Method for evaluating hangover effect of baijiu after drinking

Abstract

The application discloses a method for evaluating the hangover effect of baijiu after drinking, and belongs to the technical field of detection. The method detects multiple neurobiochemical indexes of serum and brain tissue of mice after the mice are given the baijiu by gavage, and monitors the behavior parameters such as pain threshold of the mice, and compares the indexes with the indexes of mice given normal saline and edible alcohol by gavage, so as to evaluate the hangover effect of the baijiu. The method can replace the human test to evaluate the hangover effect of the baijiu, reduces the damage to the human body, and the evaluation result is more objective, and provides a basis for further studying key head-up substances in alcoholic beverages and the action mechanism.

Description

Technical Field

[0001] This invention relates to a method for evaluating the hangover effect after drinking baijiu (Chinese liquor), belonging to the field of detection technology. Background Technology

[0002] Alcohol hangover refers to a series of physiological and psychological discomforts that occur after excessive alcohol consumption. These symptoms usually appear several hours after drinking, especially when blood alcohol content is close to zero. Common symptoms include headache, dizziness, fatigue, nausea, loss of appetite, and impaired concentration. Baijiu, as China's national liquor, holds a unique position in traditional Chinese culture and is widely loved by consumers worldwide, forming a major part of China's food and beverage industry. However, excessive consumption of baijiu can lead to severe hangover symptoms such as headache, nausea, and dizziness, causing consumer concerns about its consumption. With the continuous improvement of living standards and increasing emphasis on health among Chinese people, there are higher demands for the quality and health benefits of baijiu. To meet the public's growing pursuit of drinking comfort, it is necessary to explore an effective method for assessing the hangover effects of baijiu consumption.

[0003] Currently, the evaluation of the hangover effect of alcoholic beverages mainly relies on human trials. For example, patent CN 102218096A discloses a hangover prevention composition and a hangover relief beverage containing the composition. Human trials have shown that 70-85% of participants reported improvement in hangover symptoms. However, the questionnaire-based survey is highly subjective, with significant individual differences, and excessive alcohol consumption is harmful to human health. Therefore, it is necessary to explore the use of animal models for hangover effect evaluation. Patent CN 109568538A discloses a hangover relief composition containing milk thistle, turmeric, licorice, and cassia seed. The composition's hangover-relieving effect is assessed by administering 5 mL / kg body weight of ethanol to mice via gavage and monitoring the blood ethanol-acetaldehyde metabolism status of the mice after gavage. However, this method primarily evaluates ethanol and cannot reflect the differences in hangover effects among alcoholic beverages with different compositions; furthermore, it does not evaluate other hangover indicators in mice, such as exercise capacity. Decreased motor function and anxiety-like behaviors have been observed in laboratory animals during hangovers, but it remains unknown which of these phenomena and behaviors correspond to symptoms experienced by humans after drinking alcohol. Furthermore, there are currently no suitable indicators or evaluation systems to predict human comfort levels after drinking alcohol based on hangover symptoms observed in animal experiments.

[0004] Therefore, existing technologies lack models that comprehensively evaluate the hangover effect from the perspectives of headache, dizziness, loss of appetite, and depression, making it impossible to predict the post-drinking comfort of people who consume finished wine samples. Summary of the Invention

[0005] To address the current lack of assessments of the hangover effects of alcoholic beverages, which primarily rely on human trials and have limited reports on animal model assessments that focus only on ethanol rather than alcohol samples, this invention provides a method for evaluating the hangover effects of strong-aroma baijiu (Chinese liquor) using a mouse model. The specific technical solution is as follows:

[0006] The first objective of this invention is to provide a method for evaluating the hangover effect of strong-aroma baijiu (Chinese liquor). The method uses mice as a model, administers alcohol samples to mice via gavage, and monitors the behavioral parameters of the mice after gavage, including sucrose preference, foot pain threshold, gait symmetry, and ataxia coefficient. Blood and brain tissue are collected from the mice, and blood histamine, endothelin, brain serotonin, and dopamine are detected. The above indicators are used to characterize the degree of hangover after drinking alcoholic beverages.

[0007] In one embodiment of the present invention, the method includes the following steps:

[0008] 1) Set up an experimental group, an alcohol control group, and a blank control group. The experimental group was given a sample of alcohol by gavage, the alcohol control group was given edible alcohol with the same ethanol dose by gavage, and the blank group was given physiological saline by gavage.

