Sodium alginate-polylysine composite hydrogel alcoholism microspheres, and preparation method and application thereof

Abstract

This invention belongs to the field of biomedical materials and functional food formulation technology, and discloses a sodium alginate-polylysine composite hydrogel hangover-relieving microsphere, its preparation method, and its application. The microspheres are prepared using sodium alginate and polylysine as raw materials through interpenetrating network interaction. They possess the function of adsorbing ethanol, preventing its absorption into the bloodstream and avoiding severe intoxication. Furthermore, the microspheres can also act as drug carriers to encapsulate poorly water-soluble hangover-relieving components such as curcumin. The adsorbed ethanol within the hydrogel enhances drug absorption, thereby improving bioavailability and strengthening the hangover-relieving and liver-protecting effects. This invention solves the problems of slow onset of action, low bioavailability, and high gastrointestinal irritation associated with traditional hangover remedies. It can be prepared as an oral hangover remedy with broad application prospects.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials and functional food formulations, specifically to a sodium alginate-polylysine composite hydrogel hangover-relieving microsphere, its preparation method, and its application. Background Technology

[0002] Alcohol poisoning and post-drinking discomfort have become common health problems. Existing oral hangover remedies are mainly in conventional dosage forms such as tablets, capsules, and oral liquids. Their core hangover remedy ingredients (such as acetaldehyde dehydrogenase, glutathione, curcumin, puerarin, etc.) are easily degraded and inactivated in the acidic environment of the human stomach. Furthermore, their rapid release in the stomach can easily cause gastrointestinal irritation. At the same time, due to low intestinal absorption efficiency, the hangover remedy is slow to take effect and has poor efficacy.

[0003] Microsphere formulations, as novel targeted drug delivery carriers, can effectively protect active ingredients and improve bioavailability. Sodium alginate, due to its good biocompatibility and biodegradability, has become a commonly used natural polysaccharide carrier for encapsulating active ingredients. It also has ethanol adsorption properties, preventing ethanol absorption into the bloodstream and avoiding severe intoxication, thus acting as an ethanol barrier. Microspheres can also serve as drug carriers to encapsulate poorly water-soluble hangover-relieving components such as curcumin. The adsorption of ethanol within the hydrogel enhances drug absorption, thereby improving bioavailability and strengthening the hangover-relieving and liver-protecting effects. However, calcium alginate microspheres have poor mechanical strength and are easily broken in the intestines, affecting ethanol adsorption. Polylysine, as a natural cationic polypeptide, can form a stable polyelectrolyte composite structure with sodium alginate through interpenetrating network interactions, significantly improving the mechanical strength of the microspheres and their ethanol adsorption and release performance. Therefore, developing a sodium alginate-polylysine composite hydrogel hangover-relieving microsphere with a mild process, controllable particle size, and high ethanol adsorption efficiency is crucial to solving the problems of slow onset and poor efficacy of traditional hangover-relieving products. Summary of the Invention

[0004] To overcome the shortcomings of existing technologies, this invention provides sodium alginate-polylysine composite hydrogel microspheres for relieving hangovers. Using sodium alginate and polylysine as raw materials, these microspheres are constructed through interpenetrating networks. They possess the function of adsorbing ethanol, preventing its absorption into the bloodstream and avoiding severe intoxication. Furthermore, the microspheres can act as drug carriers, encapsulating poorly water-soluble hangover-relieving components such as curcumin. The adsorbed ethanol within the hydrogel enhances drug absorption, thereby improving bioavailability and strengthening the hangover-relieving and liver-protecting effects.

[0005] The above-mentioned objective of this invention is achieved through the following technical solution: A sodium alginate-polylysine composite hydrogel microsphere for hangover relief is developed. Using sodium alginate and polylysine as raw materials, the microspheres are constructed through an interpenetrating network to form a polyelectrolyte hydrogel. These microspheres adsorb ethanol, preventing its absorption into the bloodstream and avoiding severe intoxication. Furthermore, the microspheres can act as drug carriers, encapsulating poorly water-soluble hangover-relieving components such as curcumin. The adsorbed ethanol within the hydrogel enhances drug absorption, thereby improving bioavailability and strengthening the hangover relief and liver protection effects.

[0006] A method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres, comprising the following steps: S1 Add sodium alginate, curcumin and CaCO3 powder to deionized water and stir at room temperature for 60 min to completely dissolve them to obtain sodium alginate suspension. S2. Add the emulsifier to the liquid paraffin and stir at room temperature for 10-30 minutes to form the oil phase. S3: The sodium alginate suspension obtained in S1 is added to the oil phase and stirred at high speed for 10-30 min to form a W / O emulsion. Glacial acetic acid is then added dropwise to dissolve CaCO3 and release Ca2+. 2+ Ions form calcium alginate hydrogel microspheres. The microspheres are collected by centrifugation and washed 2-4 times with a 1.0% (w / v) Tween 80 solution to remove residual liquid paraffin. S4. The calcium alginate microspheres obtained in S3 were added to a polylysine solution to carry out a cross-linking reaction. Sodium alginate and polylysine formed a polyelectrolyte structure through interpenetrating network interaction. After the reaction was completed, the microspheres were separated by centrifugation to obtain composite hydrogel hangover relief microspheres.

[0007] Furthermore, the sodium alginate is a commercially available product with a molecular weight of 70.0~200.0 kDa, and the mass ratio of guluronic acid to mannuluronic acid is 1.0~3.0; the polylysine is a commercially available product with a molecular weight of 5.0~30.0 kDa; and the composite hydrogel hangover-relieving microspheres have a particle size of 300~2000 μm.

