CN108152386A - Miniature region tea-leaf producing area recognition methods and application based on fingerprint pattern technology - Google Patents
Miniature region tea-leaf producing area recognition methods and application based on fingerprint pattern technology Download PDFInfo
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- CN108152386A CN108152386A CN201711112006.8A CN201711112006A CN108152386A CN 108152386 A CN108152386 A CN 108152386A CN 201711112006 A CN201711112006 A CN 201711112006A CN 108152386 A CN108152386 A CN 108152386A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention belongs to tealeaves detection identification technology fields, disclose a kind of miniature region tea-leaf producing area recognition methods and application based on fingerprint pattern technology, it is detected using high performance liquid chromatography (HPLC) or gas-chromatography (GC), obtains the tealeaves liquid phase or gas chromatogram in particular microchannel region；Take the standard finger-print of average method structure specific region tealeaves；The critical value of specific region similarity is determined using SPSS softwares and standard finger-print progress similarity analysis；The similarity that zone of ignorance tea sample and specific region standard finger-print are compared, makes comparisons with critical value；Identify whether unknown tea sample is originated from the particular microchannel region.The present invention can successfully identify the tealeaves in more than 95% specific tea area, and tea-leaf producing area recognition result accuracy is more than 99%.The present invention equally has broad prospect of application in quality evaluation and identification of same batch tealeaves etc..
Description
Technical field
The invention belongs to tealeaves detection identification technology field more particularly to a kind of miniature regions based on fingerprint pattern technology
Tea-leaf producing area recognition methods and application.
Background technology
Tea leaf quality is to include active materials coordination with the synthesis as a result, type, quantity including active principle and each
Composition ratio between substance.Due to the difference of the factors such as geographical environment, climate characteristic, processing technology, varieties of plant, same type
The tealeaves of different zones shows different qualitative characteristics.The quality discrepancies such as flavour, the fragrance of specific region tealeaves and physics and chemistry
Ingredient, aroma component, the characteristic chemical constituent on Metabolite components or component distribution characteristics, be by Modern Scientific Instruments equipment,
Research means, analysis method accurately differentiate the basis of tea-leaf producing area.Traditional tealeaves differentiates Main Basiss sensory review, needs big
The experience accumulation of amount, and it is subjective, and there are serious individuation differences.Therefore, there is an urgent need to by different zones and
The discrimination method of objective and fair is established in the tealeaves component content quantization of specific region.
Chemical fingerprint technology is to remove those non-features being present in plant using certain extraction separable programming
Property ingredient, obtain the total extract in each plant with characteristic chemical composition, then measure this using modern analysis means
The collection of illustrative plates of feature total extract is planted to establish the general description to plant characteristics chemical composition, there is globality and ambiguity two greatly
Feature.High performance liquid chromatography (HPLC) be used as most common finger-print research method, have separation efficiency height, high selectivity,
Detect it is sensitive it is high, with sample it is few, have a wide range of application the advantages that.HPLC fingerprint pattern technologies are the foundation by standard finger-print,
With reference to clustering methodology (Cluster Analysis, CA), Principal Component Analysis (Principal Analysis, PCA) and phase
The Chinese herbal medicine place of production, tea-leaf producing area are identified like analysis methods such as degree analytic approach (Similarity Analysis, SA).
Metabolism group be before and after being stimulated or disturb by investigating biosystem (such as by some specific genetic mutation or
After environmental change) metabolite collection of illustrative plates and its dynamic change study a kind of technology of the metabolism network of biosystem, research pair
As being mainly endogenous small molecule of the relative molecular mass below 1000.Gas chromatography-mass spectrum (GC-MS) and liquid chromatogram-matter
It is most widely used, most effective metabolism group investigative technique to compose (LC-MS).Gas-chromatography (GC) has separation efficiency height, choosing
The advantages that selecting property is strong, dosage is few and separating rate is fast is studied especially suitable for tea aroma component analysis and finger-print.Liquid phase
Spectrometry is more highly polar, the compound of higher relative molecular mass and thermal stability difference.Using liquid chromatography-mass spectrography
The detection to metabolites such as Tea Polyphenols in Tea, catechin, water-soluble sugar, amino acid, purine base, organic acids can be completed,
It is studied suitable for the signature analysis of tealeaves component content.By building the technologies such as chemical fingerprint, HPLC, LC-MS and GC-MS
Available for tea sample fidelity, region differentiates, tea quality is evaluated and identity authentication.
