Yellow wine quality detection and analysis method

Abstract

The invention discloses a yellow rice wine quality detection and analysis method, which comprises the following steps: (a) grouping a plurality of yellow rice wine samples according to whether caramel color is added or not, and the years in each group are the same in a one-to-one correspondence manner; a plurality of parallel samples are taken from each wine sample, detection is performed through a QTOF high-resolution mass spectrometer, effective information of TOF MS and TOF MS / MS is obtained through one-time sample introduction in a DBS-IDA collection mode, and high-resolution first-level and second-level mass spectrum data are obtained; (b) analyzing by adopting SCIEX OS software, and identifying a plurality of compounds according to an SCIEX database and literatures; the structure is identified through a GNPS molecular network technology, and SCIEX OS is used for further verification and confirmation of a plurality of other compounds; and (c) carrying out difference analysis through PLS-DA. According to the method, the influence of the caramel color additive and the aging year on the yellow wine quality can be scientifically and comprehensively analyzed, and a basis is provided for yellow wine quality evaluation and quality control.

Description

Technical Field

[0001] The present invention relates to the technical field of food detection, and more specifically, to a method for detecting and analyzing the quality of yellow rice wine. Background Art

[0002] Caramel color is one of the natural food additives with the largest dosage and the widest range of use in edible pigments. It can significantly improve the color, flavor and other qualities of food, and is mainly applied to yellow rice wine, condiments (such as soy sauce, vinegar), beverages (such as cola), and candies. Caramel color has unique properties, which have a certain impact on the color, aroma, taste and body of yellow rice wine, and is also an important factor affecting the quality of yellow rice wine. In addition, during the aging process of yellow rice wine, various components in the wine body interact with each other, resulting in a large number of chemical and physical changes, which improve the taste and stability of the wine body. Therefore, the age change of yellow rice wine is also the main indicator of its quality.

[0003] However, the analysis of the influence of caramel color additives and aging years on the quality differences of yellow rice wine is a complex and difficult task. At present, there is no practical and effective quality evaluation index for these differences, resulting in difficulty in establishing a scientific and unified objective measurement standard. Summary of the Invention

[0004] The purpose of the present invention is to overcome the above-mentioned deficiencies of the prior art, and provide a method for detecting and analyzing the quality of yellow rice wine, which can scientifically and comprehensively analyze the influence of caramel color additives and aging years on the quality of yellow rice wine, and provide a basis for the quality evaluation and quality control of yellow rice wine.

[0005] To achieve the above purpose, the present invention adopts the following technical solutions:

[0006] A method for detecting and analyzing the quality of yellow rice wine, comprising the following steps:

[0007] (a) Divide multiple yellow rice wine samples into a caramel color group composed of yellow rice wine with different aging years and a non-caramel color group composed of yellow rice wine with different aging years according to whether caramel color additives are added in the samples, and the years of yellow rice wine in the caramel color group correspond one-to-one with the years of yellow rice wine in the non-caramel color group; take multiple parallel samples for each yellow rice wine sample, dilute each sample with water respectively, and then detect them through a QTOF high-resolution mass spectrometer. Obtain effective information of TOF MS and TOF MS / MS by DBS-IDA acquisition method, and obtain high-resolution first-level and second-level mass spectrometry data of compounds in the yellow rice wine samples.

[0008] (b) Data processing and compound identification: Use SCIEX OS software for data processing and analysis, and identify multiple compounds according to the SCIEX database and literature; identify the compound structure through GNPS molecular network technology and further verify it with SCIEX OS software to confirm the acquisition of multiple other compounds.

[0009] (c) Difference analysis: Through PLS-DA analysis, analyze the differential compounds in yellow rice wine of the same aging year with and without the addition of caramel color, and the differential compounds in yellow rice wine of different aging years without the addition of caramel color.

[0010] The present invention is further configured that in step (b), when further verifying by using SCIEX OS software, it accesses the Chemspider chemical structure database through the SCIEX OS software, automatically retrieves the structure by using the Chemspider chemical structure database, and performs secondary mass spectrometry matching through the Fragment panel of the fragmentation automation matching function of the SCIEX OS software, thereby confirming the compound.