[0008] Furthermore, in the method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres, in step S1, the concentration of sodium alginate solution is 10.0~40.0 mg / mL, the content of curcumin is 0.1~1.0 mg / mL, and the content of CaCO3 is 1.0~10.0 mg / mL.

[0009] Furthermore, in the method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres, in step S2, the dispersion medium is liquid paraffin, or one or more mixtures of edible oils such as soybean oil or olive oil; the emulsifier is one or more combinations of dehydrated sorbitan fatty acid ester surfactants such as Span-80, and the emulsifier concentration is 0.5~5.0 wt% (w / v); the volume ratio of the aqueous phase to the oil phase is 1:5~20; and the stirring speed is 300~1500 rpm.

[0010] Furthermore, in the method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres, in step S3, the mass ratio of glacial acetic acid to CaCO3 is 2~4:1, the dropping rate is 0.5~2.0 mL / min, the stirring speed is maintained at 1000~3000 rpm, and the calcification reaction time is 1~4h.

[0011] Furthermore, in the method for preparing sodium alginate-polylysine composite gel hangover-relieving microspheres, in step S4, the concentration of polylysine solution is 1.0~4.0 mg / mL, and the volume ratio of microspheres to polylysine is 1:5~20.

[0012] Furthermore, the composite hydrogel hangover-relieving microspheres are used in the preparation of hangover-relieving beverages, hangover-relieving jellies, and hangover-relieving gummies.

[0013] Compared with traditional hangover remedies, the beneficial effects of this invention include: 1. Innovative Dosage Form and Drug Release Characteristics: Sodium alginate-polylysine composite hydrogel microspheres for hangover relief were prepared. These microspheres have the function of adsorbing ethanol, which can prevent ethanol from being absorbed into the blood and avoid severe drunkenness. Furthermore, the microspheres can also act as drug carriers to encapsulate poorly water-soluble hangover relief components such as curcumin. The adsorbed ethanol in the hydrogel can improve the drug absorption rate, thereby improving bioavailability and enhancing the hangover relief and liver protection effects.

[0014] 2. Material composite and structural optimization: By cross-linking polylysine on calcium alginate microspheres through an interpenetrating network strategy to form a dense composite structure, the mechanical strength and stability of the microspheres are improved, the ethanol adsorption is enhanced, and the diffusion resistance of the alcohol-degrading components is increased, thereby achieving the sustained release of ethanol.

[0015] 3. Advantages in maintaining the antibacterial properties and strength of microspheres: Polylysine has antibacterial properties, so the prepared microspheres do not require high-temperature sterilization, which can avoid the impact of high temperature on the strength of hydrogel.

[0016] 4. Excellent biocompatibility: The sodium alginate and polylysine used are both natural biomaterials, non-toxic and biodegradable. Polylysine can be broken down into lysine in the human body and absorbed and utilized. Sodium alginate can be degraded by intestinal flora, meeting the biosafety requirements of oral preparations and having no risk of accumulation in the body.

[0017] 5. Simple preparation and synergistic effects: The jelly production method provided by this invention is simple and easy to use, and can enable the components to work synergistically to enhance the effect. It has both anti-drunkenness and liver protection effects, and can produce good hangover relief and liver protection effects. Attached Figure Description

[0018] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0019] Figure 1 These are composite hydrogel microspheres with different particle sizes for alcohol detoxification: a) sodium alginate concentration of 1.0%, b) sodium alginate concentration of 2.0%, c) sodium alginate concentration of 3.0%, and d) sodium alginate concentration of 4.0%. Detailed Implementation

[0020] The present invention is described in detail below through specific embodiments, but this does not limit the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and the experimental equipment, materials, reagents, etc. used can all be obtained commercially.

[0021] Example 1: Preparation of Composite Microspheres 1. Experimental Procedure A certain amount of sodium alginate was weighed and added to 10 mL of deionized water. The mixture was stirred at room temperature for 60 min (200 rpm) until completely dissolved. 100 mg of CaCO3 and 10 mg of curcumin were added to obtain a sodium alginate suspension. 100 mL of liquid paraffin was added to a beaker, along with 1.0 g of Span 80. The mixture was stirred at room temperature for 30 min (300 rpm) to form an oil phase. The sodium alginate suspension was then added to the liquid paraffin oil phase and stirred at 1000 rpm for 30 min to form a stable W / O emulsion. 0.3 mL of glacial acetic acid was added dropwise to the emulsion at a rate of 0.5 mL / min, with continuous stirring (1000 rpm). The cross-linking reaction was carried out for 2 h to completely calcify the sodium alginate into calcium alginate hydrogel microspheres. After the reaction, the microspheres were centrifuged and separated using a solution containing 1.0% (w / v) Tween. The microspheres were washed three times with 80% deionized water to remove the liquid paraffin on the surface of the microspheres. The calcium alginate microspheres were added to 10 mL of polylysine solutions of different concentrations and crosslinked at room temperature for 1 h. Sodium alginate and polylysine formed a polyelectrolyte structure through interpenetrating network. After the reaction was completed, the microspheres were centrifuged to obtain composite hydrogel hangover relief microspheres.

[0022] 2. Performance Testing (1) Measuring the particle size of microspheres At least 100 microsphere images were obtained by randomly selecting 10 fields of view under an inverted microscope. The particle size of each microsphere was read using ImageJ software, and the average value was calculated as D0.