In conclusion problem of the existing technology is：
The prior art, which is focused primarily upon, sieves the grade of tealeaves with quality using electronic nose sensor or fluorescence spectrum
Choosing；The tea of tealeaves is identified and is concentrated mainly on specific several tealeaves such as Xihu Longjing Tea and Wuyi cliff tea, is mostly using joint
Isotope and micro- method of inspection or aroma component combination genetic algorithm are detected tea-leaf producing area；Detection method is complicated,
And it is confined to several specific tealeaves.The present invention is based on the standards of the metabolic components structure particular microchannel region tealeaves of tealeaves itself
Finger-print is compared by similarity, and Production area recognition is carried out to tealeaves, has the characteristics that easy to operate and accuracy is high, can be wide
The general Production area recognition suitable for specific type tealeaves in miniature region.
Invention content
In view of the problems of the existing technology, the present invention provides a kind of miniature region tealeaves based on fingerprint pattern technology
Production area recognition method and application.
The invention is realized in this way a kind of miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology, institute
The miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology is stated to pass through to the same type of tealeaves in particular microchannel region
Sample pre-treatments are detected using high performance liquid chromatography (HPLC) or gas-chromatography (GC), obtain the tea in particular microchannel region
Leaf liquid chromatogram；Take the standard finger-print of average method structure specific region tealeaves；
By the same type of specific region and adjacent domain and different kind tealeaves, using SPSS softwares and standard fingerprint
Collection of illustrative plates carries out the critical value that similarity analysis determines specific region similarity；By zone of ignorance tea sample and specific region standard fingerprint
The similarity that collection of illustrative plates compares, makes comparisons with critical value；Identify whether unknown tea sample is originated from the particular microchannel region.
Further, the miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology specifically includes following step
Suddenly：
Step 1, the collection of particular microchannel region tea sample：Collect the tea sample in same type of known particular microchannel region, tea
Sample sample size is more than 20；
Step 2, high performance liquid chromatography (HPLC) detection：It weighs 3g and grinds sample in 500mL conical flasks, boiling is added to distill
Water 450mL moves into boiling water bath, extracts 45min；It is filtered under diminished pressure after extraction and is settled to 500mL；Millet paste is through 0.45um water systems
Sample introduction after membrane filtration, sample size 10ul；
Chromatographic condition is：Mobile phase is acetonitrile and the mixed solution of 0.2% acetic acid aqueous solution, and A phases are acetonitrile, and B phases are
0.2% acetic acid aqueous solution, gradient elution, flow velocity 1ml/min；
Chromatographic column：Agilent reverse phase Cl8 chromatographic columns；Detection wavelength is 280nm；Column temperature is 30 DEG C；
Gradient elution takes gradient elution mode：During 0min, A phases 8%；B phases 92%；
During 50min, A phases 31%, B phases 69%；After run 10min；
Step 3, the drafting of standard finger-print：It will be carried on the back before the chromatogram data analysis of particular microchannel region tea sample
Scape deducts, the pretreatment of spectral peak calibration；It is established using the Chinese medicine chromatography similarity evaluation system of CASE(Computer Aided Software Engineering) specific micro-
The common pattern of the high-efficient liquid phase color modal data of type region Tea Samples extracts the shared mould of the high-efficient liquid phase color modal data
The characteristic fingerprint pattern of formula is the standard finger-print in the particular microchannel region；
Step 4, for trying the collection of tea sample：Collect the particular microchannel region and the same type of tea sample of adjacent domain and
The different types of tea sample in particular microchannel region is several；
Step 5, for trying the chromatogram of tea sample：According to step 2, obtain it is different for examination group based on catechin and purine base
High-efficient liquid phase chromatogram；
Step 6, the establishment of particular microchannel Regional Similarity critical point：By the different chromatograms for examination group tea sample with it is specific
Miniature regional standard finger-print carries out similarity-rough set；The average value of particular microchannel region same type tea sample similarity is the place of production
The critical value of identification；By adjacent domain same type tea sample and particular microchannel region different kind tea sample and particular microchannel regional standard
Fingerprint image carries out similarity-rough set, and the average value of similarity and similarity critical value are compared；If adjacent domain with it is non-
There are significant difference p with critical value for the similarity of same type<0.05, the standard fingerprint figure in the particular microchannel region can be used for
The identification of tea-leaf producing area；
Step 7, the Production area recognition of zone of ignorance tealeaves：Zone of ignorance tealeaves according to step 2, obtain based on catechin and
The high-efficient liquid phase chromatogram of purine base；Zone of ignorance tea sample chromatogram is compared with particular microchannel regional standard fingerprint image,
Obtain the similarity value of zone of ignorance tealeaves；If similarity is greater than or equal to the critical value of the particular microchannel Regional Similarity；
The unknown tealeaves is originated from the particular microchannel region；If unknown tealeaves is less than critical value with specific region similarity value；
The unknown tealeaves is the sample that leaves a question open in the particular microchannel region.
