CN101692043A - Method for quickly and non-destructively identifying ginseng seeds by near infrared spectrum - Google Patents
Method for quickly and non-destructively identifying ginseng seeds by near infrared spectrum Download PDFInfo
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- CN101692043A CN101692043A CN200910217722A CN200910217722A CN101692043A CN 101692043 A CN101692043 A CN 101692043A CN 200910217722 A CN200910217722 A CN 200910217722A CN 200910217722 A CN200910217722 A CN 200910217722A CN 101692043 A CN101692043 A CN 101692043A
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Abstract
The invention discloses a method for quickly and non-destructively identifying ginseng seeds by near infrared spectrum, which utilizes near infrared molecular spectrum technology and adopts a hierarchical clustering method to identify and analyze the ginseng seeds. The result shows that the near infrared spectrum technology can realize the accurate identification on the ginseng seeds, and samples are correctly classified according to different varieties. Compared with the fussy and complicated conventional method, the analytic efficiency of the near infrared spectrum technology is higher. The identification of the ginseng seeds by adopting the near infrared spectrum technology has no influence on the vitality of the seeds. Original samples are adopted for direct scanning, so the pretreatment is not needed, and the identification efficiency is improved.
Description
Technical field:
The present invention relates to a kind of quick nondestructive and differentiate the method for ginseng seeds, particularly relate to a kind of method that adopts the near-infrared spectrum technique quick nondestructive to differentiate ginseng seeds, belong to plant germplasm resource authenticate technology field.
Background technology:
The evaluation of medicinal plant germplasm propagating materials is the crucial and basic of Chinese crude drug GAP enforcement, but in the introducing and planting process, the incident that panoramic medicinal plant seed is obscured happens occasionally.Genseng has kinds such as No. 1, lucky ginseng, yellow fruit genseng, and big horse bud, two horse buds, long neck, circle wing, justify farm varieties such as reed, bamboo reed, line reed, reed canary grass, output is not quite similar with, quality and adaptability, but the profile of seed is about the same, and sense organ almost can't be differentiated between kind.Therefore set up some fast, the method for non-destructively identifying ginseng seeds is most important.
Ginseng seeds width egg shape or wide obovate, slightly flat, long 4.8～7.2mm, wide 3.9～5.0mm, thick 2.1～3.4mm, surperficial yellow-white or light brown are coarse; Dorsal part is arcuate eminence, and two sides are more flat, and belly is straight or indent slightly, base portion has a little point prominent, and there is an arteries and veins (come off sometimes or partly come off) in the suction hole of last tool one little point-like, top, suction hole, through the top, reach base portion through dorsal part by the seed veutro again, arteries and veins begins to be divided into several to the seed upper end, all pulse classics are crossed the place, seed is inside nick and be the shallow ridges shape all, and exosper is wooden, thick about 0.5mm, smooth interior surfaces, glossy; Endotesta is poor, and faint yellow, adhesion is in endosperm; Flat or the indent slightly of veutro, tool one yellow or pale brown colo(u)r streak shape kind ridge often is divided into 2 (1～3) piece to the top, be connected in a little pointed seedstalk to base portion.Endosperm has oiliness, and embryo is tiny, buries the base portion of being born in kind of benevolence, mass of 1000 kernel 27g, and especially big person can reach 40g.
The someone has carried out the dna fingerprint analytical approach and has differentiated ginseng seeds research at present, but system not.Employing dna fingerprint technology such as Ma Xiaojun are analyzed 5 farmers''s types of mountain ginseng and cultivation ginseng; with RAPD and AFLP method; detect on 92 sample gene groups more than 6000 primer binding site ITS among the ginseng DNA of mountain has been carried out sequencing; obtained amount acute pyogenic infection of finger tip mark, for cultivation, breeding and the conservation of resources of genseng provides new foundation about genetic affinity between the heredity of the different germplasms of genseng and germplasm.
