Dual-algae synergistic alcohol-eliminating and liver-protecting composition and application thereof

Abstract

This invention relates to the field of biomedicine, and in particular to a dual-algae synergistic hangover relief and liver protection composition and its application. The invention provides a dual-algae synergistic hangover relief and liver protection composition comprising the following components in parts by weight: 15-25 parts of selenium-enriched Chlorella powder, 12-22 parts of kudzu root flavonoid powder, 10-22 parts of Haematococcus pluvialis powder, 12-22 parts of Hovenia dulcis powder, and 10-22 parts of corn oligopeptide powder. The full-chain hangover relief and liver protection mechanism of the dual-algae synergistic hangover relief and liver protection composition of this invention is as follows: corn oligopeptides activate alcohol dehydrogenase → B vitamins act as coenzymes to improve metabolic pathways → selenocysteine ​​and astaxanthin scavenge free radicals → kudzu root flavonoids and Hovenia dulcis alleviate symptoms, achieving multiple synergistic effects of rapid hangover relief, powerful liver protection, and liver cell repair.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a dual-algae synergistic composition for hangover relief and liver protection, and its application. Background Technology

[0002] Currently, most hangover remedies and liver protection products on the market rely on exogenous additions of kudzu root flavonoids, Japanese raisin tree fruit extract, and single B vitamins. While they do have some hangover-relieving effects, they generally have the following drawbacks:

[0003] (1) Single source of B vitamins and low bioavailability: Most are added externally by physical means, which are easily degraded during processing and storage, and lack intracellular enrichment protection, with a bioavailability of less than 30%; (2) Fragmented technical path: Selenium conversion and B vitamin enrichment in selenium-enriched Chlorella are separated, making it impossible to achieve integrated and efficient production; (3) Lack of synergistic antioxidant design: Traditional products rely only on the antioxidant effect of plant flavonoids and do not build a dual antioxidant barrier of "enzymatic + non-enzymatic"; (4) Vague quality control: There is a lack of accurate detection methods for selenocysteine ​​and full spectrum of B vitamins, resulting in poor product efficacy stability; (5) Single efficacy: Most products are only for sobering up or only for liver protection, failing to achieve full-link coverage of "sobering up-detoxification-repair".

[0004] Selenium-enriched Chlorella, as a natural green resource, can achieve high-value utilization of selenium (converted into selenocysteine) through endogenous directed biotransformation, while also possessing a natural metabolic capacity for efficiently absorbing and enriching B vitamins. However, current technologies have not organically combined these two aspects, nor have they been able to combine selenium-enriched / B-rich Chlorella with astaxanthin-rich Haematococcus pluvialis to construct a synergistic antioxidant system. Summary of the Invention

[0005] The purpose of this invention is to provide a dual-algae synergistic hangover-relieving and liver-protecting composition and its application, to solve the problems existing in the prior art. This invention provides a novel multi-target synergistic hangover-relieving and liver-protecting effect based on the endogenous selenium biotransformation and exogenous B vitamin absorption and enrichment of selenium-enriched Chlorella, the exogenous antioxidant effect of astaxanthin provided by Haematococcus pluvialis, and the traditional hangover-relieving effects of kudzu flavonoids, Hovenia dulcis seeds, and corn oligopeptides. This provides a novel dual-algae synergistic hangover-relieving and liver-protecting composition, and for the first time integrates the endogenous selenium transformation and exogenous B vitamin absorption and enrichment methods of Chlorella, along with a corresponding precise detection technology.

[0006] To achieve the above objectives, the present invention provides the following solution:

[0007] This invention provides a dual-algae synergistic hangover relief and liver protection composition, which comprises the following components in parts by weight:

[0008] 15-25 parts of selenium-enriched Chlorella powder, 12-22 parts of kudzu flavonoid powder, 10-22 parts of Haematococcus pluvialis powder, 12-22 parts of Hovenia dulcis powder, and 10-22 parts of corn oligopeptide powder.

[0009] Optionally, the method for preparing the selenium-enriched Chlorella powder includes the steps of sequentially performing B vitamin enrichment culture, selenium enrichment culture, low-temperature harvesting, and drying on the Chlorella broth obtained from propagation.

[0010] Further preferably, the B vitamin enrichment culture includes the step of mixing and culturing the Chlorella algal solution, the B vitamin mixed solution, and the absorption promoter; the food-grade B vitamin mixed solution includes vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, and folic acid; the absorption promoter includes MgSO4 and ZnSO4; the culture temperature is 28°C, and the time is 24~36h;

[0011] The selenium-enriched culture includes the step of mixing the culture medium obtained from the vitamin B enrichment culture with sodium selenite for culturing; the culturing temperature is 28℃ and the time is 48~72h.

[0012] Optionally, the B vitamin enrichment culture includes the step of mixing and culturing the Chlorella algal solution, the B vitamin mixed solution, and the absorption promoter; the food-grade B vitamin mixed solution includes vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, and folic acid; the absorption promoter includes MgSO4 and ZnSO4; the culture temperature is 28℃, the time is 30h, and the pH is 6.8~7.2, enabling Chlorella to efficiently absorb and store B vitamins in its cells through active transport;

[0013] The selenium-enriched culture includes the step of mixing the culture medium obtained from the vitamin B enrichment culture with sodium selenite for culture; the culture temperature is 28℃ and the time is 60h, to achieve the directional conversion of inorganic selenium to selenocysteine.

[0014] Optionally, the final concentration of vitamin B1 in the mixture obtained after mixing the Chlorella algae solution, the B vitamin mixed solution, and the absorption promoter is 8-12 mg / L, the final concentration of vitamin B2 is 5-8 mg / L, the final concentration of vitamin B6 is 4-6 mg / L, the final concentration of vitamin B12 is 0.5-1.2 mg / L, the final concentration of niacin is 12-15 mg / L, the final concentration of folic acid is 3-5 mg / L, the final concentration of MgSO4 is 0.2-0.4 g / L, and the final concentration of ZnSO4 is 0.05-0.1 g / L.

[0015] More preferably, the final concentration of vitamin B1 in the mixture obtained after mixing the Chlorella algae solution, the B vitamin mixed solution, and the absorption promoter is 10 mg / L, the final concentration of vitamin B2 is 6 mg / L, the final concentration of vitamin B6 is 5 mg / L, the final concentration of vitamin B12 is 0.8 mg / L, the final concentration of niacin is 15 mg / L, the final concentration of folic acid is 4 mg / L, the final concentration of MgSO4 is 0.3 g / L, and the final concentration of ZnSO4 is 0.08 g / L.

[0016] In a further preferred embodiment, the propagation culture medium uses molasses as the main organic carbon source, food-grade urea as the nitrogen source, and potassium dihydrogen phosphate as the phosphorus source.

[0017] More preferably, the commensal culture medium comprises the following components at the following concentrations:

[0018] Molasses 25-30g / L, food-grade urea 1.5-2.0g / L, potassium dihydrogen phosphate 0.8-1.2g / L, magnesium sulfate 0.3g / L and zinc sulfate 0.08g / L.

[0019] More preferably, during the propagation process, the temperature is controlled at 26-30℃, the light intensity at 8000-10000 lx, and the light-dark cycle at 14h:10h; the OD of the Chlorella algal solution obtained from the propagation is... 680 =1.0~1.5.

[0020] More preferably, the low-temperature harvesting and drying includes the steps of cooling the Chlorella liquid to 4~10℃, low-temperature centrifugation, washing, vacuum concentration, and low-temperature negative pressure spray drying (inlet air 90~110℃, outlet air 40~50℃); ensuring that the material temperature is ≤45℃ and the moisture content is ≤5%.

