CN105738313B - A kind of method and application identifying animal blood based on near-infrared spectrum technique - Google Patents
A kind of method and application identifying animal blood based on near-infrared spectrum technique Download PDFInfo
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/3577—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing liquids, e.g. polluted water
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
- G01N21/359—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light using near infrared light
Abstract
The invention discloses a kind of methods and application for identifying animal blood based on near-infrared spectrum technique, belong to the adulterated identification of blood product and test and analyze technical field.Method provided by the present invention is to animal pure blood standard sample and to the animal blood sample of adulterated inspection, acquisition original spectrum is scanned using near-infrared analyzer respectively, Pretreated spectra is carried out to resulting near infrared spectrum data is scanned, it is utilized respectively factorial analysis and extracts Effective genes number, the efficiency factor for being utilized respectively selection establishes Fisher linear discriminant analysis model, linear discriminant analysis is done to measuring samples using the Fisher linear discrimination function of pure blood standard sample, obtain prediction classification results and image, blood sample to be checked is identified by comparing the two image, it is whether adulterated to identify measuring samples.The method of the present invention identifies accuracy rate height, reaches 100% to the correct decision rate of different cultivars blood sample, is suitable for identifying animal blood product.
Description
Technical field
The present invention relates to a kind of methods and application for identifying animal blood based on near-infrared spectrum technique, belong to blood product
Adulterated identification tests and analyzes technical field.
Background technique
Since there are biggish differences in price for different types of animal blood product, and with national in recent years right
The sustainable growth of animal blood product demand, it is to reap staggering profits not stinting to adulterate that this, which allows for some illegal businessmans, with false random
Very, such as Bean curd with duck blood ox blood and pig blood are pretended to be, and are not only disrupted the market, but also cause to consumer's interests and health
Damage.The quick identification technology of research blood product can effectively ensure consumer legitimate right, ensure that the stabilization in market is strong
Kang Fazhan.
Conventional blood product reflects, and method for distinguishing is cumbersome, time-consuming, and at high cost, chemical reagent is poisonous and harmful.Near infrared technology with
Its quick, lossless, safe and environment-friendly feature is promoted to every field, and the near-infrared for studying animal blood product identifies skill
Art is significant.
The adulterated technology of detection blood is only the detection method of molecular biology at present, and the round pcr based on DNA is wide
General to be used to identify the animal source component in food, although this method accuracy is high, equipment is expensive, tests skill to operator
Art requires height, and experimental procedure is cumbersome, time-consuming and laborious, needs expensive chemical reagent, reagent is toxic to people, has dirt to environment
Dye, therefore there is an urgent need to a kind of detection methods of quick nondestructive economy.
Summary of the invention
To solve the deficiencies in the prior art, animal blood is identified based on near-infrared spectrum technique the present invention provides a kind of
Method, the technical solution adopted is as follows:
The purpose of the present invention is to provide a kind of method for identifying animal blood based on near-infrared spectrum technique, this method is
To animal pure blood standard sample and to the animal blood sample of adulterated inspection, acquisition is scanned using near-infrared analyzer respectively
Original spectrum, to the near infrared spectrum number for scanning resulting animal pure blood standard sample and the animal blood sample to adulterated inspection
According to Pretreated spectra is carried out, it is utilized respectively factorial analysis and extracts animal pure blood standard sample and the animal blood sample to adulterated inspection
The Effective genes number of product establishes animal pure blood standard sample Fisher linear discriminant analysis model using the efficiency factor of selection,
Linear discriminant point is done using the animal blood sample that the Fisher linear discriminent model of pure blood standard sample treats adulterated inspection
Analysis obtains prediction classification results and image, is identified by comparing the two image to blood sample to be checked, to identify to be checked
Whether sample is adulterated.
