CN103217408A - 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 the tobacco chemistry field, specifically a kind of method of differentiating Guizhou two class different flavor flue-cured tobaccos, the i.e. method of discriminating " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavor flue-cured tobaccos.
Background technology
Tobacco flavor is the coefficient result of multiple fragrance component, is the important indicator of evaluation tobacco and quality of item thereof.Tabacco fragrance quality and odor type are directly related with flavor matter kind and content.The substance classes that forms tobacco flavor is various, and component content is atomic, has extremely complex interactions between various aroma components.It is relevant with type, kind, ecologic environment and cultivation condition with aroma quality that difference is planted the odor type of district's tobacco leaf, and the composition of its fragrance component, content and ratio are different, cause the odor type distribution of tobacco leaf to have tangible region characteristic.Tobacco flavor material is determined jointly that by inherent cause and envirment factor the former mainly influences the character and the kind of fragrance matter, and the latter then influences the content and the proportion of composing of fragrance matter.
In tobacco quality evaluation and cigarette composition, traditionally China's tobacco leaf is divided into three types of " giving off a strong fragrance ", " delicate fragrance " and " middle fragrant ", further reaches types such as " delicate fragrance " with " giving off a strong fragrance ", " dense partially in ", " in dense partially ", " middle fragrant ", " in clear partially ", " clear partially in " again after the segmentation and segment.Exist difference to cause the different of fragrant and latent perfumery matter kind, content ratio with odor type tobacco leaf style and features owing to contained.Describe for satisfying differentiation, the continuous refinement of tobacco leaf odor type notion has proposed in succession as odor type styles such as " fresh and sweet perfume (or spice) ", " alcohol are fragrant and sweet ".Have research to think that the fragrance type of tobacco is relevant with the relative molecular mass size of flavor component in the tobacco leaf, the fragrance component that relative molecular mass is bigger in the Luzhou-flavor tobacco leaf is dominant, and the less fragrance component of relative molecular mass is dominant in the delicate fragrance type tobacco leaf; Also have research to think that odor type is relevant with nitrogen heterocycles fragrance component content, Luzhou-flavor tobacco leaf nitrogen heterocycles aroma substance content is higher, and delicate fragrance type tobacco leaf nitrogen heterocycles aroma substance content is relatively low; Discover that in addition neophytadiene, carotenoid content of degradation products in the Luzhou-flavor tobacco leaf significantly are lower than delicate fragrance type, and maillard reaction product and class west cypress alkanes content of degradation products are also low than other odor type tobacco leaf contents; Research finds that also the carotenoid content of degradation products is higher than northeast and Fujian tobacco district in the Yan Qu flue-cured tobacco of southwestern Yunnan-Guizhou, and latent odor type content of material such as Polyphenols also there are differences.
Guizhou is located in the high height above sea level of low latitude monsoon region, and the main body weather is obvious, and it is wide that tobacco planting height above sea level distributes, and ecological condition, weather are various, and unique mountain region natural conditions and ecologic environment have been brought up 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 the district and comprises two areas, four cities (Guiyang, Lupanshui City, Zunyi, Anshun) (Tongren Prefecture, Bijie Prefecture) and three autonomous prefectures (the southwest of Guizhou Province autonomous prefecture, Qiandongnan Autonomous Prefecture, the south of Guizhou Province autonomous prefecture).The style of Guizhou tobacco leaf 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 inadequately, and the zone of characteristic style remains refinement.But have in recent years odor type style that the expert proposes the Guizhou tobacco leaf also refinement be divided into two big classes, be distributed in the 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 Guizhou tobacco leaf.The fragrance type is based on hay perfume (or spice), tea perfume (or spice), herbal, Xin Xiang, fruital and nutty, and the fragrance type is abundant, and flue gas is pure and sweet abundant.The southwest of Guizhou Province in west area, territory, North Area, the west of Guizhou Province are divided into mountain region " fresh and sweet perfume (or spice) " odor type, and fragrant type is based on hay perfume (or spice), Xin Xiang, the banksia rose, animal perfume (or spice), the fresh and sweet exquisiteness of flue gas.But to further researchs that all awaits such as the chemical substance basis of the accurate description of this two odor types tobacco leaf feature, division, quantitative judgement methods.The research of Guizhou representative region tobacco leaf style and features, chemical constitution and flavor matter, particularly " middle fragrant " type tobacco leaf style characteristic and aroma substance feature Study on thinning are helped showing mountain region, Guizhou characteristic delicate fragrance style characteristic, also can for base high-quality tobacco material supply and the quality security system be set up, the in-depth resource distribution provides support.Simultaneously, can also be for adopting the control technique and the means of science in tobacco production and the process, improve the flavouring essence quality and the industrial applicability of raw tobacco material, the digital management of setting up the raw tobacco material chemical constitution creates conditions, provide foundation for the seed selection and the product zoning of tobacco new varieties.To prescription and the fragrance style of stablizing the Guizhou cigarette products, the competitive strength that improves cigarette product has important theory and realistic meaning.Tobacco leaf odor type style location and evaluation are the preconditions of plantation and development characteristic sound tobacco, also are the guarantees that makes 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 are described, and will help effective utilization of unique mountain region natural resources, the market clout and the commercial value lifting of plantation tobacco leaf.