Further, in step 1, particular microchannel region tea sample is same tea tree breed or is Different Tea Varieties.
Further, in step 2, the high-efficient liquid phase chromatogram HPLC is detected as the detection of Catechin in Tea and purine base,
Or the detection to amino acid, water-soluble sugar, aroma component, metabolic components in tealeaves；High performance liquid chromatography (HPLC) inspection
It surveys or is detected for gas-chromatography (GC).
Further, in step 3, the drafting of the standard fingerprint chromatogram is used to obtain particular microchannel region same class
The shared peak of type tealeaves and the opposite average content at shared peak；By drafting based on amino acid, water-soluble sugar, fragrance in tealeaves
The standard finger-print system of the particular microchannel region same type tealeaves of component different component, structure particular microchannel region are same
The standard database of type tealeaves.
Further, in step 4, it is more than 6 or more for examination group tea sample；Same particular microchannel region same type tea sample is used for
Determine the critical value of similarity；Adjacent domain and different kind tea sample identify for determining finger-print in tea-leaf producing area credible
Degree；
In step 6, Lin circle Zhi≤95% of particular microchannel region tealeaves similarity.
Another object of the present invention is to provide a kind of to utilize the above-mentioned miniature region tealeaves based on fingerprint pattern technology
Certain tealeaves Main Tea Area standard finger-print system of Production area recognition method structure.
Another object of the present invention is to provide a kind of to utilize the above-mentioned miniature region tealeaves based on fingerprint pattern technology
The standard finger-print system of the particular microchannel region different type tealeaves of Production area recognition method structure.Convenient for specific region tealeaves
Protection and identification.
Another object of the present invention is to provide a kind of to utilize the above-mentioned miniature region tealeaves based on fingerprint pattern technology
The digitlization large database concept of the particular microchannel region different type tealeaves of Production area recognition method structure.
Another object of the present invention is to provide a kind of to utilize the above-mentioned miniature region tealeaves based on fingerprint pattern technology
The a certain batch tealeaves of Production area recognition method structure is based on catechin, purine base, amino acid, water-soluble sugar physics and chemistry component and perfume (or spice)
The digital archive of gas component.Quality evaluation and identity authentication convenient for the batch tealeaves.
Advantages of the present invention and good effect are：
The present invention passes through high performance liquid chromatography by largely collecting same type tealeaves sample in particular microchannel tea area
(HPLC) or gas-chromatography (GC) detects, and builds specific tea area and is based on catechin, purine base, free amino acid, water-soluble sugar, perfume (or spice)
The standard finger-print of the components such as gas.Specific tea area is obtained with neighbouring tea area equivalent type with different kind tea sample through chromatography detection
Corresponding chromatogram is obtained, similarity-rough set is carried out with standard finger-print, determines the critical value of specific tea area similarity.By unknown tea
Area's tea sample carries out similarity analysis through the chromatogram that chromatography detection obtains and standard finger-print, if similarity is greater than or equal to phase
Like the critical value of degree, unknown tea sample determines to originate from specific tea area；If similarity is less than critical value, unknown tea sample not can determine that production
From in specific tea area.Miniature Cha Qu is stepped according to Yunnan Province Puer City billows Cang County Huimin township scape and the tea-leaf producing area in Mang Jingcha areas identifies
Specific implementation example, it is known that this method can successfully identify that more than 95% scape steps the tealeaves with the specific tea area such as awns scape, tea
The accuracy of leaf Production area recognition result is more than 99%.Present invention Main Tea Area in the tealeaves such as Yunnan sun withering tea, Xinyang Maojian Tea
Production area recognition is commented in the tealeaves authentication of the particular microchannels region such as the specific hilltop, stockaded village and in the quality of same batch tealeaves
Valency and identification etc. have broad prospect of application.
