WO2020031448A1 - Method for creating reference for determining classification of plant-derived material containing chlorophyll and at least one of polyphenols and terpenoids, program, and device - Google Patents
Method for creating reference for determining classification of plant-derived material containing chlorophyll and at least one of polyphenols and terpenoids, program, and device Download PDFInfo
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- WO2020031448A1 WO2020031448A1 PCT/JP2019/018743 JP2019018743W WO2020031448A1 WO 2020031448 A1 WO2020031448 A1 WO 2020031448A1 JP 2019018743 W JP2019018743 W JP 2019018743W WO 2020031448 A1 WO2020031448 A1 WO 2020031448A1
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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- the present invention relates to a method, a program, and an apparatus for creating a standard for determining a category of a plant-derived material containing chlorophyll and at least one of polyphenol and terpenoid.
- Tobacco raw materials include air-dried tobacco raw materials (for example, Burley and native varieties), forced-fired tobacco raw materials (for example, yellow varieties), and sun-dried tobacco raw materials (for example, Orient varieties). Although it exists, it is impossible to determine the classification visually.
- As a method of determining the tobacco raw material category for example, there has been proposed a method of determining the tobacco raw material category based on the reflectance when the tobacco raw material is irradiated with visible light (Patent Document 1). On the other hand, it is known to distinguish or quantify food materials using fluorescent fingerprint information (Non-Patent Document 1).
- a processing step such as drying or a storage state
- the method of determining the classification of the material using visible light as described in Patent Document 1 is simple, if the reflection data is very similar, a problem may occur in the determination ability. Since the reflectance data obtained for the yellow type and the orient type of the tobacco raw material are very similar (Patent Document 1, FIG. 2), it is difficult to discriminate between the two.
- a method for creating a standard for determining a category of a plant-derived material containing chlorophyll and at least one of a polyphenol and a terpenoid Fluorescence fingerprint information acquisition step of acquiring fluorescence fingerprint information consisting of excitation wavelength, fluorescence wavelength, and fluorescence intensity data of the material whose classification is known, Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Creating a criterion indicating the relationship between the component and the chlorophyll-derived component.
- the fluorescence fingerprint information acquiring step includes irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from at least one of polyphenol and terpenoid, or excitation of 350 to 480 nm.
- the multivariate analysis is a principal component analysis.
- the material is a tobacco raw material, and before the step of acquiring fluorescent fingerprint information, further includes a step of allowing the tobacco raw material to stand at 22 ° C. and 60% for at least 24 hours to adjust the state.
- An apparatus for creating a reference for determining a category of a material derived from a plant containing chlorophyll and at least one of a polyphenol and a terpenoid Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the presence of a component derived from at least one of polyphenol and terpenoid in the material Means for analyzing the amount and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and creating a reference indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( A2).
- An apparatus for determining a classification of a chlorophyll and a plant-derived material containing at least one of a polyphenol and a terpenoid whose classification is unknown The means (A1) and the means (A2) according to the above [13],
- the present invention it is possible to provide a criterion, a discrimination method, a program, and a device for accurately discriminating a plant-derived material containing chlorophyll and at least one of polyphenol and terpenoid.
- FIG. 7 is a diagram showing a fluorescent fingerprint in an example.
- standard using the principal component analysis in this invention The figure which shows another aspect of the reference
- FIG. 1 illustrates one embodiment of an apparatus for carrying out the present invention.
- X to Y include the end values X and Y.
- Criteria for preparing the standard Chlorophyll the standard for determining the classification of materials derived from plants containing at least one of polyphenols and terpenoids, obtains fluorescent fingerprint information of the material, at least one of the polyphenols or terpenoids in the material The abundance of the component derived from, and obtained by analyzing as an index the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component, the material category, the component derived from at least one of polyphenols or terpenoids, and the chlorophyll-derived component Created based on relationships.
- the standard is also referred to as a predetermined standard.
- Target materials Materials derived from plants containing chlorophyll and at least one of polyphenols and terpenoids are materials obtained by subjecting the plants to processing such as cutting, molding, heating, drying, or unprocessed materials.
- Terpenoids are a group of natural organic compounds formed by combining a plurality of isoprene units, and examples thereof include carotenoids. Chlorophyll and polyphenols or terpenoids contained in plants can be metabolized by processing and aging.
- the target materials include chlorophyll or chlorophyll metabolites, polyphenols or polyphenol metabolites, and terpenoids or terpenoid metabolites.
- Target materials include tobacco raw materials and tea raw materials.
- the tobacco raw material is a raw material obtained by processing tobacco leaves and stems.
- Tobacco raw materials are deboned leaves, medium bones, and regenerated tobacco (ie, tobacco materials obtained by processing leaf scraps, chopping scraps, bone scraps, fine powder, etc., generated in the work processes of factories into reusable sheets and the like). Or a mixture of these.
- Tea raw materials are raw materials obtained by processing tea leaves, tea stems and the like.
- the material category is a category based on the attribute of the material.
- the classification of the tobacco raw materials includes the classification by the drying method, that is, the air-dried tobacco raw materials (for example, Burley and native varieties), the forced-fired tobacco raw materials (for example, yellow varieties), and the sun-dried tobacco raw materials (for example, Orient species).
- a criterion for determining whether a tobacco raw material belongs to a Burley type, a yellow type, or an Orient type is included.
- a criterion for discriminating which of the green tea, the deep-steamed tea, the roasted tea, the oolong tea, the black tea and the like belongs to is provided.
- the fluorescent fingerprint information is a three-dimensional fluorescent pattern composed of data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity by measuring the fluorescence wavelength and the fluorescence intensity by irradiating the excitation light.
- the measurement is performed by (1) irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from polyphenol or terpenoid, or (2) irradiating excitation light of 350 to 480 nm to chlorophyll.
- the excitation light wavelength in (1) is preferably from 320 to 370 nm, more preferably 360 nm, and the fluorescence wavelength is preferably from 380 to 480 nm, more preferably 460 nm.
- the excitation light wavelength in 2) can be 400 nm and the fluorescence wavelength can be 680 nm.
- Fluorescence caused by chlorophyll-derived components is fluorescence caused by chlorophyll and chlorophyll metabolites. The same applies to the fluorescence caused by the polyphenol-derived component and the terpenoid-derived component.
- the properties of the target material to be subjected to the measurement are not limited, but it is preferable that the target material is pulverized into a uniform powder for the measurement. This is because each component contained in the material is uniformly dispersed, so that highly accurate measurement is possible.
- a known device can be used for the pulverization.
- the maximum particle size is preferably 1 mm or less. The maximum particle size is determined by passing through a specified size mesh. For example, as the tobacco raw material, lamina, chopped powder, and the like may be mentioned, and it is preferable that the raw material is pulverized so as to have the above-mentioned particle size, sufficiently mixed, and then subjected to measurement.
- the condition of the target material to be subjected to the measurement be adjusted in advance in order to keep the water content constant.
- the tobacco raw materials it is preferable to store the tobacco raw materials under harmony conditions (22 ° C., 60%) for 24 hours or more.
- the upper limit of the storage time is not limited, but is preferably within 30 hours.
- analysis is performed using, as an index, fluorescent fingerprint information that reflects the abundance of components derived from at least one of polyphenols and terpenoids in the material and the abundance of components derived from chlorophyll.
- a multivariate analysis such as a principal component analysis, a discriminant analysis, a decision tree analysis, a hierarchical data analysis, a non-hierarchical clustering analysis, or the like can be used.
- a principal component analysis or a PLS discriminant analysis is preferable.
- Principal component analysis is a method of extracting principal components so that the variance of the principal components is maximized. Principal component analysis is performed using only explanatory variables, and a minimum is obtained between the obtained principal components and the objective variable.
- the principal component analysis is performed on the fluorescent fingerprint information
- the first principal component (PC1) is information relating to the polyphenol-derived component, the terpenoid-derived component or both
- the third principal component (PC3) is the information relating to the chlorophyll-derived component.
- a two-dimensional plot is created from the scores of the first main component and the third component, a two-dimensional diagram showing the relationship between the material category, one or both of the polyphenol-derived component and the terpenoid-derived component, and the chlorophyll-derived component can be created.
- a reference for each material section can be created.
- FIG. 2 shows an example of a criterion using principal component analysis for determining whether a tobacco raw material belongs to a yellow type, a burley type, or an oriental type.
- the algorithm used in the process may be a more general-purpose machine learning algorithm that also supports nonlinear phenomena, such as a support vector machine (SVM), a random forest (RF), and a neural network.
- SVM support vector machine
- RF random forest
- PLS regression analysis is a method of extracting a principal component such that the covariance between the principal component and the objective variable is maximized.
- the PLS discriminant analysis refers to an analysis in which the variables are set as variables of 0 and 1.
- clusters of yellow, orient, and burley are respectively set as objective variables (1, 0, 0), (0, 1, 0), (0, 0, 1). ) And make the respective criteria.
- Step 1 Fluorescent fingerprint information including data on the excitation wavelength, fluorescence wavelength, and fluorescence intensity of the target material whose classification is unknown is acquired.
- Step 2 Analysis is performed using, as an index, fluorescent fingerprint information reflecting the abundance of components derived from at least one of polyphenols and terpenoids in the material and the abundance of components derived from chlorophyll, and is derived from at least one of polyphenols and terpenoids. To obtain the relationship between the components and the chlorophyll-derived components.
- Step 3 The reference is prepared, and the relationship between the reference and the relation obtained in step 2 is checked to determine the category of the material.
- Step 1 can be carried out as described in the method of preparing the standard.
