CN103217408B - Method for identifying two flue-cured tobaccos with different odor types in Guizhou - Google Patents
Method for identifying two flue-cured tobaccos with different odor types in Guizhou Download PDFInfo
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Abstract
The invention relates to a method for identifying two flue-cured tobaccos with different odor types in Guizhou, belonging to the field of tobacco chemistry. The method disclosed by the invention adopts a three-dimensional fluorescence spectrum technology to perform clustering identification on the two different flue-cured tobaccos, namely fresh, sweet and aromas flue-cured tobaccos planted in the southwest of Guizhou province, Liupanshui and Bijieyu, and mellow, sweet and aromas flue-cured tobaccos planted in the southeast of Guizhou province, Tongren, Zunyi and the south of Guizhou province. Relative content difference information of chemical substances in the flue-cured tobaccos of different aromas types is obtained by measuring overall characteristic spectrums of chemical components in petroleum ether extracting solutions of tobacco leaves according to positions and intensities of characteristic spectrum peaks; and clustering analysis is carried out by statistical analysis software. The characteristic spectrum detection method is capable of identifying the aromas types of the two flue-cured tobaccos in Guizhou rapidly, simply, conveniently and accurately.
Description
Technical field
The invention belongs to tobacco chemistry field, specifically a kind of method differentiating Guizhou two class different flavor flue-cured tobacco, namely differentiate the method for " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavor flue-cured tobacco.
Background technology
Tobacco flavor is the coefficient result of multiple fragrance component, is the important indicator of evaluation tobacco and quality of item thereof.Aroma Quality of Flue-cured and odor type directly related with flavor matter contamination.The substance classes forming tobacco flavor is various, and component content is atomic, there is extremely complicated interaction between various aroma component.The odor type that difference plants district's tobacco leaf is relevant with type, kind, ecologic environment and cultivation condition with aroma quality, and composition, the content of its fragrance component are different with ratio, causes the odor type of tobacco leaf to distribute and has obvious region characteristic.Tobacco flavor material is determined jointly by inherent cause and envirment factor, the character of the former major effect fragrance matter and kind, and latter affects content and the proportion of composing of fragrance matter.
In tobacco quality evaluation and cigarette composition, traditionally China's tobacco leaf is divided into " giving off a strong fragrance ", " delicate fragrance " and " middle fragrant " three types, further after segmentation again with the type segmentation such as " giving off a strong fragrance ", " dense partially in ", " in partially dense ", " middle fragrant ", " in partially clear ", " clearly partially " and " delicate fragrance ".Difference is there is owing to the contained difference causing fragrant and latent Studies of The Aromatic Substances kind, content ratio with odor type tobacco style feature.Describe for meeting differentiation, the continuous refinement of tobacco leaf odor type concept, proposes in succession as the odor type style such as " fresh and sweet perfume (or spice) ", " alcohol is fragrant and sweet ".Have research to think, the Type of aroma of tobacco is relevant with the relative molecular mass size of flavor component in tobacco leaf, and the fragrance component that in strong-flavor tobacco leaf, relative molecular mass is larger is dominant, and the fragrance component that in delicate fragrance type tobacco leaf, relative molecular mass is less is dominant; Also odor type is relevant with nitrogen heterocycles fragrance component content to have research to think, strong-flavor tobacco leaf nitrogen heterocycles Aromatic Matter Contents is higher, and delicate fragrance type tobacco leaf nitrogen heterocycles Aromatic Matter Contents is relatively low; Also studies have found that the neophytadiene in strong-flavor tobacco leaf, carotenoid degradation product content are significantly lower than delicate fragrance type, and maillard reaction product and class western cypress alkanes content of degradation products also low than other odor type tobacco leaf contents; Research also finds, in the Yan Qu flue-cured tobacco of southwestern Yunnan-Guizhou, carotenoid degradation product content is higher than northeast and northwestern Fujian, and the latent fragrant substances content such as Polyphenols also there are differences.