More rich genetic diversity is arranged in the genseng kind, analyzed the RAPD fingerprint of big horse bud, two horse buds, long neck, circle wing, 5 main farmers' of circle reed type, polymorphic site is 56.9%, proves that genseng farmers' type has more rich genetic diversity.Through detecting the AFLP of genseng farmers' type, feature polymorphism is separately closed the attitude individuality but long neck type inside has more to dye, and may more join near the mountain.The analysis showed that in addition, the hereditary difference that the mountain participates between the sibling species American Ginseng is bigger, from the analysis of its genetic distance of RAPD fingerprint, the effect in the ginseng inherent cause morphologic variation of mountain may be less than environmental factor, and this result cultivates for " mountain ginseng " and proposed theoretical foundation.Angle from Germplasm Identification, because it is the relation of " covering " that the mountain participates in the relation of garden ginsent, it is the genetic diversity of mountain ginseng, covered the genetic diversity of cultivation ginseng, cultivate the RAPD specific molecular marker that ginseng does not have so be difficult to find, we can say that the mountain participates in garden ginsent and do not have special difference in heredity for all mountain ginsengs are common.
Example remains in some problems with the dna molecular diagnostic techniques.Be subjected to all multifactor influences such as template, amplification condition as RAPD method stability, and still fail to construct database truly; When RAPD identifies minute seeds class Chinese crude drug, may contain the DNA of two or more plants in the template DNA, relevant two kinds of template DNAs exist the influence to amplification not appear in the newspapers as yet simultaneously; The medicinal material that the dna molecular diagnostic techniques is used as medicine to different parts still is difficult to distinguish; Dna marker for patent medicine, compound, extract does not appear in the newspapers yet.In addition, because of dna molecular diagnostic flag cost height in Chinese traditional medicine identification, factors such as operation experiments conditional request strictness have also hindered the popularization of this technology.Therefore, the dna molecular diagnostic techniques must experience constantly a development and a perfecting process.
Summary of the invention:
The invention provides a kind of near infrared spectrum fast, the method for non-destructively identifying ginseng seeds, have fast, characteristics such as not damaged.
Technical scheme of the present invention is achieved in that
Set up model
(1) sample
Sample comprises kind and type seeds such as No. 1, the Ji ginseng of genseng, yellow fruit genseng, big horse bud, two horse buds, long neck, circle wing, circle reed, bamboo reed, line reed, reed canary grass, purple stem, Radix Ginseng, and the extremely close with it seeds of American ginseng of outward appearance, 13 kind samples altogether.The seed of each kind respectively is divided into 3 parts, and wherein 1 part as analytic sample, and 2 parts as verification sample.
(2) collection of near infrared spectrum
Adopt the near infrared light spectrogram of diffuse reflection integrating sphere collected specimens.Built-in gold-plated diffuse reflector is done background, different types of sample is directly put into the 50cm sample cup, at 12000～4000cm
-1Between scanning, resolution is 8cm
-1, scan 64 times.Every duplicate samples is surveyed 1 time, obtains 24 near infrared light spectrograms altogether.
(3) data processing
Use Br ü kerOPUS/CLUST and OPUS/IDENT that the near infrared light spectrogram that obtains is carried out cluster analysis, and set up qualutative model.
Fig. 1 is the near infrared light spectrogram of seed samples such as genseng, American Ginseng.Principal ingredient is an amino acids in genseng and the seeds of American ginseng, moisture and different carbohydrates.From spectrogram as can be seen, at 6700cm
-1, 5800cm
-1, 5100cm
-1Near three comparatively significantly absorption peaks are arranged respectively.And 6700cm
-1Near the absorption peak one-level frequency multiplication peak of N-H key stretching vibration in the amino acid just, 5800cm
-1Near absorption peak is the one-level frequency multiplication peak of c h bond stretching vibration, 5100cm
-1Be the secondary frequency multiplication of C=O key and the sum of fundamental frequencies peak of O-H.Adopt corresponding Chemical Measurement software to come sample is analyzed discriminating.