Preferably, the method, steps are as follows:
1) to animal pure blood standard sample and to the animal blood sample of adulterated inspection, respectively using near-infrared analyzer into
Row scanning, each sample multiple scanning, sample requirement fills sample again when scanning every time, keeps the homogeneity of dress sample, acquires average
The curve of spectrum obtains the near-infrared original spectrum number of animal pure blood standard sample and the animal blood sample to adulterated inspection respectively
According to；
2) resulting animal pure blood standard sample and animal blood sample to adulterated inspection are scanned to step 1) respectively
The near-infrared original spectral data that is averaged carries out Pretreated spectra, obtains animal pure blood standard sample respectively and to the dynamic of adulterated inspection
The pre-processed spectrum data of object blood sample；The Pretreated spectra is to be handled in accordance with the following steps: 1. at first derivative
Reason；2. standard normal variable conversion process；
3) the pretreatment light of the animal pure blood standard sample to step 2) acquisition and the animal blood sample to adulterated inspection
Modal data extracts Effective genes number after carrying out factorial analysis respectively, obtains animal pure blood standard sample respectively and to adulterated inspection
The Effective genes number of animal blood sample；
4) efficiency factor for being utilized respectively the animal pure blood standard sample of step 3) selection establishes animal pure blood standard sample
Fisher linear discriminant analysis model treats the dynamic of adulterated inspection using the Fisher linear discriminent model of pure blood standard sample
Object blood sample does linear discriminant analysis, obtains prediction classification results and image, by comparing the two image to blood sample to be checked
Product are identified, so that whether identify measuring samples adulterated.
Preferably, the step 1) animal blood sample to adulterated inspection, be pure blood, pure blood product, adulterated blood or
Adulterated blood product.
It is pure duck blood, pure ox blood, pure pig blood, pure it is highly preferred that the step 1) animal blood sample to adulterated inspection
Chicken blood or their adulterated blood or adulterated blood product.
Preferably, step 1) the animal pure blood standard sample is one of duck blood, ox blood, pig blood, chicken blood or several
Kind.
Preferably, the step 1) scanning, resolution ratio 5nm.
Preferably, the step 1) scanning, scanning range are 950nm~1650nm.
Preferably, the step 3) Effective genes number is to select contribution rate of accumulative total 99.9% or more because subnumber represents
The main information of sample spectra.
It is highly preferred that the method, steps are as follows:
1) the pure blood standard sample of duck blood, ox blood, pig blood and chicken blood and the animal blood sample to adulterated inspection are chosen respectively
Product are scanned acquisition original spectrum using near-infrared analyzer respectively, and Multiple-Scan acquires average original spectrum, collect average
Original spectral data obtains the pure blood standard sample of duck blood, ox blood, pig blood and chicken blood and the animal blood to adulterated inspection respectively
The near-infrared original spectral data of sample；The scanning, resolution ratio 5nm, scanning range are 950nm~1650nm；
2) duck blood, ox blood, the minimal standards sample of pig blood and chicken blood and to the animal blood of adulterated inspection are obtained to step 1)
The near-infrared of the liquid sample original spectral data that is averaged is handled in accordance with the following steps: 1. first derivative is handled, 2. standard normal
Change of variable processing, obtains the pure blood standard sample of duck blood, ox blood, pig blood and chicken blood and the blood sample to adulterated inspection respectively
Pre-processed spectrum data；
3) duck blood that step 2) is obtained, ox blood, the pure blood standard sample of pig blood and chicken blood and to the animal of adulterated inspection
The pre-processed spectrum data of blood sample extract Effective genes number after carrying out factorial analysis respectively, and accumulation contribution rate is selected to exist
99.9% or more because subnumber representative sample spectrum main information, respectively obtain duck blood, ox blood, pig blood and chicken blood pure blood mark
The Effective genes number of quasi- sample and the animal blood sample to adulterated inspection；
4) it is utilized respectively the pure blood standard sample of duck blood, ox blood, pig blood and chicken blood that step 3) is chosen and to adulterated inspection
The efficiency factor of animal blood sample establish Fisher linear discriminant analysis model, obtain duck blood, ox blood, pig blood and chicken blood
The Fisher linear discriminant analysis model of pure blood standard sample；Duck blood, ox blood, pig blood, chicken blood are represented with number 1,2,3,4 respectively
Pure blood standard sample, the duck blood, ox blood, pig blood and chicken blood pure blood standard sample Fisher linear discriminant function model point
Not are as follows:
Y1=1433.249X1+140.856X2+170.017X3+275.968X4+562.745X5+2.535X6+41.115X7-
400.432
Y2=2308.426X1+299.230X2+137.188X3+434.840X4+1384.356X5+90.046X6+
253.564X7-801.143
Y3=2031.635X1+195.013X2+291.211X3+385.953X4+773.809X5-4.481X6+39.303X7-
846.204
Y4=5234.540X1+692.642X2+194.205X3+962.015X4+3711.533X5+258.618X6+
759.907X7-4780.0
Wherein: Y represents discriminant scores i.e. discriminant value, and X represents variable i.e. Assessing parameters；
5) using step 4) obtain duck blood, ox blood, pig blood and chicken blood pure blood standard sample Fisher linear discriminant
The animal blood sample that analysis model treats adulterated inspection does linear discriminant analysis, obtains prediction classification results, allusion quotation then differentiates letter
Number scatter plot and administrative division map identify blood sample to be checked by comparing the two image, to whether identify measuring samples
It is adulterated.