Guizhou is located in low latitude High aititude monsoon region, and main body weather is obvious, and the distribution of tobacco planting height above sea level is wide, and ecological condition, weather are various, and unique mountain region natural conditions and ecologic environment create the distinguished quality of tobacco leaf and style characteristic.Guizhou is the second largest producing region of tobacco leaf that China is only second to Yunnan, plants district and comprises two, four cities (Guiyang, Lupanshui City, Zunyi, Anshun) area (Tongren Prefecture, Bijie Prefecture) and three autonomous prefectures (Autonomous Prefecture of Southwest Guizhou Province, Qiandongnan Autonomous Prefecture, the south of Guizhou Province autonomous prefecture).The style of Tobacco Leaves in Guizhou defines continues to use traditional " middle fragrant " type between " giving off a strong fragrance " and " delicate fragrance " always, and the feature of its mountain region tobacco leaf shows clear and definite not, and the region of distinctive style needs refinement.The odor type style having expert to propose Tobacco Leaves in Guizhou in recent years also refinement can be divided into two large classes, is distributed in Guizhou Province, the south of Guizhou Province, Qian Bei, Qian Dong plant district's tobacco leaf and divide mountain region " alcohol is fragrant and sweet " type into, are the main bodys of Tobacco Leaves in Guizhou.Fragrance type is so that hay is fragrant, tea is fragrant, herbal, Xin Xiang, fruital and nutty, and Type of aroma enriches, and flue gas is pure and sweet abundant.The southwest of Guizhou Province in west area, Northwestern Guizhou Province Region dividing are mountain region " fresh and sweet perfume (or spice) " odor type, and fragrant type is fragrant based on hay perfume, Xin Xiang, the banksia rose, animal, exquisiteness that flue gas is fresh and sweet.But all need to study further to the accurate description of this two odor types tobacco leaf feature, the chemical substance basis, quantitative judgement method etc. of division.The research of Guizhou representative region tobacco style feature, chemical composition and flavor matter, particularly be conducive to showing Guizhou Mountain characteristic delicate fragrance style characteristic to " middle fragrant " type tobacco style characteristic and aroma substance feature Study on thinning, also can provide support for the raw material supply of Base environment High Quality Tobacco and the foundation of quality security system, in-depth resource distribution.Simultaneously, can also for adopting control technique and the means of science in tobacco leaf production and process, improve the flavouring essence quality of raw tobacco material and industrial applicability, the digital management of setting up raw tobacco material chemical composition creates conditions, provide foundation for the seed selection of new product of tobacco and product zoning.To formula and the aromatic style of stable Guizhou cigarette products, the competitive strength improving cigarette product has important theory and realistic meaning.Tobacco leaf odor type wind interference and evaluation are the preconditions of plantation and development characteristic sound tobacco, are also the guarantees making full use of ecological resources.Guizhou is as national characteristic sound tobacco main producing region, and the clear definition of its style characteristic and differentiation describe, and promotes being conducive to effective utilization of unique Natural Resources in Mountain Areas, the market clout of plantation tobacco leaf and commercial value.
Tobacco components is complicated, and chemical substance is numerous.As the polycomponent mixed system of complexity, the analysis of tobacco flavor material detects and mostly depends on the efficient chromatogram be separated and the high mass spectrum differentiated, the more suitable fingerprint chromatogram realizing tobacco material composition of coupling technique builds.Because tobacco bred, ecologic environment, cultivation technique, baking adjustment means are different, the composition of its fragrance component, content and ratio are not quite similar, and are only difficult to reflect its odor type feature using certain several chemical composition as evaluation tobacco aroma quality and style index comprehensively.In addition the requirement of Spectrometry checkout equipment is high, sample preparation, separation detection required time are long, can not meet demand that is quick, real-time online detection.
Infrared and fluorescence spectrum, as the Fast Detection Technique of blending ingredients, not needing under the prerequisite be separated, by means of mathematical tool and taxonomic identification determination methods, has unique advantage in tobacco components identification.
In tobacco business, near-infrared spectrum analysis has been widely used in conventional ingredients such as measuring moisture, nicotine, total reducing sugar.Along with the development of NIR technology, to differentiate and physical index measures, be applied in on-line checkingi etc. at tobacco component and flue gas analysis, Tobacco Types and the place of production.Based on the sort research of near infrared spectrum, the differentiation to tobacco variety or the place of production can be realized after mathematical modeling; Fourier transform near infrared spectrum is set up the researchs such as the near-infrared model of plant pigment mathematical model, flue-cured tobacco Content of Petroleum Ether Extract in express-analysis and prediction tobacco and is had been reported.