[0021] More preferably, the process after drying also includes low-temperature cell disruption;

[0022] The low-temperature cell wall breaking includes the step of using a low-temperature physical cell wall breaking technology at ≤4℃; the cell wall breaking rate is ≥90%.

[0023] Optionally, the dual-algae synergistic hangover relief and liver protection composition comprises the following components in parts by weight:

[0024] 20 parts of selenium-enriched Chlorella powder, 15 parts of kudzu flavonoid powder, 20 parts of Haematococcus pluvialis powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0025] Further preferably, the total selenium content of the selenium-enriched Chlorella is ≥100 μg / g, of which selenocysteine ​​accounts for ≥70% of the organic selenium; the total content of the full spectrum of B vitamins is ≥80 μg / g, of which vitamin B1 ≥15 μg / g, B2 ≥12 μg / g, B6 ≥10 μg / g, B12 ≥5 μg / g, niacin ≥25 μg / g, and folic acid ≥13 μg / g; the astaxanthin content in the Haematococcus pluvialis powder is ≥2%; the total flavonoids in the kudzu root flavonoid powder are ≥40%; and the molecular weight of the corn oligopeptide is ≤1000 Da.

[0026] This invention provides the application of the above-mentioned dual-algae synergistic hangover relief and liver protection composition in any of the following:

[0027] (1) Preparation of hangover remedies and liver-protecting drugs;

[0028] (2) Preparation of hangover remedies;

[0029] (3) Prepare hangover relief medication;

[0030] (4) Preparation of drugs to alleviate alcoholic liver injury;

[0031] (5) Preparation of drugs that enhance the metabolism of ethanol and acetaldehyde;

[0032] (6) Preparation of supplements for natural B vitamins;

[0033] (7) Preparation of drugs for repairing hepatocytes.

[0034] The present invention provides a drug comprising the above-mentioned dual algae synergistic hangover relief and liver protection composition.

[0035] Optionally, the drug is used to relieve hangovers, alleviate alcoholic liver damage, promote the metabolism of ethanol and acetaldehyde, supplement natural B vitamins, and repair liver cells.

[0036] Optionally, the drug may also include pharmaceutically acceptable excipients.

[0037] The present invention provides a method for preparing the above-mentioned drug, the method comprising the step of mixing the selenium-enriched Chlorella powder, the kudzu flavonoid powder, the Haematococcus pluvialis powder, the Hovenia dulcis powder and the corn oligopeptide powder.

[0038] More preferably, before mixing, the mixture further includes the steps of ultra-finely pulverizing the kudzu flavonoid powder, the Japanese raisin tree powder, the Haematococcus pluvialis powder, and the corn oligopeptide powder, and passing them through an 80-120 mesh sieve; controlling the moisture content to ≤5%.

[0039] More preferably, the mixing includes the step of putting the selenium-enriched Chlorella powder, the kudzu flavonoid powder, the Haematococcus pluvialis powder, the Hovenia dulcis powder and the corn oligopeptide powder into a three-dimensional mixer and mixing for 15-30 minutes.

[0040] As an additional method, the present invention also provides a method for extracting selenium-enriched chlorella selenocysteine ​​and full-spectrum B vitamins from the above-mentioned selenium-enriched chlorella powder, comprising the following steps:

[0041] (1) Determination of selenocysteine ​​(HPLC-ICP-MS method):

[0042] A. Sample pretreatment: Take selenium-enriched Chlorella powder, add proteinase K for enzymatic hydrolysis, and use ultrasound-assisted extraction. After centrifugation, take the supernatant and filter it through a filter membrane.

[0043] B. Chromatographic separation: An anion exchange column was used with phosphate buffer as mobile phase A and phosphate buffer-potassium chloride mixed solution as mobile phase B for gradient elution.

[0044] C. Mass spectrometry detection: Inductively coupled plasma mass spectrometry was used to detect selenium at specific mass-to-charge ratios (m / z=77, 78, 80), and external standard method was used for quantification.

[0045] D. Result Calculation: Calculate the selenocysteine ​​content based on the standard curve;

[0046] (2) Determination of the entire spectrum of B vitamins (HPLC-MS / MS method):

[0047] A. Sample pretreatment: Take selenium-enriched Chlorella powder, add methanol-water mixed extraction solution (volume ratio 8:2), extract with ultrasonic assistance, centrifuge to collect the supernatant, and filter through a filter membrane;

[0048] B. Chromatographic separation: A C18 reversed-phase column was used with gradient elution using 0.1% formic acid aqueous solution as mobile phase A and methanol as mobile phase B.

[0049] C. Mass spectrometry detection: A triple quadrupole mass spectrometer with an electrospray ionization (ESI) source was used. + The characteristic ion pairs of vitamins B1, B2, B6, B12, niacin, and folic acid were detected in multiple reaction monitoring (MRM) mode and quantified by external standard method.

[0050] D. Result Calculation: Calculate the content of each B vitamin according to the standard curve.

[0051] More preferably, in step (1), the enzymatic hydrolysis temperature is 35~45℃, the enzymatic hydrolysis time is 2~4h; the ultrasonic power is 200~400W, the ultrasonic time is 20~40min; and the filter membrane pore size is 0.22~0.45μm.

[0052] Further preferred, in step (2), the chromatographic column specifications are 4.6mm×250mm, 5μm; column temperature is 30℃; flow rate is 1.0mL / min; injection volume is 20μL; gradient elution program is: 0~10min, A 95%→85%; 10~25min, A 85%→70%; 25~40min, A 70%→50%; 40~50min, A 50%→10%.

[0053] This invention establishes an HPLC-ICP-MS coupled detection method for selenocysteine ​​and a simultaneous HPLC-MS / MS detection method for six B vitamins, enabling quality traceability and providing accurate detection results.

[0054] The present invention discloses the following technical effects:

[0055] This invention is the first to construct a triple-effect system of "endogenous selenium biotransformation + exogenous B vitamins absorption and enrichment + synergistic antioxidant action of two algae". This system comprises the following components by weight: 15-25 parts of selenium-enriched Chlorella powder, 12-22 parts of kudzu flavonoid powder, 10-22 parts of Haematococcus pluvialis powder, 12-22 parts of Hovenia dulcis powder, and 10-22 parts of corn oligopeptide powder. In this composition, selenium-enriched Chlorella provides selenocysteine ​​(its antioxidant mechanism is the core of glutathione peroxidase (GPx); its action is enzymatic antioxidant, scavenging hydrogen peroxide and organic peroxides), and Haematococcus pluvialis provides astaxanthin (≥2%, its antioxidant mechanism is direct scavenging of free radicals, quenching singlet oxygen; its action is non-enzymatic antioxidant, protecting cell membrane lipids), constructing a dual antioxidant barrier of "enzymatic + non-enzymatic" to synergistically scavenge free radicals and acetaldehyde produced by alcohol metabolism. The full-chain mechanism of the dual-algae synergistic hangover relief and liver protection composition of this invention is as follows: corn oligopeptides activate alcohol dehydrogenase, accelerating metabolism → B vitamins act as coenzymes to improve metabolic pathways → selenocysteine ​​and astaxanthin scavenge free radicals → kudzu root flavonoids and Japanese raisin tree fruit utilize their traditional hangover relief effects to alleviate symptoms, achieving multiple synergistic effects of rapid hangover relief, potent liver protection, and liver cell repair. The raw materials of the dual-algae synergistic hangover relief and liver protection composition of this invention are all medicinal and edible or new resource foods. Verified by both animal experiments and human trials, the composition of this invention can shorten hangover relief time by 55.8%, alleviate post-drinking discomfort by ≥88.5%, reduce serum ALT and AST by 45.2% and 41.5% respectively, increase acetaldehyde clearance rate by 62.3%, and repair alcoholic liver damage with long-term use, without any toxic side effects.