[0023] (2) Measure the reaction amount of polylysine After the above crosslinking reaction was completed, the concentration of polylysine in the reaction solution was measured using the Coomassie Brilliant Blue method, and the amount of polylysine reacted onto the microspheres was calculated using the following formula.

[0024] Polylysine reaction amount = (C0 - C) t V C0: Initial concentration of polylysine solution before reaction; C t : Concentration of polylysine after the reaction; V: Solution volume.

[0025] (3) Measuring the strength of microspheres ① Turn on the dynamic shear rheometer and preheat for 30 minutes. Then, use the instrument's built-in calibration module to calibrate the torque and temperature of the parallel plate fixture. ② Place the prepared microspheres in the center of the rheometer base plate, and slowly lower the upper plate until the sample thickness is compressed to 4 mm (ensure that the sample is completely in contact with the upper and lower plates and there are no air bubbles left). ③ Determine the storage modulus Set the scanning parameters: fix the strain to the linear viscoelastic strain determined in the pre-experiment (e.g., 1%), the angular frequency range is 0.1~100 rad / s, the scanning step size is 5 steps / decade, and the temperature is kept at 25℃.

[0026] The frequency scanning program is started, and the instrument automatically records the storage modulus data G' at different frequencies. Each sample is tested in parallel three times. A logarithmic curve is plotted with angular frequency as the abscissa and G' as the ordinate. The G' value at the characteristic frequency is read as the representative storage modulus of the sample.

[0027] 3. Experimental Results (1) Effect of sodium alginate concentration on microsphere size and storage modulus (2) Effect of polylysine crosslinking on microsphere particle size and storage modulus The experimental results show that the particle size of sodium alginate hydrogel microspheres is positively correlated with the concentration of sodium alginate; the particle size increases with increasing concentration. Furthermore, the storage modulus of the microspheres is also closely related to the sodium alginate concentration. As the sodium alginate concentration increases, the storage modulus of the hydrogel increases, especially in the low-concentration region (below 20.0 mg / mL). Above 30.0 mg / mL, the storage modulus increases slowly due to the overcrowding of molecular chains and the resulting network defects. After cross-linking calcium alginate microspheres with polylysine, the particle size decreases, indicating a more compact microsphere structure. Compared to calcium alginate microspheres, the composite microspheres exhibit a significantly increased storage modulus, which further increases with increasing polylysine concentration. At a polylysine concentration of 4.0 mg / mL, the storage modulus of the microspheres is 7.06 times higher.

[0028] Example 2: Hydrogel Ethanol Adsorption and Release Experiment 1. Experimental Procedure (1) Ethanol adsorption experiment Weigh 10 g of the composite hydrogel alcohol-degrading microspheres and place them in a 50 mL stoppered conical flask. Add 30 mL of 50% ethanol solution to the flask, ensuring the microspheres are completely submerged. Place the flask in a constant temperature water bath (set to 37℃) and allow it to adsorb by shaking at 100 r / min. Take samples at 10 min, 30 min, 1 h, 2 h, and 4 h of adsorption, 1 mL each time. Collect the supernatant and add 10 mL of distilled water. After appropriate dilution, determine the ethanol content in the liquid using gas chromatography. Detector: FID; Column: DB-FFAP (30 m × 0.25 mm × 0.25 μm); Column temperature: 45℃; Injection volume: 1 μl; Mode: splitless. Repeat the above experiment three times and take the average value.

[0029] The amount of ethanol adsorbed is calculated using the following formula: Q (g / g) = (C0V0 - C t V t ) / m C t : Ethanol concentration in the solution at time t, g / mL; C0: Ethanol concentration in the solution at the start of adsorption, g / mL; V t : Solution volume at time t; m0: Mass of the hydrogel before ethanol adsorption.

[0030] Subsequent sampling and adsorption calculations must take into account the amount of ethanol removed during sampling.

[0031] 2. Experimental Results The experimental results show that the hydrogel's ability to adsorb ethanol is positively correlated with the sodium alginate concentration, increasing with increasing sodium alginate concentration. When the sodium alginate concentration reaches above 30 mg / mL, 1g of microspheres can adsorb more than 0.4g of ethanol, and the ethanol adsorption rate is relatively fast, exceeding 0.3g of ethanol in 10 minutes and reaching saturation adsorption capacity in 30 minutes. This aligns with the need for rapid cleansing after drinking to prevent the rapid absorption of ethanol into the bloodstream. The concentration of the polylysine crosslinking solution is also closely related to the ethanol adsorption capacity, increasing with increasing polylysine concentration. Microspheres exhibit a relatively high ethanol adsorption capacity at a polylysine concentration of 3.0 mg / mL. This is attributed to the more compact hydrogel structure resulting from polylysine crosslinking, which facilitates ethanol fixation. When the sodium alginate concentration exceeds 40 mg / mL, increasing the concentration does not significantly improve the ethanol adsorption capacity of the microspheres. This is mainly because the microsphere strength does not significantly improve, and at sodium alginate concentrations above 40 mg / mL, the solution viscosity is too high, resulting in poor flowability and making microsphere preparation impossible. When the polylysine concentration reached 4.0 mg / mL, the ethanol adsorption capacity of the microspheres no longer increased, which was also due to the stagnation of microsphere strength. The main reason for this was that the cross-linking reaction between polylysine and sodium alginate reached saturation. Furthermore, as the degree of cross-linking increased, the molecular chains became overly crowded, easily agglomerating and causing network defects, resulting in a decrease in storage modulus instead of an increase. Considering the hydrogel storage modulus data, the amount of ethanol adsorbed by the hydrogel microspheres is related to their strength; higher hydrogel strength results in greater ethanol adsorption.

