Multi-dimensional biological detection method for evaluating drinking comfort of baijiu and application thereof

Abstract

The application discloses a kind of multi-dimensional biological detection method and its application for evaluating liquor drinking comfort degree, comprising the following steps: step a, male mice are selected for adaptive feeding, randomly divided into to-be-tested liquor sample group, pure alcohol control group, blank control group, set up gavage dose, using animal fine behavior analysis, obtain the moving distance ratio, moving time ratio and stagnation frequency ratio of mice after gavage to-be-tested liquor, evaluate its relaxation or excitement degree after drinking;The application relates to the field of food science and biological detection technology, and the beneficial effects of the application are that the relaxation and pleasure degree and recovery speed after drinking liquor are systematically evaluated from three dimensions of behavior, neurotransmitter and metabolism, the results are objective and quantifiable, and can be used for liquor brewing process optimization, liquor body design and product quality evaluation.

Description

Technical Field

[0001] This invention relates to the fields of food science and biological detection technology, and in particular to a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) and its application. Background Technology

[0002] Chinese baijiu consumption has entered an era of healthy drinking, with consumers highly focused on post-drinking comfort, namely "no headache, no dry mouth, quick sobering up, and a pleasant and relaxing experience." Traditional evaluation methods rely on subjective questionnaires from drinkers, resulting in significant individual differences and poor repeatability. While existing technologies include methods to infer quality by analyzing trace components in baijiu or using animal models to assess "intoxication levels," these methods suffer from limitations in their simplistic evaluation dimensions. They lack a systematic integration from overall behavior to molecular mechanisms, and have yet to establish a quantitative correlation model between "behavior, neurotransmitters, and metabolic markers," making it difficult to explain the underlying reasons for differences in comfort levels. Furthermore, a rapid and standardized testing process for baijiu design and process optimization has not yet been developed. Therefore, a scientific, objective, and quantifiable method for evaluating the comfort of baijiu consumption is urgently needed. Summary of the Invention

[0003] The purpose of this invention is to solve the above-mentioned problems by designing a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu and its application. This invention addresses the problems of existing technologies, which, although methods exist for inferring quality by analyzing trace components in baijiu or using animal models to evaluate "intoxication level," all suffer from a single evaluation dimension. They lack a systematic integration from overall behavior to molecular mechanisms, have not yet established a quantitative correlation model between "behavior-neurotransmitters-metabolic markers," and are unable to explain the intrinsic reasons for differences in comfort. Furthermore, a rapid and standardized detection process that can be used for baijiu design and process optimization has not yet been formed.

[0004] The technical solution of the present invention to achieve the above objectives is as follows: a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor), comprising the following steps: Step a: Select male mice for adaptive feeding and randomly divide them into the test wine sample group, the pure alcohol control group, and the blank control group. Set the gavage dose and use animal fine behavior analysis to obtain the ratio of movement distance, movement time, and number of pauses of the mice after gavage administration of the test wine to evaluate their relaxation or excitement level after drinking. Step b: Detect the proportion of GABAB receptor-positive neurons, dopamine concentration, and serotonin concentration in the cerebral cortex and hippocampus of mice after gavage, and evaluate their neuro-pleasurable or inhibitory effects. Step c: Detect the acetaldehyde accumulation concentration and ethanol metabolism rate in the whole blood of mice after gavage to evaluate their metabolic comfort. Based on the results of steps d and ac, calculate the overall comfort score. The higher the score, the better the drinking comfort.

[0005] Preferably, step a further includes obtaining the ratios of mice after being administered the test liquor by gavage. When the ratios of movement distance, average speed, and movement time are in the range of 0.5-1.0, it is determined to be relaxed and soothed. When they are in the range of 1.0-1.5, it is determined to be moderately excited. The closer each parameter ratio is to 1, the higher the comfort score.

[0006] Preferably, the proportion of GABAB receptor-positive neurons in step b is determined by the following criteria: the test wine sample group shows a slight but significant increase compared to the blank control group, with a statistically significant difference (P < 0.05), and there is no overactivation (positive rate < 30%).

[0007] Preferably, the dopamine concentration mentioned in step b is determined by the following criteria: the test wine sample group shows a slight but significant increase compared to the blank control group, and the difference is statistically significant, i.e., P < 0.05; the 5-hydroxytryptamine concentration is determined by the following criteria: the hippocampus region of the test wine sample group does not show a significant increase compared to the blank control group, and the difference is not statistically significant, i.e., P > 0.05.

