A method for identifying the authenticity or age of a brewed alcoholic beverage based on dynamic light scattering

Abstract

The application discloses a method for identifying the authenticity or year of brewed wine based on dynamic light scattering, comprising the following steps: (1) taking a brewed wine standard product, adding different exogenous substances into the brewed wine standard product respectively, dissolving and mixing uniformly, and then performing membrane treatment to obtain a standard product sample; (2) determining the hydrodynamic average radius of each exogenous substance in the standard product sample by dynamic light scattering, and obtaining the hydrodynamic average radius basic data of the corresponding exogenous substance in the standard product; (3) taking a to-be-detected brewed wine, and performing the same treatment as steps (1) and (2) on the to-be-detected brewed wine to obtain the hydrodynamic average radius data of the corresponding exogenous substance in the to-be-detected brewed wine; and (4) comparing the hydrodynamic average radius data of the corresponding exogenous substance in the to-be-detected brewed wine with the hydrodynamic average radius basic data of the corresponding exogenous substance in the standard product, and identifying the authenticity or year of the to-be-detected brewed wine. The application has the advantages of small sampling amount, simple sample pretreatment and analysis process, and less interference in the detection process.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of brewed wine identification, and particularly relates to a method for identifying the authenticity or age of brewed wine based on dynamic light scattering technology. BACKGROUND

[0002] Brewed wine, also known as fermented wine, is wine formed by the action of yeast on grains rich in starch and polysaccharides to produce alcohol. Traditional brewing technology is also an important technology for traditional agricultural product processing in China. With the increasing attention to the problem of "three rural issues", the development of traditional brewed wine has become one of the directions for solving the problem of "three rural issues". Yellow wine, grape wine and beer are collectively known as the world's three ancient wines, and all three belong to brewed wine. Yellow wine is more popular with consumers than the other two wines because of its soft wine nature, mellow wine body, rich nutrition and appropriate alcohol content, and has become an important development direction for the brewed wine industry in China. However, the raw materials and brewing process of wine are complex and difficult to control, and the quality of industrial production cannot be guaranteed and unified. Therefore, it is necessary to invent a method for accurately and quickly identifying brewed wine, which is also a technical problem.

[0003] Current brewed wine identification technology is not very perfect. For example, some researchers use nuclear magnetic resonance spectroscopy to detect the age of yellow wine, use near-infrared spectroscopy to detect the adulteration of glutinous rice as a raw material for yellow wine, and use voltammetric electronic tongue to identify the category of yellow wine. These methods have complex pretreatment, tedious data processing, and are relatively one-sided in wine identification. A few researchers have found that there are nanoparticles in some processed foods, but these studies also have certain limitations. First, the detection technology is complex, including ICP-MS, ELISA, AFM and other technologies, which have complex pretreatment and expensive instruments, and require professional personnel to operate. Second, it has not been applied to the field of food identification. SUMMARY

[0004] The present application provides a method for identifying the authenticity or age of brewed wine. By adding exogenous substances to disturb the originally stable food system, the light intensity fluctuation phenomenon of the sample is detected by dynamic light scattering technology, and then the hydrodynamic average data obtained by processing the light intensity data are compared to identify the authenticity or age of the brewed wine, achieving the effect of simply and quickly identifying the authenticity or age of different brewed wine varieties.

[0005] A method for identifying the authenticity or age of brewed wine based on dynamic light scattering, comprising:

[0006] (1) Take brewed wine standard products, add different exogenous substances to the brewed wine standard products and fully dissolve and mix, and then obtain standard product detection liquid by membrane treatment;

[0007] (2) The hydrodynamic average radius of the standard sample detection solution is determined by dynamic light scattering to obtain the hydrodynamic average radius basic data of the standard sample detection solution;

[0008] (3) The to-be-detected brewing wine is taken, and the to-be-detected brewing wine is subjected to the same treatment as steps (1) and (2) to obtain the hydrodynamic average radius data of the to-be-detected brewing wine detection solution;

[0009] (4) The hydrodynamic average radius data of the to-be-detected brewing wine detection solution is compared with the corresponding hydrodynamic average radius basic data in the standard sample detection solution to identify the variety authenticity or year of the to-be-detected brewing wine.