[0009] 2) Within 6-14 hours after gavage, the biochemical and behavioral indicators of the experimental subjects were tested.

[0010] In one embodiment of the invention, the alcohol sample includes spirits, whiskey, or brandy.

[0011] In one embodiment of the present invention, the selected liquor sample is a strong-aroma liquor with an alcohol content of 45%.

[0012] In one embodiment of the present invention, the experimental subjects include, but are not limited to, mice.

[0013] In one embodiment of the present invention, the selected mice are healthy male Balb / c mice, aged 6-8 weeks.

[0014] In one embodiment of the present invention, the oral gavage dose is 4.3 g / kg body weight based on the ethanol equivalent.

[0015] In one embodiment of the present invention, 10 hours after the mice were administered the sample by gavage, the plantar pain threshold of the mice was tested using a von Frey automated pain meter.

[0016] In one embodiment of the present invention, 10 hours after the mice were gavaged with the sample, the mice's sucrose preference was tested using a sucrose preference device.

[0017] In one embodiment of the present invention, 10 hours after mice were administered samples by gavage, the CatWalk XT animal gait acquisition and analysis system was used to monitor and analyze parameters such as gait symmetry and ataxia coefficient of the mice.

[0018] In one embodiment of the present invention, after the behavioral parameters were tested, the mice were euthanized, and blood and brain tissue were collected. After the blood samples were allowed to stand and centrifuged, the supernatant was taken as a serum sample and stored at -80°C. The brain tissue was flash-frozen in liquid nitrogen and stored at -80°C. Tissue homogenate was prepared before testing.

[0019] In one embodiment of the present invention, an enzyme-linked immunosorbent assay (ELISA) kit was used to test the levels of histamine, endothelin, and brain 5-hydroxytryptamine and dopamine in mouse serum.

[0020] In one embodiment of the present invention, when the test sample is baijiu (Chinese white liquor), if the serum endothelin content of mice in the baijiu sample group is 5% higher than that in the edible alcohol group, it is determined that the baijiu sample may cause more severe headache symptoms and poor comfort after drinking; if the serum endothelin content is 5% lower than that in the edible alcohol group, it is determined that the comfort after drinking baijiu is good, and the comfort after drinking is moderate if it is between the two.

[0021] In one embodiment of the present invention, when the sample is baijiu (Chinese white liquor), if the 5-hydroxytryptamine content in the brain of mice in the baijiu sample group is 8% lower than that in the edible alcohol group, it is determined that the baijiu sample may cause depressive symptoms and has poor post-drinking comfort; if it is 1% higher than that in the edible alcohol group, it is determined that the post-drinking comfort of baijiu is good, and if it is in between, it is considered that the post-drinking comfort is moderate; in addition, if the dopamine content in the brain tissue of mice in the baijiu group is 16% lower than that in the edible alcohol group, it can also be determined that the baijiu sample will not cause severe depressive symptoms and has good post-drinking comfort.

[0022] In one embodiment of the present invention, when the test sample is baijiu (Chinese white liquor), if the serum histamine content of mice in the baijiu sample group is 17% lower than that in the control group, it is determined that the baijiu sample will cause more severe anxiety symptoms; if the footprint symmetry value of mice in the experimental group is 50% higher than that in the control group, it is determined that the baijiu sample will cause more severe dizziness symptoms; if the sucrose preference value of mice in the experimental group is 37% lower than that in the blank group, it is determined that the baijiu sample can cause severe loss of appetite, apathy, and regret symptoms; if the stride length value of mice in the experimental group is 40% lower than that in the control group, it is determined that the baijiu sample will cause more severe fatigue symptoms and poor comfort after drinking.

[0023] In one embodiment of the present invention, when the test sample is baijiu (Chinese white liquor), if the foot pain threshold of mice in the baijiu sample group is 34% lower than that in the edible alcohol group, it is determined that the baijiu sample will not cause a serious decrease in alertness; or if the sucrose preference value is 24% higher than that in the edible alcohol group, it is determined that the baijiu sample will not cause a serious decrease in appetite, apathy, or regret symptoms; or if the paw angle change rate value is 29% lower than that in the edible alcohol group, it is determined that the baijiu sample will not cause a serious clumsy symptom, and it can be determined that the baijiu sample provides good comfort after drinking.