[0032] Example 3: Ethanol Release Experiment of Hydrogel Microspheres After the hydrogel microspheres reached adsorption equilibrium, they were removed, and the residual ethanol solution on the surface was blotted dry with filter paper. The mass was quickly weighed and recorded. The total mass of the gel after adsorption was recorded. The saturated ethanol hydrogel was directly placed into a stoppered conical flask containing 50 mL of pH 7.4 PBS buffer, ensuring that the gel was completely submerged, and the stopper was tightened. The flask was placed in a constant temperature water bath shaker (temperature 37℃, shaking frequency 100 r / min), and the timing was started (t=0). Samples were taken at 10 min, 30 min, 1 h, 2 h, and 4 h after release. Before each sampling, the conical flask was gently inverted 3 times to ensure uniform medium concentration. 1 mL of the release medium was pipetted as a sample. Three parallel samples were set up for each experimental condition. The supernatant was collected and 10 mL of distilled water was added. After appropriate dilution, the ethanol content in the liquid was determined by gas chromatography. Detector: FID, column: DB-FFAP (30m×0.25mm×0.25μm), column temperature: 45℃, injection volume: 1μl, mode: splitless. Repeat the above experiment 3 times and take the average value.

[0033] Ethanol release is calculated using the following formula: Single release amount Q t (g / g) = (C)t -C t-1 V t-1 / m Cumulative release Q 累积 (g / g) = C t : Ethanol concentration in the solution at time t, g / mL; C t-1 : Ethanol concentration in the solution at time t-1, g / mL; Vt: Solution volume at time point t; m: Mass of the saturated ethanol-adsorbed hydrogel. The experimental results show that the hydrogel microspheres exhibit a sustained-release effect on ethanol adsorption, and the release rate is closely related to the sodium alginate concentration. Higher sodium alginate concentrations result in slower ethanol release and a smaller amount released. At sodium alginate concentrations of 30 mg / mL and 40 mg / mL, the hydrogel release percentages after 4 hours were 44.4% and 37.3%, respectively; while at a concentration of 10 mg / mL, the hydrogel microspheres achieved a release percentage as high as 92.0% after 4 hours, and 76.0% within 30 minutes. The ethanol release rate of the hydrogel microspheres is also closely related to the concentration of the cross-linking agent polylysine; higher polylysine concentrations result in slower ethanol release and a smaller amount released. When the sodium alginate concentration was 30 mg / mL, and the polylysine concentrations were 3.0 mg / mL and 4.0 mg / mL, the hydrogel release percentages after 4 hours were 44.4% and 38.0%, respectively. However, when the polylysine concentration was 10 mg / mL, the hydrogel microspheres released as much as 57.1% after 4 hours, and reached 34.3% within 30 minutes. These experimental results are consistent with the ethanol adsorption results, demonstrating that the hydrogel has ethanol adsorption and sustained-release properties, and that polylysine cross-linking can enhance both ethanol adsorption and sustained-release capabilities, with this capability being positively correlated with the hydrogel strength. In conclusion, the aforementioned properties of the hydrogel have significant application value for developing hangover relief products.

[0034] Example 4: Animal Experiment Study of Compound Alcohol-Detoxifying Microspheres 1. Animal experimental methods (1) Laboratory animals and grouping One hundred male C57BL / 6 mice, weighing 15-30 g, were randomly divided into 10 groups after 7 days of acclimatization: blank group (n=10), model group (n=10), positive control group (commercially available hangover remedy, n=10), Example 1 group (n=10), Example 2 group (n=10), Example 3 group (n=10), Example 4 group (n=10), Example 5 group (n=10), Example 6 group (n=10), and Example 7 group (n=10).

[0035] (2) Modeling and drug administration 1) Modeling method: Fasting for 12 hours before the experiment, followed by oral administration of 42° Xinghuacun wine at a dose of 0.15mL / 10g body weight.

[0036] 2) Administration time: 30 minutes before modeling, administer once by gavage.

[0037] 3) Dosage: The model group was administered 0.25 mL / 10g of physiological saline by gavage; the positive control group was administered 0.25 mL / 10g of commercially available hangover remedy solution with a concentration of 40 mg / mL by gavage; and the experimental group was fed 0.25 g / 10g of hangover remedy microspheres.

[0038] (3) Observation 1) Observe and record the activity of mice: observe after modeling and administration.

[0039] Intoxication index: The disappearance of the righting reflex is indicated by the mouse crawling unsteadily, dragging its hindquarters on the ground, having ruffled fur, closing its eyes and remaining still, and keeping its back down for 30 seconds. Indicators of recovery from hangover: The ability to move freely, be flexible, be energetic, have smooth hair, and be able to right themselves within 30 seconds is considered a recovery of the righting reflex.

[0040] 2) Observe the time of intoxication and sobering up in mice. Intoxication latency time: the time from gavage to the disappearance of the righting reflex, taken as the average of the intoxication time of 10 mice; Sobering time: the time from the disappearance to the recovery of the righting reflex, taken as the average of the intoxication time of 10 mice.

[0041] 3) After the experiment was completed (12 hours after drinking), blood was collected from the eye sockets to measure the ethanol concentration in the blood.

[0042] ① Measure 0.1 mL of serum sample into a 2 mL PE tube, add 0.1 mL of 10% trichloroacetic acid solution, and shake to mix. ② Centrifuge at 4℃ for 10 min, with a centrifugal force of 15870×g; ③ Filter the supernatant through a 0.45 μm filter membrane and prepare for testing; ④ Gas chromatography for ethanol concentration detection Detector: FID; Column: DB-FFAP (30m × 0.25mm × 0.25μm); Column temperature: 45℃; Injection volume: 1 μl; Mode: No splitting.

