CN115389665A - Based on stable oxygen isotope (delta) 18 O) wine mixing water identification method - Google Patents
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Abstract
The invention relates to the technical field of wine identification, in particular to a stable oxygen isotope (delta) -based wine identification method 18 O) wine mixing water identification method. The method comprises the following steps: preparation of different delta 18 Water with a distribution range of O; designing fermentation experiment with exogenous water and mixing with different delta 18 Obtaining wine with different alcoholic strength by using exogenous water with O characteristic and different doping amount; measuring delta of water in grape juice before fermentation by adopting water balance instrument-stable isotope mass spectrometer 18 O sample And delta of water in fermented wine 18 O wine (ii) a Calculating the delta of the water in the wine sample to be tested and the wine sample without water 18 Judging whether water is mixed or not according to the standard deviation SD value between O; according to water delta in grape juice 18 O and externally added water delta 18 And judging the lowest water mixing amount of the wine to be detected according to the absolute value of the O difference value. The invention utilizes stable isotope mass spectrometry technologyThe method for accurately identifying whether the wine is mixed with water is established, the problem of mixing water with the wine in China is solved, the method for identifying the mixing water with the wine which is lacked in national standards is improved, and the full-juice wine is provided for consumers.
Description
Technical Field
The invention relates to the technical field of wine identification, in particular to a stable oxygen isotope (delta) -based method 18 O) identification method of wine mixing water.
Background
The wine is rich in various amino acids, minerals and vitamins, and has high nutritive value and economic value. With the progress of globalization, the wine industry develops rapidly, and the huge potential of the market is gradually developed. However, under the drive of benefits, the phenomenon of wine adulteration sometimes occurs, which is not beneficial to the stable development of the wine market. In China, the common wine adulteration modes in the market mainly comprise three modes: diluting by adding water, blending concentrated grape juice and manually blending, namely a production mode of 'three refined water and one water' wine, wherein the water adding in the wine is the most common means for manufacturing fake wine and aims to reduce the alcoholic strength or deliberately add the fake wine.
The concept of wine is defined in the national standard (GB/T15037-2006). The stipulation "the wine is made up by using fresh grape or grape juice as raw material and making them undergo the process of full or partial fermentation, and contains fermented wine with a certain alcoholic strength". Therefore, exogenous water cannot be mixed in the production process, but the standard lacks a method for identifying exogenous water in the wine matched with the standard. Therefore, a method capable of effectively identifying the wine mixing water is developed, scientific and reasonable technical means and data support can be provided for wine mixing water identification research and establishment of related standards, the market behavior can be normalized, and the healthy development of the wine industry in China can be promoted.
At present, the behavior of the wine mixed with water can be identified by methods such as sensory analysis, visible-near infrared spectrum analysis, gas chromatography analysis, high performance liquid chromatography analysis, stable isotope ratio mass spectrometry and the like. Among them, the stable isotope technique shows unique advantages because it can provide unique molecular fingerprints that cannot be copied or forged, and can be used for discriminating the water-blended before and after fermentation of wine.
Disclosure of Invention
The invention aims to provide a wine water-mixing identification method based on stable oxygen isotope, which is used for measuring the delta of water in a wine sample after fermentation 18 O value, observed to incorporate a different delta 18 Delta of water in wine after exogenous water with O characteristics and different mixing amounts 18 O value change and analysis of water delta in wine 18 Correlation of O value with water entrainment. By calculating the delta of the sample to be measured 18 O and control sample delta 18 The addition of exogenous water is judged according to the SD value between O and the delta of water in grape juice 18 O and externally added water delta 18 And judging the lowest water blending amount of the wine to be detected according to the absolute value of the O difference value, thereby perfecting the method for identifying the foreign water in the wine lacking in the standard, and promoting the healthy development of the wine industry in China.