Application of the either method described above in identification animal blood and animal blood product are adulterated.Especially identifying
Application in animal blood, adulterated animal blood, animal blood product and adulterated animal blood product.
The method of the present invention has following difference compared with the prior art:
1, it is modeled using full spectrum, it is preferred without wavelength, it saves time；
2, preprocessing procedures are different, using first derivative and standard normal variable conversion process；
3, the prior art extracts spectral signature using principal component analysis, and the present invention is extracted using factorial analysis, factorial analysis
The differentiation result obtained after extraction is 100%, and effect is more preferable；
4, the prior art is too general using the saying of discriminant analysis method, at present near infrared spectroscopy qualitative discrimination side
Method has very much, such as: PCA, FLDA, HCA, SIMCA etc., and the present invention clearly uses Fisher Fisher face, and and its
His method of discrimination has carried out effect and has compared, and differentiates accuracy rate up to 100%.
5, the method for the present invention differentiates that outcome graphic display effect is more preferable, more intuitively, clear.
Influence caused by the method for the present invention can eliminate background drift to spectroscopic data derivation, first derivative can not only disappear
Except the constant drift of background, the interference of overlapping spectra band on the other hand can also be reduced.It is right in the spectrogram processing of near infrared spectrum
The main purpose that spectrum carries out derivation processing is to eliminate the influence of the constant drift phenomenon of background, for solid sample, entire
Near infrared spectrum region has baseline drift phenomenon.Baseline drift is the principal element of influence near infrared spectrum quality, therefore right
Spectrum derivation is common near infrared spectrum preprocess method, to spectrum carry out derivation processing in addition to can eliminate baseline drift or
The influence of gentle background interference, while and profile clearer spectral information higher than original spectrum resolution ratio can also be provided.Such as
Fruit considers influence of noise, and situation will be more complicated, and since random noise is typically all high-frequency signal, derivative will be further amplified
Noise, therefore when use derivative preprocess method, usually require that original spectrum has compared with high s/n ratio.
Either solid or fluid sample are extremely difficult to ideal uniform state.The inhomogeneities of sample will cause light
By, reflected through or from sample when scattering, the scattering of light can bring error to sample spectra.Standard normal variable becomes
Changing can be used to correct the spectral error because of caused by scattering, which thinks in each spectrum, the absorbance value of each wavelength points
Certain distribution (such as normal distribution) should be met, by this hypothesis to each spectrum carry out pretreatment make its as close possible to
" ideal " spectrum (spectrum of i.e. no scattering error effect).Standard normal variable transformation is that former spectrum subtracts all light of the spectrum
It after the average value of spectrum point suction degree, then is substantially to make former spectroscopic data standard normal divided by the standard deviation of the spectroscopic data.
The method of the present invention can also identify other blood products, such as goose blood and sheep blood.It is first scanned after taking unknown sample
Then spectrum carries out Pretreated spectra, spectroscopic data is then imported into SPSS software and carries out factorial analysis, obtains best factors
Number, does linear discriminant analysis using the Fisher linear discrimination function of pure blood standard sample, obtains prediction classification results and allusion quotation
Then discriminant function scatter plot identifies blood sample to be checked by comparing the two image, to whether identify measuring samples
It is adulterated.
The invention has the advantages that:
The method for identifying animal blood at present is mainly the round pcr for utilizing molecular biology method, such as animal derived
Food duck blood, the research of pig blood DNA extraction method and double PCR detection.Although this method accuracy is high, equipment is expensive, right
Operator's experimental technique requires height, and experimental procedure is cumbersome, time-consuming and laborious, needs expensive chemical reagent, and reagent is toxic to people
Evil has pollution to environment, and method very good solution provided by the invention problem above, testing result is accurate, quickly, economical,
Environmentally protective is a kind of very useful adulteration detection method.
The method of the present invention is not necessarily to sample preparation expense, and test is quick, it is only necessary to 5 minutes, easy to operate, whole harmless, the green, peace of detection
Entirely, sampling quantity is big, and the representativeness of sample is high, avoids adulterating irregular, influences testing result.