The advantages of present invention also has is embodied in：
The present invention is used for the identification of miniature region tea-leaf producing area using standard finger-print combination similarity analysis for the first time, is
A kind of open tea-leaf producing area recognition methods.The standard finger-print based on components such as catechin, purine bases can be established, also may be used
Simultaneously structure based on catechin, purine base, amino acid, water-soluble sugar, aroma component standard finger-print system, for specific
Fidelity, the identification of particular types type tealeaves different sources and the identity authentication of same batch tealeaves of region tealeaves
Deng.With easy to operate, flexible, accuracy is high, and the features such as can constantly update optimization.
The present invention implements tea-leaf producing area identification and the Yunnan that example each provides Yunnan Province Puer City billows Cang County Jing Mai villages
The tea-leaf producing area identification of province Puer City billows Cang County Mang Jingcun.Jing Mai villages and Mang Jing villages are respectively positioned on natives iu Lancang County of Yunnan Proviuce Huimin township scape and step
In mountain, two places geographical location borders on, climatic environment is similar, tea tree breed is similar, processing technology approaches.It can be succeeded using the present invention
It identifies Jing Mai villages and the tealeaves of the different sources in Mang Jing villages, has the characteristics that accuracy is high, available for a certain hilltop, a certain village
Stockaded village or the identification of a certain specific region tealeaves.
The present invention has broad spectrum activity in the tea-leaf producing area identification in miniature tea area, available for Xinyang Maojian Tea, Yunnan sun withering hair
The main particular microchannel region of the different types tealeaves such as tea, such as the hilltop, the Production area recognition of stockaded village；Precision is had both simultaneously, also
Identification and protection available for a certain specific region tealeaves.
Description of the drawings
Fig. 1 is the miniature tea-leaf producing area recognition methods flow in region provided in an embodiment of the present invention based on fingerprint pattern technology
Figure.
Specific embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
The present invention is to collect the same type of Tea Samples in specific region, by-grinding-boiling water extraction-HPLC (efficient liquid
Phase chromatography) detection-the obtain specific region chromatogram, using similarity evaluation A editions, take
Average method generates the standard finger-print of the specific region tealeaves.Using the same type tea of same area and adjacent domain
Leaf sample carries out the chromatogram that HPLC detections obtain the same area and adjacent domain using Same Way, using SPSS softwares with
Standard finger-print carries out the critical point that similarity analysis determines the region tea sample similarity.Zone of ignorance tea sample is subjected to HPLC
Chromatogram is detected, and similarity analysis is carried out with the specific region standard finger-print, obtains similarity value.If unknown tea sample
With specific region similarity value greater than or equal to critical value, illustrate that unknown tea sample is originated from the specific region；If unknown tea sample
With specific region similarity value less than critical value, illustrate the sample that leaves a question open that unknown tea sample is the specific region.In miniature tea area
There is broad spectrum activity in tea-leaf producing area identification, available for the main specific of the different types tealeaves such as Xinyang Maojian Tea, Yunnan solar dried green tea
Miniature region, such as the hilltop, the Production area recognition of stockaded village；Precision is had both simultaneously, also the body available for a certain specific region tealeaves
Part identification and protection.
The application principle of the present invention is described in detail below in conjunction with the accompanying drawings.
Miniature tea-leaf producing area recognition methods in region provided in an embodiment of the present invention based on fingerprint pattern technology, including：
Collect the same type of Tea Samples in particular microchannel region；Through sample pre-treatments, HPLC (high performance liquid chromatography) inspections
It surveys, obtains the tealeaves liquid chromatogram in particular microchannel region；Take the standard fingerprint figure of average method structure specific region tealeaves
Spectrum.By the same type of specific region and adjacent domain and different kind tealeaves, using SPSS softwares and standard finger-print
Carry out the critical value that similarity analysis determines specific region similarity.By zone of ignorance tea sample and specific region standard finger-print
The similarity of comparison, makes comparisons with critical value.If unknown tea sample, greater than or equal to critical value, is said with specific region similarity value
Bright unknown tea sample is originated from the specific region；If unknown tea sample, less than critical value, illustrates unknown with specific region similarity value
Tea sample is the sample that leaves a question open of the specific region.
