White spirit identification method based on component dimensions

Abstract

The invention belongs to the field of detection and analysis of liquor, and particularly relates to a liquor identification method based on component dimensions. The method comprises the following steps: (1) determining the content of flavor substances in a standard Baijiu sample; (2) classifying the flavor substances into different types of flavor substances; (3) obtaining the contents of the different types of flavor substances, and determining a boundary value according to the contents of the different types of flavor substances in the standard white spirit sample; and (4) comparing the contents of different types of flavor substances in the to-be-detected wine sample with the boundary value to obtain an identification result of the to-be-detected wine sample. The component dimension-based white spirit identification method provided by the invention has the characteristics of concrete, digitization and large scale, and can be popularized and applied to product batch stability control and true and adulterated spirit identification in the white spirit industry.

Description

Technical Field

[0001] The present invention belongs to the field of wine detection and analysis, and specifically relates to a liquor identification method based on component dimensions. Background Art

[0002] 98% of liquor is water and ethanol, and other flavor substances account for about 2%. This 2% of substance content determines the quality and style of the liquor. These flavor substances include volatile components and non-volatile components, which together constitute the aroma and taste of the liquor. Different types of substances and their quantitative ratios determine the characteristics of different products.

[0003] The present invention obtains flavor substances in liquor by means of gas chromatography, gas chromatography-mass spectrometry, high performance liquid chromatography, and ultra-high performance liquid chromatography-high resolution mass spectrometry detection and analysis instruments. Then, according to the chemical properties of the substances, they are divided into acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines. The total content of the liquor reference sample in each dimension is normalized to construct a liquor material component profile map. The liquor reference sample is used as a reference to calculate the similarity values ​​of a batch of unknown wine samples. Professionals such as national judges and provincial judges determine the similarity threshold values ​​of each dimension from the senses, and then the batch stability of the wine samples is controlled according to the threshold values. For unqualified wine bodies, optimization and adjustment can be made in the dimensions of specific differences, thereby ensuring the batch stability of the product.

[0004] In the liquor identification method based on component dimension provided by the present invention, the detected components include aroma substances and flavor substances, and the evaluation results are more in line with the sensory requirements for aroma and taste. Compared with patents CN113341020A, CN117517521A and CN117805290A, which only use the aroma contribution value (OAV>1) of aroma and flavor substances to measure the similarity of liquor, it is more representative. The liquor identification method based on component dimension provided by the present invention has the characteristics of concreteness, digitization and scale, and can be promoted and applied to product batch stability control and genuine and fake liquor identification in the liquor industry. Summary of the invention

[0005] The invention provides a wine identification method, namely identifying the wine according to the composition of flavor substances, wherein the flavor substances include acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines.

[0006] In some embodiments, the flavor substances further include flavor substances other than the acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines.

[0007] In some embodiments, the acids include acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, lactic acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, oleic acid, gallic acid, ferulic acid, catechin, caffeic acid, chlorogenic acid, p-coumaric acid, vanillic acid, syringic acid, L-tartaric acid, L-malic acid, citric acid, glyceric acid, luteolin, maleic acid, or succinic acid.

[0008] In some embodiments, the esters include ethyl acetate, ethyl lactate, ethyl formate, ethyl butyrate, ethyl isovalerate, isopentyl acetate, ethyl valerate, ethyl hexanoate, ethyl hexanoate, ethyl hexanoate, butyl hexanoate, ethyl octanoate, ethyl phenylacetate, ethyl palmitate, ethyl oleate, ethyl linoleate, isobutyl acetate, ethyl 2-methylbutyrate, hexyl acetate, ethyl nonanoate, hexyl hexanoate, ethyl decanoate, diethyl succinate, ethyl undecanoate, phenylethyl acetate, ethyl dodecanoate, ethyl phenylpropionate, ethyl tridecanoate, ethyl tetradecanoate, ethyl pentadecanoate, ethyl 2-hydroxybutyrate, ethyl DL-2-hydroxyhexanoate, ethyl levulinate, ethyl furoate, ethyl 11-hexadecenoate, ethyl heptadecanoate, ethyl octadecanoate, or ethyl benzoate.

[0009] In some embodiments, the alcohol comprises methanol, propanol, sec-butanol, isobutanol, 2-pentanol, butanol, isopentanol, pentanol, hexanol, 2,3-butanediol, 1,2-propylene glycol, β-phenylethanol, 3-octanol, 1-octen-3-ol, octanol, nonanol, 2-furfuryl alcohol, benzyl alcohol, phenylethanol, 3-hexanol, 4-methylpentanol or 2-nonyl alcohol.

[0010] In some embodiments, the aldehydes include acetaldehyde, propionaldehyde, isobutyraldehyde, acetal, isovaleraldehyde, furfural, or 5-methyl-2-furfural.

[0011] In some embodiments, the ketone comprises 2-pentanone, 3-hydroxy-2-butanone, 2-nonanone, 2-undecanone, 2-tridecanone, or 2-heptanone.

[0012] In some embodiments, the phenols include guaiacol, 4-methylguaiacol, phenol, 4-ethylguaiacol, 4-methylphenol, or 4-ethylphenol.

[0013] In some embodiments, the pyrazines include 2-methylpyrazine, 2-ethylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2-ethyl-6-methylpyrazine, 2,3-dimethylpyrazine, trimethylpyrazine, 2,3-diethylpyrazine, 2,3-dimethyl-5-ethylpyrazine, or tetramethylpyrazine.

[0014] In some embodiments, the flavor substances other than the acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines include diethoxymethane, 2-acetyl-5-methylfuran, 1,1-diethoxymethane-2-methylbutane, 1,1-diethoxymethane-3-methylbutane, 2-(diethoxymethyl)furan, 2-acetylfuran, dimethyl disulfide, or dimethyl trisulfide.

[0015] In some embodiments, the present invention provides a method for identifying wine, comprising the following steps:

[0016] (1) Determining the content of the flavor substances in the standard liquor sample;

[0017] (2) classifying the flavor substances into different categories of flavor substances;

[0018] (3) obtaining the contents of the different types of flavor substances, and determining the threshold values ​​according to the contents of the different types of flavor substances in the standard liquor sample;

[0019] (4) Comparing the contents of different types of flavor substances in the wine sample to be tested with the threshold value to obtain an identification result of the wine sample to be tested.

[0020] In some embodiments, in step (3), the ratios of the contents of the different categories of flavor substances in the standard liquor sample to the total contents of the different categories of flavor substances in the standard liquor sample, i.e., the standard similarity value, are obtained respectively, and the standard similarity value is set to 1.00.

[0021] In some embodiments, the ratio of the smaller value to the larger value of the content of the different categories of flavor substances in the test wine sample and the total content of the different categories of flavor substances in the standard liquor sample is expressed as a sample similarity value.

[0022] In some embodiments, when the sample similarity values ​​of the different categories of flavor substances of the wine sample to be tested are all within the range of 0.8 to 1.00, the wine sample to be tested can be determined to be a qualified wine.

[0023] In some embodiments, when the sample similarity value of one or more of the different categories of flavor substances in the wine sample to be tested is less than 0.8, the wine sample to be tested can be determined to be unqualified wine.

[0024] In some embodiments, when the sample similarity values ​​of the different categories of flavor substances of the wine sample to be tested are all within the range of 0.85 to 1.00, the wine sample to be tested can be determined to be a qualified wine.

[0025] In some embodiments, when the sample similarity value of one or more of the different categories of flavor substances in the wine sample to be tested is less than 0.85, the wine sample to be tested can be determined to be unqualified wine.

[0026] In some embodiments, when the sample similarity values ​​of the different categories of flavor substances of the wine sample to be tested are all within the range of 0.90 to 1.00, the wine sample to be tested can be determined to be a qualified wine.

[0027] In some embodiments, when the sample similarity value of one or more of the different categories of flavor substances in the wine sample to be tested is less than 0.90, the wine sample to be tested can be determined to be unqualified wine.

[0028] In some embodiments, when the sample similarity values ​​of the different categories of flavor substances of the wine sample to be tested are all within the range of 0.92 to 1.00, the wine sample to be tested can be determined to be a qualified wine.

[0029] In some embodiments, when the sample similarity value of one or more of the different categories of flavor substances in the wine sample to be tested is less than 0.92, the wine sample to be tested can be determined to be unqualified wine.

[0030] In some embodiments, the step (4) of comparing the contents of different types of flavor substances in the wine sample to be tested with the threshold value further includes comparing them in the form of a material component profile diagram.

[0031] In some embodiments, the substance component profile is a radar chart constructed with acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines as dimensions.

[0032] In some embodiments, the numerical value of each dimension of the material component profile diagram is expressed as the similarity value of the wine sample in the category dimension.

[0033] In some embodiments, the maximum value of each dimension in the material component profile diagram is 1.