[0010] The nutrients contained in the brewing wine are mostly proteins and polysaccharides, and there are also some inorganic salt small molecules, which constitute the basic framework of the nanostructure of the brewing wine. Processed foods are stored for a period of time before being put on the market, and the products on the shelf are generally a relatively stable system. The present application found that the addition of an external substance disturbs the originally stable system, and the substances existing in the food system interact with the exogenous substances. The content and types of substances originally existing in different types of brewing wine are different, and the types and types of interactions will be different, and there will also be differences between counterfeit and genuine products.

[0011] Because the hydrodynamic radius of the nano-particles in the wine is only about 3-4 nm, the difference between different varieties is very small, about 1 nm. In the present application, the difference between different varieties of wine is shown by adding an exogenous substance to promote the interaction between the substances originally existing in the wine and the exogenous substance, so as to achieve the purpose of identification and recognition.

[0012] In a more specific scheme, the identification method of the present application first collects different types of brewing wine samples known to be true, adds an exogenous substance to fully dissolve, and then mixes them to make the added substances fully dissolved in the to-be-detected sample. The to-be-detected sample after sufficient reaction is subjected to membrane treatment, and the sample after membrane treatment is loaded into a specific sample bottle; the dynamic light scattering technology is applied, the fluctuation phenomenon of light intensity signal of particles based on irregular Brownian motion is obtained, and the light intensity basic data of the standard sample added with the exogenous substance are obtained; then the corresponding light intensity basic data of the to-be-detected sample added with the exogenous substance are detected by the same method; the particle size data obtained by fitting the corresponding light intensity basic data of the to-be-detected sample and the standard sample are compared, so as to realize the identification of different brewing wine varieties. The to-be-detected sample is compared with all known basic data, and the similar data are the same variety, otherwise the difference is large, which is a counterfeit or inferior product or another variety. The present application has the advantages of small sample amount, simple sample pretreatment and analysis process, less interference in the detection process, and clear and accurate judgment basis.

[0013] The fluctuation of the light intensity signal refers to that the scattering particles are always in random motion, and the fluctuation of the scattering light intensity directly reflects the Brownian motion of the scattering particles, thereby quantitatively measuring the mobility of the scattering particles in the solution.

[0014] The hydrodynamic average radius can be understood as follows: all particles of different shapes are approximated as an equivalent sphere, and the hydrodynamic radius is the radius of the equivalent sphere, and the friction force experienced by the equivalent sphere in the Brownian motion in the solution is the same as the friction force experienced by the scattering particles.

[0015] Optionally, the brewed wine is yellow rice wine.

[0016] Further, the yellow rice wine is any one of the following common brands: Huaiji Mountain, Guyue Dragon Mountain, Taibao, Xianheng, Jimo, and Daughter Red, and any one of different years.

[0017] Optionally, the exogenous substances are at least one of acetic acid, phenylalanine, proline, glutamic acid, arginine, and citric acid.

[0018] Further preferably, the exogenous substances are acetic acid, phenylalanine, proline, glutamic acid, arginine, and citric acid, and the adding mode is to add acetic acid, phenylalanine, proline, glutamic acid, arginine, and citric acid into the standard separately and to add all the exogenous substances into the standard at the same time.

[0019] Optionally, the exogenous substances are added at 0.15-0.25 mg / mL.

[0020] Further, the exogenous substances are added at 0.18-0.22 mg / mL, and more further, the exogenous substances are added at 0.2 mg / mL.

[0021] Optionally, when all the exogenous substances are added into the standard at the same time, each exogenous substance is added at 0.15-0.25 mg / mL (preferably 0.2 mg / mL).

[0022] The appropriate proportion of the exogenous substances is obtained by referring to the relevant literature to obtain the content of various chemical substances in the sample. Since dynamic light scattering is a relatively precise instrument and is sensitive to environmental changes, the amount of the added exogenous substances is kept consistent, and the detection data is not disturbed by the amount of the added exogenous substances.

[0023] Optionally, in the process of measuring the hydrodynamic average radius basic data of the standard added with different exogenous substances and measuring the hydrodynamic average radius data of the brewed wine to be measured, the standing time after adding and mixing the exogenous substances is 2-20 h.

[0024] Optionally, the dissolving and mixing process comprises: firstly, mixing the mixture with a vortex instrument, and then further mixing the mixture with an ultrasonic instrument, and the temperature of the ultrasonic instrument is controlled at 25 DEG C to prevent the temperature from being too high to affect the sample to be measured.