[0011] The present invention is further configured that the liquid phase conditions for detection by the QTOF high-resolution mass spectrometer are:

[0012] Chromatographic column: HSS T3, with a specification of 2.1×100 mm, 1.8 μm; column temperature: 40 °C; mobile phase: using water-0.1 wt% formic acid as phase A and acetonitrile-0.1 wt% formic acid as phase B; the gradient elution program is as follows:

[0013] Time (min) Mobile Phase A (%) Mobile Phase B (%) 3.5 95 5 12 50 50 14.5 5 95 17.5 5 95 17.7 95 5 20 95 5

[0014] Injection volume: 5 μL; flow rate: 0.4 mL / min.

[0015] The present invention is further configured that the mass spectrometry conditions for detection by the QTOF high-resolution mass spectrometer are:

[0016] Mass spectrometry model: X500R QTOF; scanning mode: TOF MS-IDA-10MS / MS; ion source: electrospray ionization source ESI+ and ESI-; primary scanning range: 70-1250 Da, acquisition time is 0.1 sec; secondary scanning range: 40-1250 Da, acquisition time is 0.05 sec;

[0017] Ion source parameter ES: positive ion spray voltage: 5500 V; negative ion spray voltage: -4500 V; nebulizing gas GS1: 50 psi; nebulizing gas GS2: 50 psi; ion source temperature TEM: 500 °C; curtain gas CUR: 30 psi; collision gas CAD: 8 psi.

[0018] The beneficial effects of the present invention are:

[0019] Based on a QTOF high-resolution mass spectrometer and DBS-IDA, the present invention enables obtaining high-quality TOF MS and TOF MS / MS effective information in a sample with a single injection. Compounds in yellow rice wine with different aging years with and without caramel color addition are identified based on SCIEX OS software and GNPS technology, and the differences in yellow rice wine of different years and with or without caramel color addition are analyzed through PLS-DA analysis method, so as to scientifically and comprehensively analyze the effects of caramel color additive and aging years on the quality of yellow rice wine, providing a basis for the quality evaluation and quality control of yellow rice wine. Description of the Drawings

[0020] Figure 1 It is the data processing flow chart of the present invention;

[0021] Figure 2 It is the chromatogram of n-fructosyl isoleucine measured in Example 1;

[0022] Figure 3 It is the first-level mass spectrum of n-fructosyl isoleucine measured in Example 1;

[0023] Figure 4 It is the second-level mass spectrum of n-fructosyl isoleucine measured in Example 1;

[0024] Figure 5 It is the PLS-DA analysis result diagram of the 9-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0025] Figure 6 It is the PLS-DA analysis result diagram of the 11-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0026] Figure 7 It is the PLS-DA analysis result diagram of the 14-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0027] Figure 8 It is the PLS-DA analysis result diagram of the 18-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0028] Figure 9 It is the PLS-DA analysis result diagram of the 21-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0029] Figure 10 It is the variable importance in projection (VIP) diagram of the 14-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0030] Figure 11 It is the clustering heat map of the 14-year-aged yellow rice wine with caramel color and the yellow rice wine without caramel color in Example 1;

[0031] Figure 12 It is a box plot of the differential compound guanosine between the 14-year-old caramel-colored yellow rice wine and the non-caramel-colored yellow rice wine in Example 1;

[0032] Figure 13 It is a box plot of the differential compound 1-naphthylamine between the 14-year-old caramel-colored yellow rice wine and the non-caramel-colored yellow rice wine in Example 1;

[0033] Figure 14 It is a box plot of the differential compound harmaline between the 14-year-old caramel-colored yellow rice wine and the non-caramel-colored yellow rice wine in Example 1;

[0034] Figure 15 It is a box plot of the differential compound norharmane between the 14-year-old caramel-colored yellow rice wine and the non-caramel-colored yellow rice wine in Example 1;

[0035] Figure 16 It is a PLS-DA analysis result diagram of the non-caramel-colored yellow rice wine with different aging years in Example 1;

[0036] Figure 17 It is a VIP diagram of the yellow rice wine with different aging years in Example 1;