[0034] In some embodiments, the maximum value of each dimension in the material component profile map is the standard similarity value.

[0035] In some embodiments, the material composition profile map also includes a similarity threshold.

[0036] In some embodiments, the flavor substances in step (1) are detected by gas chromatography, gas chromatography-mass spectrometry, high performance liquid chromatography, or ultra performance liquid chromatography-high resolution mass spectrometry.

[0037] In some embodiments, the flavor substances detected by the gas chromatography include acetaldehyde, propionaldehyde, isobutyraldehyde, ethyl formate, diethoxymethane, ethyl acetate, acetal, methanol, isovaleraldehyde, 2-pentanone, sec-butanol, ethyl butyrate, propanol, ethyl isovalerate, isobutanol, isoamyl acetate, ethyl valerate, 2-pentanol, butanol, isoamyl alcohol, ethyl hexanoate, amyl alcohol, 3-hydroxy-2-butanone, ethyl heptanoate, ethyl lactate, hexanol, butyl hexanoate, ethyl octanoate, acetic acid, furfural, propionic acid, isobutyric acid, 2,3-butanediol, 1,2-propylene glycol, butyric acid, isovaleric acid, valeric acid, ethyl phenylacetate, hexanoic acid, β-phenylethanol, heptanoic acid, octanoic acid, ethyl palmitate, ethyl oleate or ethyl linoleate.

[0038] In some embodiments, the flavor substances detected by the gas chromatography-mass spectrometry include isobutyl acetate, ethyl 2-methylbutyrate, hexyl acetate, 2-nonanone, 3-octanol, 1-octen-3-ol, benzaldehyde, ethyl nonanoate, octanol, 5-methyl-2-furfural, 2-undecanone, hexyl hexanoate, 2-acetyl-5-methylfuran, ethyl decanoate, phenylacetaldehyde, nonanol, 2-furfuryl alcohol, diethyl succinate, ethyl undecanoate, 2-tridecanone, phenylethyl acetate, ethyl dodecanoate, benzyl alcohol, ethyl phenylpropionate, phenylethyl alcohol, ethyl tridecanoate, ethyl tetradecanoate, ethyl pentadecanoate, 1,1-diethoxymethane-2-methylbutane, 1,1-diethoxymethane-3-methylbutane, 2-heptanone, 3-hexanol , 4-methylpentanol, 2-ethyl-6-methylpyrazine, ethyl 2-hydroxybutyrate, 2-(diethoxymethyl)furan, 2-acetylfuran, 2-nonyl alcohol, ethyl DL-2-hydroxyhexanoate, ethyl levulinate, ethyl furoate, ethyl 11-hexadecenoate, ethyl heptadecanoate, ethyl octadecanoate, dimethyl disulfide, 2-methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2-ethylpyrazine, 2,3-dimethylpyrazine, dimethyl trisulfide, trimethylpyrazine, 2,3-diethylpyrazine, 2,3-dimethyl-5-ethylpyrazine, tetramethylpyrazine, ethyl benzoate, guaiacol, 4-methylguaiacol, phenol, 4-ethylguaiacol, 4-methylphenol, or 4-ethylphenol.

[0039] In some embodiments, the flavor detected by the liquid chromatography includes lactic acid.

[0040] In some embodiments, the flavor substances detected by the ultra performance liquid chromatography-high resolution mass spectrometry include lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, oleic acid, gallic acid, ferulic acid, catechin, caffeic acid, chlorogenic acid, p-coumaric acid, vanillic acid, syringic acid, L-tartaric acid, L-malic acid, citric acid, glyceric acid, luteolin, maleic acid or succinic acid.

[0041] In some embodiments, the detection conditions of the gas chromatography are: gas chromatography column, chromatographic column heating program: initial temperature 20-50°C, hold for 1-10 min, increase to 100°C at 3.5°C / min, increase to 200°C at 7°C / min, hold for 5-30 min; injection port temperature 200-300°C; detector temperature 200-300°C; injection volume 1.0-5.0 μL; split ratio 5-60:1; column flow 0.5-2.0 mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35.

[0042] In some embodiments, the detection conditions of the gas chromatography are: DB-WAX UI chromatographic column, chromatographic column heating program: initial temperature 30-40°C, hold for 2-5 min, increase to 100°C at 3.5°C / min, increase to 200°C at 7°C / min, hold for 10-20 min; injection port temperature 230-260°C; detector temperature 240-260°C; injection volume 1.0-3.0 μL; split ratio 10-50:1; column flow 0.6-1.2 mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35.

[0043] In some embodiments, the detection conditions of the gas chromatography are: DB-WAX UI chromatographic column (30m×0.25mm×0.25μm), chromatographic column heating program: initial temperature 35°C, hold for 3min, increase to 100°C at 3.5°C / min, increase to 200°C at 7°C / min, hold for 15min; injection port temperature 250°C; detector temperature 250°C; injection volume 1.0μL; split ratio 30:1; column flow 0.8mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35.

[0044] In some embodiments, the substances detected by the gas chromatography are qualitatively identified using retention indices and standard substances.

[0045] In some embodiments, the substances detected by the gas chromatography are quantified using an internal standard method combined with peak area.

[0046] In some embodiments, the wine sample processing method before gas chromatography detection is to add an internal standard solution to the wine sample, vortex mix, filter, and prepare for gas chromatography detection.

[0047] In some embodiments, the vortex mixing time is 0.5 to 5 minutes.

[0048] In some embodiments, the vortex mixing time is 1 to 3 minutes.

[0049] In some embodiments, the filtering is performed using a filter membrane.

[0050] In some embodiments, the filter membrane is a 0.22 μm microporous filter membrane.

[0051] In some embodiments, the internal standard solution includes tert-amyl alcohol, n-amyl acetate, and 2-ethylbutyric acid.

[0052] In some embodiments, the final concentration of tert-amyl alcohol is 100.00-500.00 mg / L.

[0053] In some embodiments, the final concentration of tert-amyl alcohol is 389.08 mg / L.

[0054] In some embodiments, the final concentration of n-pentyl acetate is 100.00-500.00 mg / L.

[0055] In some embodiments, the final concentration of n-pentyl acetate is 382.25 mg / L.

[0056] In some embodiments, the final concentration of 2-ethylbutyric acid is 100.00 to 500.00 mg / L.

[0057] In some embodiments, the final concentration of 2-ethylbutyrate is 396.36 mg / L.

[0058] In some embodiments, the detection conditions of the gas chromatography-mass spectrometry are as follows: (1) Gas chromatography conditions: gas chromatography column; injection port temperature 200-300°C, carrier gas is He, flow rate 0.5-2.0 mL / min, non-split mode; temperature program: start at 20-50°C, hold for 0.5-10 min, then heat to 100°C at 2.5°C / min, hold for 0.5-10 min; then heat to 160°C at 3°C / min, do not hold; then heat to 230°C at 5°C / min, hold for 5-60 min; (2) Mass spectrometry conditions: transmission line temperature 200-300°C; EI ion source; ion source temperature: 200-300°C; electron energy: 10-100 eV; quadrupole temperature 100-200°C.

[0059] In some embodiments, the detection conditions of the gas chromatography-mass spectrometry are as follows: (1) gas chromatography conditions: capillary column; injection port temperature 230-260°C, carrier gas is He, flow rate 0.8-1.2 mL / min, non-split mode; temperature program: start at 30-45°C, hold for 1-3 min, then heat to 100°C at 2.5°C / min, hold for 1-3 min; then heat to 160°C at 3°C / min, do not hold; then heat to 230°C at 5°C / min, hold for 5-30 min; (2) mass spectrometry conditions: transmission line temperature 230-260°C; EI ion source; ion source temperature: 230-250°C; electron energy: 20-80 eV; quadrupole temperature 140-160°C.

[0060] In some embodiments, the detection conditions of the gas chromatography-mass spectrometry are as follows: (1) gas chromatography conditions: DB-FFAP capillary column (60m×0.25mm×0.25μm); injection port temperature 250°C, carrier gas is He, flow rate 1mL / min, non-split mode; temperature program: start at 40°C, hold for 1min, then increase the temperature to 100°C at 2.5°C / min, hold for 1min; then increase the temperature to 160°C at 3°C / min, do not hold; then increase the temperature to 230°C at 5°C / min, hold for 20min; (2) mass spectrometry conditions: transmission line temperature 250°C; EI ion source; ion source temperature: 230°C; electron energy: 70eV; quadrupole temperature 150°C.

[0061] In some embodiments, the substances detected by the gas chromatography-mass spectrometry are qualitatively identified using a spectral library, retention indexes, and standard substances.

[0062] In some embodiments, the substances detected by the gas chromatography-mass spectrometry are quantified using an internal standard method combined with peak area.