- step 2 for example, when principal component analysis is used as an analysis method, the analysis is performed as described in the method of creating a criterion, and the first principal component (PC1) and the third principal component (PC1) for the target material whose classification is unknown. The score of PC3) is obtained. Then, in step 3, the score is plotted on the reference and collated, whereby the material category of the target material whose category is unknown can be specified.
- Program A program is a data processing method described based on an arbitrary language or description method, and does not ask the format of a source code, a binary code, or the like.
- the program may be configured in a single form, but may be configured in a distributed manner as a plurality of modules or libraries, and may be configured to achieve its functions in cooperation with other existing programs. May be used.
- Apparatus for creating a standard for determining the category of the target material Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the abundance of components derived from at least one of polyphenols and terpenoids in the material And means for analyzing the fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and creating a criterion indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component (A2 ).
- FIG. 4 shows an embodiment of the device.
- A1 is a means for acquiring fluorescent fingerprint information of a material whose category is known
- A10 is fluorescent fingerprint information obtained by the means
- A2 is a means for creating a reference indicating a relationship between a material category and a component derived from polyphenol or the like
- A20 is a standard obtained by the means.
- FIG. 5 shows one mode of the device.
- A1 and A2 are as defined in FIG. 3
- B1 is means for acquiring fluorescent fingerprint information of a material whose classification is unknown
- B10 is fluorescent fingerprint information obtained by the means
- B2 is material classification.
- B20 is a unit for acquiring the relationship between components derived from polyphenols and the like
- C is a unit for collating the relationship with the standard
- C is a unit for collating the relationship with the reference.
- the device may be configured as hardware, but may be configured as a combination of function realizing means for realizing various functions by software of a computer.
- the function realizing means may include a program module.
- Example 1 The following were prepared as tobacco raw materials of known categories. Yellow (US, Africa, Italy, Moscow, India, China, Brazil) Burley (Brazil) Orient (Izmir, Greek, Thai, Chinese)
- FIG. 1 is a peak attributed to a component derived from polyphenol or carotenoid
- 3 is a peak attributed to a component derived from chlorophyll.
- the measurement conditions are as follows. Measuring instrument: F-7000 (Fluorescent fingerprint measuring device, manufactured by Hitachi High-Tech Science Corporation) Measurement method: Reflection method (FrontFace) Measurement conditions: excitation light 200 to 600 nm, fluorescence 200 to 900 nm, resolution 5 nm, slit width 5 nm, photomultiplier sensitivity 700
- Example 2 For the preprocessed fluorescent fingerprint information obtained in Example 1, PLS discriminant analysis was performed using software (“PLS TOOLBOX” manufactured by Eigenvector Research, Inc.), and the clusters of yellow, orient, and burley were determined as objective variables ( The analysis was performed as (1, 0, 0), (0, 1, 0), (0, 0, 1), and respective standards were created. Further, in order to verify the validity of the criterion, 25 samples whose classification was known were determined by the criterion, and it was examined whether or not the sample matched the actual sample classification. As a result, a match was found for 24 samples (discrimination result: 96%), which revealed that the criterion had high accuracy.
- PLS TOOLBOX manufactured by Eigenvector Research, Inc.
- Example 3 Black tea, roasted tea, and sencha were prepared as tea raw materials of which classifications were known. Each raw material was pulverized using a pulverizer so that the maximum particle size became 1 mm or less. Fluorescent fingerprint information of the material was obtained under the same conditions as in Example 1.
- Example 4 PLS discriminant analysis was performed on the preprocessed fluorescent fingerprint information obtained in Example 3 using software (“PLS TOOLBOX” manufactured by Eigenvector Research, Inc.), and clusters of black tea, roasted green tea, and sencha were determined as objective variables ( The analysis was performed as (1,0,0), (0,1,0), (0,0,1), and respective standards were created (FIG. 3). Furthermore, in order to verify the validity of the standard, 21 samples whose divisions were known were discriminated by the standard, and it was examined whether or not the sample matched the actual sample. As a result, a match was found for 20 samples (discrimination score: 96%), which revealed that the standard had high accuracy.
- B10 Fluorescent fingerprint information B2 Means for acquiring the relationship between material categories and components derived from polyphenols, etc.
- B20 Relationship C means for collating criteria and relationships
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Abstract
Provided is a method for creating a reference for determining the classification of a plant-derived material containing chlorophyll and at least one of polyphenols and terpenoids. The method includes: a fluorescence fingerprint information acquisition step in which fluorescence fingerprint information comprising data regarding the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of a sample of which the classification is already known is acquired; and a step in which analysis is performed using the abundance of a component derived from at least one of polyphenols and terpenoids within the sample and fluorescence fingerprint information reflecting the abundance of a chlorophyll-derived component as indices, then a reference indicating the relationship between the classification of the material, the component derived from at least one of polyphenols and terpenoids, and the chlorophyll-derived component is created.
Description
本発明は、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別する基準の作成方法、プログラム、および装置に関する。
(4) The present invention relates to a method, a program, and an apparatus for creating a standard for determining a category of a plant-derived material containing chlorophyll and at least one of polyphenol and terpenoid.
たばこ原料には、空気乾燥たばこ原料（例えば、バーレー種、および在来種）、強制火力乾燥たばこ原料（例えば、黄色種）、および天日乾燥たばこ原料（例えば、オリエント種）等の原料区分が存在するが、目視にて区分を判別することは不可能である。たばこ原料区分を判別する方法として、例えばたばこ原料に可視光を照射したときの反射率に基づいてたばこ原料区分を判別する方法が提案されている（特許文献１）。一方、蛍光指紋情報を用いて食品材料等の鑑別や定量を行うことが知られている（非特許文献１）。
Tobacco raw materials include air-dried tobacco raw materials (for example, Burley and native varieties), forced-fired tobacco raw materials (for example, yellow varieties), and sun-dried tobacco raw materials (for example, Orient varieties). Although it exists, it is impossible to determine the classification visually. As a method of determining the tobacco raw material category, for example, there has been proposed a method of determining the tobacco raw material category based on the reflectance when the tobacco raw material is irradiated with visible light (Patent Document 1). On the other hand, it is known to distinguish or quantify food materials using fluorescent fingerprint information (Non-Patent Document 1).
たばこ原料や茶原料等の、クロロフィルとポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料は乾燥等の加工工程や保存状態によって色調が変化するため、目視による区分の判別は困難である。特許文献１に記載されたような可視光を用いた当該材料の区分の判別方法は簡便であるが、反射データが酷似する場合は判別能力に問題が生じうる。実際にたばこ原料の黄色種とオリエント種において得られる反射率データは酷似するので（特許文献１図２）、両者の判別は困難である。かかる事情を鑑み、本発明はクロロフィルおよびポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を精度よく判別するための基準および判別方法、プログラム、および装置を提供することを課題とする。
材料 A material derived from a plant containing chlorophyll and at least one of polyphenols and terpenoids, such as a tobacco raw material and a tea raw material, changes in color depending on a processing step such as drying or a storage state, and thus it is difficult to visually determine the classification. Although the method of determining the classification of the material using visible light as described in Patent Document 1 is simple, if the reflection data is very similar, a problem may occur in the determination ability. Since the reflectance data obtained for the yellow type and the orient type of the tobacco raw material are very similar (Patent Document 1, FIG. 2), it is difficult to discriminate between the two. In view of such circumstances, it is an object of the present invention to provide a criterion and a discrimination method, a program, and an apparatus for accurately discriminating a classification of a material derived from a plant containing chlorophyll and at least one of polyphenol and terpenoid. .
発明者らは蛍光指紋情報を用いることで前記課題を解決できることを見出した。すなわち、前記課題は以下の本発明によって解決される。
［１］クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別するための基準を作成する方法であって、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得工程、
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する工程を含む、方法。
［２］前記材料が、たばこ原料または茶原料である、［１］に記載の方法。
［３］前記材料がたばこ原料であり、前記区分がバーレー種、黄色種、およびオリエント種である、［２］に記載の方法。
［４］前記材料が粉末である、［１］～［３］のいずれかに記載の方法。
［５］前記材料の最大粒径が１ｍｍ以下である、［４］に記載の方法。
［６］前記蛍光指紋情報取得工程が、２５０～３７０ｎｍの励起光を照射してポリフェノールまたはテルペノイドの少なくとも一方に由来する成分に起因する３００～４８０ｎｍの蛍光を測定すること、または３５０～４８０ｎｍの励起光を照射してクロロフィル由来成分に起因する６７０～７００ｎｍの蛍光を測定すること、あるいはその双方を含む、［１］～［５］に記載の方法。
［７］前記解析が多変量解析である、［１］～［６］のいずれかに記載の方法。
［８］前記多変量解析が主成分分析である、［７］に記載の方法。
［９］前記多変量解析がＰＬＳ判別分析である、［７］に記載の方法。
［１０］前記材料がたばこ原料であり、蛍光指紋情報を取得する工程の前に、当該たばこ原料を２２℃、６０％で２４時間以上静置して状態調整する工程をさらに含む、［１］～［９］のいずれかに記載の方法。
［１１］区分が未知である、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する工程１、
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する工程２、
前記［１］～［１０］のいずれかに記載の基準を準備して、当該基準と工程２で得た関係を照合して当該材料の区分を判別する工程３、を含む、
前記材料の区分を判別する方法。
［１２］コンピュータに前記［１］～［１１］のいずれかに記載の方法を実行させるためのプログラム。
［１３］クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別するための基準を作成するための装置であって、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する手段（Ａ１）、ならびに
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方とに由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する手段（Ａ２）、を備える装置。
［１４］区分が未知である、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方を含む植物に由来する材料の区分を判別する装置であって、
前記［１３］に記載の手段（Ａ１）および手段（Ａ２）、
前記区分が未知である材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得手段（Ｂ１）、ならびに前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する手段（Ｂ２）、ならびに
手段（Ａ２）で得た基準と、手段（Ｂ２）で得た関係とを照合して、前記区分が未知である材料の区分を判別する手段（Ｃ）を備える、装置。 The inventors have found that the above problem can be solved by using fluorescent fingerprint information. That is, the above problem is solved by the present invention described below.
