Method for distinguishing feng-flavor liquor and sesame-flavor liquor based on metabonomics

Abstract

The invention discloses a metabonomics-based method for discriminating feng-flavor liquor and sesame-flavor liquor. The specific value of the content of the D-arabinopyranose to the content of the 5-methyl-2-acetylfuran in the Baijiu is used for distinguishing Feng-flavor Baijiu and sesame-flavor Baijiu in metabonomics research, and the Baijiu with the specific value of the D-arabinopyranose to the content of the 5-methyl-2-acetylfuran in the to-be-detected sample being smaller than 2 is the Feng-flavor Baijiu. The white spirit with the ratio of the D-arabinopyranose to the 5-methyl-2-acetylfuran being more than 8 is the sesame-flavor white spirit. The method is simple to operate and reliable in detection result, and can provide a reference basis for Baijiu metabonomics research.

Description

Technical Field

[0001] The invention belongs to the field of analytical chemistry and is a method for distinguishing phoenix-flavor liquor and sesame-flavor liquor based on metabolomics. Background Art

[0002] As one of the world's six major distilled spirits, baijiu (Chinese liquor) holds a pivotal position in Chinese culinary culture. Its brewing process is complex, involving multiple key steps, including saccharification, fermentation, distillation, and aging. Even subtle differences in each step significantly influence the flavor profile of the final product. Fengxiang-style baijiu and sesame-style baijiu, two distinctive aroma styles, differ significantly in flavor and brewing techniques. Fengxiang-style baijiu possesses a soft, complex aroma, combining the elegant purity of light-fragrance baijiu with the rich, full-bodied flavor of strong-fragrance baijiu. Its rich, elegant aroma, with a subtle medicinal undertone, is intoxicating. Sesame-style baijiu draws upon the craftsmanship of sauce-flavor, light-fragrance, and strong-flavor baijiu, resulting in a unique "four highs and one high" process: high nitrogen ingredients, high-temperature stacking, high-temperature fermentation, high-temperature distillation, and a high amount of koji. This combination of techniques is key to its distinctive style.

[0003] However, due to the complex composition of baijiu (white liquor), current flavor identification still relies primarily on subjective sensory evaluation and empirical judgment, lacking objective, scientific, and quantitative analytical methods. Although previous studies have employed technologies such as electronic tongues, liquid array taste biomimetic sensors, and infrared spectroscopy for flavor identification, these methods remain limited in accuracy and comprehensiveness. In recent years, the rapid development of metabolomics has provided new research insights into baijiu flavor identification. By comprehensively analyzing metabolites in baijiu, this technology can systematically reveal the chemical composition differences between different flavors, thereby establishing a more objective basis for discrimination. This study specifically focused on two characteristic metabolites: D-arabinopyranose and 5-methyl-2-acetylfuran. This is based on two main reasons: First, previous studies have shown significant differences in the content of these two substances between phoenix- and sesame-flavored baijiu; second, no studies have yet used the absolute content or relative ratio of these two substances as discriminant indicators for baijiu flavor identification. This innovative finding provides an important theoretical basis and technical entry point for the development of a new identification method in this study. Summary of the Invention

[0004] The purpose of the present invention is to provide a method for distinguishing Fengxiang-flavor liquor and sesame-flavor liquor based on metabolomics, and to distinguish Fengxiang-flavor liquor and sesame-flavor liquor in metabolomics research by using the ratio of D-arabinopyranose and 5-methyl-2-acetylfuran content in liquor, so as to provide a basis for sample discrimination in metabolomics research.

[0005] In order to achieve the above object, the technical solution provided by the present invention is as follows: A metabolomics-based method for distinguishing Fengxiang-flavor liquor from sesame-flavor liquor was developed. The method detects the contents of D-arabinopyranose and 5-methyl-2-acetylfuran in liquor and calculates their ratio. This method distinguishes Fengxiang-flavor liquor from sesame-flavor liquor in metabolomics research: liquors with a D-arabinopyranose to 5-methyl-2-acetylfuran ratio below 2 are Fengxiang-flavor liquors, while liquors with a D-arabinopyranose to 5-methyl-2-acetylfuran ratio above 8 are sesame-flavor liquors.