Detailed description of the invention
Fig. 1 is that factorial analysis combination Fisher linear discriminant analysis dissipates the discriminant analysis result of different cultivars blood sample
Point diagram；
(1, duck blood；2, ox blood；3, pig blood；4, chicken blood).
Fig. 2 is discriminatory analysis result scatterplot of the factorial analysis combination Fisher linear discriminant analysis to adulterated duck blood sample
Figure；
(1, duck blood；2 ox bloods；3, pig blood；4, chicken blood；5,20 duck bloods mix 40% ox blood and 20 duck bloods mix 20% N
Blood).
Fig. 3 is discriminatory analysis knot of the factorial analysis combination Fisher linear discriminant analysis to the duck blood sample of different adulterated rates
Fruit scatter plot；
(1,100% duck blood；2, duck blood mixes 20% ox blood；3, duck blood mixes 40% ox blood；4, duck blood mixes 60% ox blood；5, duck
Blood mixes 80% ox blood；6,100% ox blood).
Specific embodiment
The present invention will be further described combined with specific embodiments below, but the present invention should not be limited by the examples.
Embodiment 1:
The method for identifying animal blood at present is mainly the round pcr for utilizing molecular biology method, such as animal derived
Food duck blood, the research of pig blood DNA extraction method and double PCR detection.This method is using chloroform-sodium acetate (NaAc) extraction method
DNA is extracted from solid block duck blood with KI extraction method, by synthesis amplification duck source property and pig derived component primer, is carried out dual
PCR amplification obtains the purpose extension band of duck and pig blood, thus judge whether contain pig in animal derived blood sample,
Duck blood ingredient.
This method instrument investment has ten thousand yuan of about 6-20 of EppenDorf brand PCR instrument；61 electrophoresis apparatus of Beijing (band electrophoresis tank)
About 0.4 ten thousand yuan；The Univerisal HooD II gel imager of BIORAD (is furnished with WHITE and UV lamp, Quantity One figure
As analysis process system) about 10-15 ten thousand；About 0.3 ten thousand yuan of TGL-16B table model high speed centrifuge.Sample preparation expense about 10-15 member/sample,
Test time-consuming most about 3 hours short in skilled situation, or so about 10 hours of other traditional method for extracting and detection time.And this
Inventive method instrument expense is 150,000, and is not necessarily to sample preparation expense, and test is quick, it is only necessary to which 5 minutes, be a kind of economical, quick
Detection mode.
This method operating process is complicated, needs professional training personnel, is otherwise likely to occur DNA and extracts and does not come out or PCR amplification
The problem of failure, sample need to use liquid nitrogen to carry out freeze grinding in sample making course, this walks dangerous property.And it uses to people
The organic reagent (such as imitative/isoamyl alcohol) of the harmful property of body.And operation of the present invention is simple, substantially without error operation, and examines
Surveying whole process does not have any harm operation or reagent to staff, is a kind of green, safety detection mode.
The sample extraction amount of this method is 0.4g, and sample amount is small, when blood doping is irregular, may cause testing result
Error.In the measurement of sample sensitivity and when establishing dual-PCR method, the DNA of duck, pig blood is first extracted respectively, then
DNA is diluted inspection or its method sensitivity is surveyed in mixing, without carrying out the setting of various concentration, such case in the sample
Under, extraction factor is not taken into account, this also will affect the sensitivity of actual sample detection.And sampling quantity of the present invention about 20g, sample
The representativeness of product is high, even if doping is irregular, also can guarantee the accuracy of detection, is suitble to actual detection case；And this hair
Bright all to take into account all influence factors, the sensitivity and accuracy rate when actual sample detects are all high.
This method only identifies pig blood and duck blood simultaneously, and there is competition between the primer of different blood samples, with duck
Primer is compared, and the specificity of pig primer is stronger.If further carrying out the identification of more blood samples simultaneously, competed between primer
Influence may be bigger, it will reduce the recall rate of duck blood.And the doping of existing market duck blood product is with ox blood and pig blood
Based on, there are also other animal bloods, this method does not establish other blood PCR detection methods such as ox simultaneously.And the present invention is built simultaneously
The detection method of duck blood, ox blood, pig blood and chicken blood has been found, the lossless inspection of blood sample disposably can quickly, be comprehensively carried out
It surveys, it is easier to promote.
