CN111999371A - Method for determining stable hydrogen isotope ratio of ethanol methyl site in alcoholic beverage - Google Patents
Method for determining stable hydrogen isotope ratio of ethanol methyl site in alcoholic beverage Download PDFInfo
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- 239000001257 hydrogen Substances 0.000 title claims abstract description 64
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 title claims abstract description 55
- 238000000034 method Methods 0.000 title claims abstract description 50
- 235000013334 alcoholic beverage Nutrition 0.000 title claims abstract description 43
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 194
- 239000000243 solution Substances 0.000 claims description 24
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- 239000000126 substance Substances 0.000 claims description 15
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 14
- 238000000855 fermentation Methods 0.000 claims description 11
- 230000004151 fermentation Effects 0.000 claims description 11
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 8
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- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 6
- 230000000813 microbial effect Effects 0.000 claims description 6
- 150000007522 mineralic acids Chemical class 0.000 claims description 6
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims description 5
- 239000000920 calcium hydroxide Substances 0.000 claims description 5
- 229910001861 calcium hydroxide Inorganic materials 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 238000007789 sealing Methods 0.000 claims description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 3
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 235000013405 beer Nutrition 0.000 claims description 3
- 235000013532 brandy Nutrition 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 238000013375 chromatographic separation Methods 0.000 claims description 3
- 235000019990 fruit wine Nutrition 0.000 claims description 3
- 150000002431 hydrogen Chemical class 0.000 claims description 3
- 229910017604 nitric acid Inorganic materials 0.000 claims description 3
- 230000003647 oxidation Effects 0.000 claims description 3
- 238000007254 oxidation reaction Methods 0.000 claims description 3
- 235000013533 rum Nutrition 0.000 claims description 3
- 235000013522 vodka Nutrition 0.000 claims description 3
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 230000003472 neutralizing effect Effects 0.000 claims description 2
- 238000000643 oven drying Methods 0.000 claims description 2
- 238000011160 research Methods 0.000 abstract description 5
- 238000001514 detection method Methods 0.000 abstract description 2
- 235000011054 acetic acid Nutrition 0.000 description 52
- 239000000047 product Substances 0.000 description 13
- -1 methyl hydrogen Chemical class 0.000 description 12
- 239000000523 sample Substances 0.000 description 10
- 238000005259 measurement Methods 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 description 4
- 238000004821 distillation Methods 0.000 description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 description 3
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 229910052805 deuterium Inorganic materials 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- YZCKVEUIGOORGS-NJFSPNSNSA-N Tritium Chemical compound [3H] YZCKVEUIGOORGS-NJFSPNSNSA-N 0.000 description 2
- 241000219095 Vitis Species 0.000 description 2
- 235000009754 Vitis X bourquina Nutrition 0.000 description 2
- 235000012333 Vitis X labruscana Nutrition 0.000 description 2
- 235000014787 Vitis vinifera Nutrition 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 238000002307 isotope ratio mass spectrometry Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 238000005361 D2 NMR spectroscopy Methods 0.000 description 1
- YZCKVEUIGOORGS-IGMARMGPSA-N Protium Chemical compound [1H] YZCKVEUIGOORGS-IGMARMGPSA-N 0.000 description 1
- 241000720974 Protium Species 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
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- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000011005 laboratory method Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 125000001434 methanylylidene group Chemical group [H]C#[*] 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
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- 230000008520 organization Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 229910052722 tritium Inorganic materials 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/02—Food
- G01N33/14—Beverages
- G01N33/146—Beverages containing alcohol
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Abstract
The invention provides a method for measuring the stable hydrogen isotope ratio of an ethanol methyl site in alcoholic beverages, belonging to the technical field of stable isotopes. The method comprises the following steps: a) converting ethanol in the alcoholic beverage to be tested into acetic acid; b) measuring the hydrogen isotope ratio of acetic acid in the alcoholic beverage obtained in the step a); c) calibrating the hydrogen isotope ratio of the acetic acid measured in step b) with an acetic acid standard. The method is mainly used for determining the stable hydrogen isotope ratio of the ethanol methyl sites in the alcoholic beverage and the research and application in the field of alcoholic beverage adulteration detection.
Description
Technical Field
The invention belongs to the technical research field of stable isotopes, and particularly relates to a method for determining a stable hydrogen isotope ratio of an ethanol methyl site in alcoholic beverages.