The technical scheme adopted by the invention is as follows: based on stable oxygen isotope (delta) 18 O) wine watering identification method, which is characterized in that: the method comprises the following steps:
the method comprises the following steps: preparation of different delta 18 Water with a distribution range of O;
step two: designing fermentation experiment with exogenous water and mixing with different delta 18 Obtaining wine with different alcoholic strength by using exogenous water with O characteristic and different doping amount;
step three: measuring delta of water in grape juice before fermentation by adopting water balance instrument-stable isotope mass spectrometer 18 O sample And delta of water in fermented wine 18 O wine ；
Step four: calculating the wine to be measuredDelta of water in samples and unblended wine samples 18 Standard deviation SD value between O, if SD>0.1 indicates that the sample to be detected is mixed with water, otherwise, the instrument cannot judge whether the wine sample is mixed with water;
step five: according to water delta in grape juice 18 O and externally added water delta 18 And judging the lowest water mixing amount of the wine to be detected according to the absolute value of the O difference value.
As a further improvement, the step one is to realize the delta in the water by reduced pressure rotary evaporation 18 Fractional distillation of O to different degrees to produce different deltas 18 Exogenous water with an O value in the range of: -17.74% o to-3.80% o.
As a further improvement, in the second step, the alcoholic strength of the fermented wine is measured by adopting a gas chromatograph.
As a further improvement, in the third step, when the wine samples are measured by using a water balance instrument-stable isotope mass spectrometer, each sample is measured in parallel for 5 times, and the standard deviation SD value among 5 data is calculated, wherein SD <0.1 indicates that the instrument stability is good and the data has reliability.
As a further improvement, the fourth step is that the grape juice is watered before and after delta 18 And the O difference value is small, the judgment cannot be carried out by utilizing the conventional significance difference analysis, and whether the wine is mixed with water or not is judged by utilizing the measurement precision of an instrument as a reference value, wherein the standard deviation SD =0.1.
As a further improvement, the invention is realized by the following steps: collecting a grape sample and a water sample in a target production area; (2) pretreatment of the grape sample: squeezing → mixing → filtering → split charging → storage; (3) Different stable delta 18 Preparing exogenous water with distributed O values: the different degrees of fractionation of stable oxygen isotopes in water is realized by reduced pressure rotary evaporation, and delta is prepared 18 O values are respectively-2.33 permillage (SD = 0.10), -4.35 permillage (SD = 0.09), -6.32 permillage (SD = 0.03), -8.36 permillage (SD = 0.08), -10.34 permillage (SD = 0.01), -12.33 permillage (SD = 0.10), -14.34 permillage (SD = 0.05), -16.33 permillage (SD = 0.08), -18.35 permillage (SD = 0.06), and the delta of tap water in different regions of the country is basically covered by 9 exogenous water samples 18 The O range; (4) Simulated fermentation wateringWine sample: aiming at the behavior of doping water with deliberate adulteration, 12.0 percent, 17.0 percent, 22.5 percent, 26.0 percent and 30.5 percent of 9 delta are doped into grape juice with the theoretical alcoholic strength of 8.0 percent (v/v, the same is applied below), 8.5 percent, 9.0 percent, 9.5 percent and 10.0 percent 18 Fermenting the wine with the expected alcohol content of 7.0% (the lowest alcohol content of the wine specified by national standard) by using the exogenous water with the O value; adding 7.0%, 13.0%, 18.5% of 9 kinds of delta into grape juice with theoretical alcoholicity of 14.0%, 15.0%, 16.0%, respectively 18 Exogenous water with an O value, fermenting the wine with an expected alcohol content of 13.0 percent (common alcohol content of the commercial wine); adding 5.5%, 11.5%, 15.5% of 9 kinds of delta into grape juice with theoretical alcohol content of 17.0%, 18.0%, 19.0% 18 Exogenous water with an O value, fermenting the wine with the expected alcohol content of 16.0 percent (the highest alcohol content tolerated by common yeast); (5) Measuring the alcoholic strength of different wine samples by adopting a gas chromatograph; (6) Method for measuring delta of water in wine by adopting water balance instrument-stable isotope mass spectrometer 18 O value, calculating the delta of water in the blended and unblended wine samples 18 SD value of standard deviation between O; (7) Analysis of different delta 18 Incorporation of exogenous water with distribution of O-value and delta of water in wine sample 18 Establishing an identification model of the exogenous water doping in the wine based on the stable oxygen isotope by the correlation between the O values; (8) verifying the validity of the model; (9) According to water delta in grape juice 18 O and exogenous water delta 18 And establishing a model of the lowest detection limit value of the water mixing amount of the wine according to the absolute value of the O difference value.
