CN101825572A - Method for differentiating Chinese spirits with different flavor types with fluorescein - Google Patents
Method for differentiating Chinese spirits with different flavor types with fluorescein Download PDFInfo
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Abstract
The invention provides a method for differentiating Chinese spirits with different flavor types with fluorescein, which includes the preparation of fluorescein reagent and solution to be tested, the setting of spectral parameters, the acquisition of spectral data and the pattern identification of an extracted spectral characteristic intensity value. The method first uses absolute ethanol to prepare the fluorescein reagent, the concentration of which is 3.00 multiplied by 10 minus 3mol/L, the fluorescein reagent is kept in the darkness under the temperature of 4 DEG C, the prepared fluorescein reagent is taken, the solution to be tested is prepared with the fluorescein reagent and Chinese spirit to be tested according to the volume ratio of 1:100, and is kept still for a period of time, a fluorescence spectrometer is then used for acquiring the emission spectrum of the solution to be tested, the spectral characteristic intensity value is extracted and inputted into SPSS software, which carries out hierarchical cluster analysis and principal component analysis, and by checking the fluorescence spectrum, the Chinese spirit with a different flavor type can be differentiated. The method is quick and highly efficient, and can be fulfilled within a few minutes, and moreover, the sample does not need to be preprocessed; the method can accurately differentiate Chinese spirits with different flavor types at low cost; and the method can be applied to continuous production, and is highly applicable.
Description
Technical field
The present invention relates to a kind of method of distinguishing different flavor liquor, relate in particular to a kind of method of distinguishing the identical or approaching different flavor liquor of alcoholic strength by fluorescein.
Background technology
Liquor is the characteristic drinks of China, and it is brewageed production technology and belongs to traditional industries, and long historical culture is arranged.The development of liquor has been created the magnificent culture of drinking when enriching our life.China's liquor odor type is rich and varied, is a focus of liquor industry research always.The discriminating of dialogue aroma type in the past mainly relies on people's sense organ to differentiate, but this method not only needs long term experience accumulation, and it is bigger influenced by people's the sense organ and the state of mind, and stability and accuracy are difficult to assurance.In recent years, development and application along with gas chromatographic technique, hundreds of materials such as the alcohols in the liquor, ester class, acids, aldehydes have all been carried out qualitative, quantitative research, but, often can not assign to define a kind of liquor with several even tens kinds of one-tenth because of the difference of micro constitutent content in the liquor is very little.Existing Electronic Nose analysis and detection technology, mass-spectrometric technique, near-infrared spectrum technique etc. also are widely used in distinguishing different flavor liquor, but loaded down with trivial details because of handling early stage, cost an arm and a leg, shortcoming such as length consuming time, enterprise is difficult to popularization and application at system wine.
Summary of the invention
At weak point of the prior art, the invention provides the efficient height, distinguish a kind of method of distinguishing different flavor liquor by fluorescein that cost is low and applicability is strong.
A kind of method provided by the invention by fluorescein differentiation different flavor liquor, the key step of this method comprises:
1) preparing concentration with absolute ethyl alcohol is 2.5 * 10
-3～3.5 * 10
-3The luciferase reagent of mol/L keeps in Dark Place under 3～4 ℃;
2) get the luciferase reagent that configures in the step 1), the liquor different to be measured that luciferase reagent is identical or approaching with alcoholic strength respectively disposes solution to be measured by 1: 80～1: 120 volume ratio, shakes up under the lucifuge condition of back and leaves standstill a period of time T, T＞30min;
3) from different solution to be measured, get 2.5～3.5mL, be contained in respectively in the quartz colorimetric utensil, seal;
4) excitation wavelength with fluorescence spectrophotometer is set at 490～500nm, and incident and emission slit are 2.5～10.0nm; Adopt fluorescence spectrophotometer that the solution to be measured in each quartz colorimetric utensil is scanned, sweep velocity is 800～1200nm/min, and writes down the fluorescence emission spectrum in 200～800nm wavelength coverage;
5) be to choose several corresponding fluorescence peak intensity levels in 300～380nm and the 505～530nm scope from the fluorescence emission spectrum medium wavelength, input SPSS software, carry out pattern recognition analysis by SPSS software, demonstrate the dendrogram and the two-dimentional scatter diagram of different flavor liquor correspondence, different flavor liquor is distinguished by dendrogram and two-dimentional scatter diagram.
Further, in the described step 4), the excitation wavelength of fluorescence spectrophotometer is set at 495nm.