[0008] Preferably, the acetaldehyde accumulation concentration mentioned in step b is determined by the following criteria: the whole blood acetaldehyde concentration is less than 1.5 ppm 1 hour after gavage.

[0009] Preferably, the oral dose for mice is 0.10-0.20 mL / mouse, and the oral alcohol content is 35%-45% vol.

[0010] Preferably, the liquor to be tested includes light-aroma, strong-aroma, and sauce-aroma liquors, or liquors brewed from organic highland barley through a four-stage steaming process.

[0011] Preferably, the blank control is distilled water, and the pure alcohol control is an isobaric solution prepared with edible alcohol.

[0012] The present invention provides a multi-dimensional biological detection method for evaluating the comfort of drinking baijiu (Chinese liquor) and its application. It systematically evaluates the degree of relaxation and pleasure and the speed of recovery after drinking baijiu from three dimensions: behavior, neurotransmitters and metabolism. The results are objective and quantifiable and can be used for baijiu brewing process optimization, liquor design and product quality evaluation. Attached Figure Description

[0013] Figure 1 This invention presents a multi-dimensional biological detection method for evaluating the comfort of drinking baijiu (Chinese liquor) and its application, as shown in the bar chart of the ratio of movement distance of each group of mice after gavage administration of different types of alcohol.

[0014] Figure 2 The bar chart shows the ratio of the number of pauses in each group of mice after gavage administration of different types of alcohol, as described in this invention, for a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu.

[0015] Figure 3 This is an immunofluorescence image of GABAB receptor-positive neuronal cells in a liquor sample 1, which is part of a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu as described in this invention and its application.

[0016] Figure 4 This is an immunofluorescence image of GABAB receptor-positive neuronal cells in a wine sample 2, representing a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu as described in this invention and its application.

[0017] Figure 5 This is an immunofluorescence image of GABA B receptor-positive neuronal cells in a liquor sample, representing a multi-dimensional biological detection method for evaluating the drinking comfort of baijiu as described in this invention and its application.

[0018] Figure 6 This is an immunofluorescence image of GABAB receptor-positive neuronal cells in a wine sample, representing a multidimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) as described in this invention and its application.

[0019] Figure 7 This is an immunofluorescence image of GABAB receptor-positive neuronal cells in a wine sample, representing a multidimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) as described in this invention and its application.

[0020] Figure 8 This invention relates to a multi-dimensional biological detection method for evaluating the comfort of drinking baijiu (Chinese liquor) and its application, specifically the whole blood acetaldehyde concentration-time curve after gavage administration of different types of baijiu. Detailed Implementation

[0021] The present invention will now be described in detail with reference to the accompanying drawings, such as... Figure 1-8 As shown, a multi-dimensional biological detection method for evaluating the comfort of drinking baijiu (Chinese liquor) and its application are presented. Through detailed animal behavior analysis, detection of neurotransmitters in brain regions, and quantification of blood alcohol metabolism markers, the method comprehensively evaluates the degree of relaxation / pleasure and recovery speed after drinking baijiu.

