜碱盐,其特征在于,其中甜菜碱与3-羟基丁酸盐的当量比为1:0.3-3。2. The 3-hydroxybutyric acid betaine salt according to claim 1, wherein the equivalent ratio of betaine to 3-hydroxybutyrate is 1:0.3-3. 3.如权利要求1所述的3-羟基丁酸甜菜碱盐,其特征在于,其中甜菜碱与3-羟基丁酸盐的当量比为1:1。3. The 3-hydroxybutyrate betaine salt according to claim 1, wherein the equivalent ratio of betaine to 3-hydroxybutyrate is 1:1. 4.如权利要求1所述的3-羟基丁酸甜菜碱盐,其特征在于,巴豆酸含量不高于0.2wt%。4. The 3-hydroxybutyric acid betaine salt according to claim 1, wherein the crotonic acid content is not higher than 0.2 wt%. 5.一种制备如权利要求1-4中任一项所述的3-羟基丁酸甜菜碱盐的方法,其特征在于,5. A method for preparing the 3-hydroxybutyric acid betaine salt according to any one of claims 1 to 4, characterized in that: 当所述盐是钠盐、钾盐或镁盐时,包括以下步骤:When the salt is a sodium salt, a potassium salt or a magnesium salt, the method comprises the following steps: (1)将预定当量比的甜菜碱和3-羟基丁酸金属盐溶解于水,得水溶液;(1) dissolving betaine and 3-hydroxybutyric acid metal salt in a predetermined equivalent ratio in water to obtain an aqueous solution; (2)将水溶液加热搅拌,待体系溶清后,继续搅拌反应1-5小时;(2) heating and stirring the aqueous solution until the system is dissolved and clear, and continuing to stir and react for 1-5 hours; (3)趁热过滤,除去体系中的不溶性杂质;(3) Filter while hot to remove insoluble impurities in the system; (4)对步骤(3)中的滤液进行浓缩,除去水分,形成复合盐;(4) concentrating the filtrate in step (3) to remove water and form a composite salt; (5)在步骤(4)中所得的复合盐中加入有机溶剂,直至析出晶体;(5) adding an organic solvent to the composite salt obtained in step (4) until crystals are precipitated; (6)静置冷却,直至结晶体析出基本完全;(6) standing and cooling until the crystals are almost completely precipitated; (7)过滤,烘干,得到3-羟基丁酸甜菜碱盐,并且(7) filtering and drying to obtain 3-hydroxybutyric acid betaine salt, and 当所述盐是钙盐时,包括以下步骤:When the salt is a calcium salt, the method comprises the following steps: (1)将预定当量比的甜菜碱和3-羟基丁酸钙溶解于水,得水溶液;(1) dissolving betaine and calcium 3-hydroxybutyrate in a predetermined equivalent ratio in water to obtain an aqueous solution; (2)将水溶液加热搅拌,待体系溶清后,继续搅拌反应1-5小时;(2) heating and stirring the aqueous solution until the system is dissolved and clear, and continuing to stir and react for 1-5 hours; (3)趁热过滤,除去体系中的不溶性杂质;(3) Filter while hot to remove insoluble impurities in the system; (4)对步骤(3)中的滤液进行浓缩,直至析出晶体;(4) concentrating the filtrate in step (3) until crystals are precipitated; (6)静置冷却,直至结晶体析出基本完全;(6) standing and cooling until the crystals are almost completely precipitated; (7)过滤,烘干,得到3-羟基丁酸甜菜碱钙盐。(7) filtering and drying to obtain 3-hydroxybutyric acid betaine calcium salt. 6.如权利要求5所述的方法,其特征在于,步骤(5)中所述的有机溶剂选自:乙醇、异丙醇、异戊醇、丙酮、乙酸乙酯或者它们两种以上的混合物。6. The method according to claim 5, characterized in that the organic solvent described in step (5) is selected from the group consisting of ethanol, isopropanol, isopentanol, acetone, ethyl acetate or a mixture of two or more thereof. 7.如权利要求5所述的方法,其特征在于,步骤(2)中的搅拌反应温度为65℃以下。7. The method according to claim 5, characterized in that the stirring reaction temperature in step (2) is below 65°C. 8.如权利要求5所述的方法,其特征在于,还包括下述步骤:(8)通过蒸馏或精馏,回收有机溶剂。8. The method according to claim 5, further comprising the step of: (8) recovering the organic solvent by distillation or rectification. 9.如权利要求1-4中任一项所述的3-羟基丁酸甜菜碱盐在食品和药品中的用途。9. Use of the 3-hydroxybutyric acid betaine salt according to any one of claims 1 to 4 in food and medicine. 10.如权利要求9所述的用途,其特征在于,所述食品用于减轻或者预防脂肪肝或者保护肾脏,所述药品用于治疗脂肪肝或者保护肾脏。10. The use according to claim 9, characterized in that the food is used to alleviate or prevent fatty liver or protect kidneys, and the medicine is used to treat fatty liver or protect kidneys.
CN202310975344.3A 2023-08-03 2023-08-03 Preparation method of 3-hydroxybutyric acid betaine salt Pending CN119431175A (en)

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