[0024] Beneficial effects:

[0025] This invention provides a method for evaluating the hangover effect after alcohol consumption. It analyzes multiple neurobiochemical indicators in serum and brain tissue samples from mice after oral administration of alcohol, and monitors several behavioral parameters of the mice. The results show that: serum histamine levels in mice after alcohol consumption can evaluate the degree of anxiety in humans after drinking; serum endothelin levels in mice after alcohol consumption can evaluate the degree of headache in humans after drinking; brain 5-HT and dopamine levels in mice after alcohol consumption can evaluate the degree of depression in humans after drinking; foot pain threshold in mice after alcohol consumption can evaluate the degree of decreased alertness in humans after drinking; sucrose preference in mice after alcohol consumption can evaluate the degree of decreased appetite, regret, and apathy in humans after drinking; footprint symmetry and ataxia coefficient in mice after alcohol consumption can evaluate the degree of dizziness in humans after drinking; stride length in mice after alcohol consumption can evaluate the degree of fatigue in humans after drinking; and the rate of change in paw angle in mice after alcohol consumption can evaluate the degree of clumsiness in humans after drinking.

[0026] The method for comprehensively evaluating the hangover effect of alcoholic beverages based on animal models provided by this invention can provide a basis for the identification of key hangover substances in alcoholic beverages. Attached Figure Description

[0027] Figure 1 Serum histamine and endothelin levels 10 hours after each group of mice were administered the corresponding samples via gavage;

[0028] Figure 2 The levels of 5-HT and dopamine in brain tissue of mice in each group were determined 10 hours after gavage administration of the corresponding samples.

[0029] Figure 3 The results of foot pain threshold detection were obtained for mice in each group 10 hours after gavage administration of the corresponding samples.

[0030] Figure 4 Results of sucrose preference test 10 h after gavage administration of corresponding samples to mice in each group;

[0031] Figure 5 Gait monitoring results of mice in each group 10 hours after gavage administration of the corresponding samples;

[0032] Figure 6 Correlation analysis of mouse parameters after oral administration of alcohol samples with human hangover symptom scores;

[0033] Figure 7 Correlation analysis was performed on serum prostaglandin E2, methionine enkephalin, serum 5-HT, brain histamine levels, periorbital pain threshold, and headache symptom scores of mice in each group at 10 h. Detailed Implementation

[0034] The present invention will be further described below with reference to embodiments, but the implementation of the present invention is not limited thereto.

[0035] To better illustrate the present invention, a population assessment experiment was conducted on the hangover effect of the alcohol samples used in the following embodiments to verify the method described in the present invention. This experiment is not required during application.

[0036] The two baijiu samples selected for this invention are both strong-aroma baijiu produced in Luzhou, Sichuan Province, with an alcohol content of 45% vol. The main brewing raw materials are sorghum, rice and wheat.

[0037] In this invention, the detection time for hangover symptoms in mice was 10 hours after gavage, at which point the ethanol content in the mouse serum was 0 g / L. The changes in ethanol content in mouse serum within 10 hours after gavage are shown in Table 1. Ethanol content was obtained using a blood ethanol detection kit (Nanjing Jiancheng Bioengineering Institute).

[0038] Table 1. Ethanol content in the blood of mice given different alcohol samples at different times after gavage.

[0039]

[0040] Example 1: Evaluation of the hangover effect of commercially available baijiu sample A

[0041] 1) Conduct a sensory evaluation of a commercially available strong-aroma baijiu (45% alcohol content) among a population.

[0042] The participants were primarily healthy adults, and the specific selection criteria were as follows:

[0043] (1) No serious alcohol abuse problem (score <5 on the Michigan Short-Term Alcohol Screening Test (SMAST)) and no history of alcohol therapy or counseling; the SMAST test was conducted according to the content reported by Selzer et al. in "A self-administered Short Michigan Alcoholism Screening Test (SMAST)".

[0044] (2) No health problems or current medications that do not contain alcohol;

[0045] (3) Fluent in expression;

[0046] (4) If the subject is female, a negative pregnancy test is required;

[0047] (5) No alcohol, caffeine and non-prescription drugs were consumed within 24 hours before the start of the experiment, and no food was consumed within 3 hours.