Background
The wine industry is the traditional industry in China and is also an important component of the food industry, and as the biggest beverage wine production and consumption country in the world, in recent years, the wine industry in China is continuously enlarged in scale, and the economic benefit is promoted year by year. The alcoholic beverage is beverage with alcoholic strength above 0.5% vol, and includes fermented wine, distilled wine and compound wine. The alcoholic beverage has a long history, and wine seeds with different types are produced by special raw material selection, traditional brewing technology and even unique distillation technology, so that products with different qualities are produced, and meanwhile, the alcoholic beverage also provides opportunities for adulteration of lawbreakers.
The stable isotope technology is an important means for detecting whether alcoholic beverage products and raw materials are adulterated or not. Hydrogen atoms are important elements constituting living bodies, and are classified into hydrogen/protium (H) and deuterium (H)2H) And tritium (f)3H) Wherein H and2h is stable isotope, and hydrogen-containing compounds of different sources have different hydrogen isotope ratios (based on natural fractionation effect of the stable isotope2H/H, notation2H) In that respect Currently, the determination of the hydrogen isotope ratio of ethanol mainly comprises the following technical problems: firstly, the hydrogen isotope of ethanol needs to be cracked and converted into hydrogen in the determination process, however, any hydrogen-containing substance can be converted into hydrogen in the process, the content of ethanol in wine and the like is about 12 percent, and the rest is substances such as water, organic acid and the like, so that high-purity ethanol needs to be separated in advance; secondly, ethanol has three hydrogen sites which are methyl hydrogen, methylene hydrogen and hydroxyl hydrogen respectively, hydrogen atoms on the hydroxyl are easy to exchange with water according to the isotope exchange reaction principle, and because the hydrogen isotope ratios of water in the alcoholic beverage are different, the direct utilization of the hydrogen isotope ratio of the ethanol is not scientific, and only the isotope characteristics of the ethanol methyl or methine sites which can not exchange hydrogen have application value.
The currently popular method for determining the hydrogen isotope ratio of the methyl site of ethanol is the point-specific fractionation-nuclear magnetic resonance (SNIF-NMR) method originated from 1980 s. For this technique, the European Union organization organized the laboratory method alignment [ Thomas F, Jamin E.2H NMR and 13C-IRMS analysis of acetic acid from the video, 18O-IRMS analysis of water in video [ J ] analytical chip act, 2009,649(1):98-105], validated the validity of the assay technique and established the method standard [ RESOLUTION OIV-NO 527- ] 2015 Determination of the distribution of the fatty acid in acetic acid extracted from the video in Nuclear Magnetic Resonance (NMR) ]. However, this technique has two major drawbacks: firstly, the micro-distillation system needs to be specially customized to meet the requirements that the recovery rate is higher than 96.5 percent and the ethanol concentration in the fraction is higher than 95 percent v/v, the micro-distillation system is a patent product of European fins in France, 160 ten thousand RMB are needed for a set, the requirement on the proficiency of an operator in the use process is high, and the time of single distillation is also needed to be more than 4 hours; the principle of the nuclear magnetic resonance apparatus for measuring the hydrogen isotope ratio of the ethanol methyl is to measure the absolute content of deuterium at methyl sites in a quantitative sample, and because the content of deuterium only accounts for 0.02 percent of hydrogen atoms in a natural state, the measurement time of a single sample is up to more than 5 hours to obtain enough signal-to-noise ratio and stability. Therefore, the technology has low analysis efficiency, high cost and poor universality, and seriously hinders some researches and applications of China in the field of alcoholic beverages.
Disclosure of Invention
The invention provides a method for measuring the stable hydrogen isotope ratio of an ethanol methyl site in alcoholic beverages, which is simple, safe and efficient and provides a quick and effective analysis means for alcoholic beverage authenticity identification technology.
The invention provides a method for measuring the stable hydrogen isotope ratio of an ethanol methyl site in alcoholic beverages, which comprises the following steps:
a) converting ethanol in the alcoholic beverage to be tested into acetic acid;
b) measuring the hydrogen isotope ratio of acetic acid in the alcoholic beverage obtained in the step a);
c) calibrating the hydrogen isotope ratio of the acetic acid measured in step b) with an acetic acid standard.