Three-dimensional fluorescence spectrum is that fluorescence intensity is with the fluorescent technique exciting spectroscopic data 3 D stereo or the plane contour map form display obtained with emission wavelength scanning.Three-dimensional fluorescence spectrum solve fluorescence peak in plane spectrogram overlapped, cover the problems such as not easily observation, clearly can show the detailed construction of spectrogram, express complete fluorescence information, there is fingerprint characteristic.The quantification and qualification of comparatively complicated ingredient in multicomponent mixture system can be completed by three-dimensional fluorescence spectrum information, build method for quick.The analysis be successfully applied in Chinese crude drug, medicament, food and field of petrochemical industry detects.Utilize the three-dimensional fluorescence spectrum of tobacco extract, the relation between research cigarette quality, fragrance, strength etc. are jealous has been reported.
Sherwood oil effectively can extract in tobacco leaf the volatility and non-volatile chemical composition that produce fragrance, is the potpourri of the organic substances such as fat, free fatty acid, essential oil, resin, wax, lipides, sterol, pigment and organic acid.Tobacco and tobacco product PetroChina Company Limited. ether extract content more, account for 6% of total amount, and these materials are considered to the important sources forming tobacco flavor, with the fragrance of tobacco leaf with coordinate closely related, with being proportionate property of tar content in cigarette, therefore tobacco ligroin extraction total amount Chang Zuowei evaluates an index of tobacco flavor quality.Measure ligroin extraction total amount and have standard method at tobacco business.The tobacco leaf chemical composition eurythmy that Content of Petroleum Ether Extract is high, overall quality is better, and its content and tobacco incense tolerance and quality are proportionate.Once had people to bake and bank up with earth the difference of tobacco ligroin extraction under condition to different regions, Different climate condition, different cultivation condition, different genes, kind and difference to be studied.
Smokeing panel test and Quality Identification of tobacco, depends on sensation always and judges.Tobacco scholar proposes the parameters such as nicotine value, polyphenol value, values of nitrogen might and fragrance value for this reason, attempts, by the chemical composition of tobacco, its quality is described.But concerning the fragrance of tobacco, to eat hiding be the common sensation of a kind of physiological strength, pungency and sense of taste, need to give to describe and characterize from the fragrance style of general characteristic angle to tobacco, be not that certain single component mensuration just can be defined the level and Quality Identification, the sensation also depending on artificial evaluation and analysis at present judges.
Tobacco ligroin extraction is complicated multicomponent system, three-dimensional fluorescence spectrum can provide the spectrum contour map of blending ingredients entirety under unseparated condition, obtain the characteristic fingerprint hum pattern of main chemical compositions emitting fluorescence in tobacco, thus reflect tobacco odor type style and features.Therefore, may be used for replacing the sensation of people to differentiate tobacco leaf odor type aromatic style.
Cluster analysis is the reason according to " things of a kind come together, people of a mind fall into the same group ", utilizes a kind of Multielement statistical analysis method that mathematical method is classified to sample, a large amount of samples reasonably can be classified by respective characteristic.It is more objective, accurate that this numerical taxonomy utilizing mathematical tool to carry out quantitative classification is classified than dependence experience and professional knowledge.System cluster analysis is some different classifications by test variable merger, and to classify, tree-like figure represents, makes to have comparatively close relationship between all individualities in each classification, and of all categories between mutual relationship relatively compare and become estranged.The natural pedigree of close and distant relation between a reflection individuality can be obtained after hierarchial-cluster analysis, difference between each individuality describing object of classification objectively and contact, cluster analysis and three-dimensional fluorescence measure be used for tobacco odor type aromatic style differentiation at present also nobody carried out.
Tobacco odor type traditional at present, the differentiation of aromatic style are the modes of the artificial evaluation and analysis of sampling, undertaken, be difficult to provide objective, quantitative and digitized description by fragrance profile analysis.Therefore, the foundation of the analyzing detecting method of quick, easy, accurate, rational judgment tobacco odor type feature is very important.