Compared with prior art, a kind of method by fluorescein differentiation different flavor liquor of the present invention has following advantage:
1, high efficiency: the present invention can distinguish by quickly and efficiently that alcoholic strength is identical or approaching liquor different to be measured, and wine sample of gas chromatography determination needs about 1 hour, and fluorometry generally only needs time a few minutes, and sample does not need pre-treatment.
2, accuracy height: manually distinguished different flavor liquor expense height, it is many that accuracy is subjected to the artificial subjective factor influence in the past, and method of the present invention can be low-cost, and the high accuracy different flavor liquor that alcoholic strength is identical or approaching distinguishes.
3, globality: compare gas chromatography and liquid phase chromatography to single analysis of planting material in the liquor, fluorometry lays particular emphasis on the research to liquor spectrum globality.Liquor is in the process of storing, and a series of physicochemical change take place in this complicated system its micro substance, and each component concentration tends to balance, and the wine body tends towards stability, and has new material to form.And these features can obtain embodying in the fluorescence spectrum after fluorescein and the liquor effect.
4, applicability is strong: the method that this method combines with fluorescence intensity with the fluorescence spectrum shape distinguishes the liquor of different flavor, its method is applied in the continuous production, can measures the liquor fluorescence spectrum at any time, standard liquor production run, make the liquor style and liquor of producing outstanding, best in quality.
5, in the liquor because contain alcohols, aldehydes, the ester class, acids, micro substances such as phenols itself have certain fluorescent characteristic, and are overall about 2% but micro substance content only accounts for, and cause its fluorescent characteristics conspicuousness relatively poor.And the fluorescein conjugated structure big because of its molecule has that the present invention uses, and be rigid plane, and fluorescence quantum yield is higher, and fluorescence intensity is remarkable.And compare the fluorescence spectrum of liquor self, relative with liquor fluorescent spectrum curve after the fluorescein effect level and smooth simple, have certain advantage aspect data analysis and the spectrum identification.
6, the main performance of micro constitutent in ultraviolet-visible light spectrum in the liquor paid close attention in research in the past, and fluorescence spectrum analysing method is than highly sensitive 2～3 orders of magnitude of beam split photometry, so the fluorescence spectrum difference that obtains after liquor and the luciferin reaction has reflected the difference of micro substance kind and content in the liquor to a certain extent.
Description of drawings
Fig. 1 is the fluorescence emission spectrogram of Guilin three-flower wine, Lu Zhou Lao Jiao Te Qu, Fenyang wine, Xiang Quan Jiu and 5 kinds of liquor of Lang Jiu;
Fig. 2 is the fluorescence emission spectrogram of Xifeng and 2 kinds of liquor of scape sesame Chinese distillate spirits;
Fig. 3 is the fluorescence emission spectrogram of Dong Jiu and 2 kinds of liquor of four special old cellar for storing things wine;
Fig. 4 is 9 kinds of wine sample hierarchical cluster analysis figure;
Fig. 5 is to be the diffusing point of the liquor distribution plan of axle with major component 1 and major component 2.
Embodiment
A kind of method by fluorescein differentiation different flavor liquor, key step comprises:
1) preparing concentration with absolute ethyl alcohol is 2.5 * 10
-3～3.5 * 10
-3The luciferase reagent of mol/L keeps in Dark Place under 3～4 ℃;
2) get the luciferase reagent that configures in the step 1), the liquor different to be measured that luciferase reagent is identical or approaching with alcoholic strength respectively disposes solution to be measured by 1: 80～1: 120 volume ratio, shakes up under the lucifuge condition of back and leaves standstill a period of time T, T＞30min;
3) from different solution to be measured, get 2.5～3.5mL, be contained in respectively in the quartz colorimetric utensil, seal;
4) excitation wavelength with fluorescence spectrophotometer is set at 490～500nm, and incident and emission slit are 2.5～10.0nm; Adopt fluorescence spectrophotometer that the solution to be measured in each quartz colorimetric utensil is scanned, sweep velocity is 800～1200nm/min, and writes down the fluorescence emission spectrum in 200～800nm wavelength coverage;
5) be to choose several corresponding fluorescence peak intensity levels in 300～380nm and the 505～530nm scope from the fluorescence emission spectrum medium wavelength, input SPSS software, utilize hierarchical cluster analysis in the SPSS software and principal component analysis (PCA) or discriminant analysis methods to carry out pattern recognition analysis, demonstrate the dendrogram and the two-dimentional scatter diagram of different flavor liquor correspondence, different flavor liquor is distinguished by dendrogram and two-dimentional scatter diagram.
