Xanthoceras sorbifolia bunge leaf extract with liver-protecting function against alcoholism and preparation method and application thereof

Abstract

This invention relates to the field of natural products and functional foods or pharmaceuticals, and discloses a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, its preparation method, and its applications. The method involves using *Xanthoceras sorbifolium* leaves as raw material, performing water extraction and fractional extraction with different volume fractions of ethanol solvents to obtain components of different polarities, and then screening for functional effects through in vitro alcohol dehydrogenase (ADH) activity assays to obtain target extracts with high ADH-promoting activity. Further validation is achieved using an acute alcohol poisoning animal model, detecting ADH and ALDH activities, serum ethanol and acetaldehyde levels, behavioral indicators, and liver coefficient. This invention achieves a method combining multi-polar component extraction from *Xanthoceras sorbifolium* leaves with in vitro ADH activity screening and multi-indicator animal evaluation, and can be used for the development of hangover-relieving functional foods, health products, or pharmaceuticals.

Description

Technical Field

[0001] This invention relates to the field of natural product extraction and functional food and drug technology, specifically to a *Xanthoceras sorbifolium* leaf extract with hangover relief and liver protection functions, its preparation method and application. Background Technology

[0002] Alcohol poisoning and alcoholic liver injury are common public health problems worldwide. Ethanol is primarily metabolized in the liver after entering the body. Alcohol dehydrogenase (ADH) converts ethanol into acetaldehyde, which is then further metabolized into acetic acid by aldehyde dehydrogenase (ALDH). Acetaldehyde has strong cytotoxicity and is a key intermediate metabolite causing intoxication and alcohol-induced tissue damage. Therefore, regulating key enzymes involved in ethanol metabolism is considered one of the core strategies for improving alcohol load.

[0003] In existing technologies, solutions for improving alcohol-related states mainly fall into two categories: natural products and chemical drugs. Regarding natural products, traditional medicinal plants such as kudzu root and Japanese raisin tree fruit have been extensively studied. Their extracts or active ingredients have been reported in animal or in vitro experiments to potentially improve alcohol load by regulating ethanol metabolism-related processes and through antioxidant effects.

[0004] In terms of chemical drugs, drugs such as metadoxine are mainly used to promote ethanol metabolism and relieve related symptoms, but their overall effects are mostly concentrated on a single pathway or symptom improvement, and their ability to systematically regulate the ethanol metabolism process is limited.

[0005] However, existing technologies still have the following shortcomings: (1) Natural products are mostly extracted using fixed solvents and empirical screening methods, lacking systematic graded extraction and functional enrichment strategies for components with different polarities; (2) Although chemical drugs have clear effects, they are mostly limited to a single metabolic process or symptom improvement, and it is difficult to achieve synergistic regulation of ethanol and acetaldehyde metabolism; (3) Existing evaluation systems mostly rely on single-indicator detection and lack an in vitro screening and in vivo multi-indicator linkage evaluation system with key enzymes of ethanol metabolism (especially ADH) as the core. (4) Existing research mainly focuses on traditional resources such as kudzu root and Japanese raisin tree fruit, while the systematic development of other plant resources is still relatively limited; (5) The leaves of *Sapindus mukorossi* have a large yield and low price. Currently, they are mostly discarded or used only for low value-added products. In the existing technology system, a systematic development path for detoxification and liver protection based on the combination of multipolar extraction and ADH activity screening has not yet been formed.

[0006] Therefore, developing a method for preparing a *Sapindus mukorossi* leaf extract that combines multipolar extraction with ADH activity screening to systematically improve ethanol metabolism-related processes is of significant practical importance. Summary of the Invention

[0007] The purpose of this invention is to solve the problems existing in the prior art, and to propose a *Xanthoceras sorbifolium* leaf extract with hangover relief and liver protection functions, its preparation method and application.

[0008] To achieve the above objectives, the present invention adopts the following technical solution: A method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions includes the following steps: S1. Select mature leaves of *Sapindus mukorossi* as raw material, remove impurities, wash them clean, dry them to a moisture content of 8% to 12%, pulverize them and pass them through a 40 to 80 mesh sieve to obtain *Sapindus mukorossi* leaf powder. S2. The *Sapindus mukorossi* leaf powder was subjected to gradient fraction extraction using water extraction and ethanol solvents of different volume fractions to obtain extracts of different polarities. S3. Filter and concentrate the extracts of different polarities under reduced pressure to obtain extract concentrates of different polarities; S4. The in vitro alcohol dehydrogenase (ADH) activity of the extracts with different polarities was determined. S5. Select extracts with relatively high alcohol dehydrogenase activity as target extracts.

[0009] As a further technical solution of the present invention, the extraction method in S2 is reflux extraction, the extraction temperature is 40-70℃, the extraction time is 1-3 hours, and the material-liquid ratio is 1:(10-30).

[0010] As a further technical solution of the present invention, the target extract is further used for verification in an animal model of acute alcohol poisoning.

[0011] As a further technical solution of the present invention, the animal model validation includes detecting the following indicators: (1) Alcohol dehydrogenase (ADH) activity; (2) Acetaldehyde dehydrogenase (ALDH) activity; (3) Serum ethanol content; (4) Serum acetaldehyde content; (5) Behavioral indicators, including time to intoxication or righting reflex time; (6) Liver coefficient.