[0006] The specific method for detecting the content of D-arabinopyranose and 5-methyl-2-acetylfuran is: First, the liquor samples were pretreated, and then the metabolomics analysis method of vortex-assisted liquid-liquid microextraction combined with gas chromatography-mass spectrometry (VA-LLME-GC-MS) was used to perform absolute quantitative analysis of D-arabinopyranose and 5-methyl-2-acetylfuran in the liquor samples.

[0007] The sample pretreatment method is as follows: the finished wine is diluted into 45 ml of 10% vol diluted wine sample according to the dilution ratio, about 12 g of NaCl and 12 ml of dichloromethane are added, and the mixture is vortex-mixed on a vortex mixer for 5 minutes. The mixed solution is poured into a separatory funnel, allowed to stand for 7 minutes to separate the layers, and the lower organic phase is collected in a centrifuge tube. Pivalic acid is added as an internal standard to a final concentration of 50 μg / ml, 1 g of anhydrous sodium sulfate is added, and the sample is dried at -20°C overnight. The next day, the sample is taken out of the refrigerator and concentrated to 1 ml using a nitrogen blowdown device. The sample is filtered through a 0.22 μm organic filter and then subjected to GC-MS analysis.

[0008] The gas chromatography conditions were as follows: Inert Cap Wax column (30 m×0.25 mm×0.25 μm), inlet temperature of 250°C, carrier gas He, flow rate of 1 mL / min, splitless mode, injection volume of 1 μL, temperature program: 50°C for 2 min, heating to 250°C at a rate of 6°C / min, and holding for 3 min.

[0009] The mass spectrometry conditions were as follows: EI ionization source, electron energy 70 eV, ion source temperature 240° C., mass scan range 35-350 amu, and solvent delay time 3 min.

[0010] Compared with the prior art, the present invention has the following beneficial effects: This method uses the ratio of D-arabinopyranose to 5-methyl-2-acetylfuran in liquor to distinguish between phoenix-flavor and sesame-flavor liquors. The method is simple and rapid to operate, produces accurate and reliable results, and requires minimal sample volume. This method can provide a reference for analyzing liquor metabolic characteristics and related metabolomics research on Chinese liquor. DETAILED DESCRIPTION

[0011] The present invention will be described in further detail below with reference to specific examples, but the embodiments of the present invention include but are not limited to the scope represented by the following examples.

[0012] A metabolomics-based method for distinguishing Fengxiang-flavored and sesame-flavored liquors was developed. By measuring the contents of D-arabinopyranose and 5-methyl-2-acetylfuran in liquor and calculating their ratio, the metabolomics study identified Fengxiang-flavored and sesame-flavored liquors. Liquors with a D-arabinopyranose / 5-methyl-2-acetylfuran ratio below 2 were classified as Fengxiang-flavored liquors, while those with a D-arabinopyranose / 5-methyl-2-acetylfuran ratio above 8 were classified as sesame-flavored liquors.

[0013] The Fengxiang-style liquor samples were collected from the National Flower Porcelain 20 Years, National Flower Porcelain 12 Years, Xifeng 15 Years and Huashan Lunjian 10 Years on the market and purchased on December 26, 2024.

[0014] The sesame-flavored liquor sample was collected from Yipin Jingzhi on the market and purchased on December 26, 2024.

[0015] The specific method is: Liquor samples were pretreated, and then metabolomics analysis using vortex-assisted liquid-liquid microextraction coupled with gas chromatography-mass spectrometry (VA-LLME-GC-MS) was performed to determine the absolute quantification of D-arabinopyranose and 5-methyl-2-acetylfuran in the liquor samples. The ratio of D-arabinopyranose to 5-methyl-2-acetylfuran was calculated. Samples with a D-arabinopyranose to 5-methyl-2-acetylfuran ratio below 2 were considered Fengxiang-style liquors, while those with a D-arabinopyranose to 5-methyl-2-acetylfuran ratio above 8 were considered Zhima-style liquors.