[0048] The subjects (n=5) were divided into groups of 5 participants each. Each group consumed their own sample (one type of spirit or a control of edible alcohol with the same alcohol content) between 8:00 PM and 10:00 PM. The dosage was 1.2 g / kg body weight for male subjects and 1.1 g / kg body weight for female subjects (based on ethanol). Researchers were divided into two groups: one group prepared the alcohol sample and control, while the other group cared for the subjects. Before drinking, participants completed the necessary data collection (personal information, recent drinking history). After drinking, the researchers took them to their rooms to rest. Eight hours later, after the subjects regained consciousness, they completed a questionnaire, rating their symptoms from 0 to 10. Referring to Hogewoning et al.'s report "Characteristics of social drinkers with and without ahangover after heavy alcohol consumption," the rating was from weakest to strongest, categorized as 0 (no symptoms), 1-2 (mild), 3-4 (symptoms present), 5-6 (significant symptoms), 7-8 (strong symptoms), and 9-10 (extremely strong symptoms), further subdivided. (Completed before 10:00 AM). Hangover symptoms mainly include one overall rating and 24 hangover symptoms. The overall rating criteria are the same as those for individual hangover symptoms. For non-drinkers, the default score for each hangover symptom is 0. The scoring results are shown in Table 1:

[0049] Table 2. Hangover symptom scores of liquor sample A in the population.

[0050]

[0051]

[0052] Note: Significant differences in hangover symptom scores between different sample groups in the same row are marked by lowercase letters "a" and "b". The same letter indicates no significant difference between the two groups (p>0.05), while different letters indicate a significant difference between the two groups (p<0.05).

[0053] Subjects who consumed sample A of baijiu reported generally milder hangovers, and their scores for symptoms such as headache, dizziness, and loss of appetite were significantly lower than those for the control group who consumed alcohol.

[0054] 2) Use a mouse model to evaluate the hangover effect of baijiu sample A.

[0055] Male Balb / c mice aged 6-8 weeks were housed in an SPF (Special Purpose Facility) environment. After 7 days of acclimatization, they were divided into three groups: a control group, a edible alcohol group, and a liquor group, with 10 mice in each group. The edible alcohol and liquor groups were administered the corresponding sample via gavage at a dose of 12 mL / kg body weight (equivalent to 4.3 g / kg body weight of ethanol). The control group was administered the same dose of physiological saline via gavage. The housing environment was 23±2℃ with 12-hour circulating light. Mice had free access to water and were fed a standard maintenance diet. The experimental protocol complied with the standards of the Animal Ethics Committee of Jiangnan University.

[0056] Timing began after gavage. Ten hours after gavage, five mice from each group were removed for a sucrose preference test, while the remaining five mice underwent pain threshold and gait monitoring tests. After the behavioral tests, the mice in each group were sacrificed, and blood was collected from the eyeballs. The blood was then transferred to centrifuge tubes, mixed by inversion, and centrifuged at 4°C and 3000 rpm for 15 minutes. The supernatant was collected and stored at -80°C for later testing. Brain tissue was also collected, flash-frozen in liquid nitrogen, and stored at -80°C for the detection of neurotransmitters and other indicators.

[0057] Serum histamine (HIS) and endothelin (ET) concentrations were determined using enzyme-linked immunosorbent assay (ELISA). After the ELISA kit (Jiangsu Enzyme Labeling) was equilibrated to room temperature, 10 μL of the sample to be tested was added to the reaction plate, followed by 40 μL of sample diluent. After gentle shaking, 100 μL of horseradish peroxidase-labeled detection antibody was added, and the plate was incubated at 37°C for 60 min. After washing, the chromogenic reagent was added, and the plate was incubated at 37°C in the dark for 15 min. Finally, the stop solution was added, and the plate was zeroed using a blank control well. The absorbance value at 450 nm was measured in each well, and quantification was performed using the standard curve method.

[0058] Determination of 5-hydroxytryptamine (5-HT) and dopamine (DA) concentrations in brain tissue: Brain tissue samples were added to 0.4 mol / L perchloric acid solution at a ratio of 1:9 (m:v) and allowed to stand for 10 min to precipitate proteins. The samples were then homogenized using a high-throughput tissue homogenizer, centrifuged at 12000×g at 4℃ for 15 min, and the supernatant was collected for determination using enzyme-linked immunosorbent assay (ELISA).