Cluster analysis result
Clustering algorithm adopts the most frequently used Ward ' s algorithm (Cluster Analysis) data pre-treating method commonly used that method of derivation (derivative) is arranged, comprise first derivation (first derivative), second order differentiate (secondderivative), vector normalization methods (vector normalization) etc. are normally selected the best preprocess method of effect according to experimental result
[4]Usually, preprocessing procedures and spectrum segment might not be unique in the cluster analysis, and the stability of the wide model of spectral range is high.Consider the drift and the rotation of spectrum, and feature class seemingly, through condition optimizing, the final preprocess method of selecting is first order derivative+vector normalization, and spectral range is 8890.5cm-1～4000cm
-1
Set up and identify model
Utilize OPUS/IDENT to come sample is classified.At first utilize known sample to set up discriminating model, the 2nd, the spectrogram in the spectrogram of unknown sample and the discriminating storehouse is compared, obtain analysis report.The foundation of differentiating the storehouse is the key of qualitative analysis, and it is analyzed quality and depends on sample representativeness to a great extent, the method for data pre-treatment, suitable frequency range and qualitative analysis algorithm
[5]The qualitative analysis rudimentary algorithm has two kinds: canonical algorithm and factorization method.Canonical algorithm is to calculate spectrum intervals with Euclidean distance D, and formula is:
By comparing the matching degree that Euclidean distance is come the comparison spectrogram, a
(k)And b
(k)Be the value of the ordinate of spectrogram a, b, i.e. spectral intensity.Factorization method is the linear combination that earlier spectrogram is expressed as factor spectrum, a=T
1a* f
1+ T
2a* f
2+ T
3a* f
3+ ..., f
iBe factor spectrum, each factor spectrum is orthogonal, and T is the coefficient of each factor spectrum, the corresponding factor spectrum of its numerical representation method offering the sign collection of illustrative plates.Use factor T (also claiming score) to calculate spectrum intervals then, formula is:
Calculate the distance (Hit value) of each sample spectra in all kind training sets then, and calculate the threshold value (D of each kind according to this to this kind averaged spectrum
T): D
T=Maximum (Hit)+X*SDev, wherein Maximum (Hit) is for arriving the ultimate range of this sample average spectrum in the sample spectra of measuring; SDev is the standard deviation of Hit value; X is a coefficient, uses above-mentioned formula to come the matching degree of comparison spectrogram.The result of model sees that on the one hand all modeling spectrum whether all by unique evaluation, also will see the selectivity between the different sink on the other hand, generally uses the S value representation:
Wherein D is the distance of two groups of storehouse averaged spectrum, T
1And T
2The threshold value in two groups of storehouses.If S＜1, illustrating has intersection between the storehouse; S=1 represents just to distinguish; S＞1 library representation separates.Obviously the accuracy rate of the big more explanation model prediction of S can be high more.According to experiment effect, choose suitable algorithm.In the very similar usage factor method of spectrogram, effect can be better than standard law generally speaking.By analysis, selecting the spectrogram preprocess method is first order derivative+vector normalization, and spectral range is 4358.5cm
-1～8913.7cm
-1, generally rule of thumb X is chosen as 0.2500, and algorithm is a factorization method.Analysis result is that 13 reference spectra can be by unique evaluation, and spectrogram is divided into 3 classes according to given kind, and the S value is all greater than 1.
Table 1 is the result of qualutative model.
Table 1 qualutative model result
(4) checking
Verification sample is put into sample cup respectively scan, get the near infrared light spectrogram, spectrogram is identified by institute's established model, draws sample seed conclusion.
The present invention adopts the near infrared molecular spectroscopy techniques, can carry out discriminatory analysis and has the following advantages ginseng seeds:
1. a lot of materials are little at the absorption coefficient of near infrared region, make analytic process become simple: as the absorption spectrum of molecular vibrational energy order transition generation, the frequency multiplication or the sum of fundamental frequencies absorption coefficient of near infrared region are very little, generally more infrared fundamental frequency absorbs low 1 ~ 3 order of magnitude, can directly measure so sample need not dilution, be convenient to the The real time measure of production run.
2. be applicable to the diffuse reflection technology:
Light scattering effect is big in the near-infrared region, and penetration depth is big, makes near-infrared spectrum technique can be used for the diffuse reflection technology sample is directly measured.
3. near infrared light can penetrate in glass or quartz medium: the wavelength of near-infrared region is short, is not absorbed by glass or quartz medium.Thereby can directly measure through container to sample.The more important thing is to make general glass optical fiber or silica fibre can be used for near-infrared spectrum technique, and then make traditional near-infrared spectrum technique expand to online detection under industrial processes analysis and the poisonous and harmful situation.
4. operation cost is low: sample does not need pre-service, and operation cost is low, the increasingly automated technical ability requirement that has reduced simultaneously the operator of instrument.