The Fisher linear discriminant analysis of 2.4 kinds of typical blood samples of embodiment
A kind of method for identifying animal blood based on near-infrared spectrum technique is present embodiments provided, main contents are such as
Under:
1) it acquires the original spectrum of different blood products and carries out spectroscopic data pretreatment:
A chooses representative animal blood sample, takes four duck blood, ox blood, pig blood, chicken blood different cultivars respectively；
B is evenly laid out in the specimen cup that diameter is 75mm by obtained blood sample, using near-infrared analyzer with 5nm
Resolution scan 60 times, spectral scanning range be 950~1650nm, each sample multiple scanning 3 times, every time scanning when sample
It is required that filling sample again, the homogeneity of dress sample is kept, obtains sample average primary light spectrogram.
C makees following Pretreated spectra to resulting near infrared spectrum data is scanned:
1. first derivative is handled；2. standard normal variable conversion process；
2) Effective genes number is extracted after carrying out factorial analysis to the preprocessed data of acquisition.
3) the accumulation contribution rate of preceding 8 factors can reach 100%, both may include 100% information of original variable.
Therefore the main information of preceding 8 factors representative sample spectrum is selected.
4) their Fisher linear discriminant function model is established using preceding 8 factors.It is represented respectively with number 1,2,3,4
Duck blood, ox blood, pig blood, chicken blood.4 typical linear discriminant functions are obtained, functional relation is as follows:
Y1=-62.238X1+3.341X2+383.683X3-372.688X4+40.438X5-149.269X6-3.959X7+
11.514X8-203.418
Y2=36.938X1-56.159X2+50.939X3-78.173X4-19.428X5+7.944X6+11.421X7+
9.968X8-73.632
Y3=-98.964X1+141.898X2+60.632X3-518.018X4+77.012X5-252.255X6+0.761X7+
26.484X8-463.078
Y4=124.264X1-89.079X2-1040.949X3+968.879X4-98.023X5393.579X6-8.223X7-
47.967X8-1226.976
(Y represents discriminant scores i.e. discriminant value in formula, and X represents variable i.e. Assessing parameters)
5) in SPSS software, the Fisher using the duck blood of acquisition, ox blood, pig blood and chicken blood pure blood standard sample is linear
Discriminant analysis model carries out linear discriminant analysis to above-mentioned pure blood sample, obtains the prediction classification knot of 4 kinds of Standard blood samples
Fruit, allusion quotation then discriminant function scatter plot and administrative division map predict classification results, allusion quotation then discriminant function scatter plot and administrative division map according to gained
Carry out comprehensive distinguishing, allusion quotation then discriminant function scatter plot as shown in Figure 1, as can be seen from Figure 1 four known class sample mass centers
Distribution, from result images it can be concluded that, factorial analysis combination linear discriminant analysis to different cultivars blood sample correctly sentence
Not rate reaches 100%.
Embodiment 3: the discrimination test of adulterated blood sample
Discriminatory analysis, adulterated sample are carried out to the duck blood sample of two kinds of adulterated rates of difference using the modeling method of embodiment 2
The blood sample used is duck blood and ox blood sample.
Method is as follows:
1) original spectrum and the progress spectroscopic data for acquiring 4 kinds of Standard blood products and the adulterated ratio sample of two kinds of differences are pre-
Processing:
A chooses fresh duck blood, ox blood, pig blood and chicken blood sample, is represented respectively with number 1,2,3,4.In addition two are prepared
The duck blood of the different adulterated ratios of kind, ox blood mixing blood sample, adulterated ratio are respectively as follows :+60% ox blood of 40% duck blood, 20% duck
+ 80% ox blood of blood, altogether six kinds of blood samples；
B is evenly laid out in the specimen cup that diameter is 75mm by obtained blood sample, using near-infrared analyzer with 5nm
Resolution scan 60 times, spectral scanning range be 950~1650nm, each sample multiple scanning 3 times, every time scanning when sample
It is required that filling sample again, the homogeneity of dress sample is kept, obtains sample average primary light spectrogram.
C makees following data processing to resulting near infrared spectrum data is scanned:
1. first derivative is handled；2. standard normal variable conversion process；
2) near infrared spectrum data obtained after processing is input in SPSS software and Factor minute is carried out to spectroscopic data
Effective genes number is extracted in analysis；
3) the accumulation contribution rate of preceding 7 factors can reach 99.999%, both may include the 99.999% of original variable
Information.Therefore the main information of preceding 7 factors representative sample spectrum is selected.