[0059] The mechanical withdrawal threshold (MWT) of the foot was determined according to the method reported in the literature. Experimental mice were placed in a 22cm×22cm×30cm transparent wire mesh cage for 30 minutes to acclimatize. While in a quiet state, the mechanical withdrawal threshold of the mouse's foot was measured using a von Frey pain threshold meter. The mouse's hind paw was gently touched vertically with the meter's fiber optic cable. A positive response was indicated by the mouse withdrawing its hind paw or attempting to escape. The instrument automatically recorded the minimum stimulus intensity required to elicit a positive response in the mouse. The average of three measurements was taken as the mechanical withdrawal threshold.

[0060] Sucrose preference test (SPT): SPT was performed at the same time before and after gavage administration, and SPT data were collected. In each experiment, mice were acclimatized to a 1% sucrose solution for 1 hour. Two water bottles were placed in each cage: one for 1% sucrose solution and one for pure water. To prevent mice from favoring one side of the solution, the bottles were switched after half an hour, and the consumption of the sucrose solution and pure water was recorded. Mice had free access to food and drink before the experiment, and water and sucrose consumption were calculated by weighing the bottles. Sucrose preference was calculated as a percentage of total fluid consumption relative to sucrose consumption.

[0061] Sucrose preference (SP) = (Sucrose consumption / (Sucrose consumption + Water consumption)) × 100%.

[0062] Gait analysis: 10 hours after gavage, the behavior of the mice was monitored and analyzed using the CatWalk XT animal gait acquisition and analysis system. (1) Training phase: The cages of the mice were placed in the cage holder at the rear end of the CatWalk gait analyzer for 5 minutes to establish a familiar environment for the mice. Then, one mouse was placed on the track at the front of the instrument and allowed to explore freely until it ran the entire course and entered the dark box at the end of the instrument. This was considered a successful training session. If the mouse did not explore forward on the track for a long time, a certain stimulus was given to the mouse from behind until the mouse entered the dark box. Each mouse was trained 3 times a day for 7 consecutive days until all mice could walk to the end of the track without stopping and at a constant speed without external stimulation. (2) Data acquisition phase: After the training was completed, the gait of each group of mice was analyzed. The relevant indicators were: gait symmetry, ataxia coefficient, stride length, and paw angle variability.

[0063] 3) Results Analysis

[0064] like Figure 1 As shown, 10 hours after gavage administration of baijiu sample A, the serum histamine level in mice showed a slight decreasing trend compared to the blank group and the edible alcohol group, but the difference was not significant. 10 hours after gavage administration, the serum endothelin level in the baijiu sample A group was significantly higher than that in the blank group (blank group: 40.16 ng / L, baijiu sample A group: 42.50 ng / L), but significantly lower than that in the edible alcohol group by 5.7% (edible alcohol group: 45.05 ng / L, baijiu sample A group: 42.50 ng / L). This indicates that gavage administration of baijiu sample A had a smaller effect on increasing the serum endothelin level in mice than edible alcohol, consistent with the results in human experiments showing a milder headache.

[0065] like Figure 2 As shown, oral administration of edible alcohol significantly reduced the 5-HT content in the brain tissue of mice, while the 5-HT content in the brain tissue of mice treated with baijiu sample A was only slightly lower than that in the control group, and significantly higher than that in the edible alcohol group by 1.4% (control group: 232.36 ng / L, edible alcohol group: 208.75 ng / L, baijiu sample A group: 211.76 ng / L). Ten hours after oral administration, the dopamine content in the brain tissue of mice in the edible alcohol group was significantly higher than that in the control group and the baijiu sample A group, while oral administration of baijiu sample A did not affect the dopamine content in the brain tissue of mice (it was 16% lower than that in the alcohol control group). This is consistent with the results observed in human experiments with milder levels of depression.

[0066] like Figure 3 As shown, both oral administration of edible alcohol and liquor sample A significantly increased the plantar pain threshold in mice, indicating that the mice were less alert to external stimuli. The plantar pain threshold of mice in liquor sample A group was 35% lower than that in the edible alcohol group.

[0067] like Figure 4 As shown, the sucrose preference of mice in the edible alcohol group and the baijiu sample A group was significantly lower than that in the control group, indicating that alcohol caused a decrease in appetite in mice. The sucrose preference of mice in the baijiu sample A group was 24% higher than that of mice in the edible alcohol group (edible alcohol group: 49.59%, baijiu sample A group: 61.50%), indicating that the baijiu sample A had a milder effect on reducing taste and would not cause obvious apathy or regret.