[0037] Figure 18 It is a box plot of the differential compound threonine-tyrosine of the non-caramel-colored yellow rice wine with different aging years in Example 1;

[0038] Figure 19 It is a box plot of the differential compound phenylpropane-leucine of the non-caramel-colored yellow rice wine with different aging years in Example 1;

[0039] Figure 20 It is a box plot of the differential compound salicylic acid of the non-caramel-colored yellow rice wine with different aging years in Example 1;

[0040] Figure 21 It is a box plot of the differential compound L-tyrosine-L-tyrosine of the non-caramel-colored yellow rice wine with different aging years in Example 1. Detailed implementation manners

[0041] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0042] The detection and analysis method for the quality of the yellow rice wine of the present invention includes the following steps:

[0043] (a) Multiple yellow rice wine samples were evenly divided into a caramel color group composed of yellow rice wines with different aging years and a non-caramel color group composed of yellow rice wines with different aging years according to whether caramel color additive was added or not in the wine samples, and each year of the yellow rice wine in the caramel color group corresponded one-to-one and was the same as each year of the yellow rice wine in the non-caramel color group; multiple parallel samples were taken for each yellow rice wine sample, and each yellow rice wine sample was diluted with water and then detected by a QTOF high-resolution mass spectrometer. Through the DBS-IDA acquisition method, TOF MS and TOF MS / MS effective information were obtained in one injection, and high-resolution first-level and second-level mass spectrometry data of the compounds in the yellow rice wine samples were obtained.

[0044] The liquid phase conditions for detection by the QTOF high-resolution mass spectrometer were as follows:

[0045] Chromatographic column: HSS T3, with a specification of 2.1×100 mm, 1.8 μm; column temperature: 40 °C; mobile phase: using water-0.1 wt% formic acid as phase A and acetonitrile-0.1 wt% formic acid as phase B; injection volume: 5 μL; flow rate: 0.4 mL / min; the gradient elution program is shown in the following table:

[0046] Time (min) Mobile Phase A (%) Mobile Phase B (%) 3.5 95 5 12 50 50 14.5 5 95 17.5 5 95 17.7 95 5 20 95 5

[0047] The mass spectrometry conditions for detection by the QTOF high-resolution mass spectrometer were as follows:

[0048] Mass spectrometry model: X500R QTOF; scanning mode: TOF MS-IDA-10MS / MS; ion source: electrospray ionization source ESI+ and ESI-; first-level scanning range: 70-1250 Da, acquisition time: 0.1 sec; second-level scanning range: 40-1250 Da, acquisition time: 0.05 sec; ion source parameter ES: positive ion spray voltage: 5500 V; negative ion spray voltage: -4500 V; nebulizing gas GS1: 50 psi; nebulizing gas GS2: 50 psi; ion source temperature TEM: 500 °C; curtain gas CUR: 30 psi; collision gas CAD: 8 psi.

[0049] (b) Data processing and compound identification: The SCIEX OS software was used for data processing and analysis, and multiple compounds were identified according to the SCIEX database and literature; the compound structures were identified by the GNPS molecular network technology and further verified by the SCIEX OS software (that is, accessing the Chemspider chemical structure database through the SCIEX OS software, automatically retrieving the structure using the Chemspider chemical structure database, and performing second-level mass spectrometry diagram matching through the Fragment panel of the automatic fragmentation matching function of the SCIEX OS software), and then multiple other compounds were confirmed. The data processing flow is shown in Figure 1 , and this data processing method belongs to the prior art and will not be elaborated here.

[0050] (c) Differential analysis: Through PLS-DA (Partial Least Squares Discriminant Analysis), analyze the differential compounds between yellow rice wines with and without added caramel color of the same aging year, and the differential compounds of yellow rice wines of different aging years without added caramel color.