[0016] The sample pretreatment method is as follows: the finished wine is diluted into 45 ml of 10% vol diluted wine sample according to the dilution ratio, about 12 g of NaCl and 12 ml of dichloromethane are added, and the mixture is vortex-mixed on a vortex mixer for 5 minutes. The mixed solution is poured into a separatory funnel, allowed to stand for 7 minutes to separate the layers, and the lower organic phase is collected in a centrifuge tube. Pivalic acid is added as an internal standard to a final concentration of 50 μg / ml, 1 g of anhydrous sodium sulfate is added, and the sample is dried at -20°C overnight. The next day, the sample is taken out of the refrigerator and concentrated to 1 ml using a nitrogen blowdown device. The sample is filtered through a 0.22 μm organic filter and then subjected to GC-MS analysis.

[0017] The gas chromatography conditions were as follows: Inert Cap Wax column (30 m×0.25 mm×0.25 μm), inlet temperature of 250°C, carrier gas He, flow rate of 1 mL / min, splitless mode, injection volume of 1 μL, temperature program: 50°C for 2 min, heating to 250°C at a rate of 6°C / min, and holding for 3 min.

[0018] The mass spectrometry conditions were as follows: EI ionization source, electron energy 70 eV, ion source temperature 240° C., mass scan range 35-350 amu, and solvent delay time 3 min. Example

[0019] The contents of D-arabinopyranose and 5-methyl-2-acetylfuran in liquor samples were determined by metabolomics analysis using vortex-assisted liquid-liquid microextraction coupled with gas chromatography-mass spectrometry (VA-LLME-GC-MS).

[0020] The samples to be tested include Fengxiang-flavored liquor 1, Fengxiang-flavored liquor 2, Fengxiang-flavored liquor 3, Fengxiang-flavored liquor 4, Fengxiang-flavored liquor 5, Fengxiang-flavored liquor 6, Fengxiang-flavored liquor 7, Fengxiang-flavored liquor 8, Fengxiang-flavored liquor 9, Fengxiang-flavored liquor 10, Fengxiang-flavored liquor 11, Fengxiang-flavored liquor 12, sesame-flavored liquor 1, sesame-flavored liquor 2, sesame-flavored liquor 3, and unknown-flavored liquor 1, unknown-flavored liquor 2, and unknown-flavored liquor 3.

[0021] The sample analysis steps are as follows: 1. Pre-treatment of liquor samples: The finished wine was diluted into 45 ml of 10% vol diluted wine sample according to the dilution ratio, and about 12 g of NaCl and 12 ml of dichloromethane were added. The mixture was vortex-mixed on a vortex mixer for 5 minutes. The mixture was poured into a separatory funnel and allowed to stand for 7 minutes. The lower organic phase was collected in a centrifuge tube, and pivalic acid was added as an internal standard to a final concentration of 50 μg / ml. 1 g of anhydrous sodium sulfate was added and the mixture was dried at -20°C overnight.

[0022] 2. Determination of D-arabinopyranose and 5-methyl-2-acetylfuran: The wine sample was placed at -20°C and dried overnight. The next day, the sample was taken out of the refrigerator and concentrated to 1 ml using a nitrogen blowdown device. The sample was filtered through a 0.22 μm organic filter and then analyzed by GC-MS.

[0023] The gas chromatography conditions were as follows: Inert Cap Wax column (30 m×0.25 mm×0.25 μm), inlet temperature of 250°C, carrier gas He, flow rate of 1 mL / min, splitless mode, injection volume of 1 μL, temperature program: 50°C for 2 min, heating to 250°C at a rate of 6°C / min, and holding for 3 min.

[0024] The mass spectrometry conditions were as follows: EI ionization source, electron energy 70 eV, ion source temperature 240° C., mass scan range 35-350 amu, and solvent delay time 3 min.

[0025] Under these analytical conditions, the contents of D-arabinopyranose and 5-methyl-2-acetylfuran in the above-mentioned liquor were obtained, and it was found that the ratio of D-arabinopyranose to 5-methyl-2-acetylfuran changed significantly, as shown in Table 1.