[0068] like Figure 5 As shown, oral administration of edible alcohol induced gait disorder in mice. The edible alcohol group exhibited significantly higher footprint symmetry, ataxia coefficient, and paw angle change rate compared to the control group, while stride length was significantly reduced. In contrast, the baijiu sample A group showed no significant difference in footprint symmetry and paw angle change rate compared to the control group, but was 29% lower than the edible alcohol group. The ataxia coefficient and stride length were also improved compared to the edible alcohol group, indicating that baijiu sample A caused less damage to the motor function of mice. This is consistent with the results of milder dizziness observed in human experiments.

[0069] The above results indicate that the evaluation of Baijiu sample A using the method disclosed in this invention yields results consistent with population evaluations, demonstrating good post-drinking comfort. This confirms that the method described in this invention can be used to evaluate the post-drinking hangover effect of Baijiu samples.

[0070] Example 2: Evaluation of the hangover effect of commercially available baijiu sample B

[0071] 1) Conduct a sensory evaluation of a commercially available strong-aroma baijiu (45% alcohol content) among a population.

[0072] The population trial protocol for evaluating the hangover effect of alcohol samples was referenced in Example 1, and the scoring results are shown in Table 2.

[0073] Table 3. Hangover symptom scores of Baijiu sample B in the population.

[0074]

[0075]

[0076] Note: Significant differences in hangover symptom scores between different sample groups in the same row are marked by lowercase letters "a" and "b". The same letter indicates no significant difference between the two groups (p>0.05), while different letters indicate a significant difference between the two groups (p<0.05).

[0077] Subjects who consumed baijiu (sample B) reported a higher overall level of hangover than those who consumed alcohol, and their scores for symptoms such as headache, apathy, and depression were significantly higher than those of the alcohol control group.

[0078] 2) Use a mouse model to evaluate the hangover effect of baijiu sample B.

[0079] The method for assessing the hangover effect of Baijiu sample B is the same as in Example 1, and the animal feeding process and detection indicators are as described in Example 1.

[0080] 3) Results Analysis

[0081] like Figure 1 As shown, 10 hours after gavage administration of baijiu sample B, the serum histamine level in mice was significantly lower than that in the control group and the edible alcohol group, with a decrease of 17% (control group: 15.88 ng / L, edible alcohol group: 15.34 ng / L, baijiu sample B group: 12.65 ng / L). 10 hours after gavage, the serum endothelin level in the baijiu sample B group was significantly higher than that in the control group (control group: 40.16 ng / L, baijiu sample B group: 47.45 ng / L), and significantly higher than that in the edible alcohol group by 5% (edible alcohol group: 45.05 ng / L, baijiu sample B group: 47.45 ng / L), indicating that gavage administration of baijiu sample B had a greater effect on increasing serum endothelin levels in mice than edible alcohol. This is consistent with the results in human experiments where the headache intensity was higher in the baijiu sample B group than in the alcohol group.

[0082] like Figure 2As shown, gavage administration of edible alcohol significantly reduced the 5-HT content in mouse brain tissue (a decrease of 1.4%), with the most significant decrease observed in baijiu sample B mice, which was lower than both the control group and the edible alcohol group (control group: 232.36 ng / L, edible alcohol group: 208.75 ng / L, baijiu sample B group: 191.03 ng / L). Ten hours after gavage, the dopamine content in the brain tissue of mice in the edible alcohol group was significantly higher than that in the control group, while gavage administration of baijiu sample B caused an almost identical increase in dopamine content in mouse brain tissue. The changes in 5-HT and dopamine levels were consistent with the trend of increased depression scores observed in human experiments.

[0083] like Figure 3 As shown, both oral administration of edible alcohol and liquor sample B significantly increased the foot pain threshold in mice, indicating a decrease in the mice's alertness to external stimuli. Among them, the increase in foot pain threshold in the liquor sample B group was the most significant, higher than that in the blank group and the alcohol control group, which is consistent with the trend of decreased alertness scores in human experiments.

[0084] like Figure 4 As shown, the sucrose preference of mice in the alcohol group and the baijiu sample B group was significantly lower than that of the control group, indicating that alcohol reduced the appetite of mice. The sucrose preference of mice in the baijiu sample B group was close to that of mice in the alcohol group, but 37% lower than that of the control group, indicating that baijiu sample B had a significant effect on reducing taste and may have caused anhedonia. This is consistent with the results in human experiments, where the levels of loss of appetite, regret, and indifference were higher in the alcohol group than in the alcohol group.