Further, the step b) comprises:
b1) removing carboxyl hydrogen atoms of acetic acid in the product obtained in the step a);
b2) removing water from the original solution of the product obtained in the step b 1);
b3) acidifying the product of step b2) with a dilute acid solution;
b4) determining the hydrogen isotope ratio of the acetic acid in the product obtained in step b3) by using a stable isotope ratio mass spectrometer.
Further, in the step a), the alcoholic beverage to be tested is diluted by water until the volume concentration of ethanol is 3-5% and then is converted;
in the step a), the conversion adopts a microbial fermentation method or an ethanol oxidation method.
Further, in step b1), neutralizing with an alkaline agent to remove the carboxyl hydrogen atoms of acetic acid;
preferably, the alkaline agent comprises calcium hydroxide, sodium hydroxide or potassium hydroxide.
Further, in the step b2), removing water in the original solution by adopting an oven drying mode, a chromatographic separation mode or a freeze drying mode;
further, in the step b3), the dilute acid solution is an aqueous solution of inorganic acid;
preferably, the inorganic acid includes dilute sulfuric acid, dilute hydrochloric acid, dilute nitric acid, and dilute phosphoric acid.
Further, in the step b4), after the product obtained in the step b3) is cracked at a high temperature and is converted into hydrogen, a stable isotope ratio mass spectrometer is used for measuring the hydrogen isotope ratio of the hydrogen;
preferably, the high-temperature constant temperature is 1300-1500 ℃.
Further, in step c), when the acetic acid standard substance is corrected, the acetic acid standard substance is processed by the step b) before being measured.
Further, in the step a), when the microorganism fermentation method is adopted, the acetic acid bacteria culture solution is added into the alcoholic beverage, oxygen is added, sealing is carried out, and fermentation treatment is carried out at the constant temperature of 30 ℃.
Further, in the step a), the alcoholic beverage comprises distilled liquor, fermented liquor and prepared liquor;
preferably, the fermented wine comprises beer and wine; the distilled liquor comprises Chinese liquor, brandy, whisky, vodka and rum; the compound wine comprises fruit wine.
The invention has the following advantages:
the invention provides a method for analyzing the hydrogen isotope ratio of an ethanol methyl site based on metabonomics and a stable hydrogen isotope fractionation mechanism, which has the advantages of simple operation, safe experiment and quick analysis, is suitable for the analysis and test of a large batch of samples, and is beneficial to the research and application of a hydrogen isotope technology in the field of alcoholic beverage adulteration detection.
Detailed Description
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
An embodiment of the invention provides a method for determining a stable hydrogen isotope ratio of an ethanol methyl site in alcoholic beverages, which comprises the following steps:
a) converting ethanol in the alcoholic beverage to be tested into acetic acid;
b) measuring the hydrogen isotope ratio of acetic acid in the alcoholic beverage obtained in the step a);
c) calibrating the hydrogen isotope ratio of the acetic acid measured in step b) with an acetic acid standard.
The method for determining the stable hydrogen isotope ratio of the methyl site of the ethanol in the alcoholic beverage comprises the steps of firstly converting the ethanol in the alcoholic beverage into acetic acid, then eliminating the influence of carboxyl hydrogen acetate and water in a sample, determining the hydrogen isotope ratio of the acetic acid, and finally correcting the methyl hydrogen isotope ratio of the acetic acid by using an acetic acid standard substance to obtain the hydrogen isotope ratio of the methyl site of the ethanol. The method is simple to operate, safe in experiment, rapid in analysis and suitable for analysis and test of large-batch samples.
In an embodiment of the present invention, the step b) includes:
b1) removing carboxyl hydrogen atoms of acetic acid in the product obtained in the step a);
b2) removing water from the original solution of the product obtained in the step b 1);
b3) acidifying the product of step b2) with a dilute acid solution;
b4) determining the hydrogen isotope ratio of the acetic acid in the product obtained in step b3) by using a stable isotope ratio mass spectrometer.
In an embodiment of the present invention, the first and second substrates are,
in the step a), the alcoholic beverage to be tested is diluted by water until the volume concentration of ethanol is 3-5% and then is converted;
in the step a), the conversion adopts a microbial fermentation method or an ethanol oxidation method. Preferably, the transformation is by microbial fermentation.
In one embodiment of the present invention, in step b1), the carboxylic hydrogen atoms of acetic acid are neutralized and removed with an alkaline agent.