The tobacco components complexity, chemical substance is numerous.As the polycomponent mixed system of complexity, the analyzing and testing of tobacco flavor material depends on the chromatogram and the high mass spectrum of differentiating of high efficiency separation mostly, and the fingerprint spectrogram of the more suitable realization of coupling technique tobacco material composition makes up.Because the means difference is adjusted in tobacco bred, ecologic environment, cultivation technique, baking, the composition of its fragrance component, content and ratio are not quite similar, and only are difficult to reflect its odor type feature with certain several chemical constitution as estimating tobacco aroma quality and style index comprehensively.Chromatography-mass spectroscopy analyzing and testing equipment requirements height, sample preparation, separation detection required time are long in addition, the demand that can not satisfy fast, real-time online detects.
Infrared and fluorescence spectrum under the prerequisite that need not separate, by means of mathematical tool and taxonomic identification determination methods, is having special advantages aspect tobacco components analysis and the evaluation as the Fast Detection Technique of blending ingredients.
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, differentiate in tobacco component and flue gas analysis, tobacco leaf type and the place of production and aspects such as physical index mensuration, online detection are applied.Based on the sort research of near infrared spectrum, behind mathematical modeling, can realize differentiation to the tobacco leaf kind or the place of production; Fourier transform near infrared spectrum is set up the existing report of researchs such as near-infrared model of plant pigment mathematical model, flue-cured tobacco ligroin extraction content in express-analysis and the prediction tobacco.
Three-dimensional fluorescence spectrum is that fluorescence intensity scans the fluorescent technique of the spectroscopic data of acquisition with 3 D stereo or the demonstration of plane contour map form with exciting with emission wavelength.Three-dimensional fluorescence spectrum solved in the spectrogram of plane fluorescence peak overlapped, cover problems such as being difficult for observation, can clearly show the detailed structure of spectrogram, the fluorescence information of The expressed, have the fingerprint characteristic.Can finish the comparatively qualitative and quantitative analysis of complicated ingredient in the multicomponent mixture system with three-dimensional fluorescence spectrum information, make up method for quick.The successful analyzing and testing that is applied in Chinese crude drug, medicament, food and the field of petrochemical industry.Utilize the three-dimensional fluorescence spectrum of tobacco extract, the existing report of relation between research cigarette quality, fragrance, strength etc. are jealous.
Sherwood oil can effectively extract volatility and the non-volatile chemical constitution that produces fragrance in the tobacco leaf, is the potpourri of 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 are more, account for 6% of total amount, and these materials are considered to form the important source of tobacco flavor, closely related with the fragrance and the coordination of tobacco leaf, with being proportionate property of tar content in cigarette, so tobacco ligroin extraction total amount Chang Zuowei estimates an index of tobacco flavor quality.Measure the ligroin extraction total amount and standard method is arranged at tobacco business.The tobacco leaf chemical composition eurythmy that ligroin extraction content is high, overall quality is better, and its content and tobacco incense tolerance and quality are proportionate.Once there was the people that different regions, different weather conditions, different cultivation condition, different genes, kind and different differences of baking and banking up with earth tobacco ligroin extraction under the condition are studied.