[1] A method for creating a standard for determining a category of a plant-derived material containing chlorophyll and at least one of a polyphenol and a terpenoid,
Fluorescence fingerprint information acquisition step of acquiring fluorescence fingerprint information consisting of excitation wavelength, fluorescence wavelength, and fluorescence intensity data of the material whose classification is known,
Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Creating a criterion indicating the relationship between the component and the chlorophyll-derived component.
[2] The method according to [1], wherein the material is a tobacco raw material or a tea raw material.
[3] The method according to [2], wherein the material is a tobacco raw material, and the classification is a Burley type, a yellow type, and an Orient type.
[4] The method according to any one of [1] to [3], wherein the material is a powder.
[5] The method according to [4], wherein the material has a maximum particle size of 1 mm or less.
[6] The fluorescence fingerprint information acquiring step includes irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from at least one of polyphenol and terpenoid, or excitation of 350 to 480 nm. The method according to any one of [1] to [5], which comprises irradiating light to measure fluorescence at 670 to 700 nm caused by a component derived from chlorophyll, or both.
[7] The method according to any one of [1] to [6], wherein the analysis is a multivariate analysis.
[8] The method according to [7], wherein the multivariate analysis is a principal component analysis.
[9] The method according to [7], wherein the multivariate analysis is a PLS discriminant analysis.
[10] The material is a tobacco raw material, and before the step of acquiring fluorescent fingerprint information, further includes a step of allowing the tobacco raw material to stand at 22 ° C. and 60% for at least 24 hours to adjust the state. The method according to any one of to [9].
[11]Step 1 of acquiring fluorescent fingerprint information comprising excitation wavelength, fluorescence wavelength, and fluorescence intensity data of a plant-derived material containing chlorophyll and at least one of polyphenol and terpenoid, whose classification is unknown,
Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Step 2 of obtaining the relationship between the component and the component derived from chlorophyll,
Preparing a criterion according to any one of the above [1] to [10], collating the relationship obtained in step 2 with the criterion to determine the classification of the material,
A method for determining a category of the material.
[12] A program for causing a computer to execute the method according to any one of [1] to [11].
[13] An apparatus for creating a reference for determining a category of a material derived from a plant containing chlorophyll and at least one of a polyphenol and a terpenoid,
Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the presence of a component derived from at least one of polyphenol and terpenoid in the material Means for analyzing the amount and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and creating a reference indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( A2).
[14] An apparatus for determining a classification of a chlorophyll and a plant-derived material containing at least one of a polyphenol and a terpenoid whose classification is unknown,
The means (A1) and the means (A2) according to the above [13],
A fluorescent fingerprint information acquiring means (B1) for acquiring fluorescent fingerprint information comprising data of excitation wavelength, fluorescence wavelength, and fluorescence intensity of the material whose classification is unknown, and at least one of polyphenol and terpenoid in the material Means for analyzing the abundance of the component and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and acquiring the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( B2), and an apparatus comprising means (C) for comparing the reference obtained by the means (A2) with the relationship obtained by the means (B2) to determine the category of the material whose category is unknown.
［１］クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別するための基準を作成する方法であって、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得工程、
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する工程を含む、方法。
［２］前記材料が、たばこ原料または茶原料である、［１］に記載の方法。
［３］前記材料がたばこ原料であり、前記区分がバーレー種、黄色種、およびオリエント種である、［２］に記載の方法。
［４］前記材料が粉末である、［１］～［３］のいずれかに記載の方法。
［５］前記材料の最大粒径が１ｍｍ以下である、［４］に記載の方法。
［６］前記蛍光指紋情報取得工程が、２５０～３７０ｎｍの励起光を照射してポリフェノールまたはテルペノイドの少なくとも一方に由来する成分に起因する３００～４８０ｎｍの蛍光を測定すること、または３５０～４８０ｎｍの励起光を照射してクロロフィル由来成分に起因する６７０～７００ｎｍの蛍光を測定すること、あるいはその双方を含む、［１］～［５］に記載の方法。
［７］前記解析が多変量解析である、［１］～［６］のいずれかに記載の方法。
［８］前記多変量解析が主成分分析である、［７］に記載の方法。
［９］前記多変量解析がＰＬＳ判別分析である、［７］に記載の方法。
［１０］前記材料がたばこ原料であり、蛍光指紋情報を取得する工程の前に、当該たばこ原料を２２℃、６０％で２４時間以上静置して状態調整する工程をさらに含む、［１］～［９］のいずれかに記載の方法。
［１１］区分が未知である、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する工程１、
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する工程２、
前記［１］～［１０］のいずれかに記載の基準を準備して、当該基準と工程２で得た関係を照合して当該材料の区分を判別する工程３、を含む、
前記材料の区分を判別する方法。
［１２］コンピュータに前記［１］～［１１］のいずれかに記載の方法を実行させるためのプログラム。
［１３］クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別するための基準を作成するための装置であって、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する手段（Ａ１）、ならびに
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方とに由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する手段（Ａ２）、を備える装置。
［１４］区分が未知である、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方を含む植物に由来する材料の区分を判別する装置であって、
前記［１３］に記載の手段（Ａ１）および手段（Ａ２）、
前記区分が未知である材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得手段（Ｂ１）、ならびに前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する手段（Ｂ２）、ならびに
手段（Ａ２）で得た基準と、手段（Ｂ２）で得た関係とを照合して、前記区分が未知である材料の区分を判別する手段（Ｃ）を備える、装置。 The inventors have found that the above problem can be solved by using fluorescent fingerprint information. That is, the above problem is solved by the present invention described below.
[1] A method for creating a standard for determining a category of a plant-derived material containing chlorophyll and at least one of a polyphenol and a terpenoid,
Fluorescence fingerprint information acquisition step of acquiring fluorescence fingerprint information consisting of excitation wavelength, fluorescence wavelength, and fluorescence intensity data of the material whose classification is known,
Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Creating a criterion indicating the relationship between the component and the chlorophyll-derived component.
[2] The method according to [1], wherein the material is a tobacco raw material or a tea raw material.
[3] The method according to [2], wherein the material is a tobacco raw material, and the classification is a Burley type, a yellow type, and an Orient type.
[4] The method according to any one of [1] to [3], wherein the material is a powder.
[5] The method according to [4], wherein the material has a maximum particle size of 1 mm or less.
[6] The fluorescence fingerprint information acquiring step includes irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from at least one of polyphenol and terpenoid, or excitation of 350 to 480 nm. The method according to any one of [1] to [5], which comprises irradiating light to measure fluorescence at 670 to 700 nm caused by a component derived from chlorophyll, or both.
[7] The method according to any one of [1] to [6], wherein the analysis is a multivariate analysis.
[8] The method according to [7], wherein the multivariate analysis is a principal component analysis.
[9] The method according to [7], wherein the multivariate analysis is a PLS discriminant analysis.
[10] The material is a tobacco raw material, and before the step of acquiring fluorescent fingerprint information, further includes a step of allowing the tobacco raw material to stand at 22 ° C. and 60% for at least 24 hours to adjust the state. The method according to any one of to [9].
[11]
Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Step 2 of obtaining the relationship between the component and the component derived from chlorophyll,
Preparing a criterion according to any one of the above [1] to [10], collating the relationship obtained in step 2 with the criterion to determine the classification of the material,
A method for determining a category of the material.
[12] A program for causing a computer to execute the method according to any one of [1] to [11].
[13] An apparatus for creating a reference for determining a category of a material derived from a plant containing chlorophyll and at least one of a polyphenol and a terpenoid,
Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the presence of a component derived from at least one of polyphenol and terpenoid in the material Means for analyzing the amount and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and creating a reference indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( A2).
[14] An apparatus for determining a classification of a chlorophyll and a plant-derived material containing at least one of a polyphenol and a terpenoid whose classification is unknown,
The means (A1) and the means (A2) according to the above [13],
A fluorescent fingerprint information acquiring means (B1) for acquiring fluorescent fingerprint information comprising data of excitation wavelength, fluorescence wavelength, and fluorescence intensity of the material whose classification is unknown, and at least one of polyphenol and terpenoid in the material Means for analyzing the abundance of the component and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and acquiring the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( B2), and an apparatus comprising means (C) for comparing the reference obtained by the means (A2) with the relationship obtained by the means (B2) to determine the category of the material whose category is unknown.
本発明によってクロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を精度よく判別するための基準および判別方法、プログラム、および装置を提供できる。
According to the present invention, it is possible to provide a criterion, a discrimination method, a program, and a device for accurately discriminating a plant-derived material containing chlorophyll and at least one of polyphenol and terpenoid.
以下、本発明を詳細に説明する。本発明においてＸ～Ｙはその端値であるＸおよびＹを含む。
Hereinafter, the present invention will be described in detail. In the present invention, X to Y include the end values X and Y.
１．基準の作成方法
クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方を含む植物に由来する材料の区分を判別するための基準は、当該材料の蛍光指紋情報を取得し、前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析して得た、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係に基づいて作成される。当該基準を予め定められた基準ともいう。 1. Criteria for preparing the standard Chlorophyll, the standard for determining the classification of materials derived from plants containing at least one of polyphenols and terpenoids, obtains fluorescent fingerprint information of the material, at least one of the polyphenols or terpenoids in the material The abundance of the component derived from, and obtained by analyzing as an index the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component, the material category, the component derived from at least one of polyphenols or terpenoids, and the chlorophyll-derived component Created based on relationships. The standard is also referred to as a predetermined standard.
クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方を含む植物に由来する材料の区分を判別するための基準は、当該材料の蛍光指紋情報を取得し、前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析して得た、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係に基づいて作成される。当該基準を予め定められた基準ともいう。 1. Criteria for preparing the standard Chlorophyll, the standard for determining the classification of materials derived from plants containing at least one of polyphenols and terpenoids, obtains fluorescent fingerprint information of the material, at least one of the polyphenols or terpenoids in the material The abundance of the component derived from, and obtained by analyzing as an index the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component, the material category, the component derived from at least one of polyphenols or terpenoids, and the chlorophyll-derived component Created based on relationships. The standard is also referred to as a predetermined standard.
クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料（以下単に「対象材料」ともいう）とは、当該植物に切断、成形、加熱、乾燥等の加工を施した材料または未加工の植物である。テルペノイドとはイソプレン単位が複数個結合してできた天然有機化合物群であり、その例としてはカロテノイドが挙げられる。植物に含まれるクロロフィル、およびポリフェノールまたはテルペノイドは、加工や経時変化によって代謝されうる。したがって対象材料はクロロフィルまたはクロロフィル代謝物、ポリフェノールまたはポリフェノール代謝物、ならびにテルペノイドまたはテルペノイド代謝物を含む。対象材料としては、たばこ原料や茶原料が挙げられる。たばこ原料とは、たばこの葉や茎等を加工して得られた原料である。たばこ原料は、除骨葉、中骨、再生たばこ（すなわち、工場の作業工程で生じる葉屑、刻み屑、中骨屑、細粉などを再使用可能なシート等の形状に加工したたばこ材料）、またはこれらの混合物のいずれからなるものであってもよい。茶原料とは茶葉や茶茎等を加工して得られた原料である。材料区分とは、その材料の属性による区分である。たばこ原料の区分としては、乾燥方法による区分、すなわち空気乾燥たばこ原料（例えば、バーレー種、および在来種）、強制火力乾燥たばこ原料（例えば、黄色種）、および天日乾燥たばこ原料（例えば、オリエント種）のいずれかに属するかの区分が挙げられる。したがって、一態様として、たばこ原料がバーレー種、黄色種、およびオリエント種のいずれの区分に属するかを判別するための基準が挙げられる。また、茶原料に関しては、一態様として、煎茶、深蒸し茶、ほうじ茶、ウーロン茶、紅茶等のいずれの区分に属するかを判別するための基準が提供される。
Materials derived from plants containing chlorophyll and at least one of polyphenols and terpenoids (hereinafter also simply referred to as "target materials") are materials obtained by subjecting the plants to processing such as cutting, molding, heating, drying, or unprocessed materials. Plant. Terpenoids are a group of natural organic compounds formed by combining a plurality of isoprene units, and examples thereof include carotenoids. Chlorophyll and polyphenols or terpenoids contained in plants can be metabolized by processing and aging. Thus, the target materials include chlorophyll or chlorophyll metabolites, polyphenols or polyphenol metabolites, and terpenoids or terpenoid metabolites. Target materials include tobacco raw materials and tea raw materials. The tobacco raw material is a raw material obtained by processing tobacco leaves and stems. Tobacco raw materials are deboned leaves, medium bones, and regenerated tobacco (ie, tobacco materials obtained by processing leaf scraps, chopping scraps, bone scraps, fine powder, etc., generated in the work processes of factories into reusable sheets and the like). Or a mixture of these. Tea raw materials are raw materials obtained by processing tea leaves, tea stems and the like. The material category is a category based on the attribute of the material. The classification of the tobacco raw materials includes the classification by the drying method, that is, the air-dried tobacco raw materials (for example, Burley and native varieties), the forced-fired tobacco raw materials (for example, yellow varieties), and the sun-dried tobacco raw materials (for example, Orient species). Therefore, in one embodiment, a criterion for determining whether a tobacco raw material belongs to a Burley type, a yellow type, or an Orient type is included. In addition, with respect to the tea raw material, in one embodiment, a criterion for discriminating which of the green tea, the deep-steamed tea, the roasted tea, the oolong tea, the black tea and the like belongs to is provided.
［蛍光指紋情報取得工程］
本工程では、区分が既知である対象材料の蛍光指紋情報を取得する。蛍光指紋情報とは、励起光を照射して蛍光波長と蛍光強度を測定し、励起波長、蛍光波長、および蛍光強度のデータから構成される３次元蛍光パターンである。当該測定は、（１）２５０～３７０ｎｍの励起光を照射してポリフェノールまたはテルペノイド由来成分に起因する３００～４８０ｎｍの蛍光を測定すること、または（２）３５０～４８０ｎｍの励起光を照射してクロロフィル由来成分に起因する６７０～７００ｎｍの蛍光を測定すること、あるいは（１）および（２）の双方を含むことが好ましい。各波長は対象物の特性によって適宜調整される。例えば、たばこ原料に対しては（１）における励起光波長は、好ましくは３２０～３７０ｎｍ、より好ましくは３６０ｎｍであり、蛍光波長は好ましくは３８０～４８０ｎｍ、より好ましくは４６０ｎｍとすることができ、（２）における励起光波長は４００ｎｍ、蛍光波長は６８０ｎｍとすることができる。クロロフィル由来成分に起因する蛍光とは、クロロフィルおよびクロロフィル代謝物に起因する蛍光である。ポリフェノール由来成分およびテルペノイド由来成分に起因する蛍光についても同様である。 [Fluorescent fingerprint information acquisition process]
In this step, the fluorescent fingerprint information of the target material whose classification is known is acquired. The fluorescent fingerprint information is a three-dimensional fluorescent pattern composed of data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity by measuring the fluorescence wavelength and the fluorescence intensity by irradiating the excitation light. The measurement is performed by (1) irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from polyphenol or terpenoid, or (2) irradiating excitation light of 350 to 480 nm to chlorophyll. It is preferable to measure the fluorescence at 670 to 700 nm due to the origin component, or to include both (1) and (2). Each wavelength is appropriately adjusted according to the characteristics of the object. For example, for a tobacco raw material, the excitation light wavelength in (1) is preferably from 320 to 370 nm, more preferably 360 nm, and the fluorescence wavelength is preferably from 380 to 480 nm, more preferably 460 nm. The excitation light wavelength in 2) can be 400 nm and the fluorescence wavelength can be 680 nm. Fluorescence caused by chlorophyll-derived components is fluorescence caused by chlorophyll and chlorophyll metabolites. The same applies to the fluorescence caused by the polyphenol-derived component and the terpenoid-derived component.
本工程では、区分が既知である対象材料の蛍光指紋情報を取得する。蛍光指紋情報とは、励起光を照射して蛍光波長と蛍光強度を測定し、励起波長、蛍光波長、および蛍光強度のデータから構成される３次元蛍光パターンである。当該測定は、（１）２５０～３７０ｎｍの励起光を照射してポリフェノールまたはテルペノイド由来成分に起因する３００～４８０ｎｍの蛍光を測定すること、または（２）３５０～４８０ｎｍの励起光を照射してクロロフィル由来成分に起因する６７０～７００ｎｍの蛍光を測定すること、あるいは（１）および（２）の双方を含むことが好ましい。各波長は対象物の特性によって適宜調整される。例えば、たばこ原料に対しては（１）における励起光波長は、好ましくは３２０～３７０ｎｍ、より好ましくは３６０ｎｍであり、蛍光波長は好ましくは３８０～４８０ｎｍ、より好ましくは４６０ｎｍとすることができ、（２）における励起光波長は４００ｎｍ、蛍光波長は６８０ｎｍとすることができる。クロロフィル由来成分に起因する蛍光とは、クロロフィルおよびクロロフィル代謝物に起因する蛍光である。ポリフェノール由来成分およびテルペノイド由来成分に起因する蛍光についても同様である。 [Fluorescent fingerprint information acquisition process]
In this step, the fluorescent fingerprint information of the target material whose classification is known is acquired. The fluorescent fingerprint information is a three-dimensional fluorescent pattern composed of data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity by measuring the fluorescence wavelength and the fluorescence intensity by irradiating the excitation light. The measurement is performed by (1) irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from polyphenol or terpenoid, or (2) irradiating excitation light of 350 to 480 nm to chlorophyll. It is preferable to measure the fluorescence at 670 to 700 nm due to the origin component, or to include both (1) and (2). Each wavelength is appropriately adjusted according to the characteristics of the object. For example, for a tobacco raw material, the excitation light wavelength in (1) is preferably from 320 to 370 nm, more preferably 360 nm, and the fluorescence wavelength is preferably from 380 to 480 nm, more preferably 460 nm. The excitation light wavelength in 2) can be 400 nm and the fluorescence wavelength can be 680 nm. Fluorescence caused by chlorophyll-derived components is fluorescence caused by chlorophyll and chlorophyll metabolites. The same applies to the fluorescence caused by the polyphenol-derived component and the terpenoid-derived component.