The invention has the beneficial effects that: the invention analyzes the incorporation of different delta by stable isotope technology 18 O characteristic of exogenous water and delta of its mixing amount to water in wine 18 Influence of the O value, delta of water in grape juice before and after fermentation was found 18 O has small change, and only has 0.03 per mill difference; exogenous water doped with different stable oxygen isotopes, the amount of water doped being delta to that of water in wine 18 O is in good correlation and is directed against 9 different deltas 18 Establishing 9 identification models for the external source water of O, and inverting the estimated average relative error between the water doping amount and the actual water doping amount by using the models to be less than 0.1; according to water delta in grape juice 18 O and exogenous water delta 18 Difference of oxygenThe absolute value of the value can judge the lowest doping amount of the exogenous water in the wine. Delta of water in grape juice 18 O and exogenous water delta 18 The water mixing identification of the wine can be realized when the O phase difference is more than or equal to 4.48 per thousand and the percentage of the exogenous water is more than or equal to 5.5 percent.
The invention establishes a method capable of accurately identifying whether the wine is mixed with water by using a stable isotope mass spectrometry technology, promotes the solving of the problem of mixing water with the wine in China, perfects the wine mixing identification method lacking in national standards, and provides the full-juice wine for consumers.
Drawings
FIG. 1 shows a diagram of the present invention based on a stable oxygen isotope (. Delta.) 18 O) is used for analyzing a correlation between the stable oxygen isotope value of the water in the wine and the water doping amount;
FIG. 2 shows a diagram of the present invention based on a stable oxygen isotope (. Delta.) 18 O) analysis chart of the lowest detection limit result of the water addition amount of the wine water-adding identification method.
Detailed Description
The present invention is not limited by the following examples, and specific embodiments may be determined according to the technical solutions and practical situations of the present invention.
Based on stable oxygen isotope (delta) 18 O) the wine watering identification method is characterized in that: the method comprises the following steps:
the method comprises the following steps: preparation of different delta 18 Water with a distribution range of O;
step two: designing fermentation experiment with exogenous water and mixing with different delta 18 Obtaining wine with different alcoholic strength by using exogenous water with O characteristic and different doping amount;
step three: measuring delta of water in grape juice before fermentation by adopting water balance instrument-stable isotope mass spectrometer 18 O sample And delta of water in fermented wine 18 O wine ；
Step four: calculating the delta of water in the wine sample to be tested and the wine sample without water 18 Standard deviation SD value between O, if SD>0.1 indicates that the sample to be detected is mixed with water, otherwise, the instrument cannot judge whether the wine sample is mixed with water or notWater;
step five: according to water delta in grape juice 18 O and externally added water delta 18 And judging the lowest water mixing amount of the wine to be detected according to the absolute value of the O difference value.
In the invention, step one, the delta in water is realized by reduced pressure rotary evaporation 18 Fractional distillation of O to different degrees to produce different deltas 18 Exogenous water with an O value in the range of: -17.74% o to-3.80% o.
In the second step, the alcoholic strength of the fermented wine is measured by adopting a gas chromatograph.
In the third step, when the wine sample is measured by adopting the water balance instrument-stable isotope mass spectrometer, each sample is measured in parallel for 5 times, and the standard deviation SD value among 5 data is calculated, wherein SD <0.1 shows that the instrument has good stability and the data has reliability.
In the invention, the fourth step is that the grape juice is watered before and after delta 18 And the O difference value is small, the judgment cannot be carried out by utilizing the conventional significance difference analysis, and whether the wine is mixed with water or not is judged by utilizing the measurement precision of an instrument as a reference value, wherein the standard deviation SD =0.1.
Example 1: delta of Water in wine 18 O measurement
The method comprises the following steps: determination of the Delta of Water in grape juice before fermentation 18 O(δ 18 O sample ) And water delta in fermented wine 18 O(δ 18 O wine )；
Step two: as can be seen from Table 1, δ 18 O wine Is relatively delta 18 O sample Minus 0.03 per mill indicates that no obvious fractionation effect occurs in the fermentation process.