Preferably, the alkaline agent comprises calcium hydroxide, sodium hydroxide, potassium hydroxide. The original carboxyl hydrogen is removed by neutralization with an alkaline reagent in the form of acetate.
In an embodiment of the present invention, in step b2), removing water in the original solution by drying, chromatographic separation or freeze drying; preferably freeze-dried.
In an embodiment of the present invention, in the step b3), the dilute acid solution is an aqueous solution of an inorganic acid.
Preferably, the inorganic acid comprises dilute sulfuric acid, dilute hydrochloric acid, dilute nitric acid and dilute phosphoric acid; preferably, the dilute acid solution is an aqueous solution of sulfuric acid. In dilute acid solutions, the hydrogen isotope ratio of water is known.
In an embodiment of the present invention, in the step b4), after the product obtained in the step b3) is cracked at a high temperature and is converted into hydrogen, a stable isotope ratio mass spectrometer is used to determine the hydrogen isotope ratio of the hydrogen. The high-temperature constant temperature is 1300-1500 ℃. Preferably, the constant temperature is 1420 ℃.
In one embodiment of the present invention, in the step c), the calibration of the acetic acid standard substance is performed before the measurement of the acetic acid standard substance by the step b). In the examples of the present invention, the methyl hydrogen isotope ratio of acetic acid standard substance2HMethyl radicalIt is known that the acetic acid standard substance is measured after the treatment of steps b1) to b4) before the measurement of the acetic acid standard substance2HMeasuringMeasuring the value, and then establishing the methyl site of the acetic acid standard substance2HMethyl radicalWith acetic acid standard2HMeasuringModels of the relationship between measured values, i.e.2HMethyl radicalValue a2HMeasuring+ b. Measuring the position of acetic acid methyl group in alcoholic beverage2Substituting measured values of H into the relational model2HMethyl radicalValue a2HMeasuring+ b, calculating the methyl acetate sites in the alcoholic beverage2HMethyl radicalThe result is that of the ethanolmethyl site2And H value.
In an embodiment of the present invention, in the step a), when the microbial fermentation method is adopted, the fermentation treatment is performed at a constant temperature of 30 ℃ by adding the acetic acid bacteria culture solution to the alcoholic beverage, adding oxygen, sealing the alcoholic beverage.
In an embodiment of the present invention, in step a), the alcoholic beverage includes distilled liquor, fermented liquor and compound liquor.
Preferably, the fermented wine comprises beer and wine; the distilled liquor comprises Chinese liquor, brandy, whisky, vodka and rum; the compound wine comprises fruit wine. The classification selection of the alcoholic beverage GB/T17204-.
The present invention will be described in detail with reference to examples.
Example oneMethyl acetate site stable hydrogen isotope ratio (2H) Model of relationship with measured values
1) Selecting three acetic acid working standard substances, marked as 1#, 2#, 3#, and the stable hydrogen isotope ratio of methyl site thereof (A:)2H) Respectively 223.93 per mill, -61.02 per mill and 141.65 per mill;
2) taking 2mL of the three acetic acid standard substances in the step 1), and preparing aqueous solutions with the volume concentration of 3% acetic acid content by using purified water respectively;
3) taking 25mL of the three acetic acid aqueous solutions obtained in the step 2), adding 1g of calcium hydroxide, performing vortex oscillation for 2h, centrifuging at 8000rpm for 30min, and taking supernatant for later use;
4) putting the supernatant obtained in the step 3) into a freeze dryer for freeze drying to remove water, and keeping the powder for later use;
5) adding 15mL of sulfuric acid solution (1moL/L) of water into the powder obtained in the step 4)2H is-88.51 per mill, and vortex shaking is carried out for 1H;
6) adding anhydrous acetone into the sample treated by the dilute sulfuric acid in the step 5) until the concentration of acetic acid is about 4g/L, standing, and taking supernatant to be analyzed;
7) determination of the acetic acid obtained in step 6) by means of a stable isotope ratio mass spectrometer2H values, results are shown in Table 1.