[0025] Optionally, the membrane treatment uses a 0.22-micron organic membrane.

[0026] The membrane treatment is used to avoid the influence of dust and bubbles on the detection data, and a 0.22-micron organic membrane is generally used for filtering the brewed wine.

[0027] The dynamic light scattering measurement uses a special sample bottle for detection, and in order to further ensure the accuracy of the detection data, the sample bottle is washed with acetone for about 15 minutes before the sample is loaded and detected, and then the sample bottle is sealed with tin foil paper and dried for standby use.

[0028] Optionally, the dynamic light scattering measurement uses the following parameters: the temperature condition is 25 DEG C, the solvent condition is water (i.e., the viscosity is 0.89 mPas, and the refractive index is 1.33), the measurement time is 30 seconds, the measurement angle is 25 DEG, and the measurement wavelength is 632.8 nm.

[0029] Optionally, the dynamic light scattering measurement is performed in an ALV / CGS integrated dynamic and static laser light scattering instrument, and the hydrodynamic average radius is calculated by the software (ALV-Correlator Software V.3.0) of the ALV / CGS integrated dynamic and static laser light scattering instrument.

[0030] The dynamic light scattering measurement obtains light intensity data, and the average value of 10 effective data is taken, and the light intensity data is considered as effective data when the light intensity data fluctuates within a fixed value range (the fluctuation amplitude is about 20%) and then the hydrodynamic average radius is obtained according to the light intensity data. Optionally, the light intensity data obtained by the ALV / CGS integrated dynamic and static laser light scattering instrument is converted into a light intensity autocorrelation equation by a hardware device called a correlator, and then the hydrodynamic average radius is obtained by fitting in the CONTIN mode of the fitting program.

[0031] The light intensity signal fluctuation phenomenon refers to that the scattering particles are always in random motion, and the fluctuation of the scattering light intensity with time directly reflects the Brownian motion of the scattering particles, so as to quantitatively measure the mobility of the scattering particles in the solution.

[0032] In step (4), the sample to be measured is compared with all known basic data, and the similar data are the same variety, and otherwise the large difference is a counterfeit or inferior product or another variety.

[0033] One rule for judging similar data is that the difference between the hydraulic average radius data of the test liquor and the corresponding basic data of the hydraulic average radius in the standard test liquor is within 5%.

[0034] Difference error = |r0-r1|×100% / r 0；

[0035] Where r1 is the average hydraulic radius of the wine being tested;

[0036] r0 is the basic data of the corresponding hydraulic average radius in the standard test solution.

[0037] The term "corresponding" can be understood as: comparing the average hydraulic radius data of the test liquid of the brewed wine to be tested with that of the standard test liquid after adding the same exogenous substance and standing for the same time.

[0038] Compared with the prior art, this application has the following beneficial effects:

[0039] Compared to statistical identification of chemical components and spectral characterization, this method, which alters the stable environment of brewed wine by adding external substances, transforms complex data into a single data indicator detection method. The sample is not damaged, and the detection environment is less affected by interference, making it difficult for merchants to artificially add substances to achieve the purpose of counterfeiting, thus greatly improving the efficiency of brewed wine identification. Attached Figure Description

[0040] Figure 1 A schematic diagram illustrating the fluctuations in the light intensity signal of the standard sample from Kuaiji Mountain;

[0041] Figure 2 According to Figure 1 The correlation function curve obtained by converting light intensity data;

[0042] Figure 3 A diagram showing the basic data of the average hydraulic radius of the standard sample from Kuaiji Mountain;

[0043] Figure 4 A diagram showing the basic data of the average hydraulic radius of the standard product of Gu Yue Long Shan;

[0044] Figure 5 A basic data diagram of the average hydraulic radius of the standard product of Tower Brand;

[0045] Figure 6 A diagram showing the basic data of the average hydraulic radius of the Xianheng standard product;

[0046] Figure 7 Basic data diagram of the average hydraulic radius of the three-year aged standard sample from Kuaiji Mountain;

[0047] Figure 8Basic data diagram of the average hydraulic radius of the five-year aged standard sample from Kuaiji Mountain;

[0048] Figure 9 Basic data diagram of the average hydraulic radius of the standard sample aged eight years in Kuaiji Mountain;

[0049] Figure 10 Basic data diagram of the average hydraulic radius of the ten-year-aged standard sample from Kuaiji Mountain;