In the present embodiment, the excitation wavelength of fluorescence spectrophotometer is set at 490～500nm.Fluorescein is in ethanolic solution, and maximum excitation wavelength is near the 495nm; First absorption band is between 440～520nm, and maximum emission peak is about 515nm; Select emission wavelength emission peak after 200～800nm is can write down fluorescein and liquor effect fully basically because of this wavelength coverage, the spectrum performance of crests such as diffraction peak helps the parsing of spectrum and the research of liquor character.Choosing near the characteristic exponent of 300～380nm is because this wave band is the fluorescence peak of ethanol and associated matter thereof, and itself and near the fluorescence peak of 505～530nm are worked in coordination with and can be distinguished different flavor liquor.
According to the China white wine industry standard, liquor can be divided into different flavors such as Luzhou-flavor, Maotai-flavor, delicate fragrance type, rice-fragrant type, and the present invention is an experimental subjects with 9 kinds of aromatic white spirits of China, and its alcoholic strength is distributed near 53%, 50%, 45%.Listed alcoholic strength, odor type and the place of production of 9 kinds of aromatic white spirits in the following table, be commercial with the sample of going with wine.
Embodiment 1
With Guilin three-flower wine, Lu Zhou Lao Jiao Te Qu, Fenyang wine, Xiang Quan Jiu and 5 kinds of liquor of Lang Jiu is object to be measured.
1) be 2.5 * 10 with the absolute ethyl alcohol configuration concentration
-3The luciferase reagent of mol/L keeps in Dark Place under 3 ℃ of temperature.
2) get the luciferase reagent that configures in the step 1), luciferase reagent is disposed 5 kinds of solution to be measured with Guilin three-flower wine, Lu Zhou Lao Jiao Te Qu, Fenyang wine, Xiang Quan Jiu and Lang Jiu by 1: 80 volume ratio respectively, shake up under the lucifuge condition of back and leave standstill 35min.
3) from 5 kinds of solution to be measured, respectively get 2.5mL solution to be measured respectively, be contained in respectively in the quartz colorimetric utensil, seal.
4) excitation wavelength with fluorescence spectrophotometer is set at 490nm, and incident and emission slit are 2.5nm; Respectively 5 kinds of solution to be measured in the quartz colorimetric utensil are scanned with fluorescence spectrophotometer, sweep velocity is 800nm/min, and writes down the fluorescence emission spectrum in 200～800nm wavelength coverage.
Fig. 1 is the fluorescence emission spectrogram of Guilin three-flower wine, Lu Zhou Lao Jiao Te Qu, Fenyang wine, Xiang Quan Jiu and 5 kinds of liquor of Lang Jiu, and 1 represents the Guilin three-flower wine among the figure, and 2 represent the Lu Zhou Lao Jiao Te Qu, and 3 represent Fenyang wine, and 4 represent Xiang Quan Jiu, and 5 represent Lang Jiu.At the maximum emission peak place, the relative intensity of fluorescence of Guilin three-flower wine is the strongest, and under the close situation of wine degree, fluorescence intensity and liquor micro constitutent kind and amount are than relevant.The relative intensity of fluorescence of Lang Jiu is the most weak.The fluorescence intensity of Lu Zhou Lao Jiao Te Qu, Fenyang wine and Xiang Quan Jiu thereof falls between.Embodiment shows that micro constitutent kind and amount are than having incidence relation with its fluorescence intensity in the different flavor liquor.
With Xifeng and 2 kinds of liquor of scape sesame Chinese distillate spirits is object to be measured.
1) be 3.0 * 10 with the absolute ethyl alcohol configuration concentration
-3The luciferase reagent of mol/L keeps in Dark Place under 3.5 ℃ of temperature.
2) get the luciferase reagent that configures in the step 1), luciferase reagent is disposed 2 kinds of solution to be measured with Xifeng and scape sesame white spirit by 1: 100 volume ratio respectively, shake up under the lucifuge condition of back and leave standstill 60min.
3) from 2 kinds of solution to be measured, respectively get 3mL solution to be measured respectively, be contained in respectively in the quartz colorimetric utensil, seal.
4) excitation wavelength with fluorescence spectrophotometer is set at 495nm, and incident and emission slit are 4.5nm; Respectively 2 kinds of solution to be measured in the quartz colorimetric utensil are scanned with fluorescence spectrophotometer, sweep velocity is 1000nm/min, and writes down the fluorescence emission spectrum in 200～800nm wavelength coverage.