Smokeing panel test and the quality evaluation of tobacco mainly depends on sensation always and judges.The tobacco scholar has proposed parameters such as nicotine value, polyphenol value, nitrogen value and fragrance value for this reason, attempts with the chemical constitution of tobacco its quality to be described.But to the fragrance of tobacco, eat that to hide be the common sensation of a kind of physiological strength, pungency and sense of taste, need describe and characterize the fragrance style of tobacco from the general characteristic angle, be not that certain single component mensuration just can be defined the level and quality is identified, the sensation that also mainly depends on artificial evaluation and analysis is at present judged.
The tobacco ligroin extraction is complicated multicomponent system, three-dimensional fluorescence spectrum can provide the spectrum contour map of blending ingredients integral body under unseparated condition, obtain the characteristic fingerprint hum pattern of main chemical compositions emitting fluorescence in the tobacco, thereby reflect tobacco odor type style and features.Therefore, may be used for replacing people's sensation to differentiate tobacco leaf odor type fragrance style.
Cluster analysis is the reason according to " things of a kind come together, people of a mind fall into the same group ", and a kind of multivariate statistical analysis method of utilizing mathematical method that sample is classified can reasonably be classified a large amount of samples by characteristic separately.This numerical taxonomy of utilizing mathematical tool to carry out quantitative classification is more objective, accurate than dependence experience and professional knowledge classification.The hierarchial-cluster analysis method is to be some different classifications with the test variable merger, represents with the tree-like figure that classifies, and make between all individualities in each classification to have than confidential relation, and the mutual relationship between of all categories is relatively relatively become estranged.Through obtaining the natural pedigree of close and distant relation between a reflection individuality behind the hierarchial-cluster analysis, difference and contact between each individuality of object of classification have been described objectively, cluster analysis and three-dimensional fluorescence measure be used for tobacco odor type fragrance style differentiation at present also nobody carried out.
At present traditional tobacco odor type, the differentiation of fragrance style be the mode of the artificial evaluation and analysis of sampling, undertaken by the fragrance profile analysis, is difficult to provide objective, quantitative and digitized description.Therefore, foundation quick, easy, accurate, that quantitatively judge the analyzing detecting method of tobacco odor type feature is very important.
Summary of the invention
The objective of the invention is to set up a kind of quick, easy, accurately differentiate the method for Guizhou two class flue-cured tobacco odor type styles, purpose of the present invention reaches by the following method:
A kind of method of differentiating Guizhou two class different flavor flue-cured tobaccos of the present invention, it is characterized in that utilizing the three-dimensional fluorescence spectrum technology, chemical constitution general characteristic spectrum in the petroleum ether extract of mensuration Guizhou cured tobacco leaf, position and intensity according to the general characteristic spectrum peak, obtain characterizing the difference of the chemical substance relative content of different flavor style flue-cured tobacco, carry out cluster analysis with statistical analysis software, set up the method for quick of differentiating Guizhou " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavor style flue-cured tobaccos.The indication general characteristic spectrum of three-dimensional fluorescence spectrum technical measurement tobacco leaf petroleum ether extract, be to be respectively peak I: 297/326nm to excite with emission wavelength, peak II: 250/330nm, 3 fluorescence peaks of peak III: 225/336nm are characteristic peak, record tobacco leaf petroleum ether extract total emission spectral 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 tree-shaped dendrogram, wherein D
1, D
2, D
3The fluorescence intensity mark of representing 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, can differentiate Guizhou flue-cured tobacco two class different flavors.