Summary of the invention
The object of the invention is to the method setting up a kind of quick, easy, accurate discriminating Guizhou two class flue-cured tobacco flavor types style, object of the present invention reaches by the following method:
A kind of method differentiating Guizhou two class different flavor flue-cured tobacco of the present invention, it is characterized in that utilizing three-dimensional fluorescence spectrum technology, measure chemical composition general characteristic spectrum in the petroleum ether extract of Guizhou Flue-cured Tobacco tobacco leaf, according to position and the intensity of general characteristic spectrum peak, obtain the difference of the chemical substance relative content characterizing different flavor style flue-cured tobacco, carry out cluster analysis with statistical analysis software, set up the method for quick differentiating Guizhou " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavor style flue-cured tobacco.Indication three-dimensional fluorescence spectrum technology measures the general characteristic spectrum of tobacco leaf petroleum ether extract, excite to be respectively peak I: 297/326nm with emission wavelength, peak II: 250/330nm, 3 fluorescence peaks at peak III: 225/336nm are characteristic peak, record tobacco leaf petroleum ether extract overall emission spectrum intensity F I, F II and F III respectively, with fluorescence intensity mark D
1, D
2, D
3or fluorescence intensity ratio R
1, R
2, R
3, carry out cluster analysis respectively, obtain Cluster tree, wherein D
1, D
2, D
3represent the fluorescence intensity mark at peak I, peak II and peak III respectively; R
1, R
2, R
3represent 3 fluorescence peak intensity rates respectively; D
1=F I/∑ F, D
2=F II/∑ F, D
3=F III/∑ F, ∑ F represent three fluorescence peak intensity and; R
1=F I/F II, R
2=F II/F III, R
3=F III/F I; During with fluorescence intensity mark cluster, get distance coefficient 5 ~ 7; During with fluorescence intensity ratio cluster, get distance coefficient 2 ~ 4, Guizhou Flue-cured Tobacco two class different flavor can be differentiated.
The inventive method follows these steps to differentiate:
(1) sample preparation: label taking standard " alcohol is fragrant and sweet " type, " fresh and sweet perfume (or spice) " type and flue-cured tobacco sample to be measured respectively, by after sample number into spectrum in 40 DEG C of oven dryings, pulverize, cross 40 mesh sieves, precise each sample, leave standstill after 2 hours with the infiltration of chromatographically pure sherwood oil, vibration, supernatant is sherwood oil flue-cured tobacco extract;
(2) general characteristic spectroscopic assay: get step (1) supernatant, 20 times are diluted with chromatographically pure sherwood oil, obtain analytic liquid, be transferred to unstressed configuration quartz colorimetric utensil, excitation wavelength is set at 200 ~ 500 nm, emission wavelength is in 200 ~ 600 nm scopes, with 2.0 nm for wavelength increases progressively, outgoing and entrance slit are 5.0 nm, three-dimensional fluorescence spectrum mensuration is carried out under the condition of detecting device multiplier voltage 720V, the excitation/emission wavelength obtaining each sample analytic liquid is respectively respectively peak I: 297/326 nm, peak II: 250/330 nm, the emission spectrum fluorescence intensity F I of 3 characteristic peaks of peak III: 225/336 nm, F II and F III,
(3) data analysis: measure respectively and calculate fluorescence intensity F I, F II, the F III at 3 fluorescent characteristics peaks that each analytic liquid excitation/emission wavelength is peak I: 297/326nm, peak II: 250/330nm, peak III: 225/336nm, calculating fluorescence intensity mark D
1, D
2, D
3with fluorescence intensity ratio R
1, R
2, R
3.Utilize SPSS17.0 data statistic analysis software, definition " sample number into spectrum, D in " variable view "
1, D
2, D
3" or " sample number into spectrum, R
1, R
2, R
3" variable; wherein " sample number into spectrum " is character string type; its dependent variable is numeric type; by the fluorescence intensity mark of each sample analytic liquid and fluorescence intensity ratio data input " Data View "; select " analysis " → " classification " → " hierarchial-cluster analysis " calculation function; wherein " mark case " is sample number into spectrum, selects D respectively
1, D
2, D
3and R
1, R
2, R
33 variablees, " square Euclidean distance " module in the clustering method connected between utilization group carries out hierarchial-cluster analysis, obtain Cluster tree, known by characteristic peak intensity fraction, when distance coefficient is 5 ~ 7, each numbering testing sample can be identified genus " fresh and sweet perfume (or spice) " type style or belong to the tobacco leaf of " alcohol is fragrant and sweet " type style by fluorescence intensity fractional value; From characteristic peak intensity rate, when distance coefficient is 2 ~ 4, each numbering testing sample can be identified genus " fresh and sweet perfume (or spice) " type style or belong to the tobacco leaf of " alcohol is fragrant and sweet " type style by fluorescence intensity fractional value.
