CN102384947A - Method for distinguishing genuineness of solid-fermentation liquor - Google Patents
Method for distinguishing genuineness of solid-fermentation liquor Download PDFInfo
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Abstract
The invention relates to a method for distinguishing genuineness of solid-fermentation liquor, which belongs to the liquor analysis technical field, and mainly comprises the following steps that: (1) 13C/12C ratio of ethanol in a reference standard sample (solid-fermentation liquor) is measured by utilizing gas phase chromatography/stable isotopic mass spectrum combined technology, a solid-fermentation liquor volatile ingredient delta 13C database is established; (2) the 13C/12C ratio of the ethanol in commodity liquor is measured; (3) the 13C/12C ratio of the ethanol in the two types of liquor are compared, if on remarkable difference exists between the 13C/12C ratio of the ethanol in reference standard sample and that of the ethanol in the commodity liquor, the tested sample is proved to be the solid-fermentation liquor, and if the 13C/12C ratio of the ethanol in the commodity liquor is far different from that of the ethanol in the reference standard sample, the tested sample is not solid-fermentation liquor. By utilizing the stable isotopic spectrum technology to measure the 13C/12C ratio of the ethanol in the liquor, the method for accurately distinguishing the genuineness of the solid-fermentation liquor is established, so the benefit of the consumers can be effectively protected, and the healthy development of the solid-fermentation liquor with traditional characteristics can be promoted.
Description
Technical field
A kind of method of differentiating the white spirit by solid state method true and false is specifically related to adopt alcohol component in the stable isotope mass spectroscopy liquor
13C/
12The ratio of C is differentiated the white spirit by solid state method true and false, belongs to the liquor analysis technical field.
Background technology
White spirit by solid state method is the distinct Chinese characteristics traditional foods, has a long history, and relies on its unique solid state fermentation and solid state distillation traditional fermentation technology, lists in one of the six big Spirits in the world.Along with social development; It is lower that cost appears in liquor industry; The liquid alcohol technology that automaticity is higher is through mode production of new craft liquor such as string steaming, blending and solid-liquid combination, conventional solid-state method liquor relatively; New technology liquor allows to use materials such as edible alcohol, fragrance and fragrance to be adjusted to the quality of sampling wine, and process control, production technology, quality and cost of products all are starkly lower than conventional solid-state liquor.In order to protect conventional solid-state liquor and consumers' rights and interests, national standard explicitly calls for all liquor of producing of adopting new technology to need clearly mark execution products standards, to show the traditional aromatic white spirit of difference.But because supporting detection technique disappearance, many enterprises are for maximum benefitization on the market, and the liquor of adopting new technology pretends to be traditional odor type white spirit by solid state method in market sale, has seriously disturbed the healthy and orderly competition order of liquor industry.Therefore, setting up the effective authentication technique of the white spirit by solid state method true and false is that current liquor industrial research field has technical barrier to be solved.
Isotope is one of two or more atoms with same chemical element of same atoms ordinal number.Carbon atom mainly contains stable isotope δ
12C and δ
13C; Different plants metabolic pathway in the photosynthesis process is different, like C
3Plant Calvin cycle fixation of C O
2, δ
13The negative value of C is corresponding bigger; C
4Plant a kind ofly contains 4 carbon compounds and carries out fixation of C O through forming
2, δ
13The negative value of C is corresponding less; The part plant is through crassulacean acid metabolism (CAM) approach fixation of C O
2, its δ
13C is between C3 and C4 between the two.In the liquor fermentation process, ethanol is produced by microbial metabolism with other volatile ingredients simultaneously, has the metabolism homologous relationship.Therefore, δ in the raw material in the liquor
13δ in C and ethanol, other volatile ingredients
13Certainly exist the metabolism homologous relationship between the C.According to this principle; The present invention adopts gas chromatography/stable isotope mass spectrometric hyphenated technique (GC/IRMS) to measure alcohol component carbon isotope ratio in the liquor; Theoretical in conjunction with the metabolism homology; The carbon isotope ratio difference of finding out between white spirit by solid state method and the non-white spirit by solid state method is different, thereby realizes effective discriminating of the white spirit by solid state method true and false.
Summary of the invention
1) the quasi-solution problem of determining
The present invention is directed to the white spirit by solid state method true and false that occurs in the liquor industry and be difficult to the effectively technical barrier of discriminating; Develop a kind of technical method that can effectively differentiate the white spirit by solid state method true and false; To normalize market behavior, protection consumer and legitimate enterprise interests promote the healthy and orderly development of liquor industry.