[0043] 4) 24 hours after drug administration, the liver of mice was collected to measure the malondialdehyde (MDA) content. (The MDA content represents the concentration of lipid oxides in the liver and also represents the degree of liver damage; the higher the content, the greater the degree of liver damage.) Experimental methods: ① Rinse the liver tissue with pre-cooled PBS (0.01 M, pH 7.4) and remove any residual solution; ② Mix the shredded tissue with the corresponding volume of PBS (tissue sample:PBS = 1:9). The specific volume can be adjusted according to experimental needs. Record the weight of the tissue. ③ Add to a glass homogenizer and grind thoroughly on ice; ④ Centrifuge the homogenate at 5000 g for 5-10 min and collect the supernatant for later use; ⑤ The concentration of malondialdehyde (MDA) was determined by ELISA using a malondialdehyde (MDA) detection kit.

[0044] 2. The experimental results are shown in the table below. As shown in the table above, the hangover-relieving microspheres produced in this invention have a good hangover-relieving effect, and this effect is closely related to the sodium alginate concentration, with the hangover-relieving performance increasing with increasing sodium alginate concentration. When the sodium alginate concentration is 30 mg / mL and 40 mg / mL, the latency period of intoxication is significantly prolonged compared to the model group, by 73.3% and 74.2%, respectively. This indicates that feeding the hangover-relieving microspheres can prolong the time it takes for mice to enter a state of intoxication, meaning that the combined formula has the effect of delaying ethanol absorption. The blood ethanol concentration also proves that the hangover-relieving microspheres can prevent ethanol absorption, thereby reducing the blood ethanol concentration. The sobering-up time is significantly shortened compared to the model group, by 53.1% and 53.7%, respectively, indicating that the amount of ethanol absorbed in the body is less, thus shortening the sobering-up time. The data on malondialdehyde (MPA) in the liver shows that due to the reduced amount of ethanol absorption, the amount of free radicals and lipid peroxides generated in the liver is reduced, which can alleviate the damaging effects of ethanol on the liver. The hangover-relieving effect of the microspheres was also closely related to the concentration of polylysine in the cross-linking solution, and the hangover-relieving performance increased with increasing polylysine concentration. Under the condition of sodium alginate concentration of 30 mg / mL, the hangover latency period was significantly prolonged compared to the model group at polylysine concentrations of 3.0 mg / mL and 4.0 mg / mL, respectively, by 73.3% and 73.9%. The sobering-up time was also significantly shortened compared to the model group, by 53.1% and 54.3%, respectively, indicating that the amount of ethanol absorbed by the body was less, thus shortening the sobering-up time. The levels of ethanol in the blood and malondialdehyde (MPA) in the liver were also significantly lower than in the model group, indicating that the denser the hydrogel microsphere structure, the better the hangover-relieving performance. In summary, the preferred microsphere preparation conditions are a sodium alginate concentration of 30 mg / mL and a polylysine cross-linking reaction amount of 29.42 mg / mL microspheres, under which the microspheres exhibit the best hangover-relieving effect. The experimental data showed that the alcohol-relieving microspheres fed to the experimental group had a much higher alcohol-relieving performance than a commercially available alcohol-relieving product (positive control group). Comprehensive analysis suggests that the composite alcohol-relieving microspheres have market application value and can be used to develop alcohol-relieving functional foods.

[0045] Comparative Example 1: The hangover-relieving effect of calcium alginate microspheres 1. Preparation method of calcium alginate microspheres A certain amount of sodium alginate was weighed and added to 10 mL of deionized water. The mixture was stirred at room temperature for 60 min (200 rpm) until completely dissolved. 100 mg of CaCO3 and 10 mg of curcumin were added to obtain a sodium alginate suspension. 100 mL of liquid paraffin was added to a beaker, along with 1.0 g of Span 80. The mixture was stirred at room temperature for 30 min (300 rpm) to form an oil phase. The sodium alginate suspension was then added to the liquid paraffin oil phase and stirred at 1000 rpm for 30 min to form a stable W / O emulsion. 0.3 mL of glacial acetic acid was added dropwise to the emulsion at a rate of 0.5 mL / min, with continuous stirring (1000 rpm). The cross-linking reaction was carried out for 2 h to completely calcify the sodium alginate into calcium alginate hydrogel microspheres. After the reaction, the microspheres were centrifuged and separated using a solution containing 1.0% (w / v) Tween. The microspheres were washed three times with 80°C deionized water to remove the liquid paraffin on the surface of the microspheres, yielding calcium alginate hydrogel microspheres.

[0046] 2. Animal experimental methods (1) Laboratory animals and grouping Twenty male C57BL / 6 mice, weighing 15-30 g, were acclimatized for 7 days and then divided into groups for an alcohol detoxification experiment.

[0047] (2) Modeling and drug administration 1) Modeling method: Fasting for 12 hours before the experiment, followed by oral administration of 42° Xinghuacun wine at a dose of 0.15mL / 10g body weight.

[0048] 2) Administration time: 30 minutes before modeling, administer once by gavage.

[0049] 3) Dosage: The experimental group was fed 0.25g / 10g of calcium alginate microspheres.

[0050] (3) Observation 1) Observe and record the activity of mice: observe after modeling and administration.