[0056] The dual-algae synergistic hangover relief and liver protection composition provided by this invention also has the following advantages:

[0057] 1. Dual Functions of One Algae: This invention obtains selenium-enriched Chlorella through fasciotrophic culture and selenium-induced directional transformation, converting inorganic selenium into selenocysteine ​​(organic selenium conversion rate ≥85%, selenocysteine ​​proportion ≥70%). At the same time, it realizes the endogenous conversion of selenocysteine ​​and the exogenous absorption and enrichment of B vitamins, simplifies the production process, avoids the degradation loss of exogenously added B vitamins during processing and storage, and reduces production costs by 30%.

[0058] 2. Synergistic antioxidant barrier of two algae: selenocysteine ​​(enzymatic) + astaxanthin (non-enzymatic) synergistically scavenge free radicals, with a scavenging rate 62.5% higher than that of a single component;

[0059] 3. Bioavailability of B vitamins increased by 3.2 times: Through intracellular enrichment, degradation loss during processing and storage of exogenous additions is avoided;

[0060] 4. Significant efficacy: Animal experiments showed a 55.8% reduction in sobering-up time, a 62.3% increase in acetaldehyde clearance rate, and a 45.2% / 41.5% reduction in ALT / AST.

[0061] 5. It has extremely high industrial application value and market prospects: the process can be scaled up industrially, and the product can be made into various dosage forms such as tablets, granules, and solid beverages, which are suitable for the market of functional foods for relieving hangovers and protecting the liver;

[0062] 6. Safety and Compliance: All raw materials are food and medicine homology or new resource foods, with no Western medicine ingredients, and have been verified to be safe and non-toxic through toxicology.

[0063] This invention also provides a method for preparing selenium-enriched Chlorella powder. This method, for the first time, integrates the conversion of endogenous selenium (selenocysteine) in Chlorella with the absorption and enrichment of exogenous B vitamins in the same culture system, achieving "dual functions of one algae". Specifically, a full spectrum of B vitamins is added to the culture medium, and the active transport system of Chlorella cells is used to efficiently absorb and store the B vitamins in the cells. After enrichment, the total content of B vitamins is ≥80μg / g, which is 3.2 times higher than that of direct exogenous addition. Chlorella is used to convert inorganic selenium (sodium selenite) into selenocysteine, with an organic selenium conversion rate of ≥85% and a selenocysteine ​​content of ≥70%. Attached Figure Description

[0064] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0065] Figure 1 This is a schematic diagram illustrating the full-chain hangover relief and liver protection mechanism of the dual-algae synergistic hangover relief and liver protection composition provided by the present invention;

[0066] Figure 2 The effect of different methods of adding B vitamins on in vitro release and absorption;

[0067] Figure 3 The effect of different formulations on the sobering time of mice;

[0068] Figure 4 The effects of different formulations on serum ALT and AST in mice;

[0069] Figure 5 A statistical chart showing the scoring of adverse symptoms after drinking alcohol in human trials.

[0070] Figure 6 This is a comparison chart of the synergistic antioxidant capacity of two algae. Detailed Implementation

[0071] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0072] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0073] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0074] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0075] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0076] Unless otherwise specified, all materials used in this invention are readily available to those skilled in the art and can be purchased directly.

[0077] Example 1: Optimization of the integrated preparation method of selenium-enriched Chlorella

[0078] 1. Experimental Materials

[0079] Propagation culture medium: molasses 25-30 g / L (in this example, the concentration of molasses is 28 g / L, as an organic carbon source), food-grade urea 1.5-2.0 g / L (in this example, the concentration of food-grade urea is 1.8 g / L, as a nitrogen source), potassium dihydrogen phosphate 0.8-1.2 g / L (as a phosphorus source; in this example, the concentration of potassium dihydrogen phosphate is 1 g / L based on the concentration of P2O5), magnesium sulfate 0.3 g / L, and zinc sulfate 0.08 g / L, with water as the solvent.

[0080] 2. Grouping

[0081] Group A: Group B supplementation + selenium-induced treatment, as detailed below:

[0082] 2.1. Simultaneous breeding and propagation of primary species

[0083] Chlorella (purchased from the Freshwater Algae Culture Collection of the National Aquatic Organism Germplasm Bank, species number FACHB-415) was inoculated into propagation medium and cultured to obtain primary algal solution (OD). 680 =1.2, density is 5×10 7 The primary algal solution and propagation medium were mixed at a volume ratio of 1:80. The culture temperature was 28℃, the light intensity was 9000 lx, the light-dark ratio was 14h:10h (i.e., the light time was 14h and the dark time was 10h), and the aeration rate was 1.5 vvm.

[0084] 2.2 Absorption and enrichment of exogenous B vitamins

[0085] A food-grade B vitamin mixture (comprising vitamins B1, B2, B6, B12, niacin, and folic acid) was added to the primary algal culture to achieve final concentrations of 10 mg / L for vitamin B1, 6 mg / L for vitamin B2, 5 mg / L for vitamin B6, 0.8 mg / L for vitamin B12, 15 mg / L for niacin, and 4 mg / L for folic acid. Simultaneously, an absorption enhancer (composed of MgSO4 and ZnSO4) was added to achieve final concentrations of 0.3 g / L for MgSO4 and 0.08 g / L for ZnSO4. The pH was adjusted to 7.0, and the culture was continued at 28°C for 30 hours (to allow Chlorella to absorb and store B vitamins), resulting in an algal culture enriched with B vitamins.

[0086] 2.3 Selenium-induced biotransformation

[0087] Sodium selenite was added to the algal solution enriched with B group to achieve a final concentration of 50-100 mg / L (75 mg / L in this example). The solution was then cultured at pH 7.2 and 28°C for 60 h to achieve the directional conversion of selenocysteine ​​and obtain selenium-enriched Chlorella algal solution.

[0088] 2.4 Low-temperature harvesting and drying

[0089] The selenium-enriched Chlorella liquid was cooled to 4℃ and centrifuged at low temperature (5000 r / min, 12 min). The precipitate was collected and washed three times with sterile water at 4℃. Subsequently, it was concentrated under low temperature vacuum (30℃, -0.07 MPa) until the solid content was 18 wt%. Then, it was spray-dried under low temperature negative pressure: inlet air 100℃, outlet air 45℃, negative pressure -0.04 MPa, and material temperature ≤42℃. Then, it was subjected to low temperature cell disruption (physical cell disruption at ≤4℃) with a cell disruption rate ≥92%. Finally, it was ultra-finely pulverized through a 100-mesh sieve to obtain selenium-enriched Chlorella powder.

[0090] Group B: Only Group B is added (without selenium induction), and the preparation method is the same as Group A, except that step "3, selenium-induced biotransformation" is omitted.

[0091] Group C: Selenium-induced only (no B vitamins added), preparation method is the same as Group A, the only difference is that step "2, absorption and enrichment of exogenous B vitamins" is omitted.

[0092] Group D: Blank (no additives), prepared in the same way as Group A, except that steps "2, absorption and enrichment of exogenous B vitamins" and "3, selenium-induced biotransformation" are omitted.