Fig. 2 is the fluorescence emission spectrogram of Xifeng and 2 kinds of liquor of scape sesame Chinese distillate spirits, and 6 represent Xifeng among the figure, and 7 represent the Jing Zhi Chinese distillate spirits.Under the approaching situation of alcoholic strength, fluorescent spectrometry can accurately be distinguished this two kinds of liquor.Xifeng is at λ
Em2=513.5nm place, the fluorescence intensity of emission peak is 386.00, scape sesame Chinese distillate spirits is at λ
Em2=515.0nm place, the fluorescence intensity of emission peak is 60.50.Compare with Xifeng, furfural content is higher relatively in the scape sesame Chinese distillate spirits, causes fluorescence intensity to weaken.
Embodiment 3
With Dong Jiu and 2 kinds of liquor of four special old cellar for storing things wine is object to be measured.
1) be 3.5 * 10 with the absolute ethyl alcohol configuration concentration
-3The luciferase reagent of mol/L keeps in Dark Place under 4 ℃ of temperature.
2) get the luciferase reagent that configures in the step 1), luciferase reagent is disposed 2 kinds of solution to be measured with Dong Jiu and four special wine by 1: 120 volume ratio respectively, shake up under the lucifuge condition of back and leave standstill 80min.
3) from 2 kinds of solution to be measured, respectively get 3.5mL solution to be measured respectively, be contained in respectively in the quartz colorimetric utensil, seal.
4) excitation wavelength with fluorescence spectrophotometer is set at 500nm, and incident and emission slit are 10nm; Respectively 2 kinds of solution to be measured in the quartz colorimetric utensil are scanned with fluorescence spectrophotometer, sweep velocity is 1200nm/min, and writes down the fluorescence emission spectrum in 200～800nm wavelength coverage.
Fig. 3 is the fluorescence emission spectrogram of Dong Jiu and 2 kinds of liquor of four special old cellar for storing things wine, and 8 represent Dong Jiu among the figure, and 9 represent four special old cellar for storing things wine.Than four special old cellar for storing things wine, Dong Jiu λ
Em1=342nm, the fluorescence intensity of emission peak is 29.08, relative intensity of fluorescence maximum in 9 kinds of wine samples measuring.At the maximum emission peak place, the fluorescence peak of the two almost overlaps, and these two kinds of liquor fluorescence intensity difference between 300～380nm can be used as the main foundation of judging its odor type.
The identification of SPSS software pattern
SPSS software is existing software, and it comprises hierarchical cluster analysis (HCA) and principal component analysis (PCA) (PCA) two big analytic functions.At " Classification of brandies and wine distillates using front facefluorescence spectroscopy " Food Chemistry 117 (2009) 491-498 Jana S á deck á, JanaT ó thov á, Pavel M á jek (" using the front fluorescence spectrum to differentiate brandy and wine spirit " 2009 117 phases of Food Chemistry 491-498 page or leaf, author: Jana S á deck á, Jana T ó thov á, Pavel M á jek) fluorescence spectrum has been carried out HCA to a Wen Zhong researchist and PCA analyzes." based on the liquor cluster analysis research of three-dimensional fluorescence spectrum characteristic " (photoelectron laser alignment 20 is rolled up in April, 2009 the 4th phase, the 495-498 page or leaf, author: Yang Jianlei, Zhu Tuo, Wu Hao) a Wen Zhong researchist uses SPSS software that fluorescence spectrum has been carried out the HCA analysis.