[0051] Intoxication index: The disappearance of the righting reflex is indicated by the mouse crawling unsteadily, dragging its hindquarters on the ground, having ruffled fur, closing its eyes and remaining still, and keeping its back down for 30 seconds. Indicators of recovery from hangover: The ability to move freely, be flexible, be energetic, have smooth hair, and be able to right themselves within 30 seconds is considered a recovery of the righting reflex.

[0052] 2) Observe the time of intoxication and sobering up in mice. Intoxication latency time: the time from gavage to the disappearance of the righting reflex, taken as the average of the intoxication time of 10 mice; Sobering time: the time from the disappearance to the recovery of the righting reflex, taken as the average of the intoxication time of 10 mice.

[0053] 3) After the experiment was completed (12 hours after drinking), blood was collected from the eye sockets to measure the ethanol concentration in the blood.

[0054] ① Measure 0.1 mL of serum sample into a 2 mL PE tube, add 0.1 mL of 10% trichloroacetic acid solution, and shake to mix. ② Centrifuge at 4℃ for 10 min, with a centrifugal force of 15870×g; ③ Filter the supernatant through a 0.45 μm filter membrane and prepare for testing; ④ Gas chromatography for ethanol concentration detection Detector: FID; Column: DB-FFAP (30m × 0.25mm × 0.25μm); Column temperature: 45℃; Injection volume: 1 μl; Mode: No splitting.

[0055] 4) 24 hours after drug administration, the liver of mice was collected to measure the malondialdehyde (MDA) content. (The MDA content represents the concentration of lipid oxides in the liver and also represents the degree of liver damage; the higher the content, the greater the degree of liver damage.) Experimental methods: ① Rinse the liver tissue with pre-cooled PBS (0.01 M, pH 7.4) and remove any residual solution; ② Mix the shredded tissue with the corresponding volume of PBS (tissue sample:PBS = 1:9). The specific volume can be adjusted according to experimental needs. Record the weight of the tissue. ③ Add to a glass homogenizer and grind thoroughly on ice; ④ Centrifuge the homogenate at 5000 g for 5-10 min and collect the supernatant for later use; ⑤ The concentration of malondialdehyde (MDA) was determined by ELISA using a malondialdehyde (MDA) detection kit.

[0056] 3. The experimental results are shown in the table below. The above experimental results show that, since Comparative Example 1 did not undergo electrostatic cross-linking with polylysine, its alcohol-relieving performance was significantly different from that of Examples 5, 6, 3, and 7. The latency period of intoxication was significantly prolonged, the intoxication time was significantly shortened, and the ethanol content in the blood was also lower than that of the Examples. This indicates that the polylysine cross-linking reaction can improve the alcohol-relieving performance of the microspheres. It also proves that the density and strength of the hydrogel microsphere structure have a significant impact on the alcohol-relieving performance of the microspheres. Increased hydrogel strength enhances the alcohol-relieving performance of the microspheres.

[0057] Comparative Example 2: The hangover-relieving effects of hangover jelly and hangover oral liquid 1. Method for preparing hangover jelly Weigh 300 mg of sodium alginate and add it to 10 mL of deionized water. Stir at room temperature for 60 min (200 rpm) until completely dissolved. Add 100 mg of CaCO3 and 10 mg of curcumin to obtain a sodium alginate suspension. After sterilizing the sodium alginate suspension at 100 °C, add 0.3 mL of glacial acetic acid and quickly fill into bottles to obtain a hangover jelly.

[0058] 2. Preparation method of hangover relief oral liquid Weigh 300 mg of sodium alginate and add it to 10 mL of deionized water. Stir at room temperature for 60 min (200 rpm) until completely dissolved. Add 10 mg of curcumin to obtain a sodium alginate suspension. After sterilizing the sodium alginate suspension at 100℃, fill it into soft bags to obtain an oral solution for relieving hangovers.

[0059] 2. Animal experimental methods (1) Laboratory animals and grouping Ten male C57BL / 6 mice, weighing 15-30 g, were randomly divided into two groups after 7 days of acclimatization: a hangover jelly group (n=10) and a hangover oral liquid group (n=10).

[0060] (2) Modeling and drug administration 1) Modeling method: Fasting for 12 hours before the experiment, followed by oral administration of 42° Xinghuacun wine at a dose of 0.15mL / 10g body weight.

[0061] 2) Administration time: 30 minutes before modeling, administer once by gavage.

[0062] 3) Dosage: The hangover jelly group was fed 0.25g / 10g of calcium alginate hydrogel (CaCl2 solution was added to sodium alginate solution and left to stand for 2 hours to form a hydrogel), and the hangover oral liquid group was given 0.25mL of 30mg / mL sodium alginate solution by gavage.

[0063] (3) Observation 1) Observe and record the activity of mice: observe after modeling and administration.

[0064] Intoxication index: The disappearance of the righting reflex is indicated by the mouse crawling unsteadily, dragging its hindquarters on the ground, having ruffled fur, closing its eyes and remaining still, and keeping its back down for 30 seconds. Indicators of recovery from hangover: The ability to move freely, be flexible, be energetic, have smooth hair, and be able to right themselves within 30 seconds is considered a recovery of the righting reflex.

[0065] 2) Observe the time of intoxication and sobering up in mice. Intoxication latency time: the time from gavage to the disappearance of the righting reflex, taken as the average of the intoxication time of 10 mice; Sobering time: the time from the disappearance to the recovery of the righting reflex, taken as the average of the intoxication time of 10 mice.

[0066] 3) After the experiment was completed (12 hours after drinking), blood was collected from the eye sockets to measure the ethanol concentration in the blood.