The inventive method follows these steps to differentiate:
(1) sample preparation: get standard " alcohol is fragrant and sweet " type, " fresh and sweet perfume (or spice) " type and flue-cured tobacco sample to be measured respectively, with behind the sample number into spectrum in 40 ℃ of oven dryings, pulverize, cross 40 mesh sieves, accurate each sample of weighing, 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, with 20 times of chromatographically pure sherwood oil dilutions, get analytic liquid, be transferred to no fluorescence quartz colorimetric utensil, excitation wavelength is set at 200 ~ 500 nm, emission wavelength is in 200 ~ 600 nm scopes, with 2.0 nm is that wavelength increases progressively, outgoing and entrance slit are 5.0 nm, carrying out three-dimensional fluorescence spectrum under the condition of detecting device multiplier tube voltage 720V measures, the excitation/emission wavelength that obtains each sample analytic liquid respectively is respectively the peak I: 297/326 nm, peak II: 250/330 nm, peak III: the emission spectrum fluorescence intensity F I of 3 characteristic peaks of 225/336 nm, F II and F III;
(3) data analysis: measure and calculate fluorescence intensity F I, F II, F III that each analytic liquid excitation/emission wavelength is 3 fluorescent characteristics peaks of peak I: 297/326nm, peak II: 250/330nm, peak III: 225/336nm respectively, calculate fluorescence intensity mark D
1, D
2, D
3With fluorescence intensity ratio R
1, R
2, R
3Utilize 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 a numeric type; in the fluorescence intensity mark and fluorescence intensity ratio data input " Data View " with each sample analytic liquid; 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 that connects between the utilization group is carried out hierarchial-cluster analysis, obtain tree-shaped dendrogram, by characteristic peak intensity mark as can be known, when distance coefficient is 5 ~ 7, the fluorescence intensity fractional value can be numbered each the tobacco leaf that testing sample identifies genus " fresh and sweet perfume (or spice) " type style or belongs to " alcohol fragrant and sweet " type style; By the characteristic peak intensity rate as can be known, when distance coefficient was 2 ~ 4, the fluorescence intensity fractional value can be numbered each the tobacco leaf that testing sample identifies genus " fresh and sweet perfume (or spice) " type style or belongs to " alcohol is fragrant and sweet " type style.
The used fluorescence spectrophotometer of the present invention is the Cary Eclipse type fluorospectrophotometer of U.S.'s Varian (Varian) company, and the fluorometric assay condition is: the no fluorescence quartz colorimetric utensil of the thick four sides polishing of 1 cm, excitation wavelength E
x200 ~ 500nm, emission wavelength E
m200 ~ 600nm, wavelength increases progressively 2.00nm, slit 5.00nm, voltage 720V.
The used data processing software of the present invention is spread sheet software (Excel) assembly of Microsoft's office package software (Microsoft office) and " statistics product and service solution " middle SPSS17.0 data statistic analysis software of SPSS Inc., and institute's test sample product characteristic spectrum data are carried out hierarchial-cluster analysis.
The sample collecting treatment conditions are to gather cured tobacco leaf sample C
3Tangerine three among the F() grade, sampling amount is 1kg at least, drying, accurately take by weighing 1g left and right sides sample after pulverizing, sieving, with chromatographically pure sherwood oil 30ml soak into, vibration left standstill 24 hours after 2 hours at least, get supernatant with 20 times of volumes of chromatographically pure sherwood oil dilution after as the analytic liquid of measuring three-dimensional fluorescence spectrum.
A kind of application of differentiating the method for Guizhou two class different flavor flue-cured tobaccos of the present patent application is characterized in that " fresh and sweet perfume (or spice) " and " alcohol is fragrant and sweet " the type flue-cured tobacco stylistic category differentiation that can be used for producing Anywhere.
Utilize the present patent application method to finish the different regions, Guizhou are produced flue-cured tobacco and carry out odor type classification, the flue-cured tobacco that draws the southeast of Guizhou Province, Tongren, the south of Guizhou Province and area, Zun Yi belongs 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, and wherein " alcohol is fragrant and sweet " type of " fresh and sweet perfume (or spice) " type of Bijie and Tongren is the most obvious.The present invention in analysis with Bijie flue-cured tobacco sample as " fresh and sweet perfume (or spice) " type standard model, with Tongren flue-cured tobacco sample as " alcohol fragrant and sweet " type standard model.In characteristic peak intensity mark dendrogram, in 5 ~ 7 distance coefficient scope, all gathering with Bijie flue-cured tobacco data is " fresh and sweet perfume (or spice) " type that is of a class; All gathering with Tongren flue-cured tobacco data is " alcohol the is fragrant and sweet " type that is of a class; In characteristic peak intensity rate dendrogram, in 2 ~ 4 distance coefficient scope, all gathering with Bijie flue-cured tobacco data is " fresh and sweet perfume (or spice) " type that is of a class; All gathering with Tongren flue-cured tobacco data is " alcohol the is fragrant and sweet " type that is of a class.