測定に供する対象材料の性状は限定されないが、対象材料を粉砕して均一な粉末にして測定に供することが好ましい。材料に含まれる各成分が均一に分散するので精度の高い測定が可能になるからである。粉砕には公知の機器を使用できる。最大粒径は１ｍｍ以下であることが好ましい。最大粒径は指定サイズのメッシュを通過させることによって決定される。例えば、たばこ原料としてはラミナ、刻み、粉末が挙げられるが、これを前記粒径となるように粉砕後、十分に混合して測定に供することが好ましい。測定に供する対象材料は事前に水分量を一定化するために状態調整されることが好ましい。例えばたばこ原料の場合、調和条件（２２℃、６０％）で２４時間以上蔵置することが好ましい。蔵置する時間の上限は限定されないが、３０時間以内であることが好ましい。
性 The properties of the target material to be subjected to the measurement are not limited, but it is preferable that the target material is pulverized into a uniform powder for the measurement. This is because each component contained in the material is uniformly dispersed, so that highly accurate measurement is possible. A known device can be used for the pulverization. The maximum particle size is preferably 1 mm or less. The maximum particle size is determined by passing through a specified size mesh. For example, as the tobacco raw material, lamina, chopped powder, and the like may be mentioned, and it is preferable that the raw material is pulverized so as to have the above-mentioned particle size, sufficiently mixed, and then subjected to measurement. It is preferable that the condition of the target material to be subjected to the measurement be adjusted in advance in order to keep the water content constant. For example, in the case of tobacco raw materials, it is preferable to store the tobacco raw materials under harmony conditions (22 ° C., 60%) for 24 hours or more. The upper limit of the storage time is not limited, but is preferably within 30 hours.
［基準作成工程］
本工程では、前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量およびクロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析を行う。解析は、主成分分析、判別分析、決定木分析、階層的データ分析、非階層クラスタリング分析等の多変量解析を用いることができるが、中でも主成分分析またはＰＬＳ判別分析が好ましい。主成分分析とは、主成分の分散が最大になるように主成分を抽出する手法であり、説明変数のみを用いて主成分分析を行い、得られた主成分と目的変数との間で最小二乗法による重回帰分析を行う手法である。例えば、蛍光指紋情報に対して主成分分析を行い、第１主成分（ＰＣ１）をポリフェノール由来成分、テルペノイド由来成分またはその双方にかかる情報、第３主成分（ＰＣ３）をクロロフィル由来成分にかかる情報として採用する。第１主成分と第３成分のスコアから２次元プロットを作成すれば、材料区分、ポリフェノール由来成分またはテルペノイド由来成分の一方またはその双方、およびクロロフィル由来成分の関係を示す２次元図を作成できる。この２次元図において各材料区分を判別するためのグルーピングを行うことで各材料区分の基準を作成できる。グルーピングには統計的な手法を用いることもできるが、未知の原料サンプルと既知原料サンプルのプロット群との近似性等を考慮して実施することが好ましい。異なる区分に属する複数の対象材料を測定すれば、より精度の高い基準を作成できる。特に、たばこ原料においては、黄色種、バーレー種、およびオリエント種では、ポリフェノール由来成分、テルペノイド由来成分およびクロロフィル由来成分の存在量がそれぞれ異なるので、精度の高い基準を作成できる。図２は、たばこ原料が、黄色種、バーレー種、およびオリエント種のいずれの区分に属するかを判別するための主成分分析を用いた基準の一例を示す。当該工程に用いるアルゴリズムはより汎用的かつ非線形現象にも対応する機械学習アルゴリズム、例えばsupport vector machine (SVM), random forest (RF), neural networkなどでもよい。 [Standard creation process]
In this step, analysis is performed using, as an index, fluorescent fingerprint information that reflects the abundance of components derived from at least one of polyphenols and terpenoids in the material and the abundance of components derived from chlorophyll. As the analysis, a multivariate analysis such as a principal component analysis, a discriminant analysis, a decision tree analysis, a hierarchical data analysis, a non-hierarchical clustering analysis, or the like can be used. Among them, a principal component analysis or a PLS discriminant analysis is preferable. Principal component analysis is a method of extracting principal components so that the variance of the principal components is maximized. Principal component analysis is performed using only explanatory variables, and a minimum is obtained between the obtained principal components and the objective variable. This is a method for performing multiple regression analysis by the square method. For example, the principal component analysis is performed on the fluorescent fingerprint information, and the first principal component (PC1) is information relating to the polyphenol-derived component, the terpenoid-derived component or both, and the third principal component (PC3) is the information relating to the chlorophyll-derived component. To be adopted. If a two-dimensional plot is created from the scores of the first main component and the third component, a two-dimensional diagram showing the relationship between the material category, one or both of the polyphenol-derived component and the terpenoid-derived component, and the chlorophyll-derived component can be created. By performing grouping for discriminating each material section in this two-dimensional diagram, a reference for each material section can be created. Although a statistical method can be used for grouping, it is preferable to perform the grouping in consideration of the similarity between a plot group of an unknown raw material sample and a known raw material sample. If a plurality of target materials belonging to different categories are measured, a more accurate reference can be created. In particular, in the tobacco raw material, the yellow, burley, and orient species have different amounts of the polyphenol-derived component, the terpenoid-derived component, and the chlorophyll-derived component. FIG. 2 shows an example of a criterion using principal component analysis for determining whether a tobacco raw material belongs to a yellow type, a burley type, or an oriental type. The algorithm used in the process may be a more general-purpose machine learning algorithm that also supports nonlinear phenomena, such as a support vector machine (SVM), a random forest (RF), and a neural network.
本工程では、前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量およびクロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析を行う。解析は、主成分分析、判別分析、決定木分析、階層的データ分析、非階層クラスタリング分析等の多変量解析を用いることができるが、中でも主成分分析またはＰＬＳ判別分析が好ましい。主成分分析とは、主成分の分散が最大になるように主成分を抽出する手法であり、説明変数のみを用いて主成分分析を行い、得られた主成分と目的変数との間で最小二乗法による重回帰分析を行う手法である。例えば、蛍光指紋情報に対して主成分分析を行い、第１主成分（ＰＣ１）をポリフェノール由来成分、テルペノイド由来成分またはその双方にかかる情報、第３主成分（ＰＣ３）をクロロフィル由来成分にかかる情報として採用する。第１主成分と第３成分のスコアから２次元プロットを作成すれば、材料区分、ポリフェノール由来成分またはテルペノイド由来成分の一方またはその双方、およびクロロフィル由来成分の関係を示す２次元図を作成できる。この２次元図において各材料区分を判別するためのグルーピングを行うことで各材料区分の基準を作成できる。グルーピングには統計的な手法を用いることもできるが、未知の原料サンプルと既知原料サンプルのプロット群との近似性等を考慮して実施することが好ましい。異なる区分に属する複数の対象材料を測定すれば、より精度の高い基準を作成できる。特に、たばこ原料においては、黄色種、バーレー種、およびオリエント種では、ポリフェノール由来成分、テルペノイド由来成分およびクロロフィル由来成分の存在量がそれぞれ異なるので、精度の高い基準を作成できる。図２は、たばこ原料が、黄色種、バーレー種、およびオリエント種のいずれの区分に属するかを判別するための主成分分析を用いた基準の一例を示す。当該工程に用いるアルゴリズムはより汎用的かつ非線形現象にも対応する機械学習アルゴリズム、例えばsupport vector machine (SVM), random forest (RF), neural networkなどでもよい。 [Standard creation process]
In this step, analysis is performed using, as an index, fluorescent fingerprint information that reflects the abundance of components derived from at least one of polyphenols and terpenoids in the material and the abundance of components derived from chlorophyll. As the analysis, a multivariate analysis such as a principal component analysis, a discriminant analysis, a decision tree analysis, a hierarchical data analysis, a non-hierarchical clustering analysis, or the like can be used. Among them, a principal component analysis or a PLS discriminant analysis is preferable. Principal component analysis is a method of extracting principal components so that the variance of the principal components is maximized. Principal component analysis is performed using only explanatory variables, and a minimum is obtained between the obtained principal components and the objective variable. This is a method for performing multiple regression analysis by the square method. For example, the principal component analysis is performed on the fluorescent fingerprint information, and the first principal component (PC1) is information relating to the polyphenol-derived component, the terpenoid-derived component or both, and the third principal component (PC3) is the information relating to the chlorophyll-derived component. To be adopted. If a two-dimensional plot is created from the scores of the first main component and the third component, a two-dimensional diagram showing the relationship between the material category, one or both of the polyphenol-derived component and the terpenoid-derived component, and the chlorophyll-derived component can be created. By performing grouping for discriminating each material section in this two-dimensional diagram, a reference for each material section can be created. Although a statistical method can be used for grouping, it is preferable to perform the grouping in consideration of the similarity between a plot group of an unknown raw material sample and a known raw material sample. If a plurality of target materials belonging to different categories are measured, a more accurate reference can be created. In particular, in the tobacco raw material, the yellow, burley, and orient species have different amounts of the polyphenol-derived component, the terpenoid-derived component, and the chlorophyll-derived component. FIG. 2 shows an example of a criterion using principal component analysis for determining whether a tobacco raw material belongs to a yellow type, a burley type, or an oriental type. The algorithm used in the process may be a more general-purpose machine learning algorithm that also supports nonlinear phenomena, such as a support vector machine (SVM), a random forest (RF), and a neural network.
ＰＬＳ回帰分析は主成分と目的変数との共分散が最大になるように主成分を抽出する方法である。ＰＬＳ判別分析とは前記変数を０と１の変数として行う分析をいう。蛍光指紋情報に対してＰＬＳ判別分析を行う場合は、例えば、黄色、オリエント、バーレーのクラスターをそれぞれ、目的変数（１，０，０）、（０，１，０）、（０，０，１）として解析を行い、それぞれの基準を作成する。
PLS regression analysis is a method of extracting a principal component such that the covariance between the principal component and the objective variable is maximized. The PLS discriminant analysis refers to an analysis in which the variables are set as variables of 0 and 1. When performing PLS discriminant analysis on fluorescent fingerprint information, for example, clusters of yellow, orient, and burley are respectively set as objective variables (1, 0, 0), (0, 1, 0), (0, 0, 1). ) And make the respective criteria.