[0093] 3. Methods for detecting active ingredients

[0094] The active ingredients of the Chlorella powder obtained from groups A to D were detected using the following methods:

[0095] 3.1 Selenocysteine ​​(HPLC-ICP-MS)

[0096] Pretreatment: Mix 0.5g of Chlorella powder sample with 10mL of proteinase K (20mg / mL), enzymatically hydrolyze at 40℃ for 3h, sonicate at 300W for 30min, centrifuge at 15000rpm for 20min, and filter with a filter membrane with a pore size of 0.22~0.45μm (the pore size of the filter membrane in this example is 0.25μm).

[0097] The pretreated Chlorella powder sample was analyzed by HPLC-ICP-MS under the following conditions:

[0098] Chromatographic column: Anion exchange column (4.6×250mm, 5μm)

[0099] Column temperature: 30℃;

[0100] Flow rate: 1 mL / min;

[0101] Mobile phase A: 15 mM phosphate buffer (pH 7.2); Mobile phase B: 15 mM phosphate buffer - 100 mM KCl (i.e., 15 mM phosphate buffer and 100 mM KCl are mixed in a volume ratio of 1:1).

[0102] Elution gradient program: 0-10 min 80% A, 10-25 min 60% A, 25-35 min 10% A;

[0103] ICP-MS detection: m / z=77, 78, 80.

[0104] Each treatment was repeated 3 times.

[0105] Quantification was performed using the external standard method. The specific steps were as follows: L-selenocysteine ​​standard was weighed and prepared into a standard stock solution with a concentration of 100 μg / mL; the stock solution was serially diluted to prepare five working standard solutions with different concentrations (0.2 μg / mL, 0.5 μg / mL, 1.0 μg / mL, 2.0 μg / mL, and 5.0 μg / mL); each standard solution was measured sequentially according to the previous detection method to obtain the peak area response value; a linear regression was performed with the standard concentration as the x-axis and the response value (peak area / peak height) as the y-axis to obtain the standard curve equation: Y = 1.568 × 10⁻⁶. 4 X + 2.13 × 10 2 R 2 It is 0.9996;

[0106] At the same time, the organic selenium conversion rate and the proportion of selenocysteine ​​were calculated.

[0107] (1) Formula for calculating organic selenium conversion rate:

[0108] ;

[0109] (2) Formula for calculating the proportion of selenocysteine ​​(SeCys)

[0110] .

[0111] 3.2 Full-spectrum B vitamins (HPLC-MS / MS)

[0112] Pretreatment: Mix 0.5g of Chlorella powder sample with 10mL of methanol solution (methanol (≥99.85%) and water in a volume ratio of 8:2), sonicate at 300W for 30min, and centrifuge and filter.

[0113] The pretreated Chlorella powder sample was analyzed by HPLC-MS / MS under the following conditions:

[0114] Chromatographic column: C18 reversed-phase column (4.6×250mm, 5μm)

[0115] Mobile phase A: 0.1% formic acid in water; Mobile phase B: methanol;

[0116] Gradient elution program: 0-10 min, A 95%→85%; 10-25 min, A 85%→70%; 25-40 min, A 70%→50%; 40-50 min, A 50%→10%;

[0117] MS / MS: ESI+, MRM mode, for qualitative and quantitative detection based on the characteristic ion pairs corresponding to each vitamin.

[0118] Each treatment was repeated 3 times.

[0119] Quantification was performed using the external standard method. The specific steps were as follows: Vitamin B1, Vitamin B2, Vitamin B6, Vitamin B12, niacin, and folic acid standards were weighed, dissolved in their respective solvents, and diluted to volume to prepare standard stock solutions with a concentration of 100.0 μg / mL. The stock solutions were then serially diluted to prepare five working standard solutions of different concentrations (as shown below). Each standard solution was measured sequentially according to the previously described detection method to obtain the peak area response value. A linear regression was performed with the standard concentration as the x-axis and the response value (peak area) as the y-axis to obtain the standard curve equation and R0. 2 Standard curve equation and R 2 As shown below:

[0120] The standard working solution concentration series for vitamin B1 is 0.2 μg / mL, 0.5 μg / mL, 1.0 μg / mL, 2.0 μg / mL, and 5.0 μg / mL;

[0121] The standard working solution concentration series for vitamin B2 is 0.1 μg / mL, 0.2 μg / mL, 0.5 μg / mL, 1.0 μg / mL, and 2.0 μg / mL;

[0122] The standard working solution concentration series for vitamin B6 are 0.2 μg / mL, 0.5 μg / mL, 1.0 μg / mL, 2.0 μg / mL, and 5.0 μg / mL;

[0123] The standard working solution concentration series for vitamin B12 is 0.05 μg / mL, 0.1 μg / mL, 0.2 μg / mL, 0.5 μg / mL, and 1.0 μg / mL;

[0124] The standard working solution concentration series for nicotinic acid are 0.5 μg / mL, 1.0 μg / mL, 2.0 μg / mL, 5.0 μg / mL, and 10.0 μg / mL;

[0125] The standard working solution concentration series for folic acid are 0.1 μg / mL, 0.2 μg / mL, 0.5 μg / mL, 1.0 μg / mL, and 2.0 μg / mL;

[0126] Standard curve equation and correlation coefficient:

[0127] The standard curve equation for vitamin B1 is: Y = 5.218 × 10⁻⁶ 4 X-8.76×10 2 R 2 It is 0.9997;

[0128] The standard curve equation for vitamin B2 is: Y = 3.456 × 10⁻⁶ 4 X + 2.34 × 10 2 R 2 It is 0.9995;

[0129] The standard curve equation for vitamin B6 is: Y = 4.123 × 10⁻⁶ 4 X + 1.65 × 10 2 R 2 It is 0.9998;

[0130] The standard curve equation for vitamin B12 is: Y = 9.876 × 10⁻⁶ 3 X + 5.43 × 10 1 R 2 It is 0.9993;

[0131] The standard curve equation for niacin is: Y = 2.345 × 10⁻⁶ 4 X - 3.21×10 2 R 2 It is 0.9996;

[0132] The standard curve equation for folic acid is: Y = 2.789 × 10⁻⁶ 4 X + 1.89 × 10 2 R 2 It is 0.9994.

[0133] 4. Results

[0134] The test results are shown in Table 1. The results indicate that the method provided by group A can simultaneously achieve high enrichment of B vitamins (95.5 μg / g) and efficient conversion of selenocysteine ​​(71.8%), with no interference between B vitamin addition and selenium induction; moreover, selenium induction can also promote the enrichment of B vitamins in Chlorella. Meanwhile, the total selenium content of the selenium-enriched Chlorella prepared by method A is ≥100 μg / g, of which selenocysteine ​​accounts for ≥70% of the organic selenium; the total content of the full spectrum of B vitamins is ≥80 μg / g, including vitamin B1 ≥15 μg / g, vitamin B2 ≥12 μg / g, vitamin B6 ≥10 μg / g, vitamin B12 ≥5 μg / g, niacin ≥25 μg / g, and folic acid ≥13 μg / g.