[0085] like Figure 5 As shown, oral administration of edible alcohol induced gait disorder in mice. The edible alcohol group exhibited significantly higher gait symmetry, aberration coefficient, and paw angle change rate compared to the control group, while stride length was significantly reduced. The gait impairment was even more severe in the baijiu (Chinese liquor) sample B group, with gait symmetry significantly higher than the control group (53%), aberration coefficient 33% higher, and stride length significantly lower (41%), indicating that baijiu sample B had a stronger impairing effect on the motor function of mice. This is consistent with the results of more severe dizziness observed in human experiments.

[0086] The above results indicate that the evaluation of Baijiu sample B using the method disclosed in this invention is consistent with the population evaluation results, showing poor post-drinking comfort, thus confirming that the method described in this invention can be used to evaluate the post-drinking hangover effect of Baijiu samples.

[0087] Example 3: Analysis of Indicators for Evaluating the Hangover Effect of Baijiu (Chinese liquor)

[0088] To further illustrate the practicality and reliability of this method, Spearman correlation analysis was performed on the hangover symptom scores of the two alcohol samples in the human population and various indicators in the mice using Graphpad Prism 8.0. The indicator values ​​of each group of mice (n=5) were set as independent variables, and the hangover evaluation values ​​of each sample population (n=5) were set as dependent variables. A univariate linear regression analysis was then performed on both, and the results are as follows: Figure 6 As shown.

[0089] Serum histamine, brain 5-HT, and stride length in mice were strongly negatively correlated with anxiety, depression, and fatigue (p<0.05), while serum endothelin, brain dopamine, plantar pain threshold, footprint symmetry, ataxia coefficient, and paw angle change rate were strongly positively correlated with headache, depression, decreased alertness, dizziness, and clumsiness (p<0.05). Furthermore, sucrose preference was strongly negatively correlated with decreased appetite, apathy, and regret (p<0.05). Therefore, the animal model disclosed in this invention can replace human trials for evaluating the post-drinking comfort of alcoholic beverages.

[0090] Comparative Example 1: Evaluation of the hangover effect of commercially available baijiu samples A and B using other mouse indicators.

[0091] For specific implementation methods, refer to Examples 1 and 2. Mouse indicators were monitored 10 hours after gavage. The difference was that the hangover effects of mice on commercially available liquor samples A and B were evaluated using serum prostaglandin E2 (PGE2), methionine enkephalin (MEK), serum 5-HT, brain histamine, and periorbital pain threshold.

[0092] like Figure 7 As shown, after mice were administered alcohol samples A and B by gavage, there was no significant difference in serum PGE2 and MEK levels at 10 h. Furthermore, correlation analysis showed no correlation with headache scores in the general population (p>0.05). Therefore, compared with the mouse indicators disclosed in this invention, serum PGE2 and MEK cannot be used as effective indicators to characterize differences in hangover headaches in mice.

[0093] Ten hours after gavage, serum 5-HT levels in mice in the alcohol group and the two baijiu groups were higher than those in the blank group, but there was no significant difference between baijiu A and baijiu B groups. Correlation analysis showed that there was no correlation between serum 5-HT and human depression scores (p>0.05). Therefore, compared with the mouse indicators disclosed in this invention, serum 5-HT cannot be used as an effective indicator to characterize differences in hangover depression in mice. Similarly, there was no significant difference in brain histamine levels between baijiu A and B groups. Correlation analysis showed that there was also no correlation between brain histamine and human depression scores (p>0.05).

[0094] Furthermore, no difference was found in the periorbital pain threshold among the groups after mice were administered alcohol by gavage, and correlation analysis showed that the periorbital pain threshold could not be used as an effective indicator of decreased alertness in mice (p>0.05).

[0095] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Anyone skilled in the art can make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be determined by the claims.