[0026] Table 1 Ratio of D-arabinopyranose to 5-methyl-2-acetylfuran in liquor samples To validate the method's discriminative capabilities, this study included three baijiu samples of unknown flavor. These samples were analyzed under completely blind conditions, and the final results were compared with the actual flavor data provided by the manufacturer. The results showed that the ratio of D-arabinopyranose to 5-methyl-2-acetylfuran in the unknown flavor samples was below 2. Subsequently, verification by the manufacturer confirmed that the flavor of these unknown flavor samples was indeed phoenix-flavored.

[0027] The present invention uses the ratio of D-arabinopyranose to 5-methyl-2-acetylfuran as a criterion for distinguishing between phoenix-flavor liquor and sesame-flavor liquor. Specifically, a test sample having a ratio of D-arabinopyranose to 5-methyl-2-acetylfuran of 2 or less is considered a phoenix-flavor liquor, while a test sample having a ratio of D-arabinopyranose to 5-methyl-2-acetylfuran of 8 or more is considered a sesame-flavor liquor.

Claims

1. A method for distinguishing Fengxiang-flavor liquor from Zhima-flavor liquor based on metabolomics, characterized by: By detecting the contents of D-arabinopyranose and 5-methyl-2-acetylfuran in liquor and calculating their ratio, the metabolomics study was used to distinguish between Fengxiang-type liquor and sesame-flavor liquor: the liquor with a ratio of D-arabinopyranose to 5-methyl-2-acetylfuran below 2 was Fengxiang-type liquor, and the liquor with a ratio of D-arabinopyranose to 5-methyl-2-acetylfuran above 8 was sesame-flavor liquor.

2. The method for distinguishing Fengxiang-flavor liquor from Zhima-flavor liquor based on metabolomics according to claim 1, characterized in that: When the ratio is between 2 and 8, it is necessary to combine other metabolite indicators or sensory evaluation for auxiliary judgment.

3. The method for distinguishing Fengxiang-flavor liquor from Zhima-flavor liquor based on metabolomics according to claim 2, characterized in that: The specific method for detecting the content of D-arabinopyranose and 5-methyl-2-acetylfuran in liquor is: The liquor samples were pretreated, and then the metabolomics analysis method of vortex-assisted liquid-liquid microextraction combined with gas chromatography-mass spectrometry (VA-LLME-GC-MS) was used to perform absolute quantitative analysis of D-arabinopyranose and 5-methyl-2-acetylfuran in the liquor samples.

4. The method for distinguishing Fengxiang-flavor liquor from Zhima-flavor liquor based on metabolomics according to claim 3, characterized in that: The detection conditions of the gas chromatography are: An Inert Cap Wax column was used, with an inlet temperature of 250°C, carrier gas He, a flow rate of 1 mL / min, splitless mode, an injection volume of 1 μL, and a temperature program of 50°C for 2 min, then increasing the temperature to 250°C at a rate of 6°C / min and holding for 3 min.

5. The method for distinguishing Fengxiang-flavor liquor from Zhima-flavor liquor based on metabolomics according to claim 4, characterized in that: The mass spectrometry conditions are: The EI ionization source had an electron energy of 70 eV, an ion source temperature of 240 °C, a mass scan range of 35–350 amu, and a solvent delay time of 3 min.

6. The method for distinguishing Fengxiang-flavor liquor from Zhima-flavor liquor based on metabolomics according to claim 5, characterized in that: The pre-treatment method of the liquor sample is: The finished wine was diluted into 45 ml of 10% vol diluted wine sample according to the dilution ratio, 12 g of NaCl was added, and 12 ml of dichloromethane was added at the same time. The mixture was vortexed on a vortex mixer for 5 minutes. The mixture was poured into a separatory funnel and allowed to stand for 7 minutes to separate. The lower organic phase was collected in a centrifuge tube, and pivalic acid was added to a final concentration of 50 μg / ml. 1 g of anhydrous sodium sulfate was added and the mixture was dried at -20°C overnight. The next day, the sample was taken out of the refrigerator and concentrated to 1 ml using a nitrogen blowdown device. The sample was filtered through a 0.22 μm organic filter and then analyzed by GC-MS.

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