The present patent application scans with three-dimensional fluorescence spectrum the tobacco leaf petroleum ether extract, finds that three-dimensional fluorescence spectrum has three characteristic fluorescence peaks in the extract, is respectively peak I: E
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 I＞peak III＞peak II, sees Fig. 1 and Fig. 2, and chemical constitution general characteristic spectrum in the petroleum ether extract of cured tobacco leaf is represented at three peaks.The position of general characteristic spectrum peak, intensity characterize out the difference of the chemical substance relative content of tobacco leaf different flavor style, carry out the flue-cured tobacco odor type classification that cluster analysis just can be differentiated Guizhou " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavors fast with statistical analysis software.
Gordian technique in the present patent application is: the sherwood oil extension rate of (1) cigarette sample test fluid is that the 0.5mL supernatant dilutes 20 times with sherwood oil; (2) intensity mark and the ratio of gathering 3 characteristic fluorescence peaks carries out data analysis; (3) square Euclidean distance method in the characteristic spectrum The data hierarchial-cluster analysis.
The characteristics of the present patent application are that sample treatment is simple, spectroscopic assay is quick, easy, environmental protection and data processing are very simple, need not complex mathematical calculating and modeling, the differentiation result is objective, accurate, directly perceived.Method is simple, differentiates more objective, the accurate and quantification of the more traditional method of manually smokeing panel test of result, has practical reference value.
Description of drawings:
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 respectively peak I: E
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 comparatively near and excitation wavelength has bigger difference.Peak intensity shows as peak I＞peak III＞peak II, peak I intensity maximum wherein, and peak II, peak III intensity are lower, and each sample value 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
xThe isoline density of 3 characteristic peaks shows as peak I＞peak III＞peak II, and peak I intensity maximum is described, secondly is the peak III, and peak I intensity is minimum.
The fluorescence intensity mark dendrogram of Fig. 3 tobacco sample.With Bijie flue-cured tobacco sample (mark 1) as " fresh and sweet perfume (or spice) " type standard model, in distance coefficient is 5 ~ 7 scopes, can be with sample 1(six dish shredded tobacco for water pipes samples), sample 2(the southwest of Guizhou Province cigarette sample) to differentiate be " fresh and sweet perfume (or spice) " type that belongs to standard specimen 1 together; With Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 5 ~ 7 scopes, can be with sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) differentiate for belonging to " pure fragrant and sweet " type of standard specimen 2 together.
The fluorescence intensity ratio dendrogram of Fig. 4 tobacco sample.With Bijie flue-cured tobacco sample (mark 1) as " fresh and sweet perfume (or spice) " type standard model, in distance coefficient is 2 ~ 4 scopes, can be with sample 1(six dish shredded tobacco for water pipes samples), sample 2(the southwest of Guizhou Province cigarette sample) to differentiate be " fresh and sweet perfume (or spice) " type that belongs to standard specimen 1 together; With Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 2 ~ 4 scopes, can be with sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) differentiate for belonging to " pure fragrant and sweet " type of standard specimen 2 together.