[0063] In some embodiments, the method for processing the wine sample before detection by gas chromatography-mass spectrometry is to add saturated salt water to the wine sample, then add an internal standard solution, and then add an extraction liquid to perform liquid-liquid extraction, vortex, stand and separate the layers, take the upper organic liquid, and prepare for detection on the machine.

[0064] In some embodiments, the extract comprises diethyl ether and n-hexane.

[0065] In some embodiments, the composition of the extract is diethyl ether:n-hexane (v:v)=1:1.

[0066] In some embodiments, the vortexing time is 0.5 to 5 minutes.

[0067] In some embodiments, the vortexing time is 1 to 3 minutes.

[0068] In some embodiments, the internal standard solution selected for the internal standard method includes 2-octanol and 2-methoxy-3-methylpyrazine.

[0069] In some embodiments, the final concentration of 2-octanol is 1.00-100.00 mg / L.

[0070] In some embodiments, the final concentration of 2-octanol is 11.80 mg / L.

[0071] In some embodiments, the final concentration of 2-methoxy-3-methylpyrazine is 1.00 to 100.00 mg / L.

[0072] In some embodiments, the final concentration of 2-methoxy-3-methylpyrazine is 10.34 mg / L.

[0073] In some embodiments, the diethyl ether and n-hexane are both of chromatographic grade.

[0074] In some embodiments, the detection conditions of the HPLC are: liquid chromatography column; mobile phase: KH2PO4, pH 2.0-3.0; detection wavelength 208nm, column temperature 20-50°C, mobile phase flow rate 0.5-2.0mL / min, injection volume 1-5μL.

[0075] In some embodiments, the detection conditions of the liquid chromatography are: reverse phase chromatography column; mobile phase: 0.01-0.05 mol / L KH2PO4, pH 2.0-2.5; detection wavelength 208 nm, column temperature 30-40°C, mobile phase flow rate 0.8-1.2 mL / min, injection volume 1-3 μL.

[0076] In some embodiments, the detection conditions of the liquid chromatography are: ZORBAXSB-C 18 chromatographic column (4.6mm×150mm×5μm); mobile phase: 0.02mol / L KH2PO4, pH 2.3; detection wavelength 208nm, column temperature 35°C, mobile phase flow rate 1mL / min, injection volume 1μL.

[0077] In some embodiments, the substance detected by the HPLC is qualitatively identified based on standard substances, specific wavelengths, and retention times.

[0078] In some embodiments, the substance detected by the high performance liquid chromatography is quantified using an external standard method.

[0079] In some embodiments, the method for processing wine samples before testing by high performance liquid chromatography is to filter an appropriate amount of wine samples in preparation for testing on the machine.

[0080] In some embodiments, the filtering is performed using a filter membrane.

[0081] In some embodiments, the filter membrane is a 0.22 μm microporous filter membrane.

[0082] In some embodiments, the detection conditions of the ultra-high performance liquid chromatography-high resolution mass spectrometry are as follows: (1) ultra-high performance liquid chromatography conditions: liquid chromatography column; column temperature 20-50°C, injection volume 1-5 μL, flow rate 0.05-2 mL / min, analysis time 5-60 min, mobile phase A: formic acid (containing ammonium formate at a final concentration of 0.1-10 mmol / L), mobile phase B: pure methanol; gradient elution conditions: 0-4.0 min, 100% A (the percentage refers to the volume percentage of mobile phase A); 4. 0~8.0min, 100%~5%A; 8.0~15.0min, 5%A; 15.0~15.1min, 5%~100%A; 15.1~20.0min, 100%A; (2) High-resolution mass spectrometry conditions: heated electrospray ionization source (HESI); sheath gas flow rate: 20~50arb; auxiliary gas flow rate: 5~20arb; spray voltage: 2.0~5.0kV; ion source temperature: 300~400℃; capillary temperature: 300~400℃.

[0083] In some embodiments, the detection conditions of the ultra-high performance liquid chromatography-high resolution mass spectrometry are as follows: (1) ultra-high performance liquid chromatography conditions: Hypersil Gold liquid chromatography column; column temperature 35-45°C, injection volume 2-5 μL, flow rate 0.25-1 mL / min, analysis time 10-40 min, mobile phase A: 0.1-1% formic acid (containing final concentration 0.5-5 mmol / L ammonium formate), mobile phase B: pure methanol; gradient elution conditions: 0-4.0 min, 100% A; 4.0-8.0 min, 100%-5% A; 8.0-15.0 min, 100%-20% A; 10.0-25.0 min, 100%-30% A; 10.0-25.0 min, 100%-50% A; 10.0-25.0 min, 100%-20% A; 10.0-25.0 min, 100%-3 .... 0min, 5% A; 15.0-15.1min, 5%-100% A; 15.1-20.0min, 100% A; (2) High-resolution mass spectrometry conditions: heated electrospray ion source (HESI); sheath gas flow rate: 30-45arb; auxiliary gas flow rate: 5-10arb; spray voltage: 2.6-3.5kV; ion source temperature: 320-350°C; capillary temperature: 300-350°C.

[0084] In some embodiments, the detection conditions of the ultra-high performance liquid chromatography-high resolution mass spectrometry are: (1) ultra-high performance liquid chromatography conditions: Hypersil Gold (150mm×2.1mm, 1.9μm) liquid chromatography column; column temperature 40℃, injection volume 2μL, flow rate 0.25mL / min, analysis time 20min, mobile phase A: 0.1% formic acid (containing final concentration 2mmoL / L ammonium formate), mobile phase B: pure methanol; gradient elution conditions: 0~4.0min, 100%A; 4.0~8.0min, 100%~5%A; 8.0~15.0min, 5%A; 15.0~15.1min, 5%~100%A; 15.1~20.0min, 100%A; (2) high-resolution mass spectrometry conditions: heated electrospray ionization source (HESI); sheath gas flow rate: 38arb; auxiliary gas flow rate: 9arb; spray voltage: 3.2kV; ion source temperature: 345℃; capillary temperature: 320℃.

[0085] In some embodiments, the volume percentage of mobile phase A refers to the volume percentage of mobile phase A in the mixture of mobile phase A and mobile phase B.

[0086] In some embodiments, the substances detected by the ultra-high performance liquid chromatography-high resolution mass spectrometry are qualitatively identified using characteristic ions and peak times of standard substances.

[0087] In some embodiments, the substances detected by the ultra-high performance liquid chromatography-high resolution mass spectrometry are quantified using an external standard method combined with peak area.

[0088] In some embodiments, the method for processing the wine sample before detection by ultra-performance liquid chromatography-high-resolution mass spectrometry is to concentrate the wine sample with nitrogen, take it out after concentration, cool it to room temperature, make up to volume with pure methanol, vortex mix it, and filter it to obtain the sample to be tested.

[0089] In some embodiments, the vortex mixing time is 0.5 to 5 minutes.

[0090] In some embodiments, the vortex mixing time is 1 to 3 minutes.

[0091] In some embodiments, the filtration is performed using a 0.22 μm organic PES syringe filter membrane.

[0092] In some embodiments, the pure methanol is of chromatography grade.

[0093] The present invention also provides an application of a wine identification method in improving the quality of wine, evaluating the quality of wine, optimizing the liquor preparation process, identifying the quality of liquor, or controlling the stability of liquor batches.

[0094] In some embodiments, the identification of the wine sample to be tested includes comparing the similarity value of each dimension of the wine sample to be tested with the similarity threshold value of each dimension.

[0095] In some embodiments, the identification of the wine sample to be tested also includes comparing similarity values ​​in combination with the material component profile map.

[0096] In some implementation schemes, the similarity threshold of each dimension is 0.80-1.00.

[0097] In some implementation schemes, the similarity threshold of each dimension is 0.85-1.00.

[0098] In some implementation schemes, the similarity threshold of each dimension is 0.90-1.00.

[0099] In some implementation schemes, the similarity threshold of each dimension is 0.92-1.00.

[0100] In some embodiments, the similarity threshold of each dimension is 0.92.

[0101] In some embodiments, whether some flavor substances (such as lactic acid and propanol) are included in the corresponding dimensions has a great impact on the results of liquor identification, which means that the liquor identification method based on the component dimension provided by the present invention does not simply include as many flavor substances as possible into the component dimension, but selectively includes flavor substances that contribute to more accurate identification of liquor, that is, the combination of flavor substances selected for detection by the present invention is creative.

[0102] In some embodiments, the absence of individual component dimensions (such as phenolic component dimensions) has a significant negative impact on the results of liquor identification, which means that the combination of component dimensions set in the present invention can achieve unexpectedly good technical effects.