As shown in Figure 1, the miniature tea-leaf producing area identification side in region provided in an embodiment of the present invention based on fingerprint pattern technology
Method, including：
S101：The collection of specific region tea sample：Collect specific region same type tea sample, tea sample sample size 20 with
On.
S102:HPLC is detected：It weighs 3g and grinds sample in 500mL conical flasks, add boiling distilled water 450mL, move into immediately
Boiling water bath, extraction 45min (shake primary) every l0min.It is filtered under diminished pressure while hot immediately after extraction and is settled to 500mL.
Millet paste sample introduction, sample size 10ul after 0.45um water system membrane filtrations.Chromatographic condition is：Mobile phase is acetonitrile and 0.2% acetic acid
The mixed solution of aqueous solution, A phases be acetonitrile, B phases be 0.2% acetic acid aqueous solution, gradient elution, flow velocity 1ml/min, chromatography
Column：Agilent reverse phase Cl8 chromatographic columns (250mm × 4.6mm, 5 μm)；Detection wavelength is 280nm；Column temperature is 30 DEG C.It elutes herein
Under gradient, ideal chromatography peak can be obtained, and each peak chromatographic peak baseline separation, appearance time are distributed, and concentrate on 0-35min, have
Conducive to the Structural Identification and accurate quantitative analysis of each chromatographic peak, gradient elution takes gradient elution mode：During 0min, A phases 8%；B phases
92%；During 50min, A phases 31%, B phases 69%；After run 10min.
S103：The drafting of standard finger-print：The chromatography diagram data of specific region tea sample has been done suitably before analysis
Pretreatment, including background deduction, spectral peak calibration etc. is established using CASE(Computer Aided Software Engineering) (Chinese medicine chromatography similarity evaluation system)
The common pattern of the high-efficient liquid phase color modal data of specific region Tea Samples extracts its characteristic fingerprint pattern with this, as should
The standard finger-print of specific region.
S104：For trying the collection of tea sample：Collect the specific region and the same type of tea sample of adjacent domain and given zone
The different types of tea sample in domain is several.
S105：For trying the chromatogram of tea sample：According to S102, obtain different efficient based on catechin and purine base for examination group
Liquid chromatogram.
S106：The establishment of specific region similarity critical point：The different chromatograms for examination group tea sample and specific region are marked
Quasi- finger-print carries out similarity-rough set.The average value of specific region same type tea sample similarity is the critical value of Production area recognition.
Adjacent domain same type tea sample and specific region different kind tea sample are subjected to similarity-rough set with specific region standard fingerprint figure,
And the average value of similarity and similarity critical value are compared.If the similarity and critical value of adjacent domain and different kind
There are significant difference (p<0.05), illustrate that the standard fingerprint figure of specific region can be used for the identification of tea-leaf producing area.
S107：The Production area recognition of zone of ignorance tea sample：Zone of ignorance tealeaves is obtained according to S102 based on catechin and purine
The high-efficient liquid phase chromatogram of alkali.Zone of ignorance tea sample chromatogram with specific region standard fingerprint figure is compared, is obtained unknown
The similarity value of region tealeaves.If similarity is greater than or equal to the critical value of the specific region similarity, illustrate unknown tealeaves production
From in the specific region；If unknown tealeaves, less than critical value, illustrates that unknown tealeaves is specific for this with specific region similarity value
The sample that leaves a question open in region.
In S101, the tea sample requirement of the specific region is same type tea sample；Tea sample quantitative requirement is at 20 or more；
Tea sample can be same tea tree breed, can be the main tea tree breed of Different Tea Varieties or specific region.
In S102, the HPLC is detected as the common detection method of Catechin in Tea and purine base.Standard fingerprint figure
The foundation of spectrum is not limited only to catechin and purine alkaline constituents, also can in tealeaves amino acid, water-soluble sugar, aroma component, generation
It thanks to component to be detected using the methods of corresponding liquid chromatogram, gas-chromatography, obtains corresponding chromatogram for miniature region
The identification of tea-leaf producing area.
In S103, the drafting of the standard fingerprint chromatogram predominantly obtains being total to for specific region same type tealeaves
There is the opposite average content at peak and shared peak；By drafting based on differences such as amino acid, water-soluble sugar, aroma components in tealeaves
The standard finger-print system of the specific region same type tealeaves of component can build the standard of specific region same type tealeaves
Database improves the accuracy rate and confidence level of tea-leaf producing area identification.