The present invention's fluorescence spectrophotometer used is the Cary Eclipse type fluorospectrophotometer of U.S.'s Varian (Varian) company, and fluorometric assay condition is: the thick four mirror polish unstressed configuration quartz colorimetric utensils of 1 cm, excitation wavelength E
x200 ~ 500nm, emission wavelength E
m200 ~ 600nm, wavelength increases progressively 2.00nm, slit 5.00nm, voltage 720V.
The present invention's data processing software used is spread sheet software (Excel) assembly of Microsoft's office suites software (Microsoft office) and " statistical product and service solution " middle SPSS17.0 data statistic analysis software of SPSS Inc., carries out hierarchial-cluster analysis to institute's test sample product characteristic spectrum data.
Sample collection treatment conditions gather cured tobacco leaf sample C
3tangerine three in F() grade, sampling amount is 1kg at least, drying, pulverize, sieve after accurately take about 1g sample, with chromatographically pure sherwood oil 30ml infiltrate, vibration leaves standstill at least 24 hours after 2 hours, get supernatant chromatographically pure sherwood oil and dilute analytic liquid as mensuration three-dimensional fluorescence spectrum after 20 times of volumes.
A kind of application differentiating the method for Guizhou two class different flavor flue-cured tobacco of the present patent application, is characterized in that " the fresh and sweet perfume (or spice) " and the differentiation of " alcohol is fragrant and sweet " type flue-cured tobacco stylistic category that can be used for producing Anywhere.
Utilize the present patent application method to complete to produce flue-cured tobacco to different regions, Guizhou and carry out odor type classification, draw the southeast of Guizhou Province, Tongren, the south of Guizhou Province and Zunyi Prefecture flue-cured tobacco belong to " alcohol is fragrant and sweet " type, the flue-cured tobacco of six Pan Shui, the southwest of Guizhou Province and Bijie belongs to " fresh and sweet perfume (or spice) " type, wherein " fresh and sweet perfume (or spice) " type of Bijie and " alcohol is fragrant and sweet " type of Tongren the most obvious.The present invention in analysis using Bijie flue-cured tobacco sample as " fresh and sweet perfume (or spice) " type standard model, using Tongren flue-cured tobacco sample as " alcohol is fragrant and sweet " type standard model.In characteristic peak intensity fraction dendrogram, within the scope of the distance coefficient of 5 ~ 7, all gather with Bijie flue-cured tobacco data be a class be " fresh and sweet perfume (or spice) " type; All gather with Tongren flue-cured tobacco data be a class be " alcohol is fragrant and sweet " type; In characteristic peak intensity rate dendrogram, within the scope of the distance coefficient of 2 ~ 4, all gather with Bijie flue-cured tobacco data be a class be " fresh and sweet perfume (or spice) " type; All gather with Tongren flue-cured tobacco data be a class be " alcohol is fragrant and sweet " type.
The present patent application scans tobacco leaf petroleum ether extract three-dimensional fluorescence spectrum, and finding that in extract, three-dimensional fluorescence spectrum has three characteristic fluorescence peaks, is peak I: E respectively
x/ E
m=297/326nm, peak II: E
x/ E
m=250/330nm, peak III: E
x/ E
m=225/336nm.Each peak intensity is > peak, > peak III, peak I II, sees Fig. 1 and Fig. 2, and three peaks represent chemical composition general characteristic spectrum in the petroleum ether extract of cured tobacco leaf.The position of general characteristic spectrum peak, intensity symbolize the difference of the chemical substance relative content of tobacco leaf different flavor style, carry out with statistical analysis software the flue-cured tobacco flavor types classification that cluster analysis just can differentiate Guizhou " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavor fast.