From the fluorescence emission spectrogram medium wavelength of nine kinds of aromatic white spirits of embodiment 1, embodiment 2 and embodiment 3 is to choose several corresponding fluorescence peak intensity levels in 300～380nm and the 505～530nm scope.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 1 correspondence is two sections fluorescence peaks of 310～370nm and 515～530nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 310nm place is 6.10, and the fluorescence intensity level at 370nm place is 6.42, and the fluorescence intensity level at 333.5nm place is 16.30 to the maximum in this wave band; The fluorescence intensity level at 515nm place is 142.00, and the fluorescence intensity level at 530nm place is 110.00, and the fluorescence intensity level at 515nm place is a maximum fluorescence intensity in this wave band.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 2 correspondences is two sections fluorescence peaks of 320～380nm and 504～519nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 320nm place is 0.79, and the fluorescence intensity level of 380nm correspondence is 0.52, and the fluorescence intensity level at 335.5nm place is 1.12 to the maximum in this wave band; The fluorescence intensity level at 504nm place is 106.50, and the fluorescence intensity level at 519nm place is 124.50, and the fluorescence intensity level at 515.5nm place is 130.00 to the maximum in this wave band.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 3 correspondences is two sections fluorescence peaks of 320～380nm and 504～519nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 320nm place is 2.49, and the fluorescence intensity level at 380nm place is 2.71, and the fluorescence intensity level at 350 places is 3.15 to the maximum in this wave band; The fluorescence intensity level at 504nm place is 93.30, and the fluorescence intensity level at 519nm place is 108.50, and the fluorescence intensity level at 514.5nm place is 111.50 to the maximum in this wave band.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 4 correspondences is two sections fluorescence peaks of 320～380nm and 504～519nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 320nm place is 0.65, and the fluorescence intensity level at 380nm place is 0.90, and the fluorescence intensity level at 359 places is 1.35 to the maximum in this wave band; The fluorescence intensity level at 504nm place is 84.50, and the fluorescence intensity level at 519nm place is 98.30, and the fluorescence intensity level at 515.0nm place is 102.50 to the maximum in this wave band.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 5 correspondences is two sections fluorescence peaks of 310～370nm and 515～530nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 310nm place is 0.05, and the fluorescence intensity level at 370nm place is 0.15, and the fluorescence intensity level at 351.5nm place is 0.20 to the maximum in this wave band; The fluorescence intensity level at 515nm place is 54.50, and the fluorescence intensity level at 530nm place is 41.50, and in this wave band, the fluorescence intensity level at 515nm place is a maximum fluorescence intensity.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 6 correspondences is two sections fluorescence peaks of 320～380nm and 504～519nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 320nm place is 7.13, and the fluorescence intensity level at 380nm place is 6.40, and the fluorescence intensity level at 352nm place is 7.60 to the maximum in this wave band; The fluorescence intensity level at 504nm place is 320.50, and the fluorescence intensity level at 519nm place is 372.50, and the fluorescence intensity level at 514.5nm place is 386.00 to the maximum in this wave band.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 7 correspondences is two sections fluorescence peaks of 300～360nm and 515～530nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 300nm place is 3.25, and the fluorescence intensity level at 360nm place is 3.27, and the fluorescence intensity level at 319.5nm place is 5.36 to the maximum in this wave band; The fluorescence intensity level at 515nm place is 60.50, and the fluorescence intensity level at 519nm place is 45.50, and in this wave band, the fluorescence intensity level at 515nm place is a maximum fluorescence intensity.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 8 correspondences is two sections fluorescence peaks of 310～370nm and 515～530nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 310nm place is 6.50, and the fluorescence intensity level at 370nm place is 17.70, and the fluorescence intensity level at 342.5nm place is 28.80 to the maximum in this wave band; The fluorescence intensity level at 515nm place is 70.50, and the fluorescence intensity level at 530nm place is 41.50, and in this wave band, the fluorescence intensity level at 515nm place is a maximum fluorescence intensity.
Choosing wavelength from the fluorescence emission spectrogram of wine sample 9 correspondences is two sections fluorescence peaks of 310～370nm and 515～530nm, and chooses 152 fluorescence intensity level input SPSS softwares and carry out pattern recognition analysis in two sections fluorescence peaks.Wherein the fluorescence intensity level at 310nm place is 5.45, and the fluorescence intensity level at 370nm place is 5.80, and the fluorescence intensity level at 323.0nm place is 10.50 to the maximum in this wave band; The fluorescence intensity level at 515nm place is 73.50, and the fluorescence intensity level at 530nm place is 54.00, and in this wave band, the fluorescence intensity level at 515nm place is a maximum fluorescence intensity.
Generate Fig. 4 and Fig. 5 by the hierarchical cluster analysis in the SPSS software (HCA), principal component analysis (PCA) (PCA).
In the hierarchical cluster method, the employing Euclidean distance method is carried out cluster analysis to the characteristic fluorescence intensity level of 9 kinds of liquor fluorescence spectrums, as shown in Figure 4.Generally, 9 kinds of liquor are divided into two big classes.Wherein Xifeng is separately a class, and other 8 kinds of liquor are classified as a class.Four special old cellar for storing things wine, Dong Jiu, Guilin three-flower wine, Lu Zhou Lao Jiao Te Qu wine can be distinguished separately in the first kind.And Lang Jiu and scape sesame Chinese distillate spirits sample are got together, and Fenyang wine and Xiang Quan Jiu sample are got together, so the hierarchical cluster method is distinguished poor effect to these 4 kinds of liquor of Lang Jiu, scape sesame white spirit, Fenyang wine and Xiang Quan Jiu.