4) their Fisher linear discriminant function model is established using preceding 7 factors.It is represented respectively with number 1,2,3,4
Duck blood, ox blood, pig blood and chicken blood.4 typical linear discriminant functions are obtained, functional relation is as follows:
Y1=1433.249X1+140.856X2+170.017X3+275.968X4+562.745X5+2.535X6+41.115X7-
400.432
Y2=2308.426X1+299.230X2+137.188X3+434.840X4+1384.356X5+90.046X6+
253.564X7-801.143
Y3=2031.635X1+195.013X2+291.211X3+385.953X4+773.809X5-4.481X6+39.303X7-
846.204
Y4=5234.540X1+692.642X2+194.205X3+962.015X4+3711.533X5+258.618X6+
759.907X7-4780.012
(Y represents discriminant scores i.e. discriminant value in formula, and X represents independent variable i.e. Assessing parameters)
5) it brings the independent variable of two kinds of adulterated samples into above-mentioned discriminant function, obtains discriminant value, utilize blood known to above-mentioned 4 kinds
The Fisher linear discrimination function of liquid sample does linear discriminant analysis, obtains four kinds of known samples and two kinds to adulterated inspection sample
The prediction classification results of product, allusion quotation then discriminant function scatter plot and administrative division map, predict classification results, allusion quotation then discriminant function according to gained
Scatter plot and administrative division map carry out comprehensive distinguishing, allusion quotation then discriminant function scatter plot as shown in Fig. 2, as can be seen from Figure 2 known to 4
The mass center of classification sample mass center and 1 adulterated sample is not overlapped, and is reflected by comparing the two image to blood sample to be checked
Not, so that whether identify measuring samples adulterated.Discriminatory analysis the results show that linear discriminant analysis technology to the ducks of different adulterated rates
The correct recognition rata of the discriminatory analysis totality of blood sample is 100%.Duck blood sample of the linear discriminant analysis technology to different adulterated rates
The discriminatory analysis result of product is shown in Fig. 2.
Embodiment 4: the verification test of different adulterated ratio blood samples
Discriminatory analysis, adulterated sample are carried out to the duck blood sample of two kinds of adulterated rates of difference using the modeling method of embodiment 3
The blood sample used is duck blood and ox blood sample.
Method is as follows:
1) original spectrum of the different adulterated ratio blood products of acquisition and progress spectroscopic data pretreatment:
A chooses fresh duck blood and ox blood sample and is uniformly mixed, and represents 100% with number 1,2,3,4,5,6 respectively
Duck blood ,+20% ox blood of 80% duck blood ,+40% ox blood of 60% duck blood ,+60% ox blood of 40% duck blood ,+80% ox blood of 20% duck blood,
100% ox blood, six ratio blood samples, blood sample are mixed spare altogether；
B is evenly laid out in the specimen cup that diameter is 75mm by obtained blood sample, using near-infrared analyzer with 5nm
Resolution scan 60 times, spectral scanning range be 950~1650nm, each sample multiple scanning 3 times, every time scanning when sample
It is required that filling sample again, the homogeneity of dress sample is kept, obtains sample average primary light spectrogram.
C makees following data processing to resulting near infrared spectrum data is scanned:
1. first derivative is handled；2. standard normal variable conversion process；
2) near infrared spectrum data obtained after processing is input in SPSS software and Factor minute is carried out to spectroscopic data
Effective genes number is extracted in analysis；
3) the accumulation contribution rate of preceding 13 factors can reach 99.916%, both may include the 99.916% of original variable
Information.Therefore the main information of preceding 13 factors representative sample spectrum is selected.