Gordian technique in the present patent application is: the sherwood oil extension rate of (1) cigarette sample test fluid is that 0.5mL supernatant sherwood oil dilutes 20 times; (2) intensity fraction and the ratio that gather 3 characteristic fluorescence peaks carry out data analysis; (3) square Euclidean distance method in characteristic spectrum data acquisition hierarchial-cluster analysis.
The feature of the present patent application is that sample treatment is simple, spectroscopic assay is quick, easy, environmental protection and data processing is very simple, objective, accurate, directly perceived without the need to the mathematical computations of complexity and modeling, differentiation result.Method is simple, differentiates that the method for manually smokeing panel test that result is more traditional is more objective, accurate and quantize, have practical reference value.
accompanying drawing explanation:
Fig. 1 tobacco leaf petroleum ether extract three-dimensional fluorescence spectrum 3 dimensional drawing, wherein horizontal ordinate is excitation wavelength E
x, ordinate is emission wavelength E
m, ordinate is peak intensity F.The three-dimensional fluorescence spectrum of extract has 3 characteristic fluorescence peaks, is peak I: E respectively
x/ E
m=297/326nm, peak II: E
x/ E
m=250/330nm, peak III: E
x/ E
m=225/336nm.The emission wavelength of each characteristic peak is comparatively close and excitation wavelength has larger difference.Peak intensity shows as > peak, > peak III, peak I II, and wherein peak I intensity is maximum, and peak II, peak III intensity are lower, and various kinds performance number is more or less the same.
Fig. 2 tobacco leaf petroleum ether extract three-dimensional fluorescence spectrum isogram, wherein horizontal ordinate is emission wavelength E
m, ordinate is excitation wavelength E
x.The isoline density of 3 characteristic peaks shows as > peak, > peak III, peak I II, and illustrating that peak I intensity is maximum, is secondly peak III, and peak I intensity is minimum.
The fluorescence intensity mark dendrogram of Fig. 3 tobacco sample.As " fresh and sweet perfume (or spice) " type standard model, be 5 ~ 7 scopes at distance coefficient using Bijie flue-cured tobacco sample (mark 1), sample 1(six can be coiled shredded tobacco for water pipes sample), sample 2(the southwest of Guizhou Province cigarette sample) be determined as " fresh and sweet perfume (or spice) " type belonging to standard specimen 1 together; Using Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 5 ~ 7 scope, can by sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) be determined as " alcohol the is fragrant and sweet " type belonging to standard specimen 2 together.
The fluorescence intensity ratio dendrogram of Fig. 4 tobacco sample.As " fresh and sweet perfume (or spice) " type standard model, be 2 ~ 4 scopes at distance coefficient using Bijie flue-cured tobacco sample (mark 1), sample 1(six can be coiled shredded tobacco for water pipes sample), sample 2(the southwest of Guizhou Province cigarette sample) be determined as " fresh and sweet perfume (or spice) " type belonging to standard specimen 1 together; Using Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 2 ~ 4 scope, can by sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) be determined as " alcohol the is fragrant and sweet " type belonging to standard specimen 2 together.