TABLE 1 Delta in Water before and after fermentation of grape juice 18 O comparison
Example 2: establishment and verification of wine water-mixing identification model
The method comprises the following steps: determination of unblended wine samplesDelta of reclaimed water 18 And (4) the value of O.
Step two: determination of the incorporation difference delta 18 Delta of water in wine after exogenous water with O characteristics and different mixing amounts 18 And (4) O value.
Step three: and (3) establishing a correlation analysis of the stable oxygen isotope value of water in the wine and the water doping amount, and referring to the attached figure 1.
Step four: calculating the delta of water in a water blended wine sample 18 Delta of O value to Water in unblended wine samples 18 The standard deviation SD between O values, as shown in table 2:
TABLE 2 Water Delta in wine 18 O
Note: AVG in Table 2 represents the average of the measured data of the same sample, and SD<0.1.SD represents the delta of water in the wine sample after blending 18 Delta of O with Water in unblended wine samples 18 O standard deviation of mean of two sets of data, when SD>0.1 shows that the two groups of data have significant difference, and the sample wine is mixed with water, otherwise, the water balance instrument-stable isotope mass spectrometer cannot judge whether the sample wine is mixed with water.
As can be seen from FIG. 1 and Table 2, addition of delta 18 When O is exogenous water of-2.33 per mill, the fitting equation is delta 18 O =0.0358x-5.8632 (x is exogenous water content), R 2 =0.999, the model can determine the water content of at least 7.0%. Addition of delta 18 When O is exogenous water of-4.34 per mill, the fitting equation is delta 18 O＝0.0156x-5.8566，R 2 =0.999, model can judge the water addition of at least 13.0%. Addition of delta 18 When O is exogenous water of-6.32 per mill, the fitting equation is delta 18 O＝-0.0052x-5.8564，R 2 =0.985, the SD is still less than 0.100 when the water doping amount reaches 30.5%, and the water doping amount below 30.5% cannot be judged by a prediction model.Due to delta incorporation 18 Delta of exogenous water with O of-6.32 per mill and water in grape juice 18 The O value (-5.83 per thousand) is relatively close, and even if the water content is large, the identification is still difficult. Addition of delta 18 When O is-8.36 per mill of exogenous water, the fitting equation is delta 18 O＝-0.0240x-5.8734，R 2 =0.998, the model can judge a water loading of at least 11.5%.
Addition of delta 18 When O is exogenous water of-10.34 per mill, the fitting equation is delta 18 O＝-0.0446x-5.8620，R 2 =0.999; addition of delta 18 When O is exogenous water of-12.33 per mill, the fitting equation is delta 18 O＝-0.0644x-5.8643，R 2 =0.999; addition of delta 18 When O is exogenous water of-14.34 per mill, the fitting equation is delta 18 O＝-0.0841x-5.8681，R 2 =0.999; addition of delta 18 When O is exogenous water of-16.33 per mill, the fitting equation is delta 18 O＝-0.1044x-5.8620，R 2 =0.999; addition of delta 18 When O is exogenous water of-18.35 per mill, the fitting equation is delta 18 O＝-0.1232x-5.8664，R 2 =0.999. When the above 5 kinds of differences delta are incorporated 18 The model can judge the water content of at least 5.5% for the exogenous water with the O value.
Step five: verifying the validity of the model: randomly selecting the water content of 5.5%, 11.5%, 15.5%, 22.5%, 30.5%, and mixing with exogenous water delta 18 The wine sample with O of-6.32 per mill (the model fitting degree is lowest) is substituted into the fitting equation delta 18 O = -0.0052x-5.8564, and the wine water content is estimated by inversion.
Step six: as can be seen from Table 3, the average relative error between the actual and estimated values of the sample was only 0.075, indicating that the difference is based on the delta 18 The water mixing identification model established by the exogenous water with the O characteristic has high reliability and can meet the requirement of quantitative analysis of the water mixing of the wine.