Table 1 acetic acid working standard2H measurement and methyl group2Given value of H (‰)
As can be seen from the data in Table 1: methyl acetate site2H measurement and methyl acetate site2H given value relation model:2Hmethyl radical＝1.3513*2HMeasuring+67.455. Linear correlation coefficient R of relational model2＝0.9987。
Example twoStability of stable hydrogen isotope ratio of ethanol methyl site in alcoholic beverage
Taking wine, whisky and pure ethanol as research objects, and respectively diluting with purified water until the volume concentration of the ethanol is 3%;
adding 50mL of diluted samples obtained in the step 1) into a 500mL reagent bottle, adding 2.5mL of acetic acid bacillus culture solution into each sample, filling oxygen, sealing, and performing oscillation treatment at 30 ℃ until fermentation is finished, wherein each sample is treated by 5 parts;
taking 25mL of the fermented samples in the step 2), adding 1g of calcium hydroxide, performing vortex oscillation for 2h, centrifuging at 8000rpm for 30min, and taking the supernatant for storage for later use;
putting the supernatant obtained in the step 3) into a freeze dryer for freeze drying to remove water, and keeping the powder for later use;
adding 15mL of sulfuric acid solution (1moL/L) into the powder obtained in the step 4), wherein the water content in the sulfuric acid solution2H is-88.51 per mill, and vortex shaking is carried out for 1H;
adding anhydrous acetone into the sample treated by the dilute sulfuric acid in the step 5) until the concentration of acetic acid is about 4g/L, standing, and taking the supernatant to be analyzed;
7) determining the methyl site of the acetic acid obtained in step 6) by using a stable isotope ratio mass spectrometer2H values, results are shown in Table 2.
TABLE 2 methyl site of conversion of samples to acetic acid2H value (‰)
| Sample (I) | Repetition of-1 | Repetition of-2 | Repeat-3 | Repeat-4 | Repeat-5 |
| Conversion of wine ethanol to acetic acid | -313.028 | -312.570 | -314.928 | -310.634 | -311.454 |
| Conversion of acetic acid by whisky ethanol | -327.692 | -328.452 | -325.147 | -326.972 | -324.149 |
| Conversion of pure ethanol to acetic acid | -169.577 | -170.530 | -167.945 | -168.088 | -172.332 |
As can be seen from Table 2, 3 samples of acetic acid were convertedArticle of manufacture2The standard deviation of the H determination result is 1.64 per thousand, 1.79 per thousand and 1.83 per thousand respectively, and the standard deviation of the H determination result meets the requirement that the standard deviation of the hydrogen isotope ratio determination method is better than 3 per thousand. The methyl sites of acetic acid were measured for three samples separately2Substituting the average value of the H value into the relationship model established in the first embodiment2HMethyl radical＝1.3513*2HMeasuring+67.455 calculation of methyl sites for ethanol conversion of wine to acetic acid, whisky ethanol to acetic acid and pure ethanol to acetic acid2H values are-355.04 ‰, -373.92 ‰, and-161.95 ‰, respectively, and the result is the stable hydrogen isotope ratio of the ethanol methyl site of each sample.
EXAMPLE IIIAccuracy verification of method for determining stable hydrogen isotope ratio of ethanol methyl site in alcoholic beverage
The SNIF-NMR technique is currently the official method for determining the hydrogen isotope ratio of the methyl site of ethanol as recognized by the International grape and wine organizations. To verify the accuracy of the method of the invention, three samples of example two were assayed using SNIF-NMR techniques and the results are shown in Table 3.
TABLE 3 comparison of the SNIF-NMR technique with the results of the determination according to the invention (‰)
| Sample (I) | Grape wine | Whisky | Pure ethanol |
| SNIF-NMR measurement result (D/H, ppm) | 100.54 | 97.16 | 130.75 |
| Measurement result (. delta.) by the method of the present invention2H value,% o) | -355.04 | -373.92 | -161.95 |
As can be seen from Table 3, there is a linear positive correlation between the result of SNIF-NMR measurement (D/H, ppm) and the result of the measurement of the present invention (2H value,% o), and the correlation coefficient R2The stable hydrogen isotope ratio of the ethanol methyl site in the alcoholic beverage can be accurately determined by the method disclosed by the invention, and the method can be popularized and applied as a substitute technology of SNIF-NMR.
Finally, it should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.
Claims (10)
1. A method for determining the stable hydrogen isotope ratio of the methyl site of ethanol in alcoholic beverages comprises the following steps:
a) converting ethanol in the alcoholic beverage to be tested into acetic acid;
b) measuring the hydrogen isotope ratio of acetic acid in the alcoholic beverage obtained in the step a);
c) calibrating the hydrogen isotope ratio of the acetic acid measured in step b) with an acetic acid standard.