Specific embodiment
Embodiment one: in fluorescent spectrometry, fluorescence intensity mark differentiates Guizhou two class different flavor flue-cured tobacco
Gathering Ge Zhongyan district, Guizhou 7 grade is C
3tangerine three in F() 125, cured tobacco leaf sample, sampling quantity is respectively 1kg, in 40 DEG C of oven drying 6h, pulverizes, crosses 40 mesh sieves.Precise 1g cigarette sample, leaves standstill 24 hours with chromatographically pure sherwood oil 30ml infiltration, vibration after 2 hours.Learn from else's experience supernatant 0.5 mL after above-mentioned sample treatment process, 20 times are diluted with chromatographically pure sherwood oil, be transferred to the polishing unstressed configuration quartz colorimetric utensil at 1 cm thickness four sides, excitation wavelength is at 200 ~ 500 nm, emission wavelength is in 200 ~ 600 nm scopes, and with 2.0 nm for wavelength increases progressively, outgoing and entrance slit are 5.0 nm, carry out three-dimensional fluorescence spectrum mensuration under the condition of detecting device multiplier voltage 720V, spectrogram is shown in attached Fig. 1 and 2.Obtain exciting of petroleum ether extract and be respectively peak I: 297/326 nm with emission wavelength, peak II: 250/330 nm, the fluorescence intensity of 3 characteristic peak overall emission spectrum of peak III: 225/336 nm.MS Office Applications Excel is utilized to calculate the fluorescence intensity mark D of each characteristic peak
1, D
2, D
3.Wherein D
1=F I/∑ F, D
2=F II/∑ F, D
3=F III/∑ F, ∑ F be 3 fluorescence peak intensity and.Obtain each cigarette sample three-dimensional fluorescence intensity fraction value (table 1).Utilize SPSS17.0 data statistic analysis software, definition " sample number into spectrum, D in " variable view "
1, D
2, D
3" 4 variablees, wherein " sample number into spectrum " is character string type, and other 3 is numeric type.The fluorescence intensity fractional data of different Zhi Yan district sample number into spectrum and each department tobacco leaf petroleum ether extract is inputted in " Data View ", selects " analysis " → " classification " → " hierarchial-cluster analysis " calculation function.Wherein " mark case " is sample number into spectrum, selects D
1, D
2, D
33 variablees, " square Euclidean distance " module in the clustering method connected between utilization group carries out hierarchial-cluster analysis, obtains the Cluster tree of characteristic peak fluorescence intensity fractional value, sees accompanying drawing 3.As seen from the figure, as " fresh and sweet perfume (or spice) " type standard model using Bijie flue-cured tobacco sample (mark 1), be in 5 ~ 7 scopes at distance coefficient, sample 1(six coiled shredded tobacco for water pipes sample), sample 2(the southwest of Guizhou Province cigarette sample) be determined as and belong to " fresh and sweet perfume (or spice) " type together; Using Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 5 ~ 7 scope, by sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) be determined as and belong to " alcohol is fragrant and sweet " type together.Provincial characteristics performance is clear, and classification is clear and definite.
Embodiment two: in fluorescent spectrometry, glimmering light intensity ratio differentiates Guizhou two class different flavor flue-cured tobacco
Sample collection, process and spectrometric method are with embodiment one.Calculate the fluorescence intensity ratio R at 3 characteristic fluorescence peaks
1, R
2, R
3.Wherein R
1=F I/F II, R
2=F II/F III, R
3=F III/F I.Obtain each cigarette sample three-dimensional fluorescence intensity rate data (table 2).Utilize SPSS17.0 data statistic analysis software, at " in variable view " definition " sample number into spectrum, R
1, R
2, R
3" 4 variablees, wherein " sample number into spectrum " is character string type, and other three is numeric type.The fluorescence intensity ratio data of different Zhi Yan district sample number into spectrum and each department tobacco leaf petroleum ether extract is inputted in " Data View ", selects " analysis " → " classification " → " hierarchial-cluster analysis " calculation function.Wherein " mark case " is sample number into spectrum, selects R
1, R
2, R
33 variablees, " square Euclidean distance " module in the clustering method connected between utilization group carries out hierarchial-cluster analysis, obtains the Cluster tree of characteristic peak fluorescence intensity ratio, sees accompanying drawing 4.As seen from the figure, as " fresh and sweet perfume (or spice) " type standard model using Bijie flue-cured tobacco sample (mark 1), be in 2 ~ 4 scopes at distance coefficient, sample 1(six coiled shredded tobacco for water pipes sample), sample 2(the southwest of Guizhou Province cigarette sample) be determined as and belong to " fresh and sweet perfume (or spice) " type together; Using Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 2 ~ 4 scope, by sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) be determined as and belong to " alcohol is fragrant and sweet " type together.Provincial characteristics performance is clear, and classification is clear and definite.