[0022] The technical solution of the present invention includes the following steps: Step a: Preparation of experimental animals, setting of gavage dosage, and collection of fine behavioral data of animals; The experimental animals were selected as SPF-grade male C57BL / 6J mice, 6 weeks old, weighing 18-22g, and acclimatized for 1 week. They were randomly divided into the test wine sample group, the pure alcohol control group (with the same alcohol content as the wine sample), and the blank control group (distilled water), with at least 5 mice in each group. The gavage dose was set based on the World Health Organization's recommended moderate alcohol consumption for adult males (70kg) (12-15g alcohol) and the conversion factor between mouse and human equivalent doses (9 times), determining the gavage dose per mouse to be 0.15mL (alcohol content adjusted to 40% vol). Fine behavioral data of the animals were collected before gavage (t0), 30 min after gavage (t1), and 120 min after gavage (t2). Mice were placed in an open field behavioral apparatus (a cylindrical white-bottomed, transparent-walled structure with a diameter of 60 cm). The free movement trajectory within 1 minute was recorded and analyzed in real time using Any-maze software. The ratio of movement distance (X1t=At / A0), the ratio of average speed (X2t=Bt / B0), and the ratio of number of pauses (X6t=Ft / F0) were calculated. When X1t, X2t, and X6t were all in the range of 0.5-1.5 and the closer they were to 1, the higher the post-drinking comfort. Step b, Neurotransmitter detection: After the behavioral tests were completed (t1 and t2 time points), the mice were euthanized, and the cerebral cortex and hippocampus tissue were quickly removed; GABA was detected using immunofluorescence staining. B The proportion of receptor-positive neurons was determined by ELISA, and the concentrations of dopamine (DA) and serotonin (5-HT) were measured. The test wine sample group was compared with the blank control group for GABA content. B When the percentage of receptor-positive cells increases slightly (10%-25%) and the concentration of DA increases by 10%-30%, it is considered to have a relaxing and pleasurable effect. Step c, Detection of blood alcohol metabolism markers: Take another batch of mice with the same treatment, and collect blood from the tail vein at 0.5h, 1h, 2h and 3h after gavage. Use headspace gas chromatography (HS-GC) to determine the concentration of acetaldehyde in whole blood. The alcohol sample with a peak acetaldehyde concentration of less than 1.5 ppm and rapid metabolic clearance (<20 ppm after 3h) is judged to have excellent comfort.

[0023] Step d, Comprehensive Score: Calculate the comprehensive comfort score by weighting the behavioral parameter deviation, neurotransmitter regulation index, and peak acetaldehyde concentration according to their respective weights (40%, 30%, 30%). The higher the score, the better the comfort of drinking baijiu.

[0024] The present invention will be further described below through specific embodiments.

[0025] Example 1: Evaluation of the drinking comfort of four distilled spirits (1) Experimental wine sample: Sample 1: Tianyoude Organic Highland Barley Wine (Jinbao), 42% vol, provided by Qinghai Huzhu Highland Barley Wine Co., Ltd.

[0026] Sample 2: Ordinary highland barley liquor, 44% vol, commercially available.

[0027] Sample 3: Gold Label Whisky, 40% vol, commercially available.

[0028] Sample 4: Vodka, 96% vol, diluted with deionized water to 40% vol.

[0029] Sample 5, the blank control: distilled water.

[0030] Pure alcohol comparison: Edible alcohol is prepared with deionized water to a concentration of 40% vol.

[0031] (2) Animals and gavage: C57BL / 6J male mice, 6 mice in each group, each mouse was gavaged with 0.15 mL (40% vol alcohol sample).

[0032] (3) Behavioral measurements, i.e., step a: 30 min after gavage, the movement distance ratio of sample 1 was 0.88±0.05, the average speed ratio was 0.78±0.06, and the number of pauses ratio was 0.90±0.05. All parameter ratios were significantly (P<0.05) closer to 1, with the smallest deviation, and were judged as "relaxed and soothed". The parameter ratios of samples 2 and 3 were greater than 1, and were judged as "moderate excitement" but with a large deviation. The parameter ratio of sample 4 was less than 0.6, and was judged as "excessive inhibition". 120 min after gavage, only sample 1 recovered to 0.88-0.98, close to 1, indicating that it recovered the fastest after drinking.

[0033] (4) Detection of neurotransmitters in the brain, i.e., step b: 30 min after gavage, GABA in the cerebral cortex of mice in group 1 was measured. B The proportion of receptor-positive neurons was 18.5%±2.1%, significantly lower than that in the vodka group (48.2%±3.5%) and higher than that in the blank group (5.2%±1.0%), showing "micro-activation"; the dopamine concentration (cortex) was 39.33±0.73 pg / mL, which was 28.6% higher than that in the blank control group (30.57±0.32), reaching the pleasure threshold; the 5-HT concentration (hippocampus) was 67.99±1.3 ng / mL, which was not significantly different from that in the blank group (61.77±1.0) and did not cause drowsiness.

[0034] (5) Blood acetaldehyde determination, i.e., step c: 1 hour after gavage, the whole blood acetaldehyde concentration of sample 1 was 1.12±0.31 ppm, significantly lower than that of sample 2 (2.79±0.35) and sample 3 (2.00±0.41); 3 hours later, the acetaldehyde concentration of sample 1 decreased to 0.69±0.01 ppm, while that of sample 2 remained at 0.23±0.34 (already fast, but with a high peak value), and that of sample 3 was 0.75±0.11. Sample 1 had the lowest acetaldehyde peak value, indicating rapid metabolism.