２．区分が未知である対象材料の判別方法
前述のとおり作成された基準は、区分が未知である材料の区分の判別方法に有用である。当該方法は、以下の工程を含む。
工程１：区分が未知である対象材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する。
工程２：前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量およびクロロフィルに由来する成分の存在量を反映する蛍光指紋情報を指標として解析を行い、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分およびクロロフィル由来成分の関係を取得する。
工程３：前記基準を準備して、当該基準と工程２で得た関係を照合して当該材料の区分を判別する。 2. Method of determining target material whose classification is unknown The criterion created as described above is useful for a method of determining the classification of a material whose classification is unknown. The method includes the following steps.
Step 1: Fluorescent fingerprint information including data on the excitation wavelength, fluorescence wavelength, and fluorescence intensity of the target material whose classification is unknown is acquired.
Step 2: Analysis is performed using, as an index, fluorescent fingerprint information reflecting the abundance of components derived from at least one of polyphenols and terpenoids in the material and the abundance of components derived from chlorophyll, and is derived from at least one of polyphenols and terpenoids. To obtain the relationship between the components and the chlorophyll-derived components.
Step 3: The reference is prepared, and the relationship between the reference and the relation obtained in step 2 is checked to determine the category of the material.
前述のとおり作成された基準は、区分が未知である材料の区分の判別方法に有用である。当該方法は、以下の工程を含む。
工程１：区分が未知である対象材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する。
工程２：前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量およびクロロフィルに由来する成分の存在量を反映する蛍光指紋情報を指標として解析を行い、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分およびクロロフィル由来成分の関係を取得する。
工程３：前記基準を準備して、当該基準と工程２で得た関係を照合して当該材料の区分を判別する。 2. Method of determining target material whose classification is unknown The criterion created as described above is useful for a method of determining the classification of a material whose classification is unknown. The method includes the following steps.
Step 1: Fluorescent fingerprint information including data on the excitation wavelength, fluorescence wavelength, and fluorescence intensity of the target material whose classification is unknown is acquired.
Step 2: Analysis is performed using, as an index, fluorescent fingerprint information reflecting the abundance of components derived from at least one of polyphenols and terpenoids in the material and the abundance of components derived from chlorophyll, and is derived from at least one of polyphenols and terpenoids. To obtain the relationship between the components and the chlorophyll-derived components.
Step 3: The reference is prepared, and the relationship between the reference and the relation obtained in step 2 is checked to determine the category of the material.
工程１は、基準の作成方法で述べたとおりに実施できる。工程２では、例えは解析手法として主成分分析を用いる場合、基準の作成方法で述べたとおりに分析を行い、区分が未知である対象材料について第１主成分（ＰＣ１）と第３主成分（ＰＣ３）のスコアを求める。そして、工程３において、当該スコアを前記基準上にプロットして照合することで、区分が未知である対象材料の材料区分を特定できる。
Step 1 can be carried out as described in the method of preparing the standard. In step 2, for example, when principal component analysis is used as an analysis method, the analysis is performed as described in the method of creating a criterion, and the first principal component (PC1) and the third principal component (PC1) for the target material whose classification is unknown. The score of PC3) is obtained. Then, in step 3, the score is plotted on the reference and collated, whereby the material category of the target material whose category is unknown can be specified.
３．プログラム
プログラムとは、任意の言語や記述方法に基づき記述されたデータ処理方法であり、ソースコードやバイナリコード等の形式を問うものではない。また、プログラムは単一の形で構成されてもよいが、複数のモジュールやライブラリとして分散構成されてもよく、また、他の既存のプログラムと協働してその機能を達成するように構成されたものであってもよい。 3. Program A program is a data processing method described based on an arbitrary language or description method, and does not ask the format of a source code, a binary code, or the like. In addition, the program may be configured in a single form, but may be configured in a distributed manner as a plurality of modules or libraries, and may be configured to achieve its functions in cooperation with other existing programs. May be used.
プログラムとは、任意の言語や記述方法に基づき記述されたデータ処理方法であり、ソースコードやバイナリコード等の形式を問うものではない。また、プログラムは単一の形で構成されてもよいが、複数のモジュールやライブラリとして分散構成されてもよく、また、他の既存のプログラムと協働してその機能を達成するように構成されたものであってもよい。 3. Program A program is a data processing method described based on an arbitrary language or description method, and does not ask the format of a source code, a binary code, or the like. In addition, the program may be configured in a single form, but may be configured in a distributed manner as a plurality of modules or libraries, and may be configured to achieve its functions in cooperation with other existing programs. May be used.
４．装置
（１）対象材料の区分を判別するための基準を作成する装置
当該装置は、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する手段（Ａ１）、ならびに
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する手段（Ａ２）、を備える。 4. Apparatus (1) Apparatus for creating a standard for determining the category of the target material
Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the abundance of components derived from at least one of polyphenols and terpenoids in the material And means for analyzing the fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and creating a criterion indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component (A2 ).
（１）対象材料の区分を判別するための基準を作成する装置
当該装置は、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する手段（Ａ１）、ならびに
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する手段（Ａ２）、を備える。 4. Apparatus (1) Apparatus for creating a standard for determining the category of the target material
Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the abundance of components derived from at least one of polyphenols and terpenoids in the material And means for analyzing the fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and creating a criterion indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component (A2 ).
図４に当該装置の一態様を示す。図中、Ａ１は区分が既知である材料の蛍光指紋情報を取得する手段、Ａ１０は当該手段で得た蛍光指紋情報、Ａ２は材料区分とポリフェノール等由来成分の関係を示す基準を作成する手段、Ａ２０は当該手段で得た基準である。
(4) FIG. 4 shows an embodiment of the device. In the figure, A1 is a means for acquiring fluorescent fingerprint information of a material whose category is known, A10 is fluorescent fingerprint information obtained by the means, A2 is a means for creating a reference indicating a relationship between a material category and a component derived from polyphenol or the like, A20 is a standard obtained by the means.
（２）区分が未知である対象材料の区分を判別する装置
当該装置は、
前記手段（Ａ１）および手段（Ａ２）、
前記区分が未知である材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得手段（Ｂ１）、ならびに前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する手段（Ｂ２）、ならびに
手段（Ａ２）で得た基準と、手段（Ｂ２）で得た関係とを照合して、前記区分が未知である材料の区分を判別する手段（Ｃ）を備える。 (2) Apparatus for determining the category of the target material whose category is unknown
Said means (A1) and means (A2);
A fluorescent fingerprint information obtaining means (B1) for obtaining fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescent wavelength, and the fluorescent intensity of the material whose classification is unknown, and at least one of polyphenol or terpenoid in the material Means for analyzing the abundance of the component and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and acquiring the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( B2) and a means (C) for comparing the reference obtained by the means (A2) with the relationship obtained by the means (B2) to determine the category of the material whose category is unknown.
当該装置は、
前記手段（Ａ１）および手段（Ａ２）、
前記区分が未知である材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得手段（Ｂ１）、ならびに前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する手段（Ｂ２）、ならびに
手段（Ａ２）で得た基準と、手段（Ｂ２）で得た関係とを照合して、前記区分が未知である材料の区分を判別する手段（Ｃ）を備える。 (2) Apparatus for determining the category of the target material whose category is unknown
Said means (A1) and means (A2);
A fluorescent fingerprint information obtaining means (B1) for obtaining fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescent wavelength, and the fluorescent intensity of the material whose classification is unknown, and at least one of polyphenol or terpenoid in the material Means for analyzing the abundance of the component and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and acquiring the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( B2) and a means (C) for comparing the reference obtained by the means (A2) with the relationship obtained by the means (B2) to determine the category of the material whose category is unknown.
図５に当該装置の一態様を示す。図中、Ａ１とＡ２は図３で定義されるとおりであり、Ｂ１は区分が未知である材料の蛍光指紋情報を取得する手段、Ｂ１０は当該手段で得た蛍光指紋情報、Ｂ２は材料区分とポリフェノール等由来成分の関係を取得する手段、Ｂ２０は当該手段で得た関係、Ｃは前記基準と関係とを照合する手段である。
FIG. 5 shows one mode of the device. In the figure, A1 and A2 are as defined in FIG. 3, B1 is means for acquiring fluorescent fingerprint information of a material whose classification is unknown, B10 is fluorescent fingerprint information obtained by the means, B2 is material classification. B20 is a unit for acquiring the relationship between components derived from polyphenols and the like, C is a unit for collating the relationship with the standard, and C is a unit for collating the relationship with the reference.
本発明において装置はハードウェアとして構成されてもよいが、コンピュータのソフトウェアによって各種機能を実現する機能実現手段の組合せとして構成されてもよい。機能実現手段には、プログラムモジュールが含まれうる。
In the present invention, the device may be configured as hardware, but may be configured as a combination of function realizing means for realizing various functions by software of a computer. The function realizing means may include a program module.
［実施例１］
区分が既知であるたばこ原料として以下を準備した。
黄色種（米国産、タンザニア産、イタリア産、ジンバブエ産、インド産、中国産、ブラジル産）
バーレー種（ブラジル産）
オリエント種（イズミール産、ギリシャ産、タイ産、中国産） [Example 1]
The following were prepared as tobacco raw materials of known categories.
Yellow (US, Tanzania, Italy, Zimbabwe, India, China, Brazil)
Burley (Brazil)
Orient (Izmir, Greek, Thai, Chinese)
区分が既知であるたばこ原料として以下を準備した。
黄色種（米国産、タンザニア産、イタリア産、ジンバブエ産、インド産、中国産、ブラジル産）
バーレー種（ブラジル産）
オリエント種（イズミール産、ギリシャ産、タイ産、中国産） [Example 1]
The following were prepared as tobacco raw materials of known categories.