Claims (5)
1. differentiate a method for Guizhou two class different flavor flue-cured tobacco, it is characterized in that following these steps to successively differentiate:
(1) sample preparation: label taking standard " pure fragrant and sweet " type, " fresh and sweet perfume (or spice) " type and flue-cured tobacco sample to be measured respectively, by after sample number into spectrum in 40 DEG C of oven dryings, pulverize, cross 40 mesh sieves, precise each sample, leave standstill after 2 hours with the infiltration of chromatographically pure sherwood oil, vibration, supernatant is sherwood oil flue-cured tobacco extract;
(2) general characteristic spectroscopic assay: get step (1) supernatant, 20 times are diluted with chromatographically pure sherwood oil, obtain analytic liquid, be transferred to unstressed configuration quartz colorimetric utensil, excitation wavelength is set at 200 ~ 500 nm, emission wavelength is in 200 ~ 600 nm scopes, with 2.0 nm for wavelength increases progressively, outgoing and entrance slit are 5.0 nm, three-dimensional fluorescence spectrum mensuration is carried out under the condition of detecting device multiplier voltage 720V, the excitation/emission wavelength obtaining each sample analytic liquid is respectively respectively peak I: 297/326 nm, peak II: 250/330 nm, the emission spectrum fluorescence intensity F I of 3 characteristic peaks of peak III: 225/336 nm, F II and F III,
(3) data analysis: measure respectively and calculate 3 fluorescent characteristics peak F I, F II and the F III that each analytic liquid excitation/emission wavelength is peak I: 297/326nm, peak II: 250/330nm, peak III: 225/336nm, calculating fluorescence intensity mark D
1, D
2, D
3and fluorescence intensity ratio R
1, R
2, R
3; Utilize SPSS17.0 data statistic analysis software, definition " sample number into spectrum, D in " variable view "
1, D
2, D
3" or " sample number into spectrum, R
1, R
2, R
3" variable; wherein " sample number into spectrum " is character string type; its dependent variable is numeric type; by the fluorescence intensity mark of each sample analytic liquid and fluorescence intensity ratio data input " Data View "; select " analysis " → " classification " → " hierarchial-cluster analysis " calculation function; wherein " mark case " is sample number into spectrum, selects D respectively
1, D
2, D
3or R
1, R
2, R
33 variablees, " square Euclidean distance " module in the clustering method connected between utilization group carries out hierarchial-cluster analysis, obtain Cluster tree, from characteristic peak intensity fraction, when distance coefficient is 5 ~ 7, each numbering testing sample can be identified genus " fresh and sweet perfume (or spice) " type style or belong to the tobacco leaf of " alcohol is fragrant and sweet " type style by fluorescence intensity fractional value; From characteristic peak intensity rate, when distance coefficient is 2 ~ 4, each numbering testing sample can be identified genus " fresh and sweet perfume (or spice) " type style or belong to the tobacco leaf of " alcohol is fragrant and sweet " type style by fluorescence intensity ratio.
2. a kind of method differentiating Guizhou two class different flavor flue-cured tobacco according to claim 1, it is characterized in that fluorescence spectrophotometer used is the Cary Eclipse type fluorospectrophotometer of U.S.'s Varian (Varian) company, fluorometric assay condition is: the thick four mirror polish unstressed configuration quartz colorimetric utensils of 1 cm, excitation wavelength E
x200 ~ 500nm, emission wavelength E
m200 ~ 600nm, wavelength increases progressively 2.00nm, slit 5.00nm, voltage 720V.
3. a kind of method differentiating Guizhou two class different flavor flue-cured tobacco according to claim 1, it is characterized in that cluster analysis data processing software used is spread sheet software (Excel) assembly of Microsoft's office suites software (Microsoft office) and " statistical product and service solution " middle SPSS17.0 data statistic analysis software of SPSS Inc., hierarchial-cluster analysis is carried out to institute's test sample product characteristic spectrum data.
4. a kind of method differentiating Guizhou two class different flavor flue-cured tobacco according to claim 1, is characterized in that gathering cured tobacco leaf sample C
3tangerine three in F() grade, sampling amount is 1kg at least, drying, pulverize, sieve after accurately take about 1g sample, with chromatographically pure sherwood oil 30ml infiltrate, vibration leaves standstill at least 24 hours after 2 hours, get supernatant chromatographically pure sherwood oil and dilute analytic liquid as mensuration three-dimensional fluorescence spectrum after 20 times of volumes.
5. a kind of application differentiating the method for Guizhou two class different flavor flue-cured tobacco according to claim 1, is characterized in that " fresh and sweet perfume (or spice) " type of can be used for producing Anywhere and " alcohol is fragrant and sweet " type flue-cured tobacco type identification.
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