[0051] Example 1

[0052] Ten commercially available yellow rice wine samples were evenly divided into a caramel color group and a no-caramel color group, and each group had wine samples of five aging years: 9, 11, 14, 18, and 21; 5 parallel samples were taken from each yellow rice wine sample, and after being diluted 5 times with water, injection detection, data processing, compound identification, and differential analysis were carried out according to the above method. The ten wine samples were brewed by the same method, the difference being whether caramel color was added and the different aging years. The results are as follows:

[0053] (a) Compound identification

[0054] As shown in Table 1, 157 compounds were identified using the SCIEX database and existing literature, etc., and another 51 compounds were confirmed through GNPS identification and SCIEX OS software (shown by the serial numbers 14, 17, 20, 23, 28, 34, 43, 45, 47, 49, 50, 54, 59, 60, 65, 76, 82, 92, 102, 107, 110, 115, 126, 133, 136, 137, 139, 143, 144, 145, 146, 150, 154, 161, 164, 166, 167, 170, 174, 176, 178, 180, 181, 182, 184, 188, 195, 196, 198, 199, 207 in Table 1), and a total of 208 compounds were confirmed (including 102 amino acid-related compounds, 14 polyphenol compounds, 9 ester compounds, 25 organic acid compounds, 3 lipid compounds, 3 coumarin compounds, 12 carbohydrate compounds, 9 alkaloid compounds, 4 aldehyde compounds, 11 nucleotide compounds, and 16 other compounds). Among them, the chromatogram of n-fructosyl isoleucine is shown in Figure 2 , and the first-level mass spectrum is shown in Figure 3 , and the second-level mass spectrum is shown in Figure 4 .

[0055] Table 1

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062] (b) Difference analysis (b1) Comparison of differences with or without caramel color addition

[0063] Through PLS-DA analysis, it can be found that there are significant differences between yellow rice wines with or without caramel color addition in the same aging years. The results of PLS-DA analysis are shown in Figures 5 - 9 (In each figure, the red area on the left represents yellow rice wine in the caramel color group, and the green area on the right represents yellow rice wine in the non-caramel color group).

[0064] Taking the 14-year aging as an example to analyze the differential compounds between caramel-colored yellow rice wine and non-caramel-colored yellow rice wine, after screening for differences with P<0.05 and VIP>1, 57 differential compounds were screened out (as shown in Table 2), and the variable importance in projection (VIP) graph is shown in Figure 10 (Showing the top fifteen compounds with VIP values), and the clustering heat map is shown in Figure 11 , and the box plots of some differential compounds are shown in Figures 12 - 15 . It can be seen from the analysis results that some amino acid-related compounds increase and some decrease, and there are more up-regulated amino acids after adding caramel color; the contents of most organic acid differential compounds in caramel-colored yellow rice wine are higher than those in non-caramel-colored yellow rice wine; the contents of two polyphenols in caramel-colored yellow rice wine are lower than those in non-caramel-colored yellow rice wine.

[0065] Table 2

[0066]

[0067]

[0068] (b2) Comparison of yellow rice wines with different aging years

[0069] Through PLS-DA analysis, it can be found that there are significant differences between yellow rice wines with different aging years. The result graph of PLS-DA analysis is shown in Figure 16 ; after screening for differential compounds with ANOVA P<0.05&VIP>1, 80 differential compounds were screened out (as shown in Table 3), and the VIP graph is shown in Figure 17 ; the box plots of some differential compounds are shown in Figures 18 - 21 . It can be seen from the analysis results that amino acid-related compounds increase and decrease to some extent with the increase of aging years; the contents of most differential esters, aldehydes, sugars and organic acid compounds in the 21-year-old non-caramel-colored yellow rice wine are higher than those in the 9-year-old yellow rice wine; several differential nucleotides show opposite trends with the increase of aging years. In addition, the compounds only contained in the 21-year-old yellow rice wine are shown in Table 4.

[0070] Table 3

[0071]

[0072]

[0073]

[0074] Table 4

[0075] Sample Type Compound Chinese Name Polyphenol Tricin Luteolin Amino Acid Related L-Methionine Methionine Polyphenol Kaempferol 3-glucoside-7-xyloside Kaempferol-3-glucoside-7-xyloside Nucleotide 5'-S-Methyl-5'-thioadenosine 5'-Deoxy-5'-(methylthio)adenosine Alkaloid trans-3-indole-acrylic acid 3-Indoleacrylic Acid Coumarin 7-hydroxy-4-methyl-chromen-2-one 4-Methylumbelliferone Organic Acid Phenylacetic acid Phenylacetic Acid

[0076] In summary, it can be seen that:

[0077] (1) In this embodiment, the yellow rice wine samples were identified, and 208 compounds were obtained through identification and confirmation.