2) concrete scheme of the present invention is following:
A) reference standard sample: obtaining with the Chinese sorghum from conventional solid-state fermentation liquor enterprise is the solid state process brewing spirit sample of raw material;
B) testing sample: buying with the Chinese sorghum from market is raw material and the sample that is nominally white spirit by solid state method;
C) retention time is confirmed: with ethanol is standard items, confirms that it goes out the retention time at peak in GC/IRMS;
D) ethanol
13C/
12C measures: with sample and acetone according to 1: 70 (V: V) ratio mixing, get mixed solution in 2mL sample introduction bottle, as follows ethanol in the working sample
13C/
12C, chromatographic condition is following: capillary chromatograph specification CP-Wax (50m * 2.5mm * 0.25 μ m), 200 ℃ of GC injector temperatures; Split ratio 1: 20, carrier gas are high-purity helium (99.999%), take constant current mode (1.2mL/min); Column temperature adopts temperature programme: 40 ℃ of initial temperatures, keep 1min, and be warming up to 80 ℃ with 5 ℃/min; 25 ℃/min rises to 200 ℃, the flame tube temperature: 1030 ℃; Sample size 0.7 μ L.Mass spectrum parameter: forepump vacuum 2 * 10
-3Pa, ion gun vacuum 5 * 10
-8Pa, transmitter current 115mA.Before sample determination, use the standard model calibration instrument every day.
E) reference standard sample (white spirit by solid state method) volatile ingredient δ
13The C database makes up: a) collected specimens set by step, selecting more than at least 20 batches is the liquor of raw material by solid fermentation technology with the Chinese sorghum, measures ethanol and other volatile ingredients δ respectively according to step d)
13C sets up white spirit by solid state method volatile ingredient δ
13The C database adopts statistical technique, research volatile ingredient δ
13C numerical range and metabolism correlativity.
F) the testing sample volatile ingredient is measured: measure ethanol and other volatile ingredients δ in the working sample respectively according to step d)
13C;
G) determination step: with f) step obtains testing sample and measures volatile ingredient
13C/
12Reference standard sample (white spirit by solid state method) the volatile ingredient δ that C value and step e) obtain
13The C database carries out significant difference and judges; If there is not significant difference, prove that then testing sample is a white spirit by solid state method; If there is significant difference, then testing sample is non-white spirit by solid state method.
3) beneficial effect:
The present invention has set up a kind of technical method of fast, accurately and effectively differentiating solid spirit and solid-liquid method liquor; The detection means that the white spirit by solid state method true and false faces and the problem of technical method disappearance have been solved in market surpervision; Effectively protect conventional solid-state liquor intellecture property and consumers' rights and interests, the industry development of standard liquor.
Description of drawings:
Accompanying drawing 1: gas chromatography-isotope mass spectrometry (GC-IRMS) is measured 44,45,46 ion flow graphs of ethanol;
Accompanying drawing 2: reference standard sample liquor ethanol
13C/
12C ratio frequency descriptive statistics figure.
Embodiment:
Below in conjunction with instantiation the present invention is further specified.
Embodiment one:
Ethanol in the liquor
13C/
12C measures
A) get chromatographically pure ethanol 20 μ L in the sample introduction bottle, add the 1.4mL acetone soln, mixing;
B) ethanol δ in the working sample as follows
13C
Chromatographic condition: capillary chromatograph specification CP-Wax (50m * 2.5mm * 0.25 μ m), 200 ℃ of GC injector temperatures, split ratio 1: 20; Carrier gas is high-purity helium (99.999%), takes constant current mode (1.2mL/min), and column temperature adopts temperature programme: 40 ℃ of initial temperatures; Keep 1min; Be warming up to 80 ℃ with 5 ℃/min, back 25 ℃/min rises to 200 ℃, the flame tube temperature: 1030 ℃; Sample size 0.7 μ L.
Mass spectrum parameter: forepump vacuum 2 * 10
-3Pa, ion gun vacuum 5 * 10
-8Pa, transmitter current 115mA.
C) adopt known δ
13The ethanol of C is demarcated isotope mass spectrometry reference gas CO
2δ
13C;
D) testing sample is injected 44,45, the 46 ion flow graphs that GC/IRMS obtains liquor ethanol, see accompanying drawing 1.