[0012] An extract of *Sapindus mukorossi* leaves, which has the function of relieving hangovers and protecting the liver, can promote the activity of alcohol dehydrogenase (ADH).

[0013] As a further technical solution of the present invention, the extract contains flavonoids, saponins and polyphenols as active ingredients.

[0014] A formulation of a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, wherein the formulation is any one of granules, capsules, and oral liquid.

[0015] An application of a *Sapindus mukorossi* leaf extract with hangover-relieving and liver-protecting functions in the preparation of products that alleviate physiological changes caused by acute alcohol intake, including reducing serum ethanol levels, reducing serum acetaldehyde levels, and improving alcohol metabolism-related indicators.

[0016] The beneficial effects of this invention are as follows: (1) By using water and ethanol of different volume fractions, the systematic acquisition of different polar active ingredients in the leaves of Xanthoceras sorbifolium can be achieved, providing a multi-component material basis for subsequent activity screening, which is conducive to improving the overall functional expression of the extract. (2) By introducing an in vitro activity screening step of alcohol dehydrogenase (ADH), functional-guided screening of different extracts is achieved, which improves the targeting and effectiveness of extract screening. (3) A systematic evaluation method combining in vitro ADH screening and animal multi-index evaluation was constructed, which can comprehensively evaluate the effects of extracts on alcohol metabolism-related processes from multiple levels such as behavioral and biochemical indicators, thereby improving the integrity and reliability of the evaluation system. (4) Using the leaves of the Chinese privet as raw material, which contain natural active ingredients such as flavonoids, saponins and polyphenols, has the advantages of natural source, relatively high safety and suitability for the development of functional foods or health products compared with some chemical drug source systems. (5) The extraction method uses water and ethanol solvents of different volume fractions for parallel extraction. After soaking, filtering, concentration and drying, a stable powder extract can be obtained. The process is simple, the conditions are easy to control, and the obtained product has good stability, which is convenient for preservation and subsequent formulation processing. (6) The extract of *Sapindus mukorossi* leaves obtained by this invention can be further prepared into dosage forms such as granules, capsules or oral liquids, which are convenient for application in the development of functional foods, health products or related products, and have good application and promotion value; (7) This invention utilizes the resources of *Sapindus mukorossi* leaves for the purpose of detoxification and liver protection, thereby improving the efficiency and added value of resource utilization and having certain economic and social significance. (8) It was discovered for the first time that the extract of Xanthoceras sorbifolium leaf has a certain promoting effect on alcohol dehydrogenase (ADH) in in vitro experiments, and it is suggested that it may be related to the regulation of alcohol metabolism, providing a screening basis for functional evaluation; (9) For the first time, a graded extraction strategy based on solvents of different polarities was adopted. By setting water and ethanol extraction conditions, the multipolar active ingredients in the leaves of *Sapindus mukorossi* were systematically extracted.

[0017] (10) A systematic evaluation method was first established, which uses in vitro ADH enzyme activity screening as a preliminary step and combines animal experimental behavioral and biochemical indicators (including serum ethanol, acetaldehyde content and changes in ADH / ALDH activity) to comprehensively evaluate the alcohol metabolism regulation effect of Xanthoceras sorbifolium leaf extract. Attached Figure Description

[0018] Figure 1 This is a schematic flowchart of a method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, as proposed in this invention. Detailed Implementation

[0019] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below in conjunction with specific embodiments.

[0020] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Example 1 1. A method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, characterized by comprising the following steps: S1. Select mature leaves of *Sapindus mukorossi* as raw material, remove impurities, wash them clean, dry them to a moisture content of 8% to 12%, pulverize them and pass them through a 40 to 80 mesh sieve to obtain *Sapindus mukorossi* leaf powder. S2. The *Sapindus mukorossi* leaf powder was subjected to gradient fraction extraction using water extraction and ethanol solutions of different volume fractions to obtain extracts of different polarities. S3. Filter and concentrate the extracts of different polarities under reduced pressure to obtain extract concentrates of different polarities; then freeze-dry or dry under reduced pressure to obtain *Sapindus mukorossi* leaf extracts under different solvent conditions.

[0022] Example 2 Extracts obtained from different solvents in Example 1 were prepared into solutions of specific concentrations. The in vitro enzyme activity of each extract was evaluated using an alcohol dehydrogenase (ADH) activity assay to compare the effects of different extracts on ADH activity. Using changes in ADH activity as a screening indicator, extracts exhibiting superior ADH activity enhancement were selected as candidate functional extracts for subsequent animal studies.

[0023] Example 3 An acute alcohol poisoning mouse model was established. The candidate extracts screened in Example 2 were administered to the animals by gavage at a certain dose. A blank group, a positive control group, and a model group were set up.

[0024] The main evaluation indicators include: (1) Behavioral indicators: Observe the latency period and duration of intoxication in animals; (2) Serum biochemical indicators: Detection of serum ethanol and acetaldehyde content; (3) Enzymatic indicators: Detect the activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in liver tissue; (4) Histological indicators: Calculate the liver coefficient to evaluate the effects of alcohol on the body.