[0078] (2) Statistical analysis shows that there are significant differences in the metabolites of yellow rice wine with different aging years: the contents of some esters, aldehydes, sugars and organic acids in the 21-year-old yellow rice wine are higher than those in the 9-year-old yellow rice wine; several nucleotides show a trend opposite to the increase in aging years; the main types of differential compounds are amino acid-related compounds.

[0079] (3) Adding caramel color also has a significant impact on the metabolites in yellow rice wine: more amino acids are up-regulated after adding caramel color, two polyphenol compounds are down-regulated, and multiple organic acids are up-regulated.

[0080] The above is only the preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions falling within the concept of the present invention belong to the protection scope of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements should also be regarded as within the protection scope of the present invention.

Claims

1. A method for detecting and analyzing the quality of yellow rice wine, characterized in that, It includes the following steps: (a) Divide multiple yellow rice wine samples into a caramel color group composed of yellow rice wine with different aging years and a non-caramel color group composed of yellow rice wine with different aging years according to whether caramel color additive is added or not in the samples, and each year of the yellow rice wine in the caramel color group corresponds one-to-one with each year of the yellow rice wine in the non-caramel color group; Take multiple parallel samples for each yellow rice wine sample, dilute each sample with water respectively, and then detect them by a QTOF high-resolution mass spectrometer. Through the DBS-IDA acquisition method, obtain effective TOF MS and TOF MS / MS information in one injection to get the high-resolution first-order and second-order mass spectrometry data of the compounds in the yellow rice wine samples; (b) Data processing and compound identification: Use SCIEX OS software for data processing and analysis, and identify multiple compounds according to the SCIEX database and literature; Identify the compound structures through the GNPS molecular network technology and further verify them using the SCIEX OS software to confirm the existence of multiple other compounds; (c) Differential analysis: Through PLS-DA analysis, analyze the differential compounds in yellow rice wine with and without added caramel color in the same aging year and the differential compounds in yellow rice wine with different aging years without added caramel color.

2. The method for detecting and analyzing the quality of yellow rice wine according to claim 1, characterized in that, In step (b), when further verifying using the SCIEX OS software, access the Chemspider chemical structure database through the SCIEX OS software, automatically retrieve the structure using the Chemspider chemical structure database, and perform second-order mass spectrometry diagram matching through the Fragmentpanel of the automatic fragmentation matching function of the SCIEX OS software to confirm the compounds.

3. The method for detecting and analyzing the quality of yellow rice wine according to claim 1, wherein The liquid phase conditions for detection by the QTOF high-resolution mass spectrometer are as follows: Chromatographic column: HSS T3, with a specification of 2.1×100mm, 1.8μm; Column temperature: 40°C; Mobile phase: Use water-0.1wt% formic acid as phase A and acetonitrile-0.1wt% formic acid as phase B; The gradient elution program is as follows: Injection volume: 5μL; Flow rate: 0.4mL / min.

4. The method for detecting and analyzing the quality of yellow rice wine according to claim 1, wherein The mass spectrometry conditions for detection by the QTOF high-resolution mass spectrometer are as follows: Mass spectrometry model: X500R QTOF; Scanning mode: TOF MS-IDA-10MS / MS; Ion source: Electrospray ionization source ESI+ and ESI-; First-order scanning range: 70-1250Da, acquisition time is 0.1sec; Second-order scanning range: 40-1250Da, acquisition time is 0.05sec; Ion source parameter ES: Positive ion spray voltage: 5500V; Negative ion spray voltage: -4500V; Nebulizing gas GS1: 50psi; Nebulizing gas GS2: 50psi; Ion source temperature TEM: 500°C; Curtain gas CUR: 30psi; Collision gas CAD: 8psi.

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