In S104, the tea sample that same specific region same type is mainly collected for sample, adjacent domain same type
The non-same type tea sample of tea sample and same specific region；It it is 6 or more for the requirement of examination group tea sample；Same specific region same type
Tea sample is mainly used for determining the critical value of similarity；Adjacent domain is mainly used for determining finger-print in tea with different kind tea sample
The confidence level of leaf Production area recognition.
In S106, the establishment of the specific region similarity critical point, it is desirable that specific region tealeaves similarity it is critical
Zhi≤95%, critical value is higher to illustrate that tealeaves more tends to be unified in specific region；It is required that adjacent domain and different kind tealeaves phase
Like degree, there are significant difference (p with specific region critical value<0.05), in order to specific region tealeaves progress Production area recognition.
In S107, in the Production area recognition of the zone of ignorance tea sample, the Production area recognition master of the zone of ignorance tea sample
The comparison of similarity is carried out with specific region standard fingerprint figure by the chromatogram of zone of ignorance tea sample, determines zone of ignorance tea
The place of production of sample：If similarity is greater than or equal to specific region critical value, unknown tealeaves, which can be considered, to be originated from specific region；Such as
Fruit similarity is less than similarity critical value, and unknown tealeaves can be considered the tea sample that leaves a question open of the specific region.For leaving a question open tea sample not
The possibility for coming from the specific region can be excluded, complete standard finger-print system can increase the range of tea-leaf producing area identification
With confidence level.Unknown tea sample can be compared with known different specific regions standard finger-print, similarity soprano, you can
It is considered as and comes from the specific region.
An embodiment of the present invention provides based on fingerprint pattern technology in Yunnan Province Puer City billows Cang County Jing Mai villages specific region
The method and flow applied in tea-leaf producing area identification.Blind comment more than 95% in tea sample is successfully identified in the implementation example
Jing Mai villages tea sample, accuracy is more than 99%.
An embodiment of the present invention provides based on fingerprint pattern technology in Yunnan Province Puer City billows Cang County Mang Jingcun specific regions
The method and flow applied in tea-leaf producing area identification.Blind comment more than 95% in tea sample is successfully identified in the implementation example
Mang Jing villages tea sample, accuracy is more than 99%.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology, which is characterized in that described to be based on fingerprint
The miniature region tea-leaf producing area recognition methods of graphical spectrum technology by Tea Samples pre-treatment same type of to particular microchannel region,
It is detected using high performance liquid chromatography (HPLC) or gas-chromatography (GC), obtains the chromatogram of particular microchannel region tealeaves；It adopts
It is averaged the standard finger-print of method structure specific region tealeaves；
By the same type of specific region and adjacent domain and different kind tealeaves, using SPSS softwares and standard finger-print
Carry out the critical value that similarity analysis determines specific region similarity；By zone of ignorance tea sample and specific region standard finger-print
The similarity of comparison, makes comparisons with critical value；Identify whether unknown tea sample is originated from the particular microchannel region.