Specific embodiment
Embodiment one: the fluorescence intensity mark is differentiated Guizhou two class different flavor flue-cured tobaccos in the fluorescent spectrometry
Gathering 7 kind cigarettes district, Guizhou grade is C
3Tangerine three among the F() 125 in cured tobacco leaf sample, sampling quantity respectively is 1kg, in 40 ℃ of oven drying 6h, pulverizes, and crosses 40 mesh sieves.Accurately weighing 1g cigarette sample left standstill 24 hours after 2 hours with chromatographically pure sherwood oil 30ml infiltration, vibration.Supernatant 0.5 mL after the above-mentioned sample treatment of learning from else's experience is handled, with 20 times of chromatographically pure sherwood oil dilutions, the polishing that is transferred to 1 cm thickness four sides does not have the fluorescence quartz colorimetric utensil, excitation wavelength is at 200 ~ 500 nm, emission wavelength is that wavelength increases progressively in 200 ~ 600 nm scopes with 2.0 nm, and outgoing and entrance slit are 5.0 nm, carry out three-dimensional fluorescence spectrum under the condition of detecting device multiplier tube voltage 720V and measure, spectrogram is seen attached Fig. 1 and 2.Obtain exciting of petroleum ether extract and be respectively the peak I: 297/326 nm, peak II: 250/330 nm, peak III: the fluorescence intensity of 3 characteristic peak total emission spectrum of 225/336 nm with emission wavelength.Utilize MS Office Applications Excel to calculate the fluorescence intensity mark D of each characteristic peak
1, D
2, D
3D wherein
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 fractional 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, other 3 is numeric type.Difference is planted in the fluorescence intensity fractional data input " Data View " of cigarette district sample number into spectrum and each department tobacco leaf petroleum ether extract, select " 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 that connects between the utilization group is carried out hierarchial-cluster analysis, obtains the tree-shaped dendrogram of characteristic peak fluorescence intensity fractional value, sees accompanying drawing 3.As seen from the figure, with Bijie flue-cured tobacco sample (mark 1) as " fresh and sweet perfume (or spice) " type standard model, in distance coefficient is 5 ~ 7 scopes, with sample 1(six dish shredded tobacco for water pipes samples), sample 2(the southwest of Guizhou Province cigarette sample) differentiation is for belonging to " fresh and sweet perfume (or spice) " type together; With Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 5 ~ 7 scopes, with sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) differentiate for belonging to " pure fragrant and sweet " type together.The provincial characteristics performance is clear, and classification is clear and definite.
Embodiment two: the strong ratio of fluorescence is differentiated Guizhou two class different flavor flue-cured tobaccos in the fluorescent spectrometry
Sample collecting, processing and spectrometric method are with embodiment one.Calculate the fluorescence intensity ratio R at 3 characteristic fluorescence peaks
1, R
2, R
3R wherein
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 the variable view " definition " sample number into spectrum, R
1, R
2, R
3" 4 variablees, wherein " sample number into spectrum " is character string type, other three is numeric type.Difference is planted in the fluorescence intensity ratio data input " Data View " of cigarette district sample number into spectrum and each department tobacco leaf petroleum ether extract, select " 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 that connects between the utilization group is carried out hierarchial-cluster analysis, obtains the tree-shaped dendrogram of characteristic peak fluorescence intensity ratio, sees accompanying drawing 4.As seen from the figure, with Bijie flue-cured tobacco sample (mark 1) as " fresh and sweet perfume (or spice) " type standard model, in distance coefficient is 2 ~ 4 scopes, with sample 1(six dish shredded tobacco for water pipes samples), sample 2(the southwest of Guizhou Province cigarette sample) differentiation is for belonging to " fresh and sweet perfume (or spice) " type together; With Tongren flue-cured tobacco sample (mark 2) as " alcohol is fragrant and sweet " type standard model, in distance coefficient 2 ~ 4 scopes, with sample 3(Zun Yi cigarette sample), sample 4(southeast of Guizhou Province cigarette sample), sample 5(the south of Guizhou Province cigarette sample) differentiate for belonging to " pure fragrant and sweet " type together.The provincial characteristics performance is clear, and classification is clear and definite.
Claims (6)
1. method of differentiating Guizhou two class different flavor flue-cured tobaccos, it is characterized in that utilizing the three-dimensional fluorescence spectrum technology, chemical constitution general characteristic spectrum in the petroleum ether extract of mensuration Guizhou cured tobacco leaf, position and intensity according to the general characteristic spectrum peak, obtain characterizing the difference of the chemical substance relative content of different flavor style flue-cured tobacco, carry out cluster analysis with statistical analysis software, set up the method for quick of differentiating Guizhou " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type two class different flavor style flue-cured tobaccos, the indication general characteristic spectrum of three-dimensional fluorescence spectrum technical measurement tobacco leaf petroleum ether extract, be to be respectively peak I: 297/326nm to excite with emission wavelength, peak II: 250/330nm, 3 fluorescence peaks of peak III: 225/336nm are characteristic peak, record tobacco leaf petroleum ether extract total emission spectral intensity F I respectively, F II and F III are 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 tree-shaped dendrogram, wherein D
1, D
2, D
3The fluorescence intensity mark of representing 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, can differentiate Guizhou flue-cured tobacco two class different flavors.