[0135] Table 1 Detection results of different treatments

[0136] Vitamin B1 (μg / g) 18.2±0.8 17.5±0.7 2.1±0.2 1.9±0.15 Vitamin B2 (μg / g) 14.5±0.6 13.8±0.6 1.5±0.03 1.4±0.1 Vitamin B6 (μg / g) 12.3±0.5 11.6±0.5 0.9±0.09 0.8±0.12 Vitamin B12 (μg / g) 6.2±0.3 5.8±0.3 0.2±0.07 0.2±0.05 Niacin (μg / g) 28.5±1.2 27.2±1.0 2.5±0.25 2.3±0.2 Folic acid (μg / g) 15.8±0.7 14.5±0.6 0.5±0.17 0.5±0.1 Total B vitamins content (μg / g) 95.5±4.1 90.4±3.7 7.7±0.7 7.1±0.75 Total selenium (μg / g) 128.5±5.2 2.1±0.2 115.2±4.5 1.8±0.2 Selenocysteine ​​(μg / g) 92.3±4.0 - 82.5±3.5 - Organic selenium conversion rate (%) 86.2 - 85.8 Selenocysteine ​​percentage (%) 71.8 - 71.6 -

[0137] Example 2: Optimization of different methods of adding B vitamins

[0138] 1. Grouping

[0139] Group 1 (intracellular enrichment): B vitamins were added, and the cells absorbed and enriched the cells. The preparation method was the same as Group A in Example 1, except that step "2.3, selenium-induced biotransformation" was omitted.

[0140] Group 2 (Exogenous Addition): Direct exogenous addition of B vitamins (without enrichment by Chlorella, directly mixed with algae powder). The preparation method is as follows: take Chlorella powder, add the corresponding amounts of vitamin B1, B2, B6, B12, niacin and folic acid standards according to the content of each B vitamin in the final product prepared by Group A in Example 1, and grind and mix them evenly in a mortar to obtain physically mixed algae powder.

[0141] Group 3: The preparation method is the same as Group A in Example 1, except that steps “2.2, Absorption and enrichment of exogenous B vitamins” and “3, Selenium-induced biotransformation” (i.e. blank control algal powder) are omitted.

[0142] 2. Experimental Methods

[0143] 2.1 Detection of simulated release rate of B vitamins

[0144] Take 1.0g of each of the three algal powders obtained in step “1, grouping”, and then mix them with 20 mL of simulated gastric juice (pH 2.0, containing 0.2% NaCl and 0.32% pepsin). Incubate at 37℃ and 100 rpm in the dark for 2 hours, then centrifuge and collect the supernatant. Determine the content of each B vitamin and calculate the release rate of B vitamins in simulated gastric juice.

[0145] Take 1.0g of each of the three algal powders obtained in step “1, grouping”, and then mix them with 20mL of simulated intestinal fluid (pH 7.4, containing 0.68% potassium dihydrogen phosphate and 1.0% trypsin). Incubate at 37℃ and 100rpm in the dark for 2h, then centrifuge and collect the supernatant. Determine the content of each B vitamin and calculate the release rate of B vitamins in the simulated intestinal fluid.

[0146] The formula for calculating the release rate is as follows:

[0147] Release rate (%) = (Vitamin content in simulated digestive fluid / Initial vitamin content in algae powder) × 100%.

[0148] 2.2 Detection of apparent permeability in Caco-2 cells

[0149] Take 1.0g of each of the three algal powders obtained in step "1, Grouping" and digest them in two steps using simulated gastrointestinal fluid. Collect the digestion supernatant containing B vitamins (i.e., the bioaccessible component) and filter it through a 0.22 μm sterile filter membrane. Add the filtrate to the top side (AP side) of a Caco-2 cell monolayer model that has been cultured and differentiated for 21 days. Incubate at 37℃ in a 5% CO2 incubator for 2 hours. Collect the solution on the basal side (BL side), determine the content of each B vitamin, and calculate the apparent permeability coefficient.

[0150] Caco-2 cell parameters: cell density was 1×10⁻⁶. 5 cells / cm 2 Manufacturer: Wuhan Punosai Life Technology Co., Ltd., Product No.: CL-0050.

[0151] The formula for calculating the apparent permeability coefficient is as follows:

[0152] Papp=(dQ / dt) / (A×C0);

[0153] Where dQ / dt is the amount of drug transported per unit time, A is the area of ​​the polycarbonate membrane, and C0 is the initial concentration on the top side.

[0154] 3. Results

[0155] The results are shown in Table 2 and Figure 2 As shown. The results indicated that the B vitamins in the intracellular enrichment group were released very little in gastric juice (pH 2.0), only 8.2%, mainly in the small intestine (85.6%), significantly lower than the exogenous supplementation group (45.3%), thus avoiding destruction by gastric acid; the release rate in intestinal juice (pH 7.4) reached 85.6%, significantly higher than the exogenous supplementation group (52.1%); the apparent permeability coefficient of the Caco-2 cell model (12.5 × 10⁻⁶) was... -6 The cm / s ratio was significantly higher than that of the exogenous supplementation group (3.9 × 10⁻⁶ cm / s). -6 The bioavailability was increased by 3.2 times (cm / s). These results indicate that intracellular enrichment can significantly improve the bioavailability of B vitamins.

[0156] Table 2 Detection results of different treatments

[0157] Release rate in gastric juice (%) 8.2±1.2 45.3±3.2 - Release rate in intestinal fluid (%) 85.6±3.5 52.1±3.0 - <![CDATA[Caco-2 Papp(×10 -6 cm / s)]]> 12.5±1.0 3.9±0.4 -

[0158] Raw material preparation: Selenium-enriched Chlorella powder: prepared from Group A in Example 1;

[0159] Kudzu root flavonoid powder: purchased from Wuhan Pushida Biotechnology Co., Ltd., product number: 20211201; total flavonoid content ≥40%.

[0160] Haematococcus pluvialis powder: purchased from Yunnan Boxin Biotechnology Co., Ltd., product number: BX-HP-001; astaxanthin content in Haematococcus pluvialis powder ≥2%;

[0161] Hovenia dulcis powder: It utilizes biological enzymes such as cellulase and pectinase to selectively hydrolyze cell walls, allowing flavonoids and other effective components to fully dissolve. The process is as follows: After crushing the fruit of the Japanese raisin tree, water is added at a material-to-liquid ratio of 1g:10mL. The mixture is heated to about 50℃ and a compound enzyme (composed of cellulase, pectinase, and papain, with cellulase activity of 4000~300000U / g, pectinase activity of 4000~300000 U / g, and papain activity of 4000~300000 U / g, and the amount of compound enzyme added is 0.01%~0.2% of the total weight of the medicinal material. In this example, the cellulase activity of the compound enzyme is 100000U / g, the pectinase activity is 100000 U / g, the papain activity is 100000 U / g, and the amount of compound enzyme added is 0.1% of the total weight of the medicinal material) is added. The mixture is extracted for about 5 hours. After filtration and centrifugation, the supernatant is concentrated under vacuum at 60℃ to a solid content of about 25%, and then vacuum dried to obtain the final product.

[0162] Corn oligopeptide powder is made from corn protein powder (corn yellow powder) through processes such as slurry preparation, enzymatic hydrolysis, separation, purification, and drying. A typical process is as follows: commercially available corn gluten powder is mixed with water to adjust the substrate concentration to 8%~12%, pretreated at 90℃ to denature the protein, cooled to 50~55℃, and the pH is adjusted to 8.0~9.0. Alkaline protease and flavor protease are added. The alkaline protease has an enzyme activity of ≥200 U / mg and is added at 2%~4% of the substrate weight (3% in this example). The flavor protease has an enzyme activity of ≥500 U / g and is added at 2%~4% of the substrate weight (3% in this example). Then, it is hydrolyzed at 50~55℃ for 4~6 hours. After the enzymatic hydrolysis, the temperature is raised to 85℃ to inactivate the enzyme for 15 minutes. The supernatant is collected by centrifugation, separated by ultrafiltration membrane (molecular weight cutoff 1000 Da) or decolorized and debittered by activated carbon, concentrated by nanofiltration, and spray-dried to obtain corn oligopeptide powder with a molecular weight ≤1000 Da. The molecular weight of corn oligopeptides is ≤1000 Da.

[0163] Example 3: Synergistic Algae Composition for Hangover Relief and Liver Protection

[0164] A dual-algae synergistic hangover relief and liver protection composition, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from Group A in Example 1), 15 parts of kudzu root flavonoid powder, 20 parts of Haematococcus pluvialis powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0165] The preparation method is as follows: Pueraria lobata flavonoid powder, Hovenia dulcis powder, Haematococcus pluvialis powder, and corn oligopeptide powder are ultra-finely pulverized and passed through an 80-120 mesh sieve, with the moisture content controlled at ≤5%; Mixing ratio: Weigh each raw material according to the previous weight proportions and put them into a three-dimensional mixer and mix for 15-30 minutes; Molding preparation: Produce solid beverages, granules, tablets, or capsules according to conventional existing technology; Sterilization and packaging: After low-temperature irradiation sterilization, seal and package in a light-proof container.

[0166] The composition's end-to-end hangover detoxification and liver protection mechanism is as follows: Figure 1 As shown, specifically: the composition provided by this invention accelerates alcohol metabolism and reduces its harm through the synergistic effect of multiple functional components: corn oligopeptides activate alcohol dehydrogenase (ADH), and B vitamins act as coenzymes (NAD). +NADH accelerates the conversion of ethanol to acetaldehyde and then to acetic acid, thereby speeding up alcohol clearance. Simultaneously, selenocysteine ​​(as a component of glutathione peroxidase GPx) and astaxanthin work together to eliminate free radicals generated during ethanol metabolism, reducing liver damage. Pueraria lobata flavonoids and Hovenia dulcis seeds are used to relieve hangover symptoms such as headache, nausea, and fatigue. In other words, the full-chain hangover-relieving and liver-protecting mechanism of the dual-algae synergistic hangover-relieving and liver-protecting composition provided by this invention is as follows: corn oligopeptides activate alcohol dehydrogenase → B vitamins act as coenzymes to improve the metabolic pathway → selenocysteine ​​+ astaxanthin eliminate free radicals → pueraria lobata flavonoids and Hovenia dulcis seeds relieve symptoms, achieving multiple synergistic effects of rapid hangover relief, powerful liver protection, and liver cell repair.

[0167] Comparative Example 1

[0168] A composition for relieving hangovers and protecting the liver, comprising, by weight: 15 parts of kudzu root flavonoid powder, 20 parts of Haematococcus pluvialis powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0169] The preparation method is the same as in Example 3.

[0170] Comparative Example 2

[0171] A composition for relieving hangovers and protecting the liver, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from Group A in Example 1), 15 parts of kudzu flavonoid powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0172] The preparation method is the same as in Example 3.

[0173] Comparative Example 3

[0174] A composition for relieving hangovers and protecting the liver, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from Group A in Example 1), 15 parts of kudzu root flavonoid powder, 20 parts of Haematococcus pluvialis powder, and 15 parts of Hovenia dulcis powder.

[0175] The preparation method is the same as in Example 3.

[0176] Comparative Example 4

[0177] A composition for relieving hangovers and protecting the liver, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from the two groups in Example 2), 15 parts of kudzu flavonoid powder, 20 parts of Haematococcus pluvialis powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0178] The preparation method is the same as in Example 3.

[0179] Example 4 Acute toxicity test

[0180] This embodiment aims to evaluate the safety of the edible composition of the dual algae synergistic hangover relief and liver protection.

[0181] 1. Experimental Methods

[0182] Forty SPF-grade Kunming mice (half male and half female, weighing (20±2) g) were randomly divided into a blank control group and three dosage groups (low, medium, and high), with 10 mice in each group. The dual-algae synergistic hangover relief and liver protection composition prepared in Example 3 was diluted with purified water to form suspensions of different concentrations. Each dosage group was administered orally via gavage at the following doses:

[0183] Blank control group: given an equal volume of purified water.

[0184] Low-dose group: The dosage was 5.0 g / kg BW.

[0185] Medium dose group: The dosage was 10.0 g / kg BW.

[0186] High-dose group: The dosage was 15.0 g / kg BW.

[0187] Mice were fasted for 12 hours before gavage, but water was allowed. After administration, mice were observed for 14 consecutive days, with daily records kept of their physical appearance, behavior, respiration, food intake, feces, symptoms of poisoning, and mortality. Mice were weighed on days 0, 7, and 14 of the experiment. At the end of the observation period, all mice were euthanized with carbon dioxide, and gross dissections were performed. The color, shape, and texture of major organs (heart, liver, spleen, lungs, and kidneys) were observed visually, and abnormal tissues were examined pathologically.

[0188] 2. Experimental Results

[0189] The results are shown in Table 3. During the 14-day observation period, no mice in any of the dosage groups died, and no symptoms of poisoning such as piloerection, decreased activity, diarrhea, or convulsions were observed. Respiration was stable, and feeding and excretion were normal. The body weight of mice in all dosage groups showed an increasing trend, with no statistically significant difference compared to the blank control group (P>0.05). Gross anatomical examination revealed no obvious abnormalities in the morphology, color, or texture of the heart, liver, spleen, lungs, and kidneys in any of the treatment groups.

[0190] Table 3. Effects of the synergistic alcohol-relieving and liver-protecting composition of dual algae on body weight in mice (x̄±s, n=10)

[0191] Blank control group 0 20.12±1.15 28.51±1.62 35.24±2.31 low-dose group 5.0 20.08±1.21 28.34±1.78 34.89±2.15 medium dose group 10.0 19.95±1.18 28.67±1.55 35.48±2.42 High-dose group 15.0 20.15±1.09 28.23±1.81 35.02±2.68

[0192] 3. Experimental Conclusions

[0193] Under the experimental conditions, the maximum dose administered to the test mice reached 15.0 g / kg BW (equivalent to several hundred times the recommended clinical dose for humans), and no adverse reactions were observed. The median lethal dose (LD50) was >15.0 g / kg BW. According to the National Food Safety Standard for Acute Oral Toxicity Test (GB 15193.3), this composition is practically non-toxic and has extremely high food safety.

[0194] Example 5: Comparison of animal experimental effects of different formulations

[0195] 1. Experimental Design

[0196] 1.1 Laboratory Animals

[0197] SPF-grade ICR male mice, weighing 18-22g, were randomly assigned to groups (n=10):

[0198] 1.2 Grouping

[0199] Control group: Distilled water was administered via gavage;

[0200] Model group: administered baijiu (15mL / kg) via gavage;

[0201] Example group: The dual algae synergistic hangover relief and liver protection composition of Example 3 was administered by gavage (2g / kg), followed by baijiu (15mL / kg) 30 minutes later.

[0202] Comparative Example 1: The composition for relieving hangover and protecting the liver in Comparative Example 1 was administered by gavage (2g / kg), followed by administration of baijiu (15mL / kg) 30 minutes later.

[0203] Comparative Example 2: The composition for relieving hangover and protecting the liver in Comparative Example 2 was administered by gavage (2g / kg), followed by administration of baijiu (15mL / kg) 30 minutes later.

[0204] Comparative Example 3: The composition for relieving hangover and protecting the liver in Comparative Example 3 was administered by gavage (2g / kg), followed by administration of baijiu (15mL / kg) 30 minutes later.

[0205] Comparative Example 4: The composition for relieving hangover and protecting the liver in Comparative Example 4 was administered by gavage (2g / kg), followed by administration of baijiu (15mL / kg) 30 minutes later.

[0206] 2. Results

[0207] The sobering-up time, shortening rate, acetaldehyde clearance rate, ALT, AST, ALT reduction rate, and AST reduction rate of mice under each treatment were measured. The results are shown in Table 3 and... Figures 3-4As shown in the figure. The results showed that the sobering-up time for mice in the model group was 245 min, while the sobering-up time in the example group was shortened to 108 min (a reduction rate of 55.8%), the sobering-up time in comparative example 1 was 185 min, in comparative example 2 it was 152 min, in comparative example 3 it was 135 min, and in comparative example 4 it was 165 min. The sobering-up effect of the composition provided by this invention was significantly better than that of the comparative examples (p<0.01). The foregoing results indicate that the synergistic effect of the components of this invention is crucial for shortening the sobering-up time.

[0208] The acetaldehyde clearance rate in the example group was 62.3%, significantly better than that in the comparative groups (p<0.01). These results indicate that the composition of the present invention significantly enhances the ability of animals to metabolize acetaldehyde, specifically by exhibiting high activity of acetaldehyde dehydrogenase (ALDH2), which rapidly converts acetaldehyde into harmless acetic acid and excretes it from the body, thereby reducing the accumulation of acetaldehyde in the body.

[0209] In the model group, ALT and AST levels increased to 125.6 U / L and 138.2 U / L, respectively, while in the example group, ALT decreased to 68.8 U / L (reduction rate of 45.2%) and AST decreased to 80.8 U / L (reduction rate of 41.5%), significantly better than the comparative groups (p<0.01). These results demonstrate that the composition of the present invention has a significant protective effect against alcoholic liver injury.

[0210] In summary, the composition provided by this invention shortens the sobering time by 55.8%, achieves an acetaldehyde clearance rate of 62.3%, and reduces ALT and AST by 45.2% and 41.5%, respectively, all of which are significantly better than the comparative examples (p<0.01). This demonstrates that the combined effects of synergistic effects of algae, endogenous selenium conversion, intracellular enrichment of B-group cells, and corn oligopeptides are indispensable.

[0211] Table 4 Results of the animal experiment effect survey

[0212] blank - - - 38.5±3.2 42.3±3.5 - - Model 245±22 - - 125.6±8.5 138.2±9.0 - - Example 108±10 55.9 62.3±4.5 68.8±5.2 80.8±5.5 45.2 41.5 Comparative Example 1 185±16 24.5 28.5±3.2 95.2±6.5 108.5±7.0 24.2 21.5 Comparative Example 2 152±12 38.0 42.3±3.8 82.5±5.8 92.5±6.2 34.3 33.1 Comparative Example 3 135±11 44.9 50.2±4.0 80.2±5.5 89.2±6.0 36.1 35.5 Comparative Example 4 165±14 32.7 35.5±3.5 88.5±6.0 100.5±6.5 29.5 27.3

[0213] Comparative Example 5

[0214] A composition for relieving hangovers and protecting the liver, comprising, by weight, 20 parts of kudzu root flavonoid powder, 20 parts of Haematococcus pluvialis powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0215] The preparation method is the same as in Example 3.

[0216] Comparative Example 6

[0217] A composition for relieving hangovers and protecting the liver, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from Group A in Example 1), 15 parts of kudzu flavonoid powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0218] The preparation method is the same as in Example 3.

[0219] Comparative Example 7

[0220] A composition for relieving hangovers and protecting the liver, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from Group A in Example 1), 15 parts of kudzu root flavonoid powder, 20 parts of Haematococcus pluvialis powder, and 15 parts of Hovenia dulcis powder.

[0221] The preparation method is the same as in Example 3.

[0222] Comparative Example 8

[0223] A composition for relieving hangovers and protecting the liver, by weight, comprises: 20 parts of selenium-enriched Chlorella powder (prepared from the two groups in Example 2), 15 parts of kudzu flavonoid powder, 20 parts of Haematococcus pluvialis powder, 15 parts of Hovenia dulcis powder, and 15 parts of corn oligopeptide powder.

[0224] The preparation method is the same as in Example 3.

[0225] Comparative Example 9

[0226] A dual-algae synergistic hangover relief and liver protection composition, by weight, comprises: 10 parts of selenium-enriched Chlorella powder (prepared from Group A in Example 1), 25 parts of kudzu root flavonoid powder, 5 parts of Haematococcus pluvialis powder, 10 parts of Hovenia dulcis powder, and 5 parts of corn oligopeptide powder.

[0227] The preparation method is the same as in Example 3.

[0228] Example 6: Human Trial Experiment

[0229] 1. Experimental Design

[0230] Healthy adult volunteers (20-50 years old) were randomly assigned to four groups: a model group (30 participants, no alcohol consumption), an example group (30 participants, who took 2 g / person of the dual-algae synergistic hangover relief and liver protection composition from Example 3 30 minutes before drinking alcohol), a comparative example group (30 participants, who took 2 g / person of the composition from Comparative Example 5 30 minutes before drinking alcohol), a comparative example group (30 participants, who took 2 g / person of the composition from Comparative Example 6 30 minutes before drinking alcohol), a comparative example group (30 participants, who took 2 g / person of the composition from Comparative Example 7 30 minutes before drinking alcohol), a comparative example group (30 participants, who took 2 g / person of the composition from Comparative Example 8 30 minutes before drinking alcohol), and a comparative example group (30 participants, who took 2 g / person of the composition from Comparative Example 9 30 minutes before drinking alcohol). All participants were given 0.5 mL / kg of baijiu (a type of Chinese liquor). The experiment was approved by the medical ethics committee, and all participants signed informed consent forms.

[0231] 2. Detection Method

[0232] (1) Symptom scoring after drinking: The Acute Hangover Scale (AHS) was used. One hour after drinking, the four symptoms of headache, nausea, fatigue and dizziness were rated from 0 to 4. 0 = no symptoms, 4 = very severe, with a full score of 16. The scores of each item and the total symptom score were calculated [Reference: Rohsenow DJ, et al. The Acute Hangover Scale: Anew measure of immediate hangover symptoms. Addict Behav, 2007, 32(6): 1314-1320.].

[0233] (2) Sobering time: The sobering time is recorded from the end of drinking until the subject's self-assessment that "the hangover symptoms have basically disappeared and the mental state has returned to normal" [Reference: Yang Yong, Yang Yan, Li Kun, et al. Jianpi Sobering Formula improves mild to moderate alcohol poisoning by inhibiting the TLR4 / NF-κB pathway: a randomized controlled study. Practical Clinical Medicine Journal, 2025, 29(22): 58-65].

[0234] (3) The incidence of headache the next day: The number of people in each group who reported headaches the morning after drinking alcohol was surveyed through questionnaires, and the percentage of each group was calculated.

[0235] (4) ALT detection: The Reitman-Frankel method was used to measure serum ALT activity by collecting fasting venous blood on the morning of the day after drinking alcohol. The normal reference range is 5-25 Kamen units [Reference: Reitman S, Frankel S. Acolorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol, 1957, 28(1): 56-63.].

[0236] (5) ALT reduction rate: ALT reduction rate (%) = [(ALT mean of model group - ALT mean of trial group) / (ALT mean of model group - 25)] × 100%.