Embodiment two:
White spirit by solid state method discriminating case one
A) obtain 30 reference standard samples of white spirit by solid state method sample from conventional solid-state zymotechnique enterprise;
B) from the market buy 5 in the sample that is nominally white spirit by solid state method;
C) ethanol in the liquor
13C/
12C measures
Get liquor 20 μ L in the sample introduction bottle, add the 1.4mL acetone soln, mixing, the δ of ethanol in the working sample as follows
13C/ δ
12C, chromatographic condition is following: capillary chromatograph specification CP-Wax (50m * 2.5mm * 0.25 μ m), 200 ℃ of GC injector temperatures; Split ratio 1: 20, carrier gas are high-purity helium (99.999%), take constant current mode (1.2mL/min); Column temperature adopts temperature programme: 40 ℃ of initial temperatures, keep 1min, and be warming up to 80 ℃ with 5 ℃/min; 25 ℃/the min in back rises to 200 ℃, reaction tube temperature: 1030 ℃; Sample size 0.7 μ L.Mass spectrum parameter: forepump vacuum 2 * 10
-3Pa, ion gun vacuum 5 * 10
-8Pa, transmitter current 115mA;
D) white spirit by solid state method sample ethanol
13C/
12C sees table 1, visible white spirit by solid state method ethanol
13C/
12The C ratio range is between-19.77 ‰～15.83 ‰;
Table 1 white spirit by solid state method sample determination descriptive statistics
| White spirit by solid state method | Sample number/individual | Minimal value/‰ | Maximum value/‰ | Average/‰ | Standard deviation |
| Ethanol 13C/ 12C | 30 | ?-19.77 | ?-15.83 | -17.17 | 0.71 |
E) testing sample ethanol
13C/
12C ratio is seen table 2
Table 2 testing sample is measured descriptive statistics
| Testing sample | Ethanol 13C/ 12C(‰) |
| Sample 1 | -13.87 |
| Sample 2 | -13.47 |
| Sample 3 | -12.55 |
| Sample 4 | -17.36 |
| Sample 5 | -17.82 |
Can know white spirit by solid state method ethanol by table 1
13C/
12The C ratio range is between-19.77 ‰～-15.99 ‰; Can know the ethanol of sample 4 and sample 5 by table 2
13C/
12C is respectively-17.36 ‰ and-17.82 ‰, between-19.77 ‰～-15.83 ‰, shows that sample 4 and sample 5 should belong to white spirit by solid state method; And the ethanol of sample 1, sample 2 and sample 3
13C/
12C ratio all departs from white spirit by solid state method ethanol
13C/
12Therefore the C ratio range, judges that sample 1, sample 2 and sample 3 are non-white spirit by solid state method.
Embodiment three:
White spirit by solid state method discriminating case 2
A) selection is that the white spirit by solid state method A of primary raw material is with reference to appearance with the Chinese sorghum;
B) with white spirit by solid state method A with the corn be the alcohol B of raw material according to 7: 3 and 5: 5 ratios, blend out 2 solid-liquid method liquor samples;
C) measure ethanol in white spirit by solid state method A, corn alcohol B, solid-liquid method liquor 1 (containing 30% corn alcohol) and the solid-liquid method liquor 2 (containing 50% corn alcohol) respectively according to case study on implementation one step
13C/
12C ratio obtains table 3
Table 3 sample determination descriptive statistics
| Testing sample | Ethanol 13C/ 12C |
| White spirit by solid state method A | -17.08‰ |
| Solid-liquid method liquor 1 (containing 30% corn alcohol) | -14.87‰ |
| Solid-liquid method liquor 2 (containing 50% corn alcohol) | -14.01‰ |
| Corn alcohol B | -10.53‰ |
D) visible from table 3, ethanol in the solid-liquid method liquor
13C/
12C ratio increases with adding corn alcohol ratio, and has departed from solid state process ethanol
13C/
12C ratio range-17.17 ‰ ± 0.71 this shows, ethanol in white spirit by solid state method and the solid-liquid method liquor
13C/
12There is comparatively evident difference in C, through ethanol in the working sample
13C/
12C ratio can effectively be differentiated white spirit by solid state method and solid-liquid method liquor, corn alcohol.
Should be noted that at last; Above embodiment only is used to explain technical scheme of the present invention and is unrestricted; Although with reference to preferred embodiment the present invention is specified, those of ordinary skill in the art should be appreciated that and can make amendment or be equal to replacement the technical scheme of invention; And not breaking away from the spirit and the scope of technical scheme of the present invention, it all should be encompassed in the claim scope of the present invention.