2. a kind of method of differentiating Guizhou two class different flavor flue-cured tobaccos according to claim 1 is characterized in that following these steps to successively differentiate:
(1) sample preparation: get standard " pure fragrant and sweet " type, " fresh and sweet perfume (or spice) " type and flue-cured tobacco sample to be measured respectively, with behind the sample number into spectrum in 40 ℃ of oven dryings, pulverize, cross 40 mesh sieves, accurate each sample of weighing, 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, with 20 times of chromatographically pure sherwood oil dilutions, get analytic liquid, be transferred to no fluorescence quartz colorimetric utensil, excitation wavelength is set at 200～500 nm, emission wavelength is in 200～600 nm scopes, with 2.0 nm is that wavelength increases progressively, outgoing and entrance slit are 5.0 nm, carrying out three-dimensional fluorescence spectrum under the condition of detecting device multiplier tube voltage 720V measures, the excitation/emission wavelength that obtains each sample analytic liquid respectively is respectively the peak I: 297/326 nm, peak II: 250/330 nm, peak III: the emission spectrum fluorescence intensity F I of 3 characteristic peaks of 225/336 nm, F II and F III;
(3) data analysis: measuring and calculate each analytic liquid excitation/emission wavelength respectively is 3 fluorescent characteristics peak F I, F II and the F III of peak I: 297/326nm, peak II: 250/330nm, peak III: 225/336nm; , calculate fluorescence intensity mark D
1, D
2, D
3And fluorescence intensity ratio R
1, R
2, R
3Utilize 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 a numeric type; in the fluorescence intensity mark and fluorescence intensity ratio data input " Data View " with each sample analytic liquid; 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 that connects between the utilization group is carried out hierarchial-cluster analysis, obtain tree-shaped dendrogram, by characteristic peak intensity mark as can be known, when distance coefficient is 5～7, the fluorescence intensity fractional value can be numbered each the tobacco leaf that testing sample identifies genus " fresh and sweet perfume (or spice) " type style or belongs to " alcohol fragrant and sweet " type style; By the characteristic peak intensity rate as can be known, when distance coefficient was 2～4, the fluorescence intensity fractional value can be numbered each the tobacco leaf that testing sample identifies genus " fresh and sweet perfume (or spice) " type style or belongs to " alcohol is fragrant and sweet " type style.
3. a kind of method of differentiating Guizhou two class different flavor flue-cured tobaccos according to claim 1 and 2, it is characterized in that used fluorescence spectrophotometer is the Cary Eclipse type fluorospectrophotometer of U.S.'s Varian (Varian) company, the fluorometric assay condition is: the no fluorescence quartz colorimetric utensil of the thick four sides polishing of 1 cm, excitation wavelength E
x200～500nm, emission wavelength E
m200～600nm, wavelength increases progressively 2.00nm, slit 5.00nm, voltage 720V.
4. a kind of method of differentiating Guizhou two class different flavor flue-cured tobaccos according to claim 1 and 2, it is characterized in that the used data processing software of cluster analysis is spread sheet software (Excel) assembly of Microsoft's office package software (Microsoft office) and " statistics 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.
5. a kind of method of differentiating Guizhou two class different flavor flue-cured tobaccos according to claim 1 and 2 is characterized in that gathering cured tobacco leaf sample C
3Tangerine three among the F() grade, sampling amount is 1kg at least, drying, accurately take by weighing 1g left and right sides sample after pulverizing, sieving, with chromatographically pure sherwood oil 30ml soak into, vibration left standstill 24 hours after 2 hours at least, get supernatant with 20 times of volumes of chromatographically pure sherwood oil dilution after as the analytic liquid of measuring three-dimensional fluorescence spectrum.
6. a kind of application of differentiating the method for Guizhou two class different flavor flue-cured tobaccos according to claim 1 and 2 is characterized in that " fresh and sweet perfume (or spice) " type and " alcohol is fragrant and sweet " type flue-cured tobacco type identification of can be used for producing Anywhere.
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