[0103] In some embodiments, in the liquor identification method based on component dimension provided by the present invention, the detected components include aroma substances and flavor substances, and the evaluation results are more in line with the sensory requirements for aroma and taste. Compared with patents CN113341020A, CN117517521A and CN117805290A, which only use the aroma contribution value (OAV>1) of aroma and flavor substances to measure the similarity of liquor, they are more representative. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] Figure 1 It is a profile diagram of each batch of wine samples in Example 1 of the present application.

[0105] Figure 2 This is a comparison of the evaluation results of liquor samples with and without lactic acid and propanol included in the corresponding dimensions in Comparative Example 1 of the present application.

[0106] Figure 3 This is a comparison of the evaluation results of liquor samples with and without the phenolic dimension included in the identification in Comparative Example 2 of the present application.

[0107] Figure 4 It is a profile diagram of each identification sample in Example 2 of the present application. DETAILED DESCRIPTION

[0108] The technical solution of the present invention is further described below by specific embodiments, which do not limit the protection scope of the present invention. Some non-essential modifications and adjustments made by others based on the concept of the present invention still fall within the protection scope of the present invention.

[0109] In this article, the professional terms in this field are explained as follows:

[0110] The similarity value refers to a value that measures the similarity between two or more objects. Usually, a reference object is set first, the ratio between the reference object and the reference object is calculated, and the value is obtained by normalization.

[0111] Flavor substances refer to the components in food that can make people's sensory organs perceive special aroma and taste.

[0112] In the embodiments of this article, "dimension" refers to classification according to chemical properties, and each classification is called a "dimension". For example, the material components detected in the wine samples in this article are divided into 8 category dimensions according to their chemical properties: "acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines."

[0113] In the embodiments of this article, the similarity value of each dimension is expressed by the normalized numerical value of each dimension, and the two have the same essential meaning.

[0114] In the embodiment of this article, the similarity threshold is the similarity value of each dimension of the wine sample to be tested when professional tasters cannot identify the difference between the wine sample to be tested and the standard liquor sample.

[0115] In the embodiments of this invention, the boundary lines appearing in Table 8, Table 10, and Table 11 are equivalent to the similarity boundary values.

[0116] In the drawings of this specification, in the description of each category dimension, "others" is used to refer to "flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines."

[0117] Herein, the "standard similarity value" refers to the ratio of the content of the different categories of flavor substances in the standard liquor sample to the total content of the different categories of flavor substances in the standard liquor sample, that is, the standard similarity value, and the standard similarity value is set to 1.00.

[0118] Herein, "sample similarity value" refers to the content of the different categories of flavor substances in the test wine sample being called A, the total content of the different categories of flavor substances in the standard liquor sample being called M, and the ratio between the smaller value of M and A and the larger value being the sample similarity value; if the content of the acid of the flavor substance in the test wine sample is A1, and its value is less than M, then the sample similarity value of the acid in the test wine sample is A1 / M, and so on.

[0119] Example 1: Qualification of wine samples to be shipped

[0120] Taking 10 batches of samples of the same type of wine to be packaged from a certain sauce wine company as an example, the specific implementation process is as follows:

[0121] 1. Reagents and Materials:

[0122] (1) Reagents:

[0123] Organic solvents: ethanol, methanol, formic acid, ether, and n-hexane, all of chromatographic grade, from Merck, Germany;

[0124] Internal standard substances: tert-amyl alcohol, n-amyl acetate, 2-ethylbutyric acid, 2-octanol, 2-methoxy-3-methylpyrazine (purity ≥ 99%): Shanghai Anpu Laboratory Technology Co., Ltd.;

[0125] Standards: Standards tested by gas chromatography, gas chromatography-mass spectrometry, liquid chromatography, and liquid chromatography-mass spectrometry were purchased from Sinopharm Chemical Reagent Co., Ltd., Sigma-Aldrich Chemical Reagent Company, and Shanghai Yuanye Biotechnology Co., Ltd.

[0126] (2) Materials: Qualified AT Xijiu as the control sample (AT-SD); 10 batches of AT Xijiu to be shipped (AT-1, AT-2, AT-3, AT-4, AT-5, AT-6, AT-7, AT-8, AT-9, AT-10) were sourced from Guizhou Xijiu Co., Ltd.

[0127] 2. Main equipment and instruments

[0128] Agilent 8890A gas chromatograph (Agilent, USA); Agilent 8890A gas chromatograph-5977B mass spectrometer (Agilent, USA); 1200 Infinity Series high performance liquid chromatograph (Agilent, USA); UItiMate 3000 ultra-high performance liquid phase-Q EXACTIVE Focus high-resolution mass spectrometer (ThermoFisher Scientific, USA); PL3002 electronic balance (Mettler Toledo, Switzerland); SI-T256 adjustable speed timed vortex mixer (Scientific Industries, USA); Aquaplore 3S ultrapure water system (Aquapro International, Taiwan, China); AutoEVA-20Plus fully automatic balance concentrator (Creator Group Co., Ltd.).

[0129] 3. Detection Methods

[0130] 3.1 Gas chromatography detection method

[0131] 3.1.1 Pretreatment of wine samples

[0132] Take 10mL of wine sample into a volumetric flask, add 100μL of internal standard solution prepared with 53% vol ethanol solution (final concentration: tert-amyl alcohol 389.08mg / L; n-amyl acetate 382.25mg / L; 2-ethylbutyric acid 396.36mg / L), vortex mix for 1min, draw 1.5mL with a syringe, pass through a 0.22μm microporous filter membrane into a brown injection vial, cover the bottle cap, and wait for detection on the machine.

[0133] 3.1.2 Gas chromatography detection conditions: DB-WAX UI chromatographic column (30m×0.25mm×0.25μm), column heating program: initial temperature 35℃, hold for 3min, increase to 100℃ at 3.5℃ / min, increase to 200℃ at 7℃ / min, hold for 15min.

[0134] Inlet temperature 250℃; detector temperature 250℃; injection volume 1.0μL; split ratio 30:1; column flow 0.8mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35.

[0135] 3.1.3 Qualitative and quantitative analysis of substances

[0136] (1) Qualitative analysis: Qualitative analysis is performed using retention index and standard substances.

[0137] (2) Quantification: Quantification was performed using the internal standard method combined with peak area.

[0138] 3.1.4 Test substances

[0139] Acetaldehyde, propionaldehyde, isobutyraldehyde, ethyl formate, diethoxymethane, ethyl acetate, acetal, methanol, isovaleraldehyde, 2-pentanone, sec-butyl alcohol, ethyl butyrate, propanol, ethyl isovalerate, isobutyl alcohol, isoamyl acetate, ethyl valerate, 2-pentanol, butanol, isoamyl alcohol, ethyl hexanoate, amyl alcohol, 3-hydroxy-2-butanone, ethyl heptanoate, ethyl lactate, hexanol, butyl hexanoate, ethyl octanoate, acetic acid, furfural, propionic acid, isobutyric acid, 2,3-butanediol, 1,2-propylene glycol, butyric acid, isovaleric acid, valeric acid, ethyl phenylacetate, hexanoic acid, β-phenylethanol, heptanoic acid, octanoic acid, ethyl palmitate, ethyl oleate, ethyl linoleate.

[0140] 3.2 Gas chromatography-mass spectrometry detection method

[0141] 3.2.1 Wine sample pretreatment

[0142] Accurately pipette 2 mL of wine sample into a 25 mL test tube, add 8.6 mL of saturated saline, add 100 μL of internal standard solution prepared with 53% vol ethanol solution (final concentration: 2-octanol 11.80 mg / L; 2-methoxy-3-methylpyrazine 10.34 mg / L), add 2 mL of extraction solution (ether: n-hexane = 1:1 (volume ratio)) for liquid-liquid extraction, vortex for 2 min, let stand to separate layers, use a pipette to pipette the upper organic phase into a brown injection vial, cover the bottle cap, and wait for detection on the machine.

[0143] 3.2.2 Gas chromatography-mass spectrometry detection conditions

[0144] (1) Gas chromatography conditions: DB-FFAP capillary column (60 m × 0.25 mm × 0.25 μm); injection port temperature 250 °C, carrier gas He, flow rate 1 mL / min, splitless mode; temperature program: start at 40 °C, hold for 1 min, then increase the temperature to 100 °C at 2.5 °C / min, hold for 1 min; then increase the temperature to 160 °C at 3 °C / min, do not hold; then increase the temperature to 230 °C at 5 °C / min, hold for 20 min.

[0145] (2) Mass spectrometry conditions: transfer line temperature 250°C; EI ion source; ion source temperature: 230°C; electron energy: 70 eV; quadrupole temperature 150°C.

[0146] 3.2.3 Qualitative and quantitative analysis of substances

[0147] (1) Qualitative analysis: The NIST 20 mass spectrometry database, retention index, and standard substances were used for qualitative analysis.

[0148] (2) Quantification: Quantification was performed using the internal standard method combined with peak area.