5. can be used for the qualitative of sample, also can obtain the very high quantitative result of precision: adopt multivariate calibration methods and one group of quantitative model that known similar sample is set up, can obtain relative error fast less than 0.5% measurement result; The discriminance analysis program is adopted in qualitative analysis, obtains the absorbance distributed model of one group of known sample earlier, records to treat the absorbance of qualitative sample under different wave length distributes, and determines with cluster principle whether sample belongs to existing model, i.e. the known sample of this class again.
6. do not destroy sample, do not use reagent, so free from environmental pollution: near-infrared spectrum analysis is just obtained the spectral signal of sample, can directly measure in former container sometimes, thereby not need other reagent, and test process can not produce any pollution.
7. test speed is fast: the information of near infrared spectrum must be carried out data processing and statistical study by computing machine, and a general sample can obtain qualitative or quantitative analysis results after obtaining spectroscopic data at once, and whole process can be finished in less than 2min.Another characteristics of near-infrared spectrum technique are the multiple composition or the character datas that can calculate sample by 1 spectrum.
Good effect of the present invention is: utilize the near infrared molecular spectroscopy techniques, adopt hierarchical clustering method, ginseng seeds has been carried out discriminatory analysis. the result shows that near-infrared spectrum technique can realize the accurate discriminating to ginseng seeds, sample is correctly sorted out according to different kinds. and compare with the classic method of very complicated, the analysis efficiency of near-infrared spectrum technique is higher.Adopt near-infrared spectrum technique to differentiate ginseng seeds, to not influence of seed vitality.Because adopt raw sample directly to scan, pre-treatment that it goes without doing has improved identification efficiency.
Description of drawings:
Fig. 1 is the near-infrared diffuse reflection spectrum figure of different ginseng seeds.
Embodiment:
For the ease of understanding the present invention, especially exemplified by following examples.Its effect is understood that it is to explaination of the present invention but not to any type of restriction of the present invention.
Embodiment 1:
Get " No. 1, lucky ginseng " ginseng seeds and directly put into the 50cm sample cup of the German Br ü ker MPA of company type near infrared spectrometer, at 12000～4000cm
-1Between scanning, resolution is 8cm
-1, get the near infrared light spectrogram.Spectrogram is identified by institute's established model, draws the conclusion of sample for " No. 1, lucky ginseng " ginseng seeds.
Embodiment 2:
The seed of getting " yellow fruit genseng " is directly put into the 50cm sample cup of the German Br ü ker MPA of company type near infrared spectrometer, at 12000～4000cm
-1Between scanning, resolution is 8cm
-1, get the near infrared light spectrogram.Spectrogram is identified by institute's established model, draws the conclusion of sample for " yellow fruit " ginseng seeds.
Embodiment 3:
Get " big horse bud " ginseng seeds and directly put into the 50cm sample cup of the German Br ü ker MPA of company type near infrared spectrometer, at 12000～4000cm
-1Between scanning, resolution is 8cm
-1, get the near infrared light spectrogram.Spectrogram is identified by institute's established model, draws the conclusion of sample for " big horse bud " ginseng seeds.
Embodiment 4:
Get " American Ginseng " seed and directly put into the 50cm sample cup of the German Br ü ker MPA of company type near infrared spectrometer, at 12000～4000cm
-1Between scanning, resolution is 8cm
-1, get the near infrared light spectrogram.Spectrogram is identified by institute's established model, draws the conclusion of sample for " American Ginseng " seed.