4) their Fisher linear discriminant function model is established because of number of words using first 13.Number 1,2,3,4 is used respectively,
5,6 represent 100% duck blood ,+20% ox blood of 80% duck blood ,+40% ox blood of 60% duck blood ,+60% ox blood of 40% duck blood, 20% duck
+ 80% ox blood of blood, 100% ox blood.6 typical linear discriminant functions are obtained, functional relation is as follows:
Y1=-- 525.871X1-566.609X2-547.343X3-33.622X4-120.102X5+44.561X6-1.070X7+
35.163X8+12.487X9-3.863X10+6.911X11+20.753X12+12.295X13-622.988
Y2=166.085X1+227.234X2+190.014X3+8.244X4+48.593X5-28.048X6-2.193X7-
14.093X8-7.439X9+2.880X10-5.403X11-9.233X12-7.524X13-112.106
Y3=236.948X1+290.552X2+268.590X3+26.811X4+58.586X5-25.849X6-1.661X7-
19.294X8-4.988X9-0.727X10-3.521X11-10.274X12-2.759X13-159.806
Y4=312.537X1+313.724X2+328.182X3+35.042X4+60.639X5-16.528X6+1.083X7-
23.624X8-2.822X9-2.230X10-1.762X11-10.525X12-3.432X13-218.110
Y5=437.110X1+417.034X2+443.036X3+37.721X4+75.055X5-30.092X6+0.626X7-
38.886X8-8.306X9-5.699X10-9.737X11-17.633X12-19.853X13-405.811
Y6=-626.810X1-681.936X2-682.478X3-74.195X4-122.771X5+55.955X6+3.214X7+
60.733X8+11.069X9+8.185X10+13.511X11+26.913X12+21.273X13-901.590
(Y represents discriminant scores i.e. discriminant value in formula, and X represents variable i.e. Assessing parameters)
5) linear discriminant analysis is done using the Fisher linear discrimination function of above-mentioned blood sample, obtains 6 kinds of samples
It predicts classification results, allusion quotation then discriminant function scatter plot and administrative division map, classification results, allusion quotation then discriminant function scatterplot is predicted according to gained
Figure and administrative division map carry out comprehensive distinguishing, allusion quotation then discriminant function scatter plot as shown in figure 3, as can be seen from Figure 36 known class
The distribution of sample mass center.Discriminatory analysis is the results show that identification of the linear discriminant analysis technology to the duck blood sample of different adulterated rates
The overall correct recognition rata of analysis is 100%.Discriminatory analysis of the linear discriminant analysis technology to the duck blood sample of different adulterated rates
As a result see Fig. 3.
Embodiment 5
Embodiment 1-4 effect is compared, comparison result is as shown in table 1:
The comparison of table 1 embodiment 1 and 2 effect of embodiment
As it can be seen from table 1 method provided by the present invention, in detection speed, detecting instrument equipment requirement, testing staff
Operating technology requirement, detection reagent, testing cost, sample loss and destruction, environmental protection etc. are substantially better than existing method,
A kind of means that can be used as the whether adulterated on-site quick screening of law enforcement agency's batch samples are also applied to blood product production
Producer's on-line checking.
Embodiment 6
Various qualitative discrimination method effects are compared by the present invention during method for building up, comparison result such as 2 institute of table
Show:
The comparison of the various qualitative discrimination method effects of table 2
| Method of discrimination | Verifying differentiates accuracy rate | Arithmetic speed |
| FLDA (Fisher linear discriminant analysis) | 100% | It is most fast |
| PCA (principal component analysis) | 91% | Comparatively fast |
| HCA (hierarchical cluster analysis) | 90% | Comparatively fast |
| PLS-DA (partial least squares discriminant analysis) | 86.67% | Comparatively fast |
From table 2 it can be seen that method provided by the present invention, is substantially better than in terms of arithmetic speed, differentiation
Other qualitative discrimination methods.
Although the present invention has been disclosed in the preferred embodiment as above, it is not intended to limit the invention, any to be familiar with this
The people of technology can do various changes and modification, therefore protection of the invention without departing from the spirit and scope of the present invention
Range should subject to the definition of the claims.
Claims (6)
1. a kind of method for identifying animal blood based on near-infrared spectrum technique, which is characterized in that steps are as follows:
1) animal pure blood standard sample and animal blood sample to adulterated inspection are laid in respectively in specimen cup, are used respectively
Near-infrared analyzer is scanned 60 times in the scanning range of 950nm~1650nm with the resolution ratio of 5nm, and each sample repeats to sweep
It retouches, sample requirement fills sample again when scanning every time, keeps the homogeneity of dress sample, acquires averaged spectrum curve, obtain animal respectively
The near-infrared original spectral data of pure blood standard sample and the animal blood sample to adulterated inspection；
2) the close red of resulting animal pure blood standard sample and animal blood sample to adulterated inspection is scanned to step 1) respectively
Averagely original spectral data carries out Pretreated spectra outside, obtains animal pure blood standard sample and the animal blood to adulterated inspection respectively
The pre-processed spectrum data of liquid sample；The Pretreated spectra is to be handled in accordance with the following steps: 1. first derivative is handled；②
Standard normal variable conversion process；
3) the pre-processed spectrum number of the animal pure blood standard sample to step 2) acquisition and the animal blood sample to adulterated inspection
According to Effective genes number is extracted after carrying out factorial analysis respectively, animal pure blood standard sample and the animal to adulterated inspection are obtained respectively
The Effective genes number of blood sample；The Effective genes number is to select contribution rate of accumulative total 99.9% or more because of subnumber representative sample
The main information of product spectrum；
4) efficiency factor for being utilized respectively the animal pure blood standard sample of step 3) selection establishes animal pure blood standard sample
Fisher linear discriminant analysis model treats the dynamic of adulterated inspection using the Fisher linear discriminent model of pure blood standard sample
Object blood sample does linear discriminant analysis, obtains prediction classification results and image, by comparing the two image to blood sample to be checked
Product are identified, so that whether identify measuring samples adulterated.