[0149] 3.2.4 Test substances

[0150] Isobutyl acetate, ethyl 2-methylbutyrate, hexyl acetate, 2-nonanone, 3-octanol, 1-octen-3-ol, benzaldehyde, ethyl nonanoate, octanol, 5-methyl-2-furfural, 2-undecanone, hexyl hexanoate, 2-acetyl-5-methylfuran, ethyl decanoate, phenylacetaldehyde, nonanol, 2-furfuryl alcohol, diethyl succinate, ethyl undecanoate, 2-tridecanone, phenylethyl acetate, ethyl dodecanoate, benzyl alcohol, ethyl phenylpropionate, phenylethyl alcohol, ethyl tridecanoate, ethyl tetradecanoate, ethyl pentadecanoate, 1,1-diethoxymethane-2-methylbutane, 1,1-diethoxymethane-3-methylbutane, 2-heptanone, 3-hexanol, 4-methylpentanol, 2-ethyl-6- Methylpyrazine, ethyl 2-hydroxybutyrate, 2-(diethoxymethyl)furan, 2-acetylfuran, 2-nonyl alcohol, ethyl DL-2-hydroxyhexanoate, ethyl levulinate, ethyl furoate, ethyl 11-hexadecenoate, ethyl heptadecanoate, ethyl octadecanoate, dimethyl disulfide, 2-methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2-ethylpyrazine, 2,3-dimethylpyrazine, dimethyl trisulfide, trimethylpyrazine, 2,3-diethylpyrazine, 2,3-dimethyl-5-ethylpyrazine, tetramethylpyrazine, ethyl benzoate, guaiacol, 4-methylguaiacol, phenol, 4-ethylguaiacol, 4-methylphenol, 4-ethylphenol.

[0151] 3.3 High performance liquid chromatography detection method

[0152] 3.3.1 Wine sample pretreatment

[0153] Use a syringe to draw 1.5 mL of wine sample, pass it through a 0.22 μm microporous filter membrane and place it in a sample injection vial. Cover the bottle cap and wait for the machine to detect.

[0154] 3.3.2 Liquid chromatography detection conditions

[0155] ZORBAXSB-C 18 chromatographic column (4.6mm×150mm×5μm); mobile phase: 0.02mol / L KH2PO4 aqueous solution, pH 2.3; detection wavelength 208nm, column temperature 35℃, mobile phase flow rate 1mL / min, injection volume 1μL.

[0156] 3.3.3 Qualitative and quantitative analysis of substances

[0157] (1) Qualitative analysis: Qualitative analysis is performed based on standard substances, specific wavelengths, and retention times.

[0158] (2) Quantification: Quantification was performed using the external standard method combined with peak area.

[0159] 3.3.4 Test substances

[0160] Lactic acid.

[0161] 3.4 Ultra-high performance liquid chromatography-high resolution mass spectrometry detection method

[0162] 3.4.1 Wine sample pretreatment

[0163] Accurately pipette 4 mL of Maotai-flavor liquor sample into a 10 mL graduated test tube, blow nitrogen at a temperature of 30°C, an air pressure of 3.5 psi, and a speed of 0.3 mm / min until about 1 mL of liquor remains, take it out and cool it to room temperature, dilute it to 2 mL with pure methanol, vortex mix for 1 min, and filter it through a 0.22 μm organic PES syringe filter membrane to obtain the sample to be tested;

[0164] 3.4.2 Ultra-high performance liquid chromatography-high resolution mass spectrometry detection conditions

[0165] (1) UPLC conditions: Hypersil Gold (150 mm × 2.1 mm, 1.9 μm) liquid chromatography column; column temperature 40°C, injection volume 2 μL, flow rate 0.25 mL / min, analysis time 20 min, mobile phase A: 0.1% formic acid (containing final concentration 2 mmol / L ammonium formate), mobile phase B: pure methanol.

[0166] The elution program is shown in Table 1 below. The volume percentage of mobile phase A refers to the volume percentage of mobile phase A in the mixture of mobile phase A and mobile phase B.

[0167] Table 1 Elution program

[0168] Serial number Time (min) Mobile phase A (v / v%) Mobile phase B (v / v%) 1 0 100 0 2 4.0 100 0 3 8.0 5 95 4 15.0 5 95 5 15.1 100 0 6 20.0 100 0

[0169] (2) High-resolution mass spectrometry conditions: heated electrospray ionization (HESI); sheath gas flow rate: 38 arb; auxiliary gas flow rate: 9 arb; spray voltage: 3.2 kV; ion source temperature: 345°C; capillary temperature: 320°C.

[0170] 3.4.3 Qualitative and quantitative analysis of substances

[0171] (1) Qualitative analysis: Qualitative analysis is performed using the characteristic ions and peak times of standard substances.

[0172] (2) Quantification: Quantification was performed using the external standard method combined with peak area.

[0173] 3.4.4 Test substances

[0174] Lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, oleic acid, gallic acid, ferulic acid, catechin, caffeic acid, chlorogenic acid, p-coumaric acid, vanillic acid, syringic acid, L-tartaric acid, L-malic acid, citric acid, glyceric acid, luteolin, maleic acid, and succinic acid.

[0175] 4. Statistical analysis of wine samples

[0176] (1) The content of the substance components in the wine samples was tested according to the above-mentioned detection method. The test results of the content of the substance components in the control sample and AT-1 to AT-10 are shown in Table 2.

[0177] Table 2 Test results of substance components in wine samples, unit: mg / L

[0178]

[0179]

[0180]

[0181]

[0182] (2) The substance components detected in the wine samples were divided into eight categories according to their chemical properties: acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines. The specific classification is shown in Table 3. The substance component contents of the eight categories of acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines of each wine sample were summarized, and the total substance content of the control sample was calculated. The results are shown in Table 4.

[0183] Table 3 Classification of material components by chemical properties

[0184]

[0185] Table 4 Classification and summary of substance contents in wine samples, unit: mg / L

[0186]

[0187] (3) Calculation of the percentage of substance content in each dimension: The substance content in the eight dimensions of each wine sample was divided by the total substance content of the control sample to obtain the percentage of substance content in each dimension. The results are shown in Table 5.

[0188] Table 5 Ratio of the content of each dimension of the wine sample to the total content of the control sample

[0189]

[0190]

[0191] (4) Normalization: The values ​​of each dimension of AT-1 to AT-10 and the control sample were normalized in the dimension to obtain the normalized data of each dimension. The results are shown in Table 6.

[0192] Table 6 Normalization results of AT-1 to AT-10 and control samples

[0193]

[0194] (5) Normalized value adjustment: adjust the normalized values ​​of each dimension of AT-1 to AT-10: if the normalized value is greater than 1, perform the inverse processing; if the normalized value is less than or equal to 1, the value remains unchanged. The normalized value adjustment results are shown in Table 7.

[0195] Table 7 Normalized value adjustment results of AT-1~AT-10 in each dimension

[0196]

[0197]

[0198] 5. Set the similarity thresholds on each dimension and construct the contour map

[0199] Ten professional tasters (five Chinese national liquor judges and five Guizhou liquor judges, with a male-female ratio of 1:1) used the control sample as a reference to perform sensory identification of AT-1 to AT-10 in various dimensions. The result was that when the similarity of each dimension of the liquor sample was ≥ 0.92, the professional tasters could not identify the difference between the liquor sample and the control sample. Therefore, 0.92 was used as the similarity threshold of each dimension to distinguish the liquor samples. Using the control sample as a reference, the profile map of each batch of liquor samples was constructed, and the results are shown in Table 8 and Figure 1 As shown in the table, the boundary line is equivalent to the similarity boundary.

[0200] Table 8 Normalized value adjustment results of AT-1~AT-10 in each dimension

[0201]

[0202] 6. Evaluation and Adjustment

[0203] Evaluation basis: (1) If the similarity values ​​of the eight dimensions of each batch of wine samples are between 0.92 and 1.00, it indicates that the wine samples are stable and qualified and can be packaged and shipped;

[0204] (2) If the similarity values ​​of each batch of wine samples in one or more dimensions are not between 0.92 and 1.00, it means that there are sensory differences between the wine samples and the control samples in one or more dimensions. In this case, the blending personnel need to make adjustments until the similarity of each dimension is within the range of 0.92 to 1.00, so as to meet the qualified requirements. For example, AT-1 needs to be optimized and adjusted in the "acid" dimension; AT-3 needs to be optimized and adjusted in the "alcohol" dimension; AT-7 and AT-10 need to be optimized and adjusted in the "phenol" dimension.

[0205] Comparative Example 1 Effect of lack of flavoring substances on evaluation results

[0206] Based on Example 1, the similarity values ​​of the corresponding dimensions of each wine sample determined by comparing whether lactic acid and propanol are included in the acid or alcohol dimension are shown in Table 9 and Figure 2 shown.