[0025] The effects of *Sapindus mukorossi* leaf extract on alcohol metabolism and bodily status were comprehensively evaluated using the above indicators.

[0026] Example 4 The *Sapindus mukorossi* leaf extract prepared by the method described in Example 1, or the extract with superior ADH activity screened in Example 2, was used as an active ingredient to prepare different dosage forms of hangover-relieving and liver-protecting preparations.

[0027] When preparing it as a granule, the *Xanthoceras sorbifolium* leaf extract powder can be processed into easily dissolved granules. Take an appropriate amount of *Xanthoceras sorbifolium* leaf extract, add suitable excipients (such as maltodextrin, microcrystalline cellulose, or soluble starch), and mix evenly in a certain proportion. Add an appropriate amount of purified water to the mixture to form a soft mass, then use a granulator to prepare wet granules. After drying, granulate to obtain the finished granule product.

[0028] When preparing capsules, the powdered extract of *Xanthoceras sorbifolium* leaves is mixed evenly with filler excipients (such as starch, lactose, or micronized silica) to ensure good flowability. The mixed powder is then filled into hard capsule shells to obtain the capsule dosage form. This capsule dosage form can be used for oral administration, facilitating dosage control and portability.

[0029] When preparing an oral liquid, take the extract of Xanthoceras sorbifolium leaf, add an appropriate amount of purified water to dissolve or disperse it, and add an appropriate amount of flavoring agent, stabilizer and preservative (such as sucrose, citric acid or sodium benzoate, etc.). After stirring evenly, filter to remove impurities, fill and sterilize to obtain the oral liquid preparation.

[0030] The above-mentioned preparations can be used in functional foods, health products, food and medicine homology products or drugs as functional ingredients with hangover relief and liver protection effects. They can be used to promote the metabolism of ethanol and acetaldehyde, increase the activity of ADH and ALDH, and improve alcohol-induced behavioral abnormalities and alcohol load-related physiological indicators.

[0031] In summary, this invention uses *Xanthoceras sorbifolium* leaves as raw material, employing water extraction and fractional extraction with different volume fractions of ethanol solvents to obtain a stable powdered extract. Combined with in vitro ADH activity evaluation and verification using an acute alcohol poisoning animal model, it has been determined that this *Xanthoceras sorbifolium* leaf extract promotes alcohol metabolism, improves abnormal behaviors during alcohol poisoning, and reduces liver damage. The extract can be further prepared into granules, capsules, oral liquids, and other dosage forms, suitable for functional foods, health products, food-medicine homologous products, or drugs for hangover relief and liver protection.

[0032] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, characterized in that, Includes the following steps: S1. Select mature leaves of *Sapindus mukorossi* as raw material, remove impurities, wash them clean, dry them to a moisture content of 8% to 12%, pulverize them and pass them through a 40 to 80 mesh sieve to obtain *Sapindus mukorossi* leaf powder. S2. The *Sapindus mukorossi* leaf powder was subjected to gradient fraction extraction using water extraction and ethanol solvents of different volume fractions to obtain extracts of different polarities. S3. Filter and concentrate the extracts of different polarities under reduced pressure to obtain extract concentrates of different polarities; S4. The in vitro alcohol dehydrogenase activity of the extracts with different polarities was determined. S5. Select extracts with relatively high alcohol dehydrogenase activity as target extracts.

2. The method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions according to claim 1, characterized in that, The extraction method in S2 is reflux extraction, with an extraction temperature of 40-70℃, an extraction time of 1-3 hours, and a material-to-liquid ratio of 1:

20.

3. The method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions according to claim 1, characterized in that, The target extract was further used for validation in an animal model of acute alcohol poisoning.

4. The method for preparing a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions according to claim 3, characterized in that, The animal model validation includes detecting the following indicators: (1) Ethanol dehydrogenase activity; (2) Acetaldehyde dehydrogenase activity; (3) Serum ethanol content; (4) Serum acetaldehyde content; (5) Behavioral indicators, including time to intoxication or righting reflex time; (6) Liver coefficient.

5. A *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, characterized in that... The extract of *Sapindus mukorossi* leaf, which has the function of relieving hangovers and protecting the liver, was prepared using the method described in any one of claims 1-4, and has the effect of promoting the activity of alcohol dehydrogenase.

6. The *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions according to claim 5, characterized in that, The extract contains flavonoids, saponins, and polyphenols as active ingredients.

7. A formulation of *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, characterized in that, The preparation of the *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions as described in any one of claims 5-6 is characterized in that the preparation is any one of granules, capsules, and oral liquid.

8. The application of a *Xanthoceras sorbifolium* leaf extract with hangover-relieving and liver-protecting functions, characterized in that... The leaf extract of *Sapindus mukorossi*, which has the function of relieving hangovers and protecting the liver, as described in any one of claims 5-6, is used to prepare products that alleviate physiological changes caused by acute alcohol intake, including reducing serum ethanol levels, reducing serum acetaldehyde levels, and improving alcohol metabolism indicators.

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