2. method according to claim 1, which is characterized in that the step 1) animal blood sample to adulterated inspection is
Pure blood, pure blood product, adulterated blood or adulterated blood product.
3. stating method according to claim 2, which is characterized in that the step 1) animal blood sample to adulterated inspection, is pure
Duck blood, pure ox blood, pure pig blood, pure chicken blood or their adulterated blood or adulterated blood product.
4. method according to claim 1, which is characterized in that step 1) the animal pure blood standard sample is duck blood, ox
One or more of blood, pig blood, chicken blood.
5. method according to claim 1, which is characterized in that steps are as follows:
1) the pure blood standard sample of duck blood, ox blood, pig blood and chicken blood and the animal blood sample to adulterated inspection are chosen respectively, point
It is not scanned acquisition original spectrum using near-infrared analyzer, Multiple-Scan acquires average original spectrum, collects average original
Spectroscopic data obtains the pure blood standard sample of duck blood, ox blood, pig blood and chicken blood and the animal blood sample to adulterated inspection respectively
Near-infrared original spectral data；The scanning, resolution ratio 5nm, scanning range are 950nm~1650nm；
2) duck blood, ox blood, the minimal standards sample of pig blood and chicken blood and to the animal blood sample of adulterated inspection are obtained to step 1)
The near-infrared of the product original spectral data that is averaged is handled in accordance with the following steps: 1. first derivative is handled, 2. standard normal variable
Conversion process obtains the pre- of duck blood, ox blood, the pure blood standard sample of pig blood and chicken blood and the blood sample to adulterated inspection respectively
Handle spectroscopic data；
3) duck blood that step 2) is obtained, ox blood, the pure blood standard sample of pig blood and chicken blood and to the animal blood of adulterated inspection
The pre-processed spectrum data of sample carry out extracting Effective genes number after factorial analysis respectively, select accumulation contribution rate 99.9% with
On because subnumber representative sample spectrum main information, obtain respectively duck blood, ox blood, pig blood and chicken blood pure blood standard sample and
The Effective genes number of animal blood sample to adulterated inspection；
4) duck blood, ox blood, pig blood and the pure blood standard sample of chicken blood that are utilized respectively step 3) selection and moving to adulterated inspection
The efficiency factor of object blood sample establishes Fisher linear discriminant analysis model, obtains the pure blood of duck blood, ox blood, pig blood and chicken blood
The Fisher linear discriminant analysis model of standard sample；Respectively with number 1,2,3,4 represent duck blood, ox blood, pig blood, chicken blood it is pure
Blood standard sample, the duck blood, ox blood, pig blood and chicken blood pure blood standard sample Fisher linear discriminant function model difference
Are as follows:
Y1=1433.249X1+140.856X2+170.017X3+275.968X4+562.745X5+2.535X6+41.115X7-
400.432
Y2=2308.426X1+299.230X2+137.188X3+434.840X4+1384.356X5+90.046X6+253.564X7-
801.143
Y3=2031.635X1+195.013X2+291.211X3+385.953X4+773.809X5-4.481X6+39.303X7-
846.204
Y4=5234.540X1+692.642X2+194.205X3+962.015X4+3711.533X5+258.618X6+759.907X7-
4780.0
Wherein: Y represents discriminant scores i.e. discriminant value, and X represents variable i.e. Assessing parameters；
5) using step 4) obtain duck blood, ox blood, pig blood and chicken blood pure blood standard sample Fisher linear discriminant analysis
The animal blood sample that model treats adulterated inspection does linear discriminant analysis, and obtaining prediction classification results, allusion quotation, then discriminant function dissipates
Point diagram and administrative division map identify blood sample to be checked by comparing the two image, so that whether identify measuring samples adulterated.
6. application of the either method described in claim 1-5 in identification animal blood and animal blood product are adulterated.
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