[0207] The results show that when lactic acid content is included in the acid dimension, the similarity value of AT-1 in the acid dimension is not between 0.92 and 1.00 compared with the standard sample AT-SD, and the wine sample is judged to be unqualified; when lactic acid content is not included in the acid dimension, the similarity of AT-1 in the acid dimension is 0.96, and between 0.92 and 1.00, and the wine sample is judged to be qualified. Similarly, when propanol content is included in the alcohol dimension, the similarity of AT-3 in the alcohol dimension is 0.88 compared with the standard sample AT-SD, and the wine sample is judged to be unqualified; when propanol content is not included in the alcohol, the similarity of AT-3 in the alcohol dimension is 0.96, and the wine sample is judged to be qualified. Therefore, whether some substances are included in the corresponding dimensions for calculation directly affects the judgment results.

[0208] Table 9 Statistical results of comparison of missing flavor substances

[0209]

[0210] Comparative Example 2: Effect of missing phenol dimension on evaluation results

[0211] Based on Example 1, the effect of the lack of phenolic dimension on the similarity evaluation of liquor was compared, and the statistical analysis is shown in Tables 10, 11 and Figure 3 shown.

[0212] As shown in Table 10, when the phenolic dimension is included in the evaluation, compared with the standard sample AT-SD, the similarity of the phenolic dimension of the wine samples of the two batches of AT-7 and AT-10 is not between 0.92 and 1.00, and the wine samples are judged to be unqualified; as shown in Table 11, when the phenolic dimension is not included in the evaluation, compared with the standard sample AT-SD, the similarity of the flavor substances other than acids, esters, alcohols, aldehydes, ketones, pyrazines and flavor substances of the wine samples of the two batches of AT-7 and AT-10 is between 0.92 and 1.00, and the wine samples are judged to be qualified. In fact, there are sensory differences between the wine samples of the two batches of AT-7 and AT-10 and the control samples, which can be judged that the division and selection of each component dimension are indispensable.

[0213] Table 10 Statistical results of phenolic dimensions

[0214]

[0215] Table 11 Unstated results of phenolic dimension

[0216]

[0217] Embodiment 2 True and false discrimination of wine samples in the market

[0218] Materials: 5 market identification samples (JD1, JD2, JD3, JD4, JD5), purchased from the circulation market. Other reagent materials, equipment, instruments, and detection methods are the same as those in Example 1.

[0219] Based on the method of Example 1, market identification samples JD1, JD2, JD3, JD4, and JD5 were tested. Figure 4 It can be seen that the similarity values ​​of JD2's seven categories of acids, esters, aldehydes, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines are not within the range of 0.92 to 1.00, and JD2 is judged to be a counterfeit wine of this product. The similarity values ​​of JD5's seven categories of acids, esters, alcohols, ketones, phenols, pyrazines, and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines are not within the range of 0.92 to 1.00, and JD5 is judged to be a counterfeit wine of this product. This method has the characteristics of visualization, digitization, and concreteness, and is suitable for the authenticity judgment of wine samples circulating in the market.

[0220] Table 12 Similarity value results of wine samples identified in the market

[0221]

[0222] The 10 professionals in Example 1 were further organized to conduct a sensory comparison of eight dimensions of acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines on the control sample and the market identification samples JD1, JD2, JD3, JD4 and JD5. As shown in Table 13, JD2 has sensory differences with the control sample in seven categories of acids, esters, aldehydes, ketones, phenols, pyrazines and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines; JD5 has sensory differences with the control sample in seven categories of acids, esters, alcohols, ketones, phenols, pyrazines and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines; indicating that JD2 and JD5 are counterfeit wines of this product. This result is consistent with the result determined by the method of the present invention in Example 2, which proves that the method of the present invention is reliable and has high accuracy.

[0223] Table 13 Comprehensive results of sensory comparison between market identification wine samples and control samples

[0224]

[0225] Note: “0” indicates no sensory difference compared with the control sample; “*” indicates sensory difference compared with the control sample; “**” indicates significant difference compared with the control sample.

[0226] It is to be understood that the present invention is described by some embodiments, and it is known to those skilled in the art that various changes or equivalent substitutions may be made to these features and embodiments without departing from the spirit and scope of the present invention. In addition, under the teachings of the present invention, these features and embodiments may be modified to adapt to specific circumstances and materials without departing from the spirit and scope of the present invention. Therefore, the present invention is not limited by the specific embodiments disclosed herein, and all embodiments falling within the scope of the claims of this application are within the scope of protection of the present invention.

Claims

1. A method for identifying wine, characterized in that: The wine is identified based on the composition of flavor substances, which include acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines.

2. The identification method according to claim 1, characterized in that The flavor substances also include flavor substances other than the acids, esters, alcohols, aldehydes, ketones, phenols, and pyrazines; Preferably, the acid comprises acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid, caproic acid, heptanoic acid, caprylic acid, lactic acid, lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, oleic acid, gallic acid, ferulic acid, catechin, caffeic acid, chlorogenic acid, p-coumaric acid, vanillic acid, syringic acid, L-tartaric acid, L-malic acid, citric acid, glyceric acid, luteolin, maleic acid or succinic acid; Preferably, the esters include ethyl acetate, ethyl lactate, ethyl formate, ethyl butyrate, ethyl isovalerate, isopentyl acetate, ethyl valerate, ethyl caproate, ethyl hexanoate, ethyl hexanoate, butyl caproate, ethyl caprylate, ethyl phenylacetate, ethyl palmitate, ethyl oleate, ethyl linoleate, isobutyl acetate, ethyl 2-methylbutyrate, hexyl acetate, ethyl nonanoate, hexyl caproate, ethyl decanoate, diethyl succinate, ethyl undecanoate, phenylethyl acetate, ethyl dodecanoate, ethyl phenylpropionate, ethyl tridecanoate, ethyl tetradecanoate, ethyl pentadecanoate, ethyl 2-hydroxybutyrate, ethyl DL-2-hydroxyhexanoate, ethyl levulinate, ethyl furoate, ethyl 11-hexadecenoate, ethyl heptadecanoate, ethyl octadecanoate or ethyl benzoate; Preferably, the alcohol comprises methanol, propanol, sec-butanol, isobutanol, 2-pentanol, butanol, isoamyl alcohol, pentanol, hexanol, 2,3-butanediol, 1,2-propylene glycol, β-phenylethanol, 3-octanol, 1-octen-3-ol, octanol, nonanol, 2-furfuryl alcohol, benzyl alcohol, phenylethyl alcohol, 3-hexanol, 4-methylpentanol or 2-nonyl alcohol; Preferably, the aldehydes include acetaldehyde, propionaldehyde, isobutylaldehyde, acetal, isovaleraldehyde, furfural or 5-methyl-2-furfural; preferably, the ketones include 2-pentanone, 3-hydroxy-2-butanone, 2-nonanone, 2-undecanone, 2-tridecanone or 2-heptanone; Preferably, the phenols include guaiacol, 4-methylguaiacol, phenol, 4-ethylguaiacol, 4-methylphenol or 4-ethylphenol; Preferably, the pyrazines include 2-methylpyrazine, 2-ethylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2-ethyl-6-methylpyrazine, 2,3-dimethylpyrazine, trimethylpyrazine, 2,3-diethylpyrazine, 2,3-dimethyl-5-ethylpyrazine or tetramethylpyrazine; Preferably, the flavor substances other than the acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines include diethoxymethane, 2-acetyl-5-methylfuran, 1,1-diethoxymethane-2-methylbutane, 1,1-diethoxymethane-3-methylbutane, 2-(diethoxymethyl)furan, 2-acetylfuran, dimethyl disulfide or dimethyl trisulfide.

3. The identification method according to claim 1, characterized in that: The following steps are involved: (1) Determining the content of the flavor substances in the standard liquor sample; (2) classifying the flavor substances into different categories of flavor substances; (3) obtaining the contents of the different types of flavor substances, and determining the threshold values ​​according to the contents of the different types of flavor substances in the standard liquor sample; (4) Comparing the contents of different types of flavor substances in the wine sample to be tested with the threshold value to obtain an identification result of the wine sample to be tested.