Yellow (US, Tanzania, Italy, Zimbabwe, India, China, Brazil)
Burley (Brazil)
Orient (Izmir, Greek, Thai, Chinese)
各原料のラミナを、粉砕機を用いて最大粒径が１ｍｍ以下になるように粉砕した。当該材料の蛍光指紋情報を取得した（図１）。図１中、１はポリフェノール由来成分またはカロテノイド由来成分に起因するピークであり、３はクロロフィル由来成分に起因するピークであった。
測定条件は以下のとおりである。
測定機器：Ｆ－７０００（蛍光指紋測定装置、日立ハイテクサイエンス社製）
測定法：反射法（ＦｒｏｎｔＦａｃｅ）
測定条件：励起光２００～６００ｎｍ、蛍光２００～９００ｎｍ、分解能５ｎｍ、スリット幅５ｎｍ、フォトマル感度７００ The lamina of each raw material was pulverized using a pulverizer so that the maximum particle size became 1 mm or less. The fluorescent fingerprint information of the material was obtained (FIG. 1). In FIG. 1, 1 is a peak attributed to a component derived from polyphenol or carotenoid, and 3 is a peak attributed to a component derived from chlorophyll.
The measurement conditions are as follows.
Measuring instrument: F-7000 (Fluorescent fingerprint measuring device, manufactured by Hitachi High-Tech Science Corporation)
Measurement method: Reflection method (FrontFace)
Measurement conditions:excitation light 200 to 600 nm, fluorescence 200 to 900 nm, resolution 5 nm, slit width 5 nm, photomultiplier sensitivity 700
測定条件は以下のとおりである。
測定機器：Ｆ－７０００（蛍光指紋測定装置、日立ハイテクサイエンス社製）
測定法：反射法（ＦｒｏｎｔＦａｃｅ）
測定条件：励起光２００～６００ｎｍ、蛍光２００～９００ｎｍ、分解能５ｎｍ、スリット幅５ｎｍ、フォトマル感度７００ The lamina of each raw material was pulverized using a pulverizer so that the maximum particle size became 1 mm or less. The fluorescent fingerprint information of the material was obtained (FIG. 1). In FIG. 1, 1 is a peak attributed to a component derived from polyphenol or carotenoid, and 3 is a peak attributed to a component derived from chlorophyll.
The measurement conditions are as follows.
Measuring instrument: F-7000 (Fluorescent fingerprint measuring device, manufactured by Hitachi High-Tech Science Corporation)
Measurement method: Reflection method (FrontFace)
Measurement conditions:
得られた蛍光指紋情報について、対象とする成分に関与しない不要な波長を削除した。具体的には、非蛍光成分の除去処理、散乱光の除去処理、低感度領域の削除処理を実施した。さらに蛍光指紋情報についてノーマライズおよびオートスケールを行った。前処理を施した蛍光指紋情報についてソフトウェア「Matlab」を用いて主成分分析を行い、ＰＣ１をポリフェノール由来成分およびカロテノイド由来成分、ＰＣ３をクロロフィル由来成分として、材料区分、ポリフェノール由来成分とカロテノイド由来成分、およびクロロフィル由来成分の関係を示す基準を作成した。結果を図２に示す。図２に示すように、同一区分のたばこ原料が近い位置にプロットされた。これらを例えば目視により各区分にグルーピングすることによって、基準を作成した。
に つ い て In the obtained fluorescent fingerprint information, unnecessary wavelengths not related to the target component were deleted. Specifically, a non-fluorescent component removal process, a scattered light removal process, and a low-sensitivity region removal process were performed. Further, normalization and auto-scale were performed on the fluorescent fingerprint information. The main component analysis is performed on the pre-processed fluorescent fingerprint information using software "Matlab", and PC1 is a polyphenol-derived component and a carotenoid-derived component, and PC3 is a chlorophyll-derived component. And a criterion showing the relationship between chlorophyll-derived components. The results are shown in FIG. As shown in FIG. 2, tobacco raw materials of the same category were plotted at close positions. The criteria were created by, for example, visually grouping these into sections.
［実施例２］
実施例１で得た前処理を施した蛍光指紋情報について、ソフトウェア（Eigenvector Research, Inc製「PLS TOOLBOX」）を用いてＰＬＳ判別分析を行い、黄色、オリエント、バーレーのクラスターをそれぞれ、目的変数（１，０，０）、（０，１，０）、（０，０，１）として解析を行い、それぞれの基準を作成した。さらに、基準の妥当性を検証するために、区分が既知の２５のサンプルを当該基準により判別し、実際のサンプルの区分と合致するか調べた。その結果、２４のサンプルについて合致がみられ（判別成績９６％）、当該基準は精度が高いことが明らかとなった。 [Example 2]
For the preprocessed fluorescent fingerprint information obtained in Example 1, PLS discriminant analysis was performed using software (“PLS TOOLBOX” manufactured by Eigenvector Research, Inc.), and the clusters of yellow, orient, and burley were determined as objective variables ( The analysis was performed as (1, 0, 0), (0, 1, 0), (0, 0, 1), and respective standards were created. Further, in order to verify the validity of the criterion, 25 samples whose classification was known were determined by the criterion, and it was examined whether or not the sample matched the actual sample classification. As a result, a match was found for 24 samples (discrimination result: 96%), which revealed that the criterion had high accuracy.
実施例１で得た前処理を施した蛍光指紋情報について、ソフトウェア（Eigenvector Research, Inc製「PLS TOOLBOX」）を用いてＰＬＳ判別分析を行い、黄色、オリエント、バーレーのクラスターをそれぞれ、目的変数（１，０，０）、（０，１，０）、（０，０，１）として解析を行い、それぞれの基準を作成した。さらに、基準の妥当性を検証するために、区分が既知の２５のサンプルを当該基準により判別し、実際のサンプルの区分と合致するか調べた。その結果、２４のサンプルについて合致がみられ（判別成績９６％）、当該基準は精度が高いことが明らかとなった。 [Example 2]
For the preprocessed fluorescent fingerprint information obtained in Example 1, PLS discriminant analysis was performed using software (“PLS TOOLBOX” manufactured by Eigenvector Research, Inc.), and the clusters of yellow, orient, and burley were determined as objective variables ( The analysis was performed as (1, 0, 0), (0, 1, 0), (0, 0, 1), and respective standards were created. Further, in order to verify the validity of the criterion, 25 samples whose classification was known were determined by the criterion, and it was examined whether or not the sample matched the actual sample classification. As a result, a match was found for 24 samples (discrimination result: 96%), which revealed that the criterion had high accuracy.
［実施例３］
区分が既知である茶原料として、紅茶、ほうじ茶、煎茶を準備した。各原料を、粉砕機を用いて最大粒径が１ｍｍ以下になるように粉砕した。実施例１と同条件で当該材料の蛍光指紋情報を取得した。 [Example 3]
Black tea, roasted tea, and sencha were prepared as tea raw materials of which classifications were known. Each raw material was pulverized using a pulverizer so that the maximum particle size became 1 mm or less. Fluorescent fingerprint information of the material was obtained under the same conditions as in Example 1.
区分が既知である茶原料として、紅茶、ほうじ茶、煎茶を準備した。各原料を、粉砕機を用いて最大粒径が１ｍｍ以下になるように粉砕した。実施例１と同条件で当該材料の蛍光指紋情報を取得した。 [Example 3]
Black tea, roasted tea, and sencha were prepared as tea raw materials of which classifications were known. Each raw material was pulverized using a pulverizer so that the maximum particle size became 1 mm or less. Fluorescent fingerprint information of the material was obtained under the same conditions as in Example 1.
［実施例４］
実施例３で得た前処理を施した蛍光指紋情報について、ソフトウェア（Eigenvector Research, Inc製「PLS TOOLBOX」）を用いてＰＬＳ判別分析を行い、紅茶、ほうじ茶、煎茶のクラスターをそれぞれ、目的変数（１，０，０）、（０，１，０）、（０，０，１）として解析を行い、それぞれの基準を作成した（図３）。さらに、基準の妥当性を検証するために、区分が既知の２１のサンプルを当該基準により判別し、実際のサンプルの区分と合致するか調べた。その結果、２０のサンプルについて合致がみられ（判別成績９６％）、当該基準は精度が高いことが明らかとなった。 [Example 4]
PLS discriminant analysis was performed on the preprocessed fluorescent fingerprint information obtained in Example 3 using software (“PLS TOOLBOX” manufactured by Eigenvector Research, Inc.), and clusters of black tea, roasted green tea, and sencha were determined as objective variables ( The analysis was performed as (1,0,0), (0,1,0), (0,0,1), and respective standards were created (FIG. 3). Furthermore, in order to verify the validity of the standard, 21 samples whose divisions were known were discriminated by the standard, and it was examined whether or not the sample matched the actual sample. As a result, a match was found for 20 samples (discrimination score: 96%), which revealed that the standard had high accuracy.