[0067] ① Measure 0.1 mL of serum sample into a 2 mL PE tube, add 0.1 mL of 10% trichloroacetic acid solution, and shake to mix. ② Centrifuge at 4℃ for 10 min, with a centrifugal force of 15870×g; ③ Filter the supernatant through a 0.45 μm filter membrane and prepare for testing; ④ Gas chromatography for ethanol concentration detection Detector: FID; Column: DB-FFAP (30m × 0.25mm × 0.25μm); Column temperature: 45℃; Injection volume: 1 μl; Mode: No splitting.

[0068] 4) 24 hours after drug administration, the liver of mice was collected to measure the malondialdehyde (MDA) content. (The MDA content represents the concentration of lipid oxides in the liver and also represents the degree of liver damage; the higher the content, the greater the degree of liver damage.) Experimental methods: ① Rinse the liver tissue with pre-cooled PBS (0.01 M, pH 7.4) and remove any residual solution; ② Mix the shredded tissue with the corresponding volume of PBS (tissue sample:PBS = 1:9). The specific volume can be adjusted according to experimental needs. Record the weight of the tissue. ③ Add to a glass homogenizer and grind thoroughly on ice; ④ Centrifuge the homogenate at 5000 g for 5-10 min and collect the supernatant for later use; ⑤ The concentration of malondialdehyde (MDA) was determined by ELISA using a malondialdehyde (MDA) detection kit.

[0069] 3. The experimental results are shown in the table below. The above experimental results show that the jelly group and oral liquid group, due to the lack of electrostatic cross-linking reaction with polylysine, exhibit significantly different alcohol-relieving performance compared to the polylysine-crosslinked microsphere group. The latency period for intoxication was significantly prolonged, the duration of intoxication was significantly shortened, and the blood ethanol content was lower than in the previous example. This indicates that the polylysine cross-linking reaction can improve the alcohol-relieving performance of the microspheres, and also demonstrates that the density and strength of the hydrogel microsphere structure have a significant impact on their alcohol-relieving performance; increased hydrogel strength enhances the alcohol-relieving performance of the microspheres. Furthermore, compared to the calcium alginate microsphere group (Comparative Example 1), these two groups have a shorter latency period for intoxication, a longer duration of intoxication, and a higher blood ethanol concentration. The reason for this is that the microspheres are uniformly dispersed in gastric juice, allowing ethanol to fully contact and be rapidly absorbed after entering the stomach. In contrast, the jelly and oral liquid are unevenly distributed in the stomach; ethanol not in contact with the hydrogel cannot be rapidly absorbed, while ethanol enters the bloodstream quickly. Therefore, compared to the microspheres, the ethanol absorption is greater, resulting in a weaker alcohol-relieving effect. Compared to oral hangover remedies, hangover jelly is more effective at relieving hangovers. This is mainly because oral hangover remedies, upon entering the stomach, form hydrogels under the influence of gastric acid. Due to the rapid gelation process, these hydrogels only form large, in-situ blocks, resulting in a less uniform distribution within the stomach compared to jelly, thus leading to a weaker hangover-relieving effect. The experimental results show that hydrogel microspheres have a better hangover-relieving effect. Furthermore, due to their good fluidity, hydrogel microspheres can be added as a core ingredient to the formulation of hangover relief products, providing a new raw material option for the research and development of hangover relief products.

[0070] Comparative Example 3: The hangover-relieving effect of curcumin 1. Animal experimental methods (1) Laboratory animals and grouping Twenty male C57BL / 6 mice, weighing 15–30 g, were randomly divided into two groups after 7 days of acclimatization: a curcumin group (n=10) and a control group (n=10).

[0071] (2) Modeling and drug administration 1) Modeling method: Fasting for 12 hours before the experiment, followed by oral administration of 42° Xinghuacun wine at a dose of 0.15mL / 10g body weight.

[0072] 2) Administration time: 30 minutes before modeling, administer once by gavage.

[0073] 3) Dosage: The curcumin group was orally administered 0.25 mL / 10 g of curcumin suspension, with a concentration of 1.0 mg / mL; the control group was fed 0.25 g / 10 g of hangover-relieving microspheres (curcumin content of 0.1 mg / mL).

[0074] (3) Observation 1) Observe and record the activity of mice: observe after modeling and administration.

[0075] Intoxication index: The disappearance of the righting reflex is indicated by the mouse crawling unsteadily, dragging its hindquarters on the ground, having ruffled fur, closing its eyes and remaining still, and keeping its back down for 30 seconds. Indicators of recovery from hangover: The ability to move freely, be flexible, be energetic, have smooth hair, and be able to right themselves within 30 seconds is considered a recovery of the righting reflex.

[0076] 2) Observe the time of intoxication and sobering up in mice. Intoxication latency time: the time from gavage to the disappearance of the righting reflex, taken as the average of the intoxication time of 10 mice; Sobering time: the time from the disappearance to the recovery of the righting reflex, taken as the average of the intoxication time of 10 mice.

[0077] 3) After the experiment was completed (12 hours after drinking), blood was collected from the eye sockets to measure the ethanol concentration in the blood.

[0078] ① Measure 0.1 mL of serum sample into a 2 mL PE tube, add 0.1 mL of 10% trichloroacetic acid solution, and shake to mix. ② Centrifuge at 4℃ for 10 min, with a centrifugal force of 15870×g; ③ Filter the supernatant through a 0.45 μm filter membrane and prepare for testing; ④ Gas chromatography for ethanol concentration detection Detector: FID; Column: DB-FFAP (30m × 0.25mm × 0.25μm); Column temperature: 45℃; Injection volume: 1 μl; Mode: No splitting.

