CN106546553A - A kind of quick nondestructive discrimination method of genetically engineered soybean oil - Google Patents
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- 239000003549 soybean oil Substances 0.000 title claims abstract description 25
- 235000012424 soybean oil Nutrition 0.000 title claims abstract description 25
- 238000012850 discrimination method Methods 0.000 title claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 30
- 238000004458 analytical method Methods 0.000 claims abstract description 24
- 238000002329 infrared spectrum Methods 0.000 claims abstract description 12
- 238000011068 loading method Methods 0.000 claims abstract description 8
- 230000009261 transgenic effect Effects 0.000 claims abstract description 6
- 239000000843 powder Substances 0.000 claims abstract description 5
- 238000000513 principal component analysis Methods 0.000 claims abstract description 5
- 238000007781 pre-processing Methods 0.000 claims abstract description 4
- 238000002203 pretreatment Methods 0.000 claims abstract description 4
- 235000013322 soy milk Nutrition 0.000 claims abstract description 4
- 244000068988 Glycine max Species 0.000 claims description 18
- 235000010469 Glycine max Nutrition 0.000 claims description 18
- 239000003921 oil Substances 0.000 claims description 8
- 235000019198 oils Nutrition 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 4
- 238000009499 grossing Methods 0.000 claims description 3
- 238000010606 normalization Methods 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 238000000605 extraction Methods 0.000 claims description 2
- 238000003825 pressing Methods 0.000 claims description 2
- 230000008961 swelling Effects 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 description 11
- 230000000694 effects Effects 0.000 description 8
- 235000013305 food Nutrition 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 230000003595 spectral effect Effects 0.000 description 5
- 244000037671 genetically modified crops Species 0.000 description 4
- 238000004611 spectroscopical analysis Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 235000003869 genetically modified organism Nutrition 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 2
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 239000003925 fat Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 210000004885 white matter Anatomy 0.000 description 2
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 206010068052 Mosaicism Diseases 0.000 description 1
- 108700019146 Transgenes Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 101150065438 cry1Ab gene Proteins 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000009659 non-destructive testing Methods 0.000 description 1
- 235000021049 nutrient content Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000003765 sex chromosome Anatomy 0.000 description 1
- 230000009329 sexual behaviour Effects 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 235000021404 traditional food Nutrition 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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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- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The present invention discloses a kind of quick nondestructive discrimination method of genetically engineered soybean oil, including step：(1) ir data of genetically engineered soybean oil and Non-transgenic soybean oil is gathered using infrared spectrometer；(2) to described ir data using the smooth pretreatments of Savitzky Golay, and carry out principal component analysis；(3) for passing through the ir data for pre-processing, characteristic wavelength is extracted using the method for PCA loading；(4) the discriminant analysis model of infrared spectrum and characteristic wavelength, including PLS DA discriminant analyses models and SVM discriminant analysis models are set up respectively；(5) for transgenosis soymilk powder to be measured and non-transgenic sample, step (1) step (4) is passed sequentially through, by the infrared spectrum of sample to be tested, classification is estimated through described discriminant analysis model.The present invention without the need for complicated sample preprocessing, and quickly, lossless, high precision.
Description
Technical field
The invention belongs to the technical field of nondestructive testing of GM food examination, and in particular to a kind of genetically engineered soybean oil
Quick nondestructive discrimination method.
Background technology
With the development of modern biotechnology, the research of transgenic technology is developed rapidly and is promoted.With transgenosis
Technology can cultivate high yield, high anti-, high-quality, and the improved seeds of maladjustment ecological environment greatly reduce the administration of agriculture chemical
Amount, is conducive to environmental protection.However, can not ignore, the uncertainty of advanced science and technology causes transgenic technology to become
One " double-edged sword ".GM food difference topmost with traditional food is that the former is imported containing useful technique for gene engineering
Foreign gene, and by its specific exogenous proteins of expression.In recent years, impact and transgenosis of the genetically modified crops to ecological environment
The edible safety sex chromosome mosaicism of food becomes focus of concern.From the point of view of international experience, administrative department of national governments is to turning base
Because product takes the attitude of discretion, it is also that one, China is urgent and great to implement system, the management of efficient Transgene-safty
Strategic Sexual behavior mode.An important content therein, exactly strengthens GMO detection technical research, with by supervision and check mark
Knowledge is regulated market order.Traditional detection GMOs are primarily directed to the foreign DNA and egg of genetically modified crops and its correlated product
What white matter was carried out, although these methods are preparing DNA and protein needed for detection with higher accuracy and sensitivity
Sample when be required to carry out destruction extraction to genetically modified crops extremely processed goods, and waste time and energy, program is complicated, cost compared with
Height, layman are difficult to be competent at.Netically modified foods are through different procedure (grinding, heating, microwave, acid-base value, micro- life
Thing fermentation etc.), its foreign protein and foreign gene can occur degraded in various degree and rupture, and exist so as to have influence on transgene component
Content and effect in final products, so need to fully take into account food in analysis and evaluation Transgenic Food Safety Issue adding
Work process.
China is the major country of production of soybean, and the quantity of soybean resource socially occupies very big ratio.Soybean is high
Protein food, with abundant nutritive value, plays critically important effect in the life of modern people.Contain in the composition of soybean
There is 40% or so protein, 17% carbohydrate, with multivitamin, and the content of fat is relatively low, and only 18%
Left and right, therefore, in daily life, high quality soybean product can supplement the nutrient content of needed by human body, bean product
Using relatively more, the fast development of soybean planting industry and bean product is also promoted to a great extent.Soybean oil is China
People is particularly one of main edible oil in northern people's lives, and genetically engineered soybean oil is from nutrient composition content and constitutes and non-turn
The almost no difference of transgenic soybean oil, also just causes fats and oils processing enterprise to be usually not described in this respect or avoid mentioning.For
The right to know and right to choose of consumer are ensured, seeking the method for quick nondestructive becomes when business differentiating genetically engineered soybean oil
It is anxious.
Infrared spectrum can capture the absorption spectrum information of the protein molecule related to genetic mutation, this is because egg
Contain substantial amounts of hydric group X-H in white matter molecule, infrared spectrