Salicornia alcoholism-preventing liver-protecting oral liquid and preparation method thereof

Abstract

The application discloses a kind of salicornia alcoholism liver protection oral liquid and preparation method thereof, the oral liquid includes: salicornia extract 40~50%, silybum marianum extract 5~10%, mulberry extract 5~10%, fructooligosaccharide 4~8%, pure water 30~40%.The flavonoids components in salicornia are extracted by ultrasonic cell crushing assisted ethanol reflux method, the active ingredients in silybum marianum and mulberry are extracted by hot water extraction method, and are compounded according to proportion, are filtered by using precision filtration equipment, to prepare salicornia alcoholism liver protection oral liquid.The oral liquid has the effects of relieving alcoholism and invigorating spleen, protecting and nourishing liver, and can improve the detoxification capacity of liver, and is suitable for liver damage caused by drinking alcohol.The research results show that the oral liquid can significantly reduce liver damage after drinking alcohol, improve mental state, and relieve the harm caused by liver damage after drinking alcohol to organism.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a Sea Sprout Fruit hangover relief and liver protection oral liquid and its preparation method. Background Technology

[0002] Alcoholic liver disease (ALD) is toxic liver damage caused by long-term heavy drinking. It initially manifests as fatty degeneration of hepatocytes, progressing to alcoholic hepatitis, and ultimately leading to liver fibrosis and alcoholic cirrhosis. Even short-term severe alcohol abuse can induce widespread hepatocyte damage or even liver failure. Due to the prevalence of excessive alcohol consumption, the number of people suffering from ALD is gradually increasing, posing a threat to human health. Currently, many methods are used to treat alcoholic liver disease, including drug therapy, nutritional support, and liver transplantation. Although these methods can have some therapeutic effect on alcoholic liver disease, there is currently no specific cure.

[0003] When alcohol is ingested, it is metabolized by the liver, producing a large amount of acetaldehyde. Acetaldehyde is absorbed in the intestines, generating a large number of free radicals. Free radicals are harmful to biological membranes, producing large amounts of O2. 2- Under normal circumstances, reactive oxygen species (ROS) are eliminated by glucose-stimulating hormone (GSH). However, the increasing consumption of antioxidants in the body weakens their ability to eliminate these free radicals. Long-term alcohol consumption leads to a large accumulation of malondialdehyde (MDA), which can cause liver necrosis or fibrosis in severe cases. Flavonoids can rapidly break down ethanol, reducing blood ethanol concentration and eliminating large amounts of free radicals generated by lipid oxidation, thus protecting liver cells from damage caused by electrophilic free radical compounds. Flavonoids are a class of compounds derived from fruits, vegetables, tea, seeds, or plant roots. They possess broad pharmacological activities and low toxicity, exhibiting varying degrees of preventative and therapeutic effects on cardiovascular, nervous, endocrine, immune, and digestive / respiratory system diseases, making them highly valuable in clinical applications. In recent years, flavonoids have made some progress in the prevention and treatment of alcoholic liver injury.

[0004] Sea sedge (Salicornia) belongs to the genus Salicornia in the family Chenopodiaceae. It grows in moist, heavily saline soils in saline-alkali lands, along salt lakes, and in coastal areas and river valleys. It is a stalked, leafless green plant, mainly distributed in Jiangsu, North China, Shandong, Northwest China, and Liaoning. Sea sedge is a healthy green food beneficial to human health. It not only has a refreshing, salty, and crisp taste but is also rich in nutrients and has high utilization value. It contains various nutrients such as fatty acids, proteins, minerals, and vitamins, and is a recognized green organic vegetable. Studies have shown that the active substances isolated from sea sedge include flavonoids, alkaloids, polysaccharides, and saponins. These active components have anti-inflammatory, antioxidant, anti-tumor, blood sugar-lowering, and blood lipid-lowering effects. Among them, flavonoids account for up to 60% of the total content of sea sedge extract, and flavonoids have a certain detoxification effect on alcohol poisoning. Therefore, sea sedge has high nutritional value and good development prospects. In addition, sea spirea has a strong salt tolerance and can absorb salt from saline-alkali land year by year. Therefore, planting sea spirea can make full use of tidal flats and coastal wastelands and improve the geology of saline-alkali land. Thus, developing sea spirea oral liquid for hangover relief and liver protection has significant application value and social benefits.

[0005] Milk thistle (scientific name: *Silybum marianum* (L.) Gaertn.) is an annual or biennial herb belonging to the genus *Silybum* in the family Asteraceae. It is bitter and cool in nature, and has the effects of clearing heat and dampness, soothing the liver and gallbladder. Clinically, it is used for liver protection, lowering blood lipids, preventing atherosclerotic plaque formation, its effects on the cardiovascular system, its protective effect against cerebral ischemia, and its anti-platelet aggregation properties. Milk thistle extract has been used as a hepatoprotective drug.

[0006] Mulberries are the ripe fruit of the mulberry tree, a member of the Moraceae family. They are sweet and juicy, and a popular fruit. Mulberries are sweet and cold in nature, entering the heart, liver, and kidney meridians. They nourish yin and replenish blood, and can treat yin deficiency, insufficient body fluids, and insomnia. Mulberries also have excellent effects in nourishing the heart, liver, and kidneys, as well as nourishing blood and dispelling wind.

[0007] However, there are currently no reports of hangover remedies or liver-protecting drugs that use sea sedge as the main ingredient and milk thistle, mulberry, and other raw materials as auxiliary ingredients. Summary of the Invention

[0008] To address the shortcomings of existing technologies, this invention provides a Sea Sprouts Decoction for Relieving Hangovers and Protecting Liver, along with its preparation method. This oral liquid has the effects of relieving hangovers, invigorating the spleen, protecting and nourishing the liver, and enhancing the liver's detoxification ability. It is suitable for liver damage caused by alcohol consumption, significantly reducing liver damage after drinking, improving mental state, and alleviating the harm caused by alcohol-induced liver damage to the body.

[0009] This invention is achieved through the following technical solution:

[0010] A type of oral liquid containing sea sedge for hangover relief and liver protection contains the following ingredients by weight percentage: 40-50% sea sedge extract, 5-10% milk thistle extract, 5-10% mulberry extract, 4-8% fructooligosaccharides, and 30-40% purified water.

[0011] A method for preparing a Sea Sprout Seed oral liquid for relieving hangovers and protecting the liver includes the following steps:

[0012] Step 1) Pre-treatment of materials: Select fresh sea sedge tender stems, wash them with clean water to remove mud and sand, drain the water and dry them, then grind them into powder for later use; dry milk thistle and mulberry, grind them into powder for later use.

[0013] Step 2) Extraction of active ingredients: Take sea spirulina powder, add ethyl acetate and let stand overnight to remove ester-soluble components, filter, and evaporate the residual ethyl acetate; use ultrasonic-assisted extraction method to dissolve the treated sea spirulina powder in 60% ethanol aqueous solution and extract by ultrasonication; repeat the extraction twice, combine the filtrates, remove the solvent, and freeze-dry to obtain sea spirulina extract;

[0014] Milk thistle and mulberry powders were weighed separately, and both were extracted by heating and reflux extraction to obtain milk thistle extract and mulberry extract, respectively. The heating and reflux extraction method is as follows: the first extraction was carried out by heating and reflux extraction at a material-to-liquid ratio of 1g:12mL for 1.5h, and the second extraction was carried out by heating and reflux extraction at a material-to-liquid ratio of 1g:10mL for 1.5h. The two filtrates were combined, the solvent was removed, and the extracts were freeze-dried to obtain the final product.

[0015] Step 3) Preparation of oral liquid: Combine sea sedge extract, milk thistle extract, mulberry extract, fructooligosaccharide and purified water in proportion, filter first, then fine filter, pre-sterilize the filling bottle, and bottle to obtain the oral liquid.

[0016] Preferably, in step 2), the ratio of sea spirulina powder to ethyl acetate is 1 g: 4 mL.

[0017] Preferably, the ratio of the treated sea spirulina powder to the ethanol aqueous solution in step 2) is 1g:20mL.

[0018] Preferably, the ultrasonic extraction in step 2) uses a power of 150W, a time of 1 hour, and an extraction temperature of 50℃.

[0019] The beneficial effects of this invention are as follows:

[0020] This invention uses sea sedge, milk thistle, and mulberry as raw materials. Ultrasonic cell disruption-assisted ethanol reflux extraction is used to extract flavonoids from sea sedge, while hot water extraction is used to extract active ingredients from milk thistle and mulberry. These are then compounded in a specific ratio and filtered using a fine filtration device to prepare a sea sedge-based hangover-relieving and liver-protecting oral liquid. Flavonoids can rapidly decompose ethanol, thereby reducing the concentration of ethanol in the blood. They can also scavenge a large number of free radicals generated by lipid oxidation that are harmful to biological membranes, achieving the effect of hangover relief and liver protection. Research results show that the production process of the sea sedge-based hangover-relieving and liver-protecting oral liquid is simple, can greatly shorten the production cycle, and can achieve continuous production, avoiding many problems associated with intermittent production. The raw materials sea sedge, milk thistle, and mulberry are easy to obtain; the product preparation process is simple and easy to implement, the production cost is moderate, and the quality is stable and controllable; furthermore, it is easy to implement mechanized assembly line operation, meeting the requirements of industrial production. Attached Figure Description

[0021] Figure 1 A pathological section of mouse liver tissue;

[0022] Figure 1 In the diagram: A represents the blank control group; B represents the model group; C represents the positive control group; and D represents the sea spirulina oral liquid group.

[0023] Figure 2 Figure 1 shows the results of quantitative real-time PCR for different inflammatory factors.

[0024] Figure 2 In the table: A represents the relative expression level of IL-1β mRNA; B represents the relative expression level of IL-6 mRNA; C represents the relative expression level of TNF-α mRNA; and D represents the relative expression level of CYP2E1 mRNA. Detailed Implementation

[0025] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0026] Example 1

[0027] A type of oral liquid for relieving hangovers and protecting the liver, made from the following ingredients in the following percentages by weight: 40-50% sea sedge extract, 5-10% milk thistle extract, 5-10% mulberry extract, 4-8% fructooligosaccharides, and 30-40% purified water.

[0028] A method for preparing a Sea Sprout (Spatholobus suberectus) oral liquid for relieving hangovers and protecting the liver, the specific steps of which are as follows:

[0029] (1) Material pretreatment:

[0030] Select fresh, tender stems of sea sedge, wash them with clean water to remove dirt and sand, drain and dry them, then grind them into powder for later use. Dry the milk thistle and mulberries, then grind them into powder for later use.

[0031] (2) Obtaining the active ingredients:

[0032] Take 100g of *Sargassum fusiforme* powder, add 400mL of ethyl acetate and let stand overnight to remove most of the ester-soluble components. Filter and evaporate the remaining ethyl acetate. Using ultrasound-assisted extraction, dissolve the powder in 60% ethanol aqueous solution at a material-to-liquid ratio of 1:20 (g / mL) and extract at 150W for 1 hour at 50℃. Repeat the extraction twice, combine the filtrates, remove the solvent using a rotary evaporator, and freeze-dry to obtain the active ingredient of *Sargassum fusiforme* (*Sargassum fusiforme* extract).

[0033] Appropriate amounts of milk thistle and mulberry powder were weighed separately, and the active components of milk thistle (milk thistle extract) and mulberry (mulberry extract) were obtained by heating and reflux extraction, respectively, for later use. The heating and reflux extraction method is as follows: the first extraction was carried out by heating and reflux extraction at a material-to-liquid ratio of 1:12 (g / mL) for 1.5 h, and the second extraction was carried out by heating and reflux extraction at a material-to-liquid ratio of 1:10 (g / mL) for 1.5 h. The two filtrates were combined, the solvent was removed, and the extracts were freeze-dried to obtain the final product.

[0034] (3) Preparation of oral liquid:

[0035] The mixture is compounded according to the following mass ratios: 40-50% sea spirulina extract, 5-10% milk thistle extract, 5-10% mulberry extract, 4-8% fructooligosaccharides, and 30-40% purified water. After filtration, the mixture is further filtered using a fine filtration device. The bottling bottles are then pre-sterilized and bottled to obtain the sea spirulina hangover relief and liver protection oral liquid product (hereinafter referred to as sea spirulina oral liquid).

[0036] The results showed that the *Hydrocotyle sibthorpioides* oral liquid for hangover relief and liver protection produced using this technology and equipment showed no sedimentation after several months. This process effectively removes bacteria and impurities, clarifies the liquid, and prevents sedimentation even after prolonged storage.

[0037] Example 2

[0038] The hepatoprotective effect of the oral liquid containing sea sedge prepared in Example 1 was studied:

[0039] Thirty-two healthy male mice were randomly divided into a blank control group, a model group, a positive control group (silymarin 200 mg / kg), and a sea sedge oral liquid dosage group (100 mg / kg), with eight mice in each group. The mice were administered the drugs by gavage. The blank control group and the model group were administered 5 mL / kg of physiological saline by gavage, while the positive control group was administered 200 mg / kg of silymarin, a hepatoprotective drug, daily by gavage for 28 days. Except for the blank control group, all other groups were given alcohol by gavage at a dose of 6 g / kg to induce an acute liver injury model in mice, repeated every 12 hours for a total of three times.

[0040] 1. Effect of Seaweed Extract (Hydrocotyle vulgaris) on Liver Factor in Mice with Alcoholic Liver Injury

[0041] Immediately after euthanasia, the mice were dissected, and the liver tissue was removed and quickly immersed in cold saline to wash away surface bloodstains. The surface moisture was then absorbed with absorbent paper, and the fatty tissue and fascia on the liver were removed. The liver weight was then accurately measured, and the liver organ index was calculated in combination with the mouse body weight.

[0042] As shown in Table 1 below, compared with the blank group, the liver coefficient of the model group mice was significantly increased (P < 0.01), indicating that the model was successfully established. Compared with the model group, the liver coefficient of the Sea Sprout Oral Liquid group was significantly decreased (P < 0.05). Compared with the positive control group, the organ coefficient of the Sea Sprout Oral Liquid group was lower than that of the positive control group, indicating that the Sea Sprout Oral Liquid can significantly inhibit hepatocyte edema.

[0043] Table 1. Changes in liver coefficient in each group of mice

[0044]

[0045] Table 1 Note: Compared with the model group # P < 0.05 ## P < 0.01; compared with the control group * P < 0.05 ** P < 0.01

[0046] 2. Effects of *Sargassum fusiforme* extract on serum ALT and AST levels and SOD and GSH activities in mice with alcoholic liver injury.

[0047] After dissection of the mice, blood was drawn from the heart using a disposable syringe, taking precautions to prevent contamination and hemolysis. The blood was placed in a heparinized centrifuge tube, incubated at room temperature for 30 minutes, and then centrifuged at 3000 rpm for 10 minutes to prepare serum. ALT, AST, SOD, and GSH levels in the serum were measured using kits purchased from Nanjing Jiancheng Biotechnology Institute, and all procedures were strictly performed according to the kit instructions.

[0048] As shown in Table 2, compared with the blank group, the serum ALT and AST levels in the model group mice were significantly increased (P < 0.01), indicating successful model establishment. Compared with the model group, the ALT and AST levels in the *Sargassum fusiforme* oral liquid group were significantly decreased (P < 0.01), with statistically significant differences. The serum SOD and GSH activities in the model group mice were decreased (P < 0.05), indicating successful model establishment. Compared with the model group, the SOD and GSH activities in the high-dose group of *Sargassum fusiforme* extract (*Sargassum fusiforme* oral liquid group were increased (P < 0.05), with statistically significant differences. Compared with the positive control group, the serum indicators in the *Sargassum fusiforme* oral liquid group were all better than those in the positive control group, indicating that *Sargassum fusiforme* oral liquid has a strong hepatoprotective effect.

[0049] Table 2. Effects of *Sargassum fusiforme* on ALT, AST, SOD, and GSH levels in mice with alcoholic liver injury.

[0050]

[0051] Table 2 Note: Compared with the model group * P<0.05 ** P<0.01; compared with the control group # P<0.05 ## P<0.01

[0052] 3. Effects of Sea Sprout Extract on Liver Morphology in Mice with Alcoholic Liver Injury

[0053] After weighing the mouse liver tissue, it was immediately immersed in 10% neutral formalin fixative for 72 hours, dehydrated and fixed, routinely sampled, embedded in paraffin, sectioned with a microtome to a thickness of 5 μm, stained with H&E, and the morphological changes of each liver tissue were observed under a light microscope.

[0054] Figure 1 This is a pathological section of mouse liver tissue. Figure 1 In the diagram: A represents the blank control group, B the model group, C the positive control group, and D the *Hydrocotyle sibthorpioides* oral liquid group. The samples were observed under a 200x microscope using the H&E staining method. Under a light microscope, it can be seen that... Figure 1 In the blank group shown in A, the liver cells had clear structures, intact nuclei, abundant cytoplasm, and were arranged relatively neatly, with the hepatic cords radiating outwards. Figure 1 The mouse hepatocytes in model group B showed severe degenerative changes, with extensive edema around the cells, cell nucleus cavitation, patchy necrosis of the hepatocytes, and disappearance of hepatic sinusoids, indicating that the mouse model of alcoholic liver injury was successfully established. Figure 1C, using silymarin administered via gavage for liver protection as a positive control, showed a significant reduction in perihepatic edema compared to the model group. Hepatocytes were more neatly arranged, with well-preserved nuclei, and the hepatic cords exhibited a radial pattern. Figure 1 Group D of seaweed extract oral liquid and Figure 1 Morphological comparison of mouse hepatocytes in model group B showed that liver tissue damage was significantly reduced in the group treated with the oral liquid of *Hylocereus undatus*, hepatocyte edema gradually subsided, hepatic cords gradually returned to a radial morphology, and cell nuclei gradually became more complete and clear. Figure 1 Compared with the silymarin-positive control group, the hepatoprotective effect of sea sedge was better than that of silymarin.

[0055] 4. Detection of different inflammatory factors by real-time PCR

[0056] Total RNA was extracted from liver tissue using the Trizol method. Approximately 50 mg of liver tissue was added to Trizol solution and dispersed using a high-speed disperser. Pre-cooled isopropanol precipitated the total RNA, and excess impurities were washed away with ethanol to obtain the final total RNA. cDNA was obtained by reverse transcription using the Thermo K1622 reverse transcription kit. Quantitative real-time PCR was performed. The RT-PCR reaction system consisted of: Water, 3 μL of PCR grade (Vial2, colorless cap), 2 μL of PCR Primer 10×conc, 12 μL of Master Mix 2×conc (Vial1, green cap), and 3 μL of the reverse transcription product, for a total reaction volume of 20 μL. β-actin was used as an internal control gene for quantitative PCR.

[0057] Figure 2 The results of quantitative PCR for different inflammatory factors. Figure 2 In the table: A represents the relative expression level of IL-1β mRNA, B represents the relative expression level of IL-6 mRNA, C represents the relative expression level of TNF-α mRNA, and D represents the relative expression level of CYP2E1 mRNA.

[0058] like Figure 2 As shown, the expression levels of IL-1β, IL-6, TNF-α, and CYP2E1 in the model group were significantly higher than those in the blank group, indicating that the model was successfully established. Compared with the model group, the expression levels of the three inflammatory factors were decreased in the positive control group. Figure 2 A, Figure 2 B Figure 2In the C group, the expression levels of inflammatory mediators IL-1β, IL-6, and TNF-α in the *Hypericum perforatum* oral liquid group all showed a decreasing trend compared to the model group. This indicates that *Hypericum perforatum* oral liquid protects the liver by inhibiting the expression of related inflammatory factors IL-1β, IL-6, and TNF-α. Increased CYP2E1 expression levels lead to enhanced oxidative stress, depletion of the body's antioxidant system, and extensive rupture of hepatocyte mitochondrial membranes, causing liver damage. Figure 2 The expression level of CYP2E1 in D was significantly reduced, indicating that sea sedge can reduce alcohol-induced liver damage by regulating the expression level of CYP2E1.