Claims

1. A method for evaluating the hangover effect after drinking alcohol, characterized in that, Includes the following steps: (1) Set up an experimental group, an alcohol control group and a blank control group. The experimental group was given a sample of wine by gavage, the alcohol control group was given edible alcohol with the same ethanol dose by gavage, and the blank group was given physiological saline by gavage. (2) After gavage, the biochemical and behavioral indicators of the experimental subjects were tested; the experimental subjects included mice; The biochemical indicators include serum histamine content, serum endothelin content, brain tissue serotonin content, and brain tissue dopamine content; the behavioral indicators include pain threshold, gait parameters, and sucrose preference, and the gait parameters include footprint symmetry, motor incoordination coefficient, stride length, and foot angle change rate; In step (2), the test is conducted when the experimental subject is in a hangover state; the hangover state refers to the state when the serum ethanol content drops to 0 after drinking alcohol; Multiple biochemical indicators in serum and brain tissue samples of experimental subjects after oral administration of alcohol were analyzed, and several behavioral indicators were monitored. Serum histamine levels were used to assess post-drinking anxiety; serum endothelin levels were used to assess post-drinking headache; brain 5-HT and dopamine levels were used to assess post-drinking depression; foot pain threshold was used to assess post-drinking decreased alertness; and sucrose preference was used to assess post-drinking decreased appetite, regret, and apathy. The degree of dizziness after drinking was evaluated by the symmetry of the participants' footprints and the ataxia coefficient; the degree of fatigue after drinking was evaluated by the stride length after drinking; and the degree of clumsiness after drinking was evaluated by the rate of change of the participants' claw angles. A comprehensive assessment of the hangover effect after drinking alcohol was conducted using animal models.

2. The method as described in claim 1, characterized in that, In step (2), the test is performed 6-14 hours after gavage.

3. The method as described in claim 2, characterized in that, In step (1), the gavage dose is 3.0-6.0 g / kg body weight based on the ethanol equivalent.

4. The method as described in claim 3, characterized in that, The alcohol samples include spirits, whiskey, or brandy.

5. The method as described in claim 4, characterized in that, If the serum endothelin content in the experimental group mice is 5% higher than that in the alcohol control group, it is determined that the alcohol sample will cause more severe headache symptoms and poor post-drinking comfort; if the serum endothelin content is 5% lower than that in the alcohol control group, it is determined that the alcohol sample will not cause more severe headache symptoms and good post-drinking comfort; if it is in between, the post-drinking comfort is moderate.

6. The method as described in claim 5, characterized in that, Depressive mood can be assessed using brain serotonin levels and / or brain tissue dopamine levels. If the 5-hydroxytryptamine level in the brain of the experimental group mice is 8% lower than that in the alcohol control group, it is determined that the alcohol sample will cause depressive mood and poor post-drinking comfort; if the 5-hydroxytryptamine level in the brain is 1% higher than that in the alcohol control group, it is determined that the alcohol sample will not cause depressive mood and good post-drinking comfort; if it is in between, it is considered that the post-drinking comfort is moderate. If the dopamine content in the brain tissue of the experimental group mice is 16% higher than that in the alcohol control group, it is determined that the alcohol sample will cause more severe depressive mood and poor post-drinking comfort; if the dopamine content in the brain tissue of the experimental group mice is 16% lower than that in the alcohol control group, it is determined that the alcohol sample will not cause more severe depressive mood and good post-drinking comfort.

7. The method as described in claim 6, characterized in that, If the serum histamine level in the experimental group mice is 17% lower than that in the alcohol control group, it is determined that the alcohol sample will cause more severe anxiety symptoms; if the footprint symmetry value of the experimental group mice is 50% higher than that in the blank control group, it is determined that the alcohol sample will cause more severe dizziness symptoms; if the sucrose preference value of the experimental group mice is 37% lower than that in the blank group, it is determined that the liquor sample can cause severe loss of appetite, apathy, and regret symptoms; if the stride length value of the experimental group mice is 40% lower than that in the alcohol control group, it is determined that the alcohol sample will cause more severe fatigue symptoms and poor post-drinking comfort.

8. The method as described in claim 7, characterized in that, If the plantar pain threshold of mice in the experimental group is 34% lower than that of the alcohol control group, it can be determined that the baijiu sample will not cause a serious decrease in alertness; or if the sucrose preference value is 24% higher than that of the alcohol control group, it can be determined that the baijiu sample will not cause a serious decrease in appetite, apathy, or regret; or if the change rate of paw angle is 29% lower than that of the alcohol control group, it can be determined that the baijiu sample will not cause a serious clumsy symptom, and it can be determined that the baijiu sample provides good comfort after drinking.

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