[0079] 4) 24 hours after drug administration, the liver of mice was collected to measure the malondialdehyde (MDA) content. (The MDA content represents the concentration of lipid oxides in the liver and also represents the degree of liver damage; the higher the content, the greater the degree of liver damage.) Experimental methods: ① Rinse the liver tissue with pre-cooled PBS (0.01 M, pH 7.4) and remove any residual solution; ② Mix the shredded tissue with the corresponding volume of PBS (tissue sample:PBS = 1:9). The specific volume can be adjusted according to experimental needs. Record the weight of the tissue. ③ Add to a glass homogenizer and grind thoroughly on ice; ④ Centrifuge the homogenate at 5000 g for 5-10 min and collect the supernatant for later use; ⑤ The concentration of malondialdehyde (MDA) was determined by ELISA using a malondialdehyde (MDA) detection kit.

[0080] 2. The experimental results are shown in the table below. The above experimental results show that curcumin alone has no effect on relieving hangovers. There were no significant differences in the latency period of intoxication, duration of intoxication, blood ethanol concentration, and malondialdehyde (MDA) content in the liver compared to the model group. This is because curcumin is insoluble in water and cannot be absorbed into the bloodstream after ingestion, resulting in low bioavailability. The curcumin content in Comparative Example 3 was lower than that in Example 3. The experimental results show no significant differences in the latency period of intoxication, duration of intoxication, and blood ethanol concentration compared to Example 3, indicating that curcumin has no obvious effect on relieving or preventing hangovers. However, the results regarding MDA content in the liver show that improving the bioavailability of curcumin can effectively reduce MDA content in the liver, indicating that curcumin has a good hepatoprotective effect.

[0081] The embodiments described above are merely preferred embodiments of the present invention, and not all feasible embodiments of the present invention. Any obvious modifications made by those skilled in the art without departing from the principles and spirit of the present invention should be considered to be included within the scope of protection of the claims of the present invention.

Claims

1. A sodium alginate-polylysine composite hydrogel hangover-relieving microsphere, characterized in that, Using sodium alginate and polylysine as raw materials, polyelectrolyte hydrogel microspheres are constructed through an interpenetrating network. These hydrogel microspheres have the function of adsorbing ethanol, which can prevent ethanol from being absorbed into the blood. They can also act as drug carriers to encapsulate poorly water-soluble hangover-relieving components such as curcumin. By using the ethanol adsorbed in the hydrogel, the drug absorption rate is improved, thereby increasing bioavailability and enhancing the hangover-relieving and liver-protecting effects.

2. The sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 1, characterized in that, The sodium alginate has a molecular weight of 70.0~200.0 kDa, and the mass ratio of guluronic acid to mannuluronic acid is 1.0~3.0; the polylysine has a molecular weight of 5.0~30.0 kDa.

3. The sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 1, characterized in that, The composite hydrogel hangover-relieving microspheres have a particle size of 300~2000μm.

4. A method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres, characterized in that, Includes the following steps: S1 Add sodium alginate, curcumin and CaCO3 powder to deionized water and stir at room temperature to completely dissolve them to obtain sodium alginate suspension. S2. Add the emulsifier to the liquid paraffin and stir at room temperature for 10-30 minutes to form the oil phase. S3: The sodium alginate suspension obtained in S1 is added to the oil phase and stirred at high speed for 10-30 min to form a W / O emulsion. Glacial acetic acid is then added dropwise to dissolve CaCO3 and release Ca2+. 2+ Ions form calcium alginate hydrogel microspheres. The microspheres are collected by centrifugation and washed 2-4 times with a 1.0% (w / v) Tween 80 solution to remove residual liquid paraffin. S4. The calcium alginate microspheres obtained in S3 were added to a polylysine solution to carry out a cross-linking reaction. Sodium alginate and polylysine formed a polyelectrolyte structure through interpenetrating network interaction. After the reaction was completed, the microspheres were separated by centrifugation to obtain composite hydrogel hangover relief microspheres.

5. The method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 4, characterized in that, In step S1, the concentration of sodium alginate solution is 10.0~40.0 mg / mL, the content of curcumin is 0.1~1.0 mg / mL, and the content of CaCO3 is 1.0~10.0 mg / mL.

6. The method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 4, characterized in that, In step S2, the dispersion medium is liquid paraffin, or one or more mixtures of edible oils such as soybean oil or olive oil; the emulsifier is one or more combinations of dehydrated sorbitan fatty acid ester surfactants, and the emulsifier concentration is 0.5~5.0 wt% (w / v); the volume ratio of the aqueous phase to the oil phase is 1:5~20.

7. The method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 4, characterized in that, In step S3, the mass ratio of glacial acetic acid to CaCO3 is 2~4:1, the glacial acetic acid dropping rate is 0.5~2.0 mL / min, and the calcification reaction time is 1~4 h.

8. The method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 4, characterized in that, In step S2, the stirring speed is 300~1500 rpm; in step S3, the stirring speed is maintained at 1000~3000 rpm.

9. The method for preparing sodium alginate-polylysine composite hydrogel hangover-relieving microspheres according to claim 4, characterized in that, In step S4, the concentration of polylysine solution is 1.0~4.0 mg / mL, and the volume ratio of microspheres to polylysine is 1:5~20.

10. The use of sodium alginate-polylysine composite hydrogel hangover relief microspheres according to any one of claims 1-3 in the preparation of hangover relief beverages, jellies, gummies or medicines.

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