[0059] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. However, these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions claimed by the present invention.

Claims

1. A Seaweed Extract-based Oral Solution for Relieving Hangovers and Protecting the Liver, characterized in that, Made from the following ingredients by weight percentage: Seaweed extract 40-50%, Milk thistle extract 5-10%, Mulberry extract 5-10%, Fructooligosaccharides 4-8%, 30-40% purified water; The sea spirulina extract is prepared by the following method: Take sea spirulina powder, add ethyl acetate and let stand overnight to remove ester-soluble components, filter, and evaporate the residual ethyl acetate; use ultrasonic-assisted extraction to dissolve the treated sea spirulina powder in 60% ethanol aqueous solution and extract ultrasonically; repeat the extraction twice, combine the filtrates, remove the solvent, and freeze-dry to obtain the product; The milk thistle extract and mulberry extract were prepared by the following method: Milk thistle and mulberry powders were weighed separately and extracted using water as a solvent by heating and reflux extraction to obtain milk thistle extract and mulberry extract, respectively. The heating and reflux extraction method was as follows: the first extraction was carried out at a material-to-liquid ratio of 1 g: 12 mL and heated and refluxed for 1.5 h; the second extraction was carried out at a material-to-liquid ratio of 1 g: 10 mL and heated and refluxed for 1.5 h. The two filtrates were combined, the solvent was removed, and the extracts were freeze-dried to obtain the final product.

2. The preparation method of the Sea Sprouts Decoction for Relieving Hangover and Protecting the Liver as described in claim 1, characterized in that, Includes the following steps: Step 1) Pre-treatment of materials: Select fresh tender stems of sea sedge, wash them with clean water to remove mud and sand, drain the water and dry them, then grind them into powder for later use; dry milk thistle and mulberry, grind them into powder for later use; Step 2) Extraction of active ingredients: Take sea spirulina powder, add ethyl acetate and let stand overnight to remove ester-soluble components, filter, and evaporate the residual ethyl acetate; use ultrasonic-assisted extraction method to dissolve the treated sea spirulina powder in 60% ethanol aqueous solution and extract by ultrasonication; repeat the extraction twice, combine the filtrates, remove the solvent, and freeze-dry to obtain sea spirulina extract; Milk thistle and mulberry powders were weighed separately and extracted using water as a solvent by heating and reflux extraction to obtain milk thistle extract and mulberry extract, respectively. The heating and reflux extraction method was as follows: the first extraction was carried out at a material-to-liquid ratio of 1 g: 12 mL and heated and refluxed for 1.5 h; the second extraction was carried out at a material-to-liquid ratio of 1 g: 10 mL and heated and refluxed for 1.5 h. The two filtrates were combined, the solvent was removed, and the extracts were freeze-dried to obtain the final product. Step 3) Preparation of oral liquid: Combine sea sedge extract, milk thistle extract, mulberry extract, fructooligosaccharide and purified water in proportion, filter first, then fine filter, pre-sterilize the filling bottle, and bottle to obtain the oral liquid.

3. The preparation method of the Sea Sprouts Decoction for Relieving Hangover and Protecting the Liver according to claim 2, characterized in that, Step 2) The ratio of sea spirulina powder to ethyl acetate is 1 g: 4 mL.

4. The preparation method of the Sea Sprouts Decoction for Relieving Hangover and Protecting the Liver according to claim 2, characterized in that, Step 2) The ratio of the treated sea spirulina powder to the ethanol aqueous solution is 1 g: 20 mL.

5. The preparation method of the Sea Sprouts Decoction for Relieving Hangover and Protecting the Liver according to claim 2, characterized in that, Step 2) The ultrasonic extraction power is 150 W, the time is 1 h, and the extraction temperature is 50℃.

Images