Claims (1)
- A near infrared spectrum fast, the method for non-destructively identifying ginseng seeds, it is characterized in that may further comprise the steps:(1) sample is selectedSelect No. 1, the Ji ginseng of genseng, yellow fruit genseng, big horse bud, two horse buds, long neck, circle wing, circle reed, bamboo reed, line reed, reed canary grass, purple stem, Radix Ginseng kind and type seed, and the extremely close with it seeds of American ginseng of outward appearance, 13 kind samples altogether; The seed of each kind respectively is divided into 3 parts, and wherein 1 part as analytic sample, and 2 parts as verification sample;(2) collection of near infrared spectrumAdopt the near infrared light spectrogram of diffuse reflection integrating sphere collected specimens; Built-in gold-plated diffuse reflector is done background, different types of sample is directly put into the 50cm sample cup, at 12000～4000cm -1Between scanning, resolution is 8cm -1, scanning 64 times, every duplicate samples is surveyed 1 time, gets the near infrared light spectrogram;(3) data processingUse Br ü kerOPUS/CLUST and OPUS/IDENT that the near infrared light spectrogram that obtains is carried out cluster analysis, and set up qualutative model; Utilize OPUS/IDENT to come sample is classified; Analysis result is that reference spectra can be by unique evaluation, and spectrogram is divided into 3 classes according to given kind, and the S value should be greater than 1;(4) checkingVerification sample is put into sample cup respectively scan,, judge identification result by this model is identified.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175648A (en) * | 2011-01-04 | 2011-09-07 | 大连理工大学 | Method for distinguishing variety of fritillaria and detecting total alkaloid content of fritillaria by virtue of near infrared spectrum |
| CN102636452A (en) * | 2012-05-03 | 2012-08-15 | 中国科学院长春光学精密机械与物理研究所 | NIR (Near Infrared Spectrum) undamaged identification authenticity method for wild ginseng |
| CN103344602A (en) * | 2013-07-04 | 2013-10-09 | 中国科学院合肥物质科学研究院 | Nondestructive testing method for rice idioplasm authenticity based on near infrared spectrum |
| CN103353443A (en) * | 2013-06-18 | 2013-10-16 | 西北农林科技大学 | Near infrared spectrum based discrimination method for Zhongning fructus lycii |
| CN104062248A (en) * | 2014-06-26 | 2014-09-24 | 中国农业大学 | Method for detecting mechanical damage of apples |
| CN104062262A (en) * | 2014-07-09 | 2014-09-24 | 中国科学院半导体研究所 | Crop seed variety authenticity identification method based on near infrared spectrum |
| CN104198428A (en) * | 2014-08-21 | 2014-12-10 | 中国农业大学 | Method and system for rapidly identifying authenticity of seeds with seed coatings |
| CN105223164A (en) * | 2015-08-04 | 2016-01-06 | 内蒙古农业大学 | Differentiate the method and system of buckwheat or the adulterated wheat flour of oatmeal |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175648A (en) * | 2011-01-04 | 2011-09-07 | 大连理工大学 | Method for distinguishing variety of fritillaria and detecting total alkaloid content of fritillaria by virtue of near infrared spectrum |
| CN102636452A (en) * | 2012-05-03 | 2012-08-15 | 中国科学院长春光学精密机械与物理研究所 | NIR (Near Infrared Spectrum) undamaged identification authenticity method for wild ginseng |
| CN102636452B (en) * | 2012-05-03 | 2015-02-18 | 中国科学院长春光学精密机械与物理研究所 | NIR (Near Infrared Spectrum) undamaged identification authenticity method for wild ginseng |
| CN103353443A (en) * | 2013-06-18 | 2013-10-16 | 西北农林科技大学 | Near infrared spectrum based discrimination method for Zhongning fructus lycii |
| CN103344602A (en) * | 2013-07-04 | 2013-10-09 | 中国科学院合肥物质科学研究院 | Nondestructive testing method for rice idioplasm authenticity based on near infrared spectrum |
| CN103344602B (en) * | 2013-07-04 | 2016-07-27 | 中国科学院合肥物质科学研究院 | A kind of rice germplasm true and false lossless detection method based near infrared spectrum |
| CN104062248A (en) * | 2014-06-26 | 2014-09-24 | 中国农业大学 | Method for detecting mechanical damage of apples |
| CN104062262A (en) * | 2014-07-09 | 2014-09-24 | 中国科学院半导体研究所 | Crop seed variety authenticity identification method based on near infrared spectrum |
| CN104198428A (en) * | 2014-08-21 | 2014-12-10 | 中国农业大学 | Method and system for rapidly identifying authenticity of seeds with seed coatings |
| CN104198428B (en) * | 2014-08-21 | 2016-08-24 | 中国农业大学 | Band seed coat agent seed authenticity rapid identification method and system |
| CN105223164A (en) * | 2015-08-04 | 2016-01-06 | 内蒙古农业大学 | Differentiate the method and system of buckwheat or the adulterated wheat flour of oatmeal |
| CN105223164B (en) * | 2015-08-04 | 2017-11-07 | 内蒙古农业大学 | Differentiate the method and system of buckwheat or the adulterated wheat flour of oatmeal |
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Application publication date: 20100407 |