2. The method of claim 1,
the step b) comprises the following steps:
b1) removing carboxyl hydrogen atoms of acetic acid in the product obtained in the step a);
b2) removing water from the original solution of the product obtained in the step b 1);
b3) acidifying the product of step b2) with a dilute acid solution;
b4) determining the hydrogen isotope ratio of the acetic acid in the product obtained in step b3) by using a stable isotope ratio mass spectrometer.
3. The method of claim 1,
in the step a), the alcoholic beverage to be tested is diluted by water until the volume concentration of ethanol is 3-5% and then is converted;
in the step a), the conversion adopts a microbial fermentation method or an ethanol oxidation method.
4. The method of claim 2,
in the step b1), neutralizing and removing carboxyl hydrogen atoms of acetic acid by using an alkaline reagent;
preferably, the alkaline agent comprises calcium hydroxide, sodium hydroxide or potassium hydroxide.
5. The method of claim 2,
in the step b2), removing the water in the original solution by adopting an oven drying mode, a chromatographic separation mode or a freeze drying mode.
6. The method of claim 2,
in the step b3), the dilute acid solution is an aqueous solution of inorganic acid;
preferably, the inorganic acid includes dilute sulfuric acid, dilute hydrochloric acid, dilute nitric acid, and dilute phosphoric acid.
7. The method of claim 2,
in the step b4), after the product obtained in the step b3) is cracked at a high temperature and a constant temperature and is converted into hydrogen, a stable isotope ratio mass spectrometer is used for measuring the hydrogen isotope ratio of the hydrogen;
preferably, the high-temperature constant temperature is 1300-1500 ℃.
8. The method of claim 2,
in the step c), when the acetic acid standard substance is corrected, the acetic acid standard substance needs to be treated by the step b) before being measured.
9. The method of claim 1,
in the step a), the alcoholic beverage comprises distilled liquor, fermented liquor and prepared liquor;
preferably, the fermented wine comprises beer and wine; the distilled liquor comprises Chinese liquor, brandy, whisky, vodka and rum; the compound wine comprises fruit wine.
10. The method of claim 3,
in the step a), when a microbial fermentation method is adopted, the acetic acid bacillus culture solution is added into the alcoholic beverage, oxygen is added, sealing is carried out, and fermentation treatment is carried out at the constant temperature of 30 ℃.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110136097A1 (en) * | 2008-05-15 | 2011-06-09 | Ivan Smajlovic | Method for determining origin of alcohol or sugar containing products |
| US20110223629A1 (en) * | 2008-10-06 | 2011-09-15 | Ivan Smajlovic | Method for determination of delta-d values of non- exchangeable hydrogen stable isotopes on ethanol' s methyl group by means of irms instrumental technique |
| CN111257452A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for detecting synthetic acetic acid added in Zhenjiang aromatic vinegar |
| CN111257449A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for determining stable hydrogen isotope ratio of acetic acid methyl sites in vinegar beverage |
| CN111257402A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for identifying synthetic acetic acid in food additive glacial acetic acid |
| CN111257447A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for analyzing stable hydrogen isotope ratio of non-exchangeable hydrogen in total sugar of food |
| CN111257403A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for measuring stable hydrogen isotope ratio of pure acetic acid methyl site |
-
2020
- 2020-07-09 CN CN202010656906.4A patent/CN111999371A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110136097A1 (en) * | 2008-05-15 | 2011-06-09 | Ivan Smajlovic | Method for determining origin of alcohol or sugar containing products |
| US20110223629A1 (en) * | 2008-10-06 | 2011-09-15 | Ivan Smajlovic | Method for determination of delta-d values of non- exchangeable hydrogen stable isotopes on ethanol' s methyl group by means of irms instrumental technique |
| CN111257452A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for detecting synthetic acetic acid added in Zhenjiang aromatic vinegar |
| CN111257449A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for determining stable hydrogen isotope ratio of acetic acid methyl sites in vinegar beverage |
| CN111257402A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for identifying synthetic acetic acid in food additive glacial acetic acid |
| CN111257447A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for analyzing stable hydrogen isotope ratio of non-exchangeable hydrogen in total sugar of food |
| CN111257403A (en) * | 2020-02-06 | 2020-06-09 | 中国食品发酵工业研究院有限公司 | Method for measuring stable hydrogen isotope ratio of pure acetic acid methyl site |
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