4. The identification method according to claim 3, characterized in that: In the step (3), the ratios of the contents of the different types of flavor substances in the standard liquor sample to the total contents of the different types of flavor substances in the standard liquor sample are respectively obtained, i.e., the standard similarity values, and the standard similarity values ​​are set to 1.00; The ratio of the content of the different types of flavor substances in the tested wine sample to the total content of the different types of flavor substances in the standard liquor sample, whichever is smaller, is greater, is expressed as a sample similarity value; When the sample similarity values ​​of the flavor substances of the wine sample to be tested are all within the range of 0.8 to 1.00, the wine sample to be tested can be determined to be a qualified wine; Preferably, when the sample similarity value of one or more of the different categories of flavor substances of the wine sample to be tested is less than 0.8, the wine sample to be tested can be determined to be unqualified wine; Preferably, when the sample similarity values ​​of the flavor substances of the different categories of the wine sample to be tested are all within the range of 0.85 to 1.00, the wine sample to be tested can be determined to be a qualified wine; Preferably, when the sample similarity value of one or more of the different categories of flavor substances of the wine sample to be tested is less than 0.85, the wine sample to be tested can be determined to be unqualified wine; Preferably, when the sample similarity values ​​of the flavor substances of the different categories of the wine sample to be tested are all within the range of 0.90 to 1.00, the wine sample to be tested can be determined to be a qualified wine; Preferably, when the sample similarity value of one or more of the different categories of flavor substances of the wine sample to be tested is less than 0.90, the wine sample to be tested can be determined to be unqualified wine; Preferably, when the sample similarity values ​​of the flavor substances of the different categories of the wine sample to be tested are all within the range of 0.92 to 1.00, the wine sample to be tested can be determined to be a qualified wine; Preferably, when the sample similarity value of one or more of the different categories of flavor substances of the wine sample to be tested is less than 0.92, the wine sample to be tested can be determined to be unqualified wine; Preferably, the step (4) of comparing the contents of different types of flavor substances in the wine sample to be tested with the threshold value further comprises comparing them in the form of a material component profile diagram; Preferably, the substance component profile is a radar chart constructed with acids, esters, alcohols, aldehydes, ketones, phenols, pyrazines and flavor substances other than acids, esters, alcohols, aldehydes, ketones, phenols and pyrazines as dimensions; Preferably, the numerical value of each dimension of the material component profile diagram is expressed as the similarity value of the wine sample in the dimension of the category; Preferably, the maximum value of each dimension value in the material component profile diagram is 1; Preferably, the maximum value of each dimension in the material component profile diagram is the standard similarity value; Preferably, the material component profile diagram also includes a similarity threshold.

5. The identification method according to claim 3, characterized in that: In the step (1), the flavor substances are detected by gas chromatography, gas chromatography-mass spectrometry, high performance liquid chromatography, or ultra-high performance liquid chromatography-high resolution mass spectrometry; Preferably, the flavor substances detected by the gas chromatography include acetaldehyde, propionaldehyde, isobutyraldehyde, ethyl formate, diethoxymethane, ethyl acetate, acetal, methanol, isovaleraldehyde, 2-pentanone, sec-butyl alcohol, ethyl butyrate, propanol, ethyl isovalerate, isobutyl alcohol, isoamyl acetate, ethyl valerate, 2-pentanol, butanol, isoamyl alcohol, ethyl hexanoate, amyl alcohol, 3-hydroxy-2-butanone, ethyl heptanoate, ethyl lactate, hexanol, butyl hexanoate, ethyl octanoate, acetic acid, furfural, propionic acid, isobutyric acid, 2,3-butanediol, 1,2-propylene glycol, butyric acid, isovaleric acid, valeric acid, ethyl phenylacetate, hexanoic acid, β-phenylethanol, heptanoic acid, octanoic acid, ethyl palmitate, ethyl oleate or ethyl linoleate; Preferably, the flavor substances detected by the gas chromatography-mass spectrometry include isobutyl acetate, ethyl 2-methylbutyrate, hexyl acetate, 2-nonanone, 3-octanol, 1-octen-3-ol, benzaldehyde, ethyl nonanoate, octanol, 5-methyl-2-furfural, 2-undecanone, hexyl hexanoate, 2-acetyl-5-methylfuran, ethyl decanoate, phenylacetaldehyde, nonanol, 2-furfuryl alcohol, diethyl succinate, ethyl undecanoate, 2-tridecanone, phenylethyl acetate, ethyl dodecanoate, benzyl alcohol, ethyl phenylpropionate, phenylethyl alcohol, ethyl tridecanoate, ethyl tetradecanoate, ethyl pentadecanoate, 1,1-diethoxymethane-2-methylbutane, 1,1-diethoxymethane-3-methylbutane, 2-heptanone, 3-hexanol, 4- methylpentanol, 2-ethyl-6-methylpyrazine, ethyl 2-hydroxybutyrate, 2-(diethoxymethyl)furan, 2-acetylfuran, 2-nonyl alcohol, ethyl DL-2-hydroxyhexanoate, ethyl levulinate, ethyl furoate, ethyl 11-hexadecenoate, ethyl heptadecanoate, ethyl octadecanoate, dimethyl disulfide, 2-methylpyrazine, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine, 2-ethylpyrazine, 2,3-dimethylpyrazine, dimethyl trisulfide, trimethylpyrazine, 2,3-diethylpyrazine, 2,3-dimethyl-5-ethylpyrazine, tetramethylpyrazine, ethyl benzoate, guaiacol, 4-methylguaiacol, phenol, 4-ethylguaiacol, 4-methylphenol, or 4-ethylphenol; Preferably, the flavor substances detected by the liquid chromatography include lactic acid; Preferably, the flavor substances detected by the ultra performance liquid chromatography-high resolution mass spectrometry include lauric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, oleic acid, gallic acid, ferulic acid, catechin, caffeic acid, chlorogenic acid, p-coumaric acid, vanillic acid, syringic acid, L-tartaric acid, L-malic acid, citric acid, glyceric acid, luteolin, maleic acid or succinic acid.

6. The identification method according to claim 5, characterized in that: The detection conditions of the gas chromatography are as follows: gas chromatography column, chromatographic column heating program: initial temperature 20-50°C, maintained for 1-10 min, increased to 100°C at 3.5°C / min, increased to 200°C at 7°C / min, maintained for 5-30 min; injection port temperature 200-300°C; detector temperature 200-300°C; injection volume 1.0-5.0 μL; split ratio 5-60:1; column flow 0.5-2.0 mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35; Preferably, the detection conditions of the gas chromatography are: DB-WAX UI chromatographic column, chromatographic column heating program: initial temperature 30-40°C, maintained for 2-5 min, increased to 100°C at 3.5°C / min, increased to 200°C at 7°C / min, maintained for 10-20 min; injection port temperature 230-260°C; detector temperature 240-260°C; injection volume 1.0-3.0 μL; split ratio 10-50:1; column flow 0.6-1.2 mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35; Preferably, the detection conditions of the gas chromatography are: DB-WAX UI chromatographic column (30m×0.25mm×0.25μm), chromatographic column heating program: initial temperature 35°C, hold for 3min, increase to 100°C at 3.5°C / min, increase to 200°C at 7°C / min, hold for 15min; injection port temperature 250°C; detector temperature 250°C; injection volume 1.0μL; split ratio 30:1; Column flow rate 0.8 mL / min; hydrogen (H2): air: tail gas (N2) = 300:30:35; Preferably, the substances detected by the gas chromatography are qualitatively characterized using retention indices and standard substances; Preferably, the substances detected by the gas chromatography are quantified using an internal standard method combined with peak area; Preferably, the wine sample processing method before gas chromatography detection is to add an internal standard solution to the wine sample, vortex mix, filter, and prepare for gas chromatography detection; Preferably, the vortex mixing time is 0.5 to 5 minutes; Preferably, the vortex mixing time is 1 to 3 minutes; Preferably, the filtration is performed using a filter membrane; Preferably, the filter membrane is a 0.22 μm microporous filter membrane; Preferably, the internal standard solution comprises tert-amyl alcohol, n-amyl acetate, and 2-ethylbutyric acid; Preferably, the final concentration of tert-amyl alcohol is 100.00-500.00 mg / L; Preferably, the final concentration of tert-amyl alcohol is 389.08 mg / L; Preferably, the final concentration of n-pentyl acetate is 100.00 to 500.00 mg / L; Preferably, the final concentration of n-pentyl acetate is 382.25 mg / L; Preferably, the final concentration of the 2-ethylbutyric acid is 100.00 to 500.00 mg / L; Preferably, the final concentration of 2-ethylbutyric acid is 396.36 mg / L.