[0035] (6) Overall score, i.e. step d: Sample 1 scored 94.5 (out of 100), Sample 2 scored 72.0, Sample 3 scored 78.5, and Sample 4 scored 45.0. The results show that Tianyoude Organic Highland Barley Wine (Jinbao) has a significantly better drinking comfort than other samples.

[0036] Example 2: Optimizing the highland barley wine blending process using the method of the present invention The method of this invention was used to evaluate barley spirits produced in different seasons (spring, summer, autumn, and winter). Results showed that autumn spirits had the highest total acid content (1410 mg / L), a relatively high total ester content, the smallest deviation in behavioral parameters (movement distance ratio 0.92), a peak acetaldehyde level of only 1.05 ppm, and a comprehensive comfort score of 92. Based on this, the distillery increased the proportion of autumn spirits in the finished product blending from 20% to 35%, while reducing the proportion of summer spirits. The improved product, after retesting using the method of this invention, showed a further 12% reduction in the deviation in behavioral parameters, a decrease in the peak acetaldehyde level to 0.95 ppm, and an improvement in the comprehensive score to 96.5. Post-market consumer satisfaction surveys showed a 23% increase in the selection rate for the "post-drinking comfort" option.

[0037] The above technical solutions only embody the preferred technical solutions of the present invention. Any modifications that may be made by those skilled in the art to certain parts thereof embody the principles of the present invention and fall within the protection scope of the present invention.

Claims

1. A multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor), characterized in that, Includes the following steps: Step a: Select male mice for adaptive feeding and randomly divide them into the test wine sample group, the pure alcohol control group, and the blank control group. Set the gavage dose and use animal fine behavior analysis to obtain the ratio of movement distance, movement time, and number of pauses of the mice after gavage administration of the test wine to evaluate their relaxation or excitement level after drinking. Step b: Detect the proportion of GABAB receptor-positive neurons, dopamine concentration, and serotonin concentration in the cerebral cortex and hippocampus of mice after gavage, and evaluate their neuro-pleasurable or inhibitory effects. Step c: Detect the acetaldehyde accumulation concentration and ethanol metabolism rate in the whole blood of mice after gavage to evaluate their metabolic comfort. Based on the results of steps d and ac, calculate the overall comfort score. The higher the score, the better the drinking comfort.

2. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, Step a also includes obtaining the ratios of mice after being given the test liquor by gavage. When the ratios of movement distance, average speed, and movement time are in the range of 0.5-1.0, it is considered relaxed and soothing. When they are in the range of 1.0-1.5, it is considered moderately excited. The closer each parameter ratio is to 1, the higher the comfort score.

3. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, The preferred criterion for determining the proportion of GABAB receptor-positive neurons in step b is: the test wine sample group shows a slight but significant increase compared to the blank control group, with a statistically significant difference (P < 0.05), and there is no overactivation (positive rate < 30%).

4. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, The preferred criterion for determining the dopamine concentration mentioned in step b is: the test wine sample group shows a slight but significant increase compared to the blank control group, and the difference is statistically significant, i.e., P < 0.05; the preferred criterion for determining the 5-hydroxytryptamine concentration is: the hippocampus region of the test wine sample group does not show a significant increase compared to the blank control group, and the difference is not statistically significant, i.e., P > 0.

05.

5. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, The preferred criterion for determining the acetaldehyde accumulation concentration mentioned in step b is: the whole blood acetaldehyde concentration is less than 1.5 ppm 1 hour after gavage.

6. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, The oral dose for mice was 0.10-0.20 mL per mouse, and the oral alcohol content was 35%-45% vol.

7. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, The liquor to be tested includes light-aroma, strong-aroma, and sauce-aroma liquors, or liquors brewed from organic highland barley through a four-stage steaming process.

8. The multi-dimensional biological detection method for evaluating the drinking comfort of baijiu (Chinese liquor) according to claim 1, characterized in that, The blank control is distilled water, and the pure alcohol control is an alcohol solution of equal concentration prepared with edible alcohol.

9. The application of the method according to any one of claims 1-8 in the optimization of baijiu brewing process, baijiu body design, product quality evaluation, or guidance on healthy baijiu consumption.

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