[0050] Figure 11 The diagram shows the detection principle of the ALV / CGS integrated dynamic and static laser light scattering instrument (the reference numerals in the diagram are: 1: light scattering sample bottle; 2: laser; 3: detector; 4: light scattering detection range). Detailed Implementation

[0051] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0053] Example 1: Different Brands of Shaoxing Wine

[0054] 1. Main instruments and reagents

[0055] ALV / CGS integrated dynamic and static laser light scattering instrument, vortex analyzer, analytical balance, rinsing device, disposable syringe, filter membrane, sample bottle, various additives, acetone, and rice wine samples.

[0056] 2. Determine the types and proportions of exogenous substances to be added.

[0057] Literature review shows that rice wine contains a variety of amino acids and organic acids, as well as some functional factors, among which arginine, phenylalanine, proline, glutamic acid, citric acid, and acetic acid are present in relatively high amounts.

[0058] Table 1 Content of various additives

[0059]

[0060] Based on the specific content of the components in Table 1, the exogenous substances were added to the rice wine at a concentration of 0.2 mg / mL, resulting in a total of 7 samples. Total matter refers to all the substances listed above being added to the rice wine samples. The solutions were thoroughly mixed using a vortex mixer until the solutes were completely dissolved. For poorly soluble substances, further mixing was performed using an ultrasonic instrument. The ultrasonic temperature was controlled at 25℃ throughout the process to obtain the test samples. The hydraulic radius data were measured after standing for 2 hours and 20 hours, respectively.

[0061] 3. Detection of dynamic light scattering

[0062] Preheat the instrument for 30 minutes before measurement to stabilize the temperature and laser (22mV helium-neon laser), reducing errors caused by the measurement environment. Use a 2.5mL disposable syringe to draw the sample (from step 2. Determine the type and proportion of the added exogenous substance), filter it through a 0.22μm organic membrane, and slowly inject it into a specific light scattering sample vial that has been rinsed with acetone. Before placing the vial into the sample cell, wipe the vial clean to ensure a clean environment that will not affect the detection data. Place the sample in the vial and close the switch to connect the laser path. The detection principle is as follows: Figure 11 As shown.

[0063] Set the required parameters for the test: measurement temperature 25℃, measurement angle 25°, solvent conditions set to water, viscosity 0.89 mPas, refractive index 1.33, laser wavelength 632.8 nm, measurement time 30 seconds per measurement, and average the 10 valid data points. The valid data is the light intensity data (e.g., ...). Figure 1 The data fluctuates within a certain range, and there are no sharp peaks or large peaks.

[0064] 4. Result Judgment

[0065] The raw light intensity data was converted into an autocorrelation equation using a correlator (ALV / LSE-5004 Light Scattering Electronics and Multiple Tau Rigital Correlator), and then further transformed according to the Stokes-Einstein equation. After analysis and processing, the relevant hydraulic mean radius data were obtained (as shown in Table 2). Figure 3 4, 5, and 6 are the basic data for four brands of rice wine (data transformation is automatically generated by the instrument's built-in software) and serve as the basis for comparison of the samples to be tested.

[0066] Table 2. Hydraulic radius of different brands of Shaoxing wine at different settling times.

[0067]

[0068] Example 2: Shaoxing wine from different years

[0069] The experimental procedure was the same as in Example 1. Different years of Kuaijishan rice wine were purchased, and exogenous substances of corresponding amounts (same as in Example 1) were added to ensure complete dissolution.

[0070] Set the required detection parameters: measurement temperature 25℃, measurement angle 25°, solvent condition set to water, viscosity 0.89 mPas, refractive index 1.33, laser wavelength 632.8 nm, measurement time 30 s per measurement, and average of 10 valid measurements. Table 3 and... Figure 7 8, 9, and 10 are the basic data for the average hydraulic radius of wines from different years.

[0071] Table 3. Hydraulic radius of different years of Kuaijishan rice wine at different settling times

[0072]

[0073] Example 3: Determination of Adulterated and Blended Shaoxing Wine

[0074] To prepare the counterfeit rice wine solution, 50 mL of a 15% ethanol solution was mixed with 1 g / L of sulfite caramel coloring, 120 μL of acetic acid, 0.8 g / L of ethyl butyrate, and 1.6 g / L of isoamyl alcohol. The 15% ethanol solution simulates the alcohol content of rice wine, the caramel coloring simulates its color and flavor, the acetic acid simulates its acidic environment, and the ethyl butyrate and isoamyl alcohol are sources of the characteristic flavor of rice wine, thus mimicking its unique taste. 5 mL of the counterfeit rice wine solution was taken, and the experimental procedure was the same as in Example 1. The same amount of exogenous substances were added, and after complete dissolution, the solution was allowed to stand for 2 hours before testing. The solution was then allowed to stand for 20 hours before further testing.

[0075] The results are shown in Table 4. By comparing the data with the 15% alcohol content in Tables 2 and 3, it can be seen that the hydraulic radius of the adulterated rice wine is significantly larger than that of the normal Kuaijishan three-year rice wine, which can accurately identify the counterfeit rice wine. The alcohol content of different rice wines is shown in Table 5.

[0076] Table 4. Hydraulic radius of adulterated rice wine (15% alcohol) after different standing times

[0077]

[0078]

[0079] Table 5. Alcohol content of different types of rice wine

[0080]

[0081] Example 4: Determination of water / alcohol adulteration in rice wine

[0082] The experimental procedure was the same as in Example 1, except that the three-year aged rice wine from Kuaijishan was replaced with a solution of three-year aged rice wine from Kuaijishan diluted twice with ultrapure water and a solution of three-year aged rice wine from Kuaijishan diluted twice with 15% ethanol. The results are shown in Table 6. Compared with the data of three-year aged rice wine in Table 3, it was found that the hydraulic radius of the rice wine mixed with water and ethanol was also significantly larger than that of the normal rice wine.

[0083] Table 6. Hydraulic radius of Kuaijishan three-year-aged rice wine at different dilution ratios and different settling times.

[0084]

[0085] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. A method for identifying the authenticity or vintage of brewed wine based on dynamic light scattering, characterized in that, include: (1) Take the brewed wine standard, add different exogenous substances to the brewed wine standard and dissolve and mix them thoroughly, and then process them through a membrane to obtain the standard test solution; (2) The basic data of the hydraulic mean radius of the standard test solution were obtained by measuring the hydraulic mean radius of the standard test solution through dynamic light scattering. (3) Take the wine to be tested and process it in the same way as steps (1) and (2) to obtain the average hydraulic radius data of the wine to be tested. (4) Compare the hydraulic mean radius data of the test liquid of the brewed wine to be tested with the corresponding basic data of the hydraulic mean radius in the test liquid of the standard to identify the authenticity or year of the brewed wine to be tested. The brewed wine is rice wine; the exogenous substance is at least one of acetic acid, phenylalanine, proline, glutamic acid, arginine, and citric acid.

2. The method according to claim 1, characterized in that, The exogenous substances are acetic acid, phenylalanine, proline, glutamic acid, arginine, and citric acid. The addition method is as follows: acetic acid, phenylalanine, proline, glutamic acid, arginine, and citric acid are added to the standard separately, and all exogenous substances are added to the standard simultaneously.

3. The method according to claim 1, characterized in that, All exogenous substances were added at a concentration of 0.15~0.25 mg / mL.

4. The method according to claim 1, characterized in that, In determining the basic data of the average hydraulic radius of the standard test solution with different added exogenous substances and the average hydraulic radius of the test wine, the standing time after adding the exogenous substances and mixing was 2~20h.

5. The method according to claim 1, characterized in that, A 0.22 μm organic membrane is selected for the membrane treatment.

6. The method according to claim 1, characterized in that, The parameters for dynamic light scattering measurement were set as follows: temperature 25℃, solvent water, measurement time 30s, measurement angle 25°, and measurement wavelength 632.8nm.

7. The method according to claim 1, characterized in that, The instrument used for the dynamic light scattering measurement is an ALV / CGS integrated dynamic and static laser light scattering instrument.

8. The method according to claim 1, characterized in that, If the difference between the hydraulic mean radius data of the tested brewed wine and the corresponding baseline hydraulic mean radius data of the standard test solution is within 5%, then the tested brewed wine and the brewed wine standard are determined to be of the same variety; otherwise, they are different. Difference error = |r0-r1| / r0; Where r1 is the average hydraulic radius of the wine being tested; r0 is the basic data of the corresponding hydraulic average radius in the standard test solution.

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