7. The identification method according to claim 5, characterized in that: The detection conditions of the gas chromatography-mass spectrometry are as follows: (1) gas chromatography conditions: gas chromatography column; injection port temperature 200-300°C, carrier gas He, flow rate 0.5-2.0 mL / min, non-split mode; temperature program: start at 20-50°C, hold for 0.5-10 min, then heat to 100°C at 2.5°C / min, hold for 0.5-10 min; then heat to 160°C at 3°C / min, do not hold; then heat to 230°C at 5°C / min, hold for 5-60 min; (2) mass spectrometry conditions: transmission line temperature 200-300°C; EI ion source; ion source temperature: 200-300°C; electron energy: 10-100 eV; quadrupole temperature 100-200°C; Preferably, the detection conditions of the gas chromatography-mass spectrometry are as follows: (1) gas chromatography conditions: capillary column; injection port temperature 230-260°C, carrier gas He, flow rate 0.8-1.2 mL / min, non-split mode; temperature program: start at 30-45°C, hold for 1-3 min, then heat to 100°C at 2.5°C / min, hold for 1-3 min; then heat to 160°C at 3°C / min, do not hold; then heat to 230°C at 5°C / min, hold for 5-30 min; (2) mass spectrometry conditions: transmission line temperature 230-260°C; EI ion source; ion source temperature: 230-250°C; electron energy: 20-80 eV; quadrupole temperature 140-160°C; Preferably, the detection conditions of the gas chromatography-mass spectrometry are as follows: (1) gas chromatography conditions: DB-FFAP capillary column (60m×0.25mm×0.25μm); injection port temperature 250°C, carrier gas He, flow rate 1mL / min, non-split mode; temperature program: start at 40°C, hold for 1min, then increase the temperature to 100°C at 2.5°C / min, hold for 1min; then increase the temperature to 160°C at 3°C / min, do not hold; then increase the temperature to 230°C at 5°C / min, hold for 20min; (2) mass spectrometry conditions: transmission line temperature 250°C; EI ion source; ion source temperature: 230°C; electron energy: 70eV; quadrupole temperature 150°C; Preferably, the substances detected by the gas chromatography-mass spectrometry are qualitatively characterized using a spectral library, retention index and standard substances; Preferably, the substances detected by the gas chromatography-mass spectrometry are quantified using an internal standard method combined with peak area; Preferably, the wine sample treatment method before detection by gas chromatography-mass spectrometry is to add saturated salt water to the wine sample, then add an internal standard solution, and then add an extraction liquid to perform liquid-liquid extraction, vortex, stand for stratification, take the upper organic liquid, and prepare for detection on the machine; Preferably, the extract comprises diethyl ether and n-hexane; Preferably, the composition of the extract is diethyl ether: n-hexane (v:v) = 1:1; Preferably, the vortexing time is 0.5 to 5 minutes; Preferably, the vortexing time is 1 to 3 minutes; Preferably, the internal standard solution selected by the internal standard method includes 2-octanol and 2-methoxy-3-methylpyrazine; Preferably, the final concentration of the 2-octanol is 1.00 to 100.00 mg / L; Preferably, the final concentration of 2-octanol is 11.80 mg / L; Preferably, the final concentration of 2-methoxy-3-methylpyrazine is 1.00 to 100.00 mg / L; Preferably, the final concentration of 2-methoxy-3-methylpyrazine is 10.34 mg / L; Preferably, the diethyl ether and n-hexane are both of chromatographic grade.

8. The identification method according to claim 5, characterized in that: The detection conditions of the high performance liquid chromatography are: liquid chromatography column; mobile phase: KH2PO4, pH 2.0-3.0; detection wavelength 208nm, column temperature 20-50°C, mobile phase flow rate 0.5-2.0mL / min, injection volume 1-5μL; Preferably, the detection conditions of the liquid chromatography are: reverse phase chromatography column; Mobile phase: 0.01-0.05 mol / L KH2PO4, pH 2.0-2.5; detection wavelength 208 nm, column temperature 30-40 °C, mobile phase flow rate 0.8-1.2 mL / min, injection volume 1-3 μL; Preferably, the detection conditions of the liquid chromatography are: ZORBAXSB-C 18 chromatographic column (4.6mm×150mm×5μm); mobile phase: 0.02mol / LKH2PO4, pH 2.3; detection wavelength 208nm, column temperature 35°C, mobile phase flow rate 1mL / min, injection volume 1μL; Preferably, the substance detected by the high performance liquid chromatography is qualitatively characterized based on standard substances, specific wavelengths and retention times; Preferably, the substances detected by the high performance liquid chromatography are quantified using an external standard method; Preferably, the wine sample processing method before the detection by high performance liquid chromatography is to filter an appropriate amount of the wine sample in preparation for the detection on the machine; Preferably, the filtration is performed using a filter membrane; Preferably, the filter membrane is a 0.22 μm microporous filter membrane.

9. The identification method according to claim 5, characterized in that: The detection conditions of the ultra-high performance liquid chromatography-high resolution mass spectrometry are as follows: (1) ultra-high performance liquid chromatography conditions: liquid chromatography column; column temperature 20-50°C, injection volume 1-5 μL, flow rate 0.05-2 mL / min, analysis time 5-60 min, mobile phase A: formic acid (containing ammonium formate at a final concentration of 0.1-10 mmol / L), mobile phase B: pure methanol; gradient elution conditions: 0-4.0 min, 100% A (the percentage refers to the volume percentage of mobile phase A); 4.0-8. 0min, 100%~5%A; 8.0~15.0min, 5%A; 15.0~15.1min, 5%~100%A; 15.1~20.0min, 100%A; (2) High-resolution mass spectrometry conditions: heated electrospray ion source (HESI); sheath gas flow rate: 20~50arb; auxiliary gas flow rate: 5~20arb; spray voltage: 2.0~5.0kV; ion source temperature: 300~400℃; capillary temperature: 300~400℃; Preferably, the detection conditions of the ultra-high performance liquid chromatography-high resolution mass spectrometry are as follows: (1) ultra-high performance liquid chromatography conditions: Hypersil Gold liquid chromatography column; column temperature 35-45°C, injection volume 2-5 μL, flow rate 0.25-1 mL / min, analysis time 10-40 min, mobile phase A: 0.1-1% formic acid (containing final concentration 0.5-5 mmol / L ammonium formate), mobile phase B: pure methanol; gradient elution conditions: 0-4.0 min, 100% A; 4.0-8.0 min, 100%-5% A; 8.0-15. 0min, 5% A; 15.0-15.1min, 5%-100% A; 15.1-20.0min, 100% A; (2) High-resolution mass spectrometry conditions: heated electrospray ion source (HESI); sheath gas flow rate: 30-45arb; auxiliary gas flow rate: 5-10arb; spray voltage: 2.6-3.5kV; ion source temperature: 320-350°C; capillary temperature: 300-350°C; Preferably, the detection conditions of the ultra-high performance liquid chromatography-high resolution mass spectrometry are as follows: (1) ultra-high performance liquid chromatography conditions: Hypersil Gold (150 mm×2.1 mm, 1.9 μm) liquid chromatography column; column temperature 40°C, injection volume 2 μL, flow rate 0.25 mL / min, analysis time 20 min, mobile phase A: 0.1% formic acid (containing final concentration 2 mmol / L ammonium formate), mobile phase B: pure methanol; gradient elution conditions: 0-4.0 min, 100% A; 4.0-8.0 min, 100%-5% A; 8.0-15.0 min, 5% A; 15.0-15.1 min, 5%-100% A; 15.1-20.0 min, 100% A; (2) High-resolution mass spectrometry conditions: heated electrospray ionization source (HESI); sheath gas flow rate: 38 arb; auxiliary gas flow rate: 9 arb; spray voltage: 3.2 kV; ion source temperature: 345°C; capillary temperature: 320°C; Preferably, the volume percentage of the mobile phase A refers to the volume percentage of the mobile phase A in the mixture of the mobile phase A and the mobile phase B; Preferably, the substances detected by the ultra-high performance liquid chromatography-high resolution mass spectrometry are qualitatively identified by using characteristic ions and peak times of standard substances; Preferably, the substances detected by the ultra-high performance liquid chromatography-high resolution mass spectrometry are quantified using an external standard method combined with peak area; Preferably, the wine sample processing method before detection by ultra-high performance liquid chromatography-high resolution mass spectrometry is to concentrate the wine sample by nitrogen blowing, take it out after concentration, cool it to room temperature, make up to volume with pure methanol, vortex mix it, and filter it to obtain the sample to be tested; Preferably, the vortex mixing time is 0.5 to 5 minutes; Preferably, the vortex mixing time is 1 to 3 minutes; Preferably, the filtration is performed using a 0.22 μm organic PES syringe filter membrane; Preferably, the pure methanol is of chromatographic grade.

10. Application of the identification method according to claim 1 in improving the quality of wine, evaluating the quality of wine, optimizing the liquor preparation process, identifying the quality of liquor, or controlling the stability of liquor batches; Preferably, the identification of the wine sample to be tested includes comparing the similarity value of each dimension of the wine sample to be tested with the similarity threshold value of each dimension; Preferably, the identification of the wine sample to be tested further comprises comparing similarity values ​​in combination with the material component profile diagram; Preferably, the similarity threshold of each dimension is 0.80-1.00; Preferably, the similarity threshold of each dimension is 0.85-1.00; Preferably, the similarity threshold of each dimension is 0.90-1.00; Preferably, the similarity threshold of each dimension is 0.92-1.00; Preferably, the similarity threshold of each dimension is 0.92.

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