In view of the limitation of hierarchical cluster method to the differentiation of part wine sample, the present invention carries out PCA to the spectral signature fluorescence intensity level of choosing and analyzes, major component 1 and major component 2 sums account for 98.07% of full detail amount, can effectively represent the information that is comprised in the former fluorescence spectrum, reach the purpose that different flavor liquor is distinguished.From Fig. 5 as seen, the two-dimentional scatter diagram that carries out obtaining after PCA analyzes with the characteristic index of different flavor liquor can be divided different flavor.
The method that the present invention combines with pattern-recognition by fluorescent spectrometry is set up the spectra database of different flavor liquor and is distinguished the different flavor Chinese sprits standard, be the control of production technology procedure quality, the differentiation of unknown aromatic white spirit and commercially available liquor character surveillance provide theoretical foundation.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although the present invention is had been described in detail with reference to preferred embodiment, those of ordinary skill in the art is to be understood that, can make amendment or be equal to replacement technical scheme of the present invention, and not breaking away from the aim and the scope of technical solution of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (2)
1. distinguish the method for different flavor liquor by fluorescein for one kind, it is characterized in that the key step of this method comprises:
1) preparing concentration with absolute ethyl alcohol is 2.5 * 10
-3～3.5 * 10
-3The luciferase reagent of mol/L keeps in Dark Place under 3～4 ℃;
2) get the luciferase reagent that configures in the step 1), the liquor different to be measured that luciferase reagent is identical or approaching with alcoholic strength respectively disposes solution to be measured by 1: 80～1: 120 volume ratio, shakes up under the lucifuge condition of back and leaves standstill a period of time T, T＞30min;
3) from different solution to be measured, get 2.5～3.5mL, be contained in respectively in the quartz colorimetric utensil, seal;
4) excitation wavelength with fluorescence spectrophotometer is set at 490～500nm, and incident and emission slit are 2.5～10.0nm; Adopt fluorescence spectrophotometer that the solution to be measured in each quartz colorimetric utensil is scanned, sweep velocity is 800～1200nm/min, and writes down the fluorescence emission spectrum in 200～800nm wavelength coverage;
5) be to choose several corresponding fluorescence peak intensity levels in 300～380nm and the 505～530nm scope from the fluorescence emission spectrum medium wavelength, input SPSS software, carry out pattern recognition analysis by SPSS software, demonstrate the dendrogram and the two-dimentional scatter diagram of different flavor liquor correspondence, different flavor liquor is distinguished by dendrogram and two-dimentional scatter diagram.
2. a kind of method by fluorescein differentiation different flavor liquor according to claim 1, it is characterized in that: in the described step 4), the excitation wavelength of fluorescence spectrophotometer is set at 495nm.
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| CN104502320A (en) * | 2014-12-29 | 2015-04-08 | 江南大学 | Method for identifying strong flavor Baijiu by combining three-dimensional fluorescence spectrum with PCA-SVM |
| CN104865360A (en) * | 2015-06-03 | 2015-08-26 | 四川理工学院 | Quality evaluation method and system for strong-flavor distiller's yeast |
| CN108444985A (en) * | 2018-04-25 | 2018-08-24 | 山西省食品药品检验所(山西省药品包装材料监测中心) | A kind of pig bezoar ICP-MS discrimination methods and application |
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| CN108489775A (en) * | 2018-05-14 | 2018-09-04 | 中国计量大学 | A kind of aerosol automatic collection and Raman spectrum detecting device |
| CN110455763A (en) * | 2019-08-22 | 2019-11-15 | 四川省绵阳市丰谷酒业有限责任公司 | The spectrogram measuring method and difference analysis method of white wine |
| CN111735804A (en) * | 2020-07-01 | 2020-10-02 | 北京百漾科技股份有限公司 | Ratio type fluorescence method for distinguishing fen-flavor raw pulp from liquid-method white spirit and solid-liquid-method white spirit |
| CN111735804B (en) * | 2020-07-01 | 2023-09-22 | 北京百漾科技股份有限公司 | Ratio type fluorescence method for distinguishing fen-flavor primary pulp from liquid-method white spirit and solid-liquid-method white spirit |
| CN114279991A (en) * | 2021-12-30 | 2022-04-05 | 宜宾五粮液股份有限公司 | Method for identifying white spirit brand |
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