実施例３で得た前処理を施した蛍光指紋情報について、ソフトウェア（Eigenvector Research, Inc製「PLS TOOLBOX」）を用いてＰＬＳ判別分析を行い、紅茶、ほうじ茶、煎茶のクラスターをそれぞれ、目的変数（１，０，０）、（０，１，０）、（０，０，１）として解析を行い、それぞれの基準を作成した（図３）。さらに、基準の妥当性を検証するために、区分が既知の２１のサンプルを当該基準により判別し、実際のサンプルの区分と合致するか調べた。その結果、２０のサンプルについて合致がみられ（判別成績９６％）、当該基準は精度が高いことが明らかとなった。 [Example 4]
PLS discriminant analysis was performed on the preprocessed fluorescent fingerprint information obtained in Example 3 using software (“PLS TOOLBOX” manufactured by Eigenvector Research, Inc.), and clusters of black tea, roasted green tea, and sencha were determined as objective variables ( The analysis was performed as (1,0,0), (0,1,0), (0,0,1), and respective standards were created (FIG. 3). Furthermore, in order to verify the validity of the standard, 21 samples whose divisions were known were discriminated by the standard, and it was examined whether or not the sample matched the actual sample. As a result, a match was found for 20 samples (discrimination score: 96%), which revealed that the standard had high accuracy.
１ ポリフェノール由来成分またはテルペノイド由来成分に起因するピーク
３ クロロフィル由来成分に起因するピーク
Ａ１ 区分が既知である材料の蛍光指紋情報を取得する手段、
Ａ１０ 蛍光指紋情報
Ａ２ 材料区分とポリフェノール等由来成分の関係を示す基準を作成する手段
Ａ２０ 基準
Ｂ１ 区分が未知である材料の蛍光指紋情報を取得する手段
Ｂ１０ 蛍光指紋情報
Ｂ２ 材料区分とポリフェノール等由来成分の関係を取得する手段
Ｂ２０ 関係
Ｃ 基準と関係とを照合する手段 1 Peak due to polyphenol or terpenoid derivedcomponent 3 Peak due to chlorophyll derived component
A1 means for acquiring fluorescent fingerprint information of a material whose category is known,
A10 Fluorescent fingerprint information
A2 Means for creating a standard indicating the relationship between material categories and components derived from polyphenols, etc. A20 Standard
B1 Means of acquiring fluorescent fingerprint information of a material whose classification is unknown B10 Fluorescent fingerprint information
B2 Means for acquiring the relationship between material categories and components derived from polyphenols, etc. B20 Relationship
C means for collating criteria and relationships
３ クロロフィル由来成分に起因するピーク
Ａ１ 区分が既知である材料の蛍光指紋情報を取得する手段、
Ａ１０ 蛍光指紋情報
Ａ２ 材料区分とポリフェノール等由来成分の関係を示す基準を作成する手段
Ａ２０ 基準
Ｂ１ 区分が未知である材料の蛍光指紋情報を取得する手段
Ｂ１０ 蛍光指紋情報
Ｂ２ 材料区分とポリフェノール等由来成分の関係を取得する手段
Ｂ２０ 関係
Ｃ 基準と関係とを照合する手段 1 Peak due to polyphenol or terpenoid derived
A1 means for acquiring fluorescent fingerprint information of a material whose category is known,
A10 Fluorescent fingerprint information
A2 Means for creating a standard indicating the relationship between material categories and components derived from polyphenols, etc. A20 Standard
B1 Means of acquiring fluorescent fingerprint information of a material whose classification is unknown B10 Fluorescent fingerprint information
B2 Means for acquiring the relationship between material categories and components derived from polyphenols, etc. B20 Relationship
C means for collating criteria and relationships
Claims (14)
- クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別するための基準を作成する方法であって、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得工程、
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する工程を含む、方法。 A method of creating a criterion for determining the classification of a material derived from a plant containing chlorophyll and at least one of a polyphenol and a terpenoid,
Fluorescence fingerprint information acquisition step of acquiring fluorescence fingerprint information consisting of excitation wavelength, fluorescence wavelength, and fluorescence intensity data of the material whose classification is known,
Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Creating a criterion indicating the relationship between the component and the chlorophyll-derived component. - 前記材料が、たばこ原料または茶原料である、請求項１に記載の方法。 The method according to claim 1, wherein the ingredient is a tobacco ingredient or a tea ingredient.
- 前記材料がたばこ原料であり、
前記区分がバーレー種、黄色種、およびオリエント種である、請求項２に記載の方法。 The material is a tobacco raw material,
3. The method of claim 2, wherein the categories are Burley, Yellow, and Orient. - 前記材料が粉末である、請求項１～３のいずれかに記載の方法。 方法 The method according to any one of claims 1 to 3, wherein the material is a powder.
- 前記材料の最大粒径が１ｍｍ以下である、請求項４に記載の方法。 The method according to claim 4, wherein the maximum particle size of the material is 1 mm or less.
- 前記蛍光指紋情報取得工程が、２５０～３７０ｎｍの励起光を照射してポリフェノールまたはテルペノイドの少なくとも一方に由来する成分に起因する３００～４８０ｎｍの蛍光を測定すること、または３５０～４８０ｎｍの励起光を照射してクロロフィル由来成分に起因する６７０～７００ｎｍの蛍光を測定すること、あるいはその双方を含む、請求項１～５に記載の方法。 The fluorescence fingerprint information acquiring step includes irradiating excitation light of 250 to 370 nm to measure fluorescence of 300 to 480 nm caused by a component derived from at least one of polyphenol and terpenoid, or irradiating excitation light of 350 to 480 nm. The method according to any one of claims 1 to 5, further comprising measuring fluorescence at 670 to 700 nm caused by the component derived from chlorophyll, or both.
- 前記解析が多変量解析である、請求項１～６のいずれかに記載の方法。 方法 The method according to any one of claims 1 to 6, wherein the analysis is a multivariate analysis.
- 前記多変量解析が主成分分析である、請求項７に記載の方法。 The method according to claim 7, wherein the multivariate analysis is a principal component analysis.
- 前記多変量解析がＰＬＳ判別分析である、請求項７に記載の方法。 The method of claim 7, wherein the multivariate analysis is a PLS discriminant analysis.
- 前記材料がたばこ原料であり、蛍光指紋情報を取得する工程の前に、当該たばこ原料を２２℃、６０％で２４時間以上静置して状態調整する工程をさらに含む、請求項１～９のいずれかに記載の方法。 The method according to any one of claims 1 to 9, wherein the material is a tobacco material, and the method further comprises a step of allowing the tobacco material to stand at 22 ° C. and 60% for at least 24 hours to adjust the state before obtaining the fluorescent fingerprint information. The method according to any of the above.
- 区分が未知である、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する工程１、
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する工程２、
請求項１～１０のいずれかに記載の基準を準備して、当該基準と工程２で得た関係を照合して当該材料の区分を判別する工程３、を含む、
前記材料の区分を判別する方法。 The category is unknown, chlorophyll, excitation wavelength of a material derived from a plant containing at least one of polyphenols and terpenoids, fluorescence wavelength, and obtaining fluorescence fingerprint information consisting of fluorescence intensity data 1,
Analysis of the abundance of components derived from at least one of polyphenols and terpenoids in the material and fluorescent fingerprint information reflecting the abundance of components derived from chlorophyll as an index, the material category, derived from at least one of polyphenols or terpenoids Step 2 of obtaining the relationship between the component and the component derived from chlorophyll,
Preparing a criterion according to any one of claims 1 to 10, collating the criterion with the relationship obtained in step 2, and determining a classification of the material;
A method for determining a category of the material. - コンピュータに請求項１～１１のいずれかに記載の方法を実行させるためのプログラム。 A program for causing a computer to execute the method according to any one of claims 1 to 11.
- クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別するための基準を作成するための装置であって、
区分が既知である前記材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する手段（Ａ１）、ならびに
前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を示す基準を作成する手段（Ａ２）、を備える装置。 A device for creating a standard for determining the classification of materials derived from plants containing chlorophyll and at least one of polyphenols and terpenoids,
Means (A1) for acquiring fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescence wavelength, and the fluorescence intensity of the material whose category is known, and the abundance of components derived from at least one of polyphenols and terpenoids in the material And means for analyzing by using the fluorescent fingerprint information reflecting the abundance of the component derived from chlorophyll as an index, and creating a reference indicating the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the component derived from chlorophyll (A2 ). - 区分が未知である、クロロフィルと、ポリフェノールまたはテルペノイドの少なくとも一方とを含む植物に由来する材料の区分を判別する装置であって、
請求項１３に記載の手段（Ａ１）および手段（Ａ２）、
前記区分が未知である材料の励起波長、蛍光波長、および蛍光強度のデータからなる蛍光指紋情報を取得する蛍光指紋情報取得手段（Ｂ１）、ならびに前記材料中のポリフェノールまたはテルペノイドの少なくとも一方に由来する成分の存在量と、クロロフィル由来成分の存在量を反映する蛍光指紋情報を指標として解析し、前記材料区分、ポリフェノールまたはテルペノイドの少なくとも一方に由来する成分、およびクロロフィル由来成分の関係を取得する手段（Ｂ２）、ならびに
手段（Ａ２）で得た基準と、手段（Ｂ２）で得た関係とを照合して、前記区分が未知である材料の区分を判別する手段（Ｃ）を備える、装置。 An apparatus for determining the classification of a material derived from a plant containing chlorophyll and at least one of polyphenols and terpenoids whose classification is unknown,
The means (A1) and the means (A2) according to claim 13,
A fluorescent fingerprint information obtaining means (B1) for obtaining fluorescent fingerprint information comprising data of the excitation wavelength, the fluorescent wavelength, and the fluorescent intensity of the material whose classification is unknown, and at least one of polyphenol or terpenoid in the material Means for analyzing the abundance of the component and the fluorescent fingerprint information reflecting the abundance of the chlorophyll-derived component as an index, and acquiring the relationship between the material category, the component derived from at least one of polyphenol or terpenoid, and the chlorophyll-derived component ( B2) and an apparatus comprising means (C) for comparing the reference obtained by the means (A2) with the relationship obtained by the means (B2) to determine the category of the material whose category is unknown.
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