[0237] Table 5. Survey Results for Each Treatment

[0238] Headache score (1 hour) 2.5±0.3 0.4±0.1 1.3±0.2 0.8±0.2 0.7±0.1 0.9±0.2 1.0±0.2 84.0 Nausea rating (1 hour) 2.2±0.3 0.3±0.1 1.2±0.2 0.7±0.1 0.6±0.1 0.8±0.2 0.9±0.1 86.4 Fatigue score (1 hour) 2.0±0.3 0.2±0.1 0.9±0.2 0.6±0.1 0.5±0.1 0.7±0.1 0.8±0.1 90.0 Dizziness score (1 hour) 2.3±0.3 0.4±0.1 1.1±0.2 0.7±0.1 0.7±0.1 0.8±0.2 0.9±0.1 82.6 Total symptom score 9.0±0.8 1.3±0.3 4.5±0.5 2.8±0.4 2.5±0.3 3.2±0.5 3.6±0.4 85.6 Decanting time (min) 210±25 95±12 159±18 130±16 116±20 141±22 136±19 54.8 Incidence of headache the following day (%) 86.7 13.3 40.0 26.7 23.3 30.0 33.3 84.6 ALT (U / L) 68.5±8.3 32.1±5.6 51.9±7.2 45.0±6.8 41.0±7.1 46.1±7.5 47.6±6.9 — ALT reduction rate (%) — 83.7 38.2 54.0 63.2 51.5 48.0 —

[0239] Note: The improvement rate is the improvement rate of the experimental group in Example 1 compared to the model group.

[0240] 3. Results

[0241] The results are shown in Table 5. The symptom scores of the experimental group in the Example were significantly lower than those of the model group and all comparative groups (P<0.05), with a total discomfort relief rate of 85.6%, a 54.8% reduction in sobering-up time, an 84.6% reduction in the incidence of headache the following day, and an 83.7% reduction in ALT. The improvement rates of the comparative groups 5-9 were lower than those of the experimental group in the Example, further verifying the synergistic effect of the complete compound of selenium-enriched Chlorella powder, Haematococcus pluvialis powder, corn oligopeptide powder, intracellularly enriched B vitamins, and Hovenia dulcis powder in the composition of this invention.

[0242] Example 7 Validation of the synergistic antioxidant mechanism of two algae

[0243] The free radical scavenging abilities of selenium-enriched Chlorella powder, Haematococcus pluvialis powder, and a combination of selenium-enriched Chlorella powder and Haematococcus pluvialis powder (mass ratio 1:1) were determined using DPPH, ABTS, and FRAP methods. The results are shown in Table 6. Figure 6 As shown in the results, when used alone, the DPPH scavenging rates of selenium-enriched Chlorella powder (containing selenocysteine) and Haematococcus pluvialis powder (containing astaxanthin) were 62.3% and 85.2%, respectively. When combined at a mass ratio of 1:1, the DPPH scavenging rate reached 92.5%, significantly higher than the theoretical summation (73.8%), with a synergistic coefficient of 1.25. ABTS scavenging rate and FRAP value also showed similar synergistic effects (synergistic coefficients of 1.17 and 1.34), indicating a significant synergistic effect between selenocysteine ​​and astaxanthin in antioxidant activity. In conclusion, the antioxidant activity of the dual-algae combination was significantly higher than that of the single components and the theoretical summation, with synergistic coefficients of 1.17-1.34, demonstrating a synergistic antioxidant effect between selenocysteine ​​and astaxanthin.

[0244] Table 6 Synergistic Antioxidant Effect of Two Algae

[0245] Selenium-enriched Chlorella powder (containing selenocysteine) 62.3±3.5 68.5±3.2 185±12 Haematococcus pluvialis powder (containing astaxanthin) 85.2±4.0 78.5±3.5 210±15 Selenium-enriched Chlorella + Haematococcus pluvialis (1:1) 92.5±4.2 86.2±3.8 265±18 Theoretical sum value 73.8 73.5 197.5 Coordination coefficient 1.25 1.17 1.34

[0246] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A dual-algae synergistic composition for hangover relief and liver protection, characterized in that, The dual-algae synergistic hangover relief and liver protection composition comprises the following components in parts by weight: 15-25 parts of selenium-enriched Chlorella powder, 12-22 parts of kudzu flavonoid powder, 10-22 parts of Haematococcus pluvialis powder, 12-22 parts of Hovenia dulcis powder, and 10-22 parts of corn oligopeptide powder.

2. The dual-algae synergistic hangover relief and liver protection composition according to claim 1, characterized in that, The method for preparing the selenium-enriched Chlorella powder includes the following steps: sequentially performing B vitamin enrichment culture, selenium enrichment culture, low-temperature harvesting, and drying on the Chlorella algal liquid obtained from propagation.

3. The dual-algae synergistic hangover relief and liver protection composition according to claim 2, characterized in that, The enrichment culture of B vitamins includes the step of mixing and culturing the Chlorella algal solution, the mixed solution of B vitamins, and the absorption promoter; the food-grade mixed solution of B vitamins includes vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, and folic acid; the absorption promoter includes MgSO4 and ZnSO4; the culture temperature is 28℃ and the time is 24~36h; The selenium-enriched culture includes the step of mixing the culture medium obtained from the vitamin B enrichment culture with sodium selenite for culturing; the culturing temperature is 28℃ and the time is 48~72h.

4. The dual-algae synergistic hangover relief and liver protection composition according to claim 3, characterized in that, The final concentration of vitamin B1 in the mixture obtained by mixing the Chlorella algae solution, the B vitamin mixed solution, and the absorption promoter is 8-12 mg / L, the final concentration of vitamin B2 is 5-8 mg / L, the final concentration of vitamin B6 is 4-6 mg / L, the final concentration of vitamin B12 is 0.5-1.2 mg / L, the final concentration of niacin is 12-15 mg / L, the final concentration of folic acid is 3-5 mg / L, the final concentration of MgSO4 is 0.2-0.4 g / L, and the final concentration of ZnSO4 is 0.05-0.1 g / L.

5. The dual-algae synergistic hangover relief and liver protection composition according to claim 1, characterized in that, The dual-algae synergistic hangover relief and liver protection composition comprises the following components in parts by weight: 20 parts selenium-enriched Chlorella powder, 15 parts kudzu flavonoid powder, 20 parts Haematococcus pluvialis powder, 15 parts Hovenia dulcis powder, and 15 parts corn oligopeptide powder.

6. The use of the dual-algae synergistic hangover relief and liver protection composition according to any one of claims 1-5 in any of the following: (1) Preparation of hangover remedies and liver-protecting drugs; (2) Preparation of hangover remedies; (3) Prepare hangover relief medication; (4) Preparation of drugs to alleviate alcoholic liver injury; (5) Preparation of drugs that enhance the metabolism of ethanol and acetaldehyde; (6) Preparation of drugs to supplement natural B vitamins; (7) Preparation of drugs for repairing hepatocytes.

7. A drug, characterized in that, The drug comprises the dual algae synergistic hangover relief and liver protection composition according to any one of claims 1-5.

8. The medicament according to claim 7, characterized in that, The medication is used to relieve hangovers, protect the liver, alleviate alcoholic liver damage, promote the metabolism of ethanol and acetaldehyde, supplement natural B vitamins, and repair liver cells.

9. The drug according to claim 7, characterized in that, The drug also includes pharmaceutically acceptable excipients.

10. The method for preparing the drug according to claim 7, characterized in that, The preparation method includes the step of mixing the selenium-enriched Chlorella powder, the kudzu flavonoid powder, the Haematococcus pluvialis powder, the Hovenia dulcis powder, and the corn oligopeptide powder.

Images