2. the miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology as described in claim 1, which is characterized in that
The miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology specifically includes following steps：
Step 1, the collection of particular microchannel region tea sample：The tea sample in same type of known particular microchannel region is collected, tea is all
Product quantity is more than 20；
Step 2, high performance liquid chromatography (HPLC) detection：It weighs 3g and grinds sample in 500mL conical flasks, add boiling distilled water
450mL moves into boiling water bath, extracts 45min；It is filtered under diminished pressure after extraction and is settled to 500mL；Millet paste is through 0.45um water system films
Sample introduction after filtering, sample size 10ul；
Chromatographic condition is：Mobile phase is acetonitrile and the mixed solution of 0.2% acetic acid aqueous solution, and A phases are acetonitrile, and B phases are 0.2% second
Aqueous acid, gradient elution, flow velocity 1ml/min；
Chromatographic column：Agilent reverse phase Cl8 chromatographic columns；Detection wavelength is 280nm；Column temperature is 30 DEG C；
Gradient elution takes gradient elution mode：During 0min, A phases 8%；B phases 92%；
During 50min, A phases 31%, B phases 69%；After run 10min；
Step 3, the drafting of standard finger-print：It is buckled background is carried out before the chromatogram data analysis of particular microchannel region tea sample
It removes, the pretreatment of spectral peak calibration；Particular microchannel area is established using the Chinese medicine chromatography similarity evaluation system of CASE(Computer Aided Software Engineering)
The common pattern of the high-efficient liquid phase color modal data of domain Tea Samples extracts the common pattern of the high-efficient liquid phase color modal data
Characteristic fingerprint pattern is the standard finger-print in the particular microchannel region；
Step 4, for trying the collection of tea sample：Collect particular microchannel region tea sample same type of with adjacent domain and specific
The miniature different types of tea sample in region is several；
Step 5, for trying the chromatogram of tea sample：According to step 2, obtain different efficient based on catechin and purine base for examination group
Liquid chromatogram；
Step 6, the establishment of particular microchannel Regional Similarity critical point：By different chromatograms and particular microchannel for examination group tea sample
Regional standard finger-print carries out similarity-rough set；The average value of particular microchannel region same type tea sample similarity is Production area recognition
Critical value；By adjacent domain same type tea sample and particular microchannel region different kind tea sample and particular microchannel regional standard fingerprint
Figure carries out similarity-rough set, and the average value of similarity and similarity critical value are compared；If adjacent domain with it is non-similar
There are significant difference p with critical value for the similarity of type<0.05, the standard fingerprint figure in the particular microchannel region can be used for tealeaves
The identification in the place of production；
Step 7, the Production area recognition of zone of ignorance tealeaves：Zone of ignorance tealeaves is obtained according to step 2 based on catechin and purine
The high-efficient liquid phase chromatogram of alkali；Zone of ignorance tea sample chromatogram with particular microchannel regional standard fingerprint image is compared, is obtained
The similarity value of zone of ignorance tealeaves；If similarity is greater than or equal to the critical value of the particular microchannel Regional Similarity；It is described
Unknown tealeaves is originated from the particular microchannel region；If unknown tealeaves is less than critical value with specific region similarity value；It is described
Unknown tealeaves is the sample that leaves a question open in the particular microchannel region.
3. the miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology as claimed in claim 2, which is characterized in that
In step 1, particular microchannel region tea sample is same tea tree breed or is Different Tea Varieties.
4. the miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology as claimed in claim 2, which is characterized in that
In step 2, the high performance liquid chromatography (HPLC) is detected as the detection of Catechin in Tea and purine base or in tealeaves
The detection of amino acid, water-soluble sugar, aroma component, metabolic components；High performance liquid chromatography (HPLC) detection is gas phase color
Compose (GC) detection.
5. the miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology as claimed in claim 2, which is characterized in that
In step 3, the drafting of the standard fingerprint chromatogram for obtain the shared peak of particular microchannel region same type tealeaves with
And the opposite average content at shared peak；By drawing the spy based on amino acid, water-soluble sugar, aroma component different component in tealeaves
The standard finger-print system of fixed miniature region same type tealeaves, the criterion numeral of structure particular microchannel region same type tealeaves
According to library.
6. the miniature region tea-leaf producing area recognition methods based on fingerprint pattern technology as claimed in claim 2, which is characterized in that
In step 4, it is more than 6 or more for examination group tea sample；Same particular microchannel region same type tea sample is used to determine the critical of similarity
Value；Adjacent domain is used to determine the confidence level that finger-print is identified in tea-leaf producing area with different kind tea sample；
In step 6, Lin circle Zhi≤95% of particular microchannel region tealeaves similarity.
It is built 7. a kind of using miniature tea-leaf producing area recognition methods in region described in claim 1 based on fingerprint pattern technology
Certain tealeaves Main Tea Area standard finger-print system.
It is built 8. a kind of using miniature tea-leaf producing area recognition methods in region described in claim 1 based on fingerprint pattern technology
The standard finger-print system of particular microchannel region different type tealeaves.
It is built 9. a kind of using miniature tea-leaf producing area recognition methods in region described in claim 1 based on fingerprint pattern technology
The digitlization large database concept of particular microchannel region different type tealeaves.
10. a kind of built using the miniature tea-leaf producing area recognition methods in region described in claim 1 based on fingerprint pattern technology
Digitlization shelves of a certain batch tealeaves based on catechin, purine base, amino acid, water-soluble sugar physics and chemistry component and aroma component
Case.
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