Claims (6)
1. a method of differentiating the white spirit by solid state method true and false is characterized in that, said method comprising the steps of:
1) employing gas chromatography/stable isotope mass spectrometric hyphenated technique is measured the middle ethanol of reference standard sample (white spirit by solid state method)
13C/
12C ratio is set up white spirit by solid state method volatile ingredient δ
13The C database;
2) measure ethanol in the testing sample liquor
13C/
12C ratio;
3) testing sample is measured alcohol component
13C/
12C ratio and white spirit by solid state method ethanol
13C/
12C ratio compares, if do not exist than big-difference, proves that then testing sample is a white spirit by solid state method; If there is notable difference, then testing sample is non-white spirit by solid state method.
2. method according to claim 1 is characterized in that, it is the white spirit by solid state method real and fake discrimination of brewing materials that said method is used for the Chinese sorghum.
3. method according to claim 1 is characterized in that, said assay method is gas chromatography/isotope mass spectrometry coupling technique (GC/IRMS).
4. method according to claim 1 is characterized in that, when working sample ethanol
13C/
12During the C value, need utilize acetone that the liquor sample is diluted, liquor and acetone volume ratio are 1: 75;
5. method according to claim 3 is characterized in that, when working sample ethanol
13C/
12During C ratio, chromatographic condition is following: capillary chromatograph specification CP-Wax (50m * 2.5mm * 0.25 μ m), 200 ℃ of GC injector temperatures; Split ratio 1: 20, carrier gas are high-purity helium (99.999%), take constant current mode (1.2mL/min); Column temperature adopts temperature programme: 40 ℃ of initial temperatures, keep 1min, and be warming up to 80 ℃ with 5 ℃/min; 25 ℃/the min in back rises to 200 ℃, the flame tube temperature: 1030 ℃; Sample size 0.7 μ L.
6. method according to claim 3 is characterized in that, when working sample ethanol
13C/
12The isotope mass spectrometry condition is during C ratio: forepump vacuum 2 * 10
-3Pa, ion gun vacuum 5 * 10
-8Pa, transmitter current 115mA.
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| CN103235030A (en) * | 2013-03-25 | 2013-08-07 | 江苏易谱恒科技有限公司 | Distillate spirit brand identification method based on support vector machine and time-of-flight mass spectrometry |
| CN103293141A (en) * | 2013-03-25 | 2013-09-11 | 江苏省质量安全工程研究院 | A liquor vintage recognition method based on a fusion technology of ion mobility spectrometry/ mass spectrometry/ Raman spectroscopy |
| CN105092756A (en) * | 2015-06-09 | 2015-11-25 | 江南大学 | Method for rapid detection of dhurrin in Baijiu brewing raw material sorghum |
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| CN106908509A (en) * | 2017-02-08 | 2017-06-30 | 北京农业质量标准与检测技术研究中心 | The quick and quantitative detecting method of doping paraffin in a kind of lard |
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Cited By (10)
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| CN102967648A (en) * | 2012-11-20 | 2013-03-13 | 中国食品发酵工业研究院 | Method for identifying organic hot pepper based on stable isotope ratio |
| CN102980937A (en) * | 2012-11-20 | 2013-03-20 | 中国食品发酵工业研究院 | Method for identifying organic coriander based on stable isotope ratio |
| CN103235030A (en) * | 2013-03-25 | 2013-08-07 | 江苏易谱恒科技有限公司 | Distillate spirit brand identification method based on support vector machine and time-of-flight mass spectrometry |
| CN103293141A (en) * | 2013-03-25 | 2013-09-11 | 江苏省质量安全工程研究院 | A liquor vintage recognition method based on a fusion technology of ion mobility spectrometry/ mass spectrometry/ Raman spectroscopy |
| CN103293141B (en) * | 2013-03-25 | 2015-03-11 | 江苏省质量安全工程研究院 | A liquor vintage recognition method based on a fusion technology of ion mobility spectrometry/ mass spectrometry/ Raman spectroscopy |
| CN103235030B (en) * | 2013-03-25 | 2015-06-03 | 江苏易谱恒科技有限公司 | Distillate spirit brand identification method based on support vector machine and time-of-flight mass spectrometry |
| CN105092756A (en) * | 2015-06-09 | 2015-11-25 | 江南大学 | Method for rapid detection of dhurrin in Baijiu brewing raw material sorghum |
| CN105424897A (en) * | 2015-11-24 | 2016-03-23 | 中国食品发酵工业研究院 | Method for detecting exogenously-added ethyl caproate in baijiu |
| CN105445429A (en) * | 2015-11-24 | 2016-03-30 | 中国食品发酵工业研究院 | Method for detecting exogenously added ethyl caproate and ethyl lactate in liquor |
| CN106908509A (en) * | 2017-02-08 | 2017-06-30 | 北京农业质量标准与检测技术研究中心 | The quick and quantitative detecting method of doping paraffin in a kind of lard |
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