TABLE 3 wine sample estimation
| Sample(s) | True value/%) | Estimated value/%) | Absolute error (absolute value) | Relative error (absolute value) |
| Sample 1 | 5.5 | 4.54 | 0.96 | 0.175 |
| Sample No. 2 | 11.5 | 10.31 | 1.19 | 0.103 |
| Sample 3 | 15.5 | 16.08 | 0.58 | 0.037 |
| Sample 4 | 22.5 | 21.85 | 0.65 | 0.029 |
| Sample 5 | 30.5 | 29.54 | 0.96 | 0.031 |
Step seven: according to water delta in grape juice 18 O(δ 18 O sample ) Delta. With exogenous water 18 O(δ 18 O water ) And establishing a model of the lowest detection limit value of the water content of the wine according to the absolute difference value, and referring to an attached figure 2.
According to the analysis of the results, when delta 18 O sample And delta 18 O water When the difference is 1.51 per mill, the model can judge the water addition amount of at least 13.0 percent. Delta 18 O sample And delta 18 O water When the difference is 2.50 per mill, the model can judge the water content of at least 11.5 percent. Delta 18 O sample And delta 18 O water When the difference is 3.53 per mill, the model can judge the water content of at least 7.0 percent. Delta 18 O sample And delta 18 O water When the difference is 4.48 per mill, the model can judge the water content of at least 5.5 percent. Thereby establishing delta 18 O sample And delta 18 O water The absolute value of the difference value and the minimum detection limit value of the amount of the doped exogenous water.
As can be seen from FIG. 2, δ 18 O sample And delta 18 O water The absolute value of the difference value and the lowest detection limit of the amount of the doped exogenous water show good linear negative correlation change, and the fitting equation is y = -2.7230x +17.4325 (x is delta) 18 O sample And delta 18 O water Absolute value of difference), R 2 =0.958, the fitting effect is good, and the method has higher reference value for identifying the water mixing of the wine in any region.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (5)
1. Based on stable oxygen isotope (delta) 18 O) wine watering identification method, which is characterized in that: the method comprises the following steps:
the method comprises the following steps: preparation of different delta 18 Water with a distribution range of O;
step two: designing fermentation experiment with exogenous water and mixing with different delta 18 Obtaining wine with different alcoholic strength by using exogenous water with O characteristic and different doping amount;
step three: measuring delta of water in grape juice before fermentation by adopting water balance instrument-stable isotope mass spectrometer 18 O sample And delta of water in fermented wine 18 O wine ；
Step four: calculating the delta of water in the wine sample to be tested and the wine sample without water 18 Standard deviation SD value between O, if SD>0.1 indicates that the sample to be detected is mixed with water, otherwise, the instrument cannot judge whether the wine sample is mixed with water or not;
step five: according to water delta in grape juice 18 O and externally added water delta 18 And judging the lowest water mixing amount of the wine to be detected according to the absolute value of the O difference value.
2. A stable oxygen isotope (δ) based on claim 1 18 O) wine watering identification method, which is characterized in that: in the first step, the delta in water is realized by reduced pressure rotary evaporation 18 Fractional distillation of O to different degrees to produce different deltas 18 Exogenous water with an O value in the range of: -17.74% o to-3.80% o.
3. A stable oxygen isotope (δ) based on claim 1 18 O) wine watering identification method, which is characterized in that: and step two, measuring the alcoholic strength of the fermented wine by adopting a gas chromatograph.
4. The method of claim 1Based on stable oxygen isotope (delta) 18 O) the wine watering identification method is characterized in that: and step three, when the wine samples are measured by adopting a water balance instrument-stable isotope mass spectrometer, each sample is measured in parallel for 5 times, and the standard deviation SD value and SD between 5 data are calculated<0.1 shows that the instrument has good stability and the data has reliability.
5. A stable oxygen isotope (δ) based on claim 1 18 O) wine watering identification method, which is characterized in that: step four, the grape juice is watered before and after delta 18 And the O difference value is small, the judgment cannot be carried out by utilizing the conventional significance difference analysis, and whether the wine is mixed with water or not is judged by utilizing the measurement precision of an instrument as a reference value, wherein the standard deviation SD =0.1.
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| CN1595126A (en) * | 2003-09-10 | 2005-03-16 | 咸嘉泉 | White spirit counterfeit deterrence verification method |
| CN105021732A (en) * | 2014-11-03 | 2015-11-04 | 中国食品发酵工业研究院 | Method for fast determination of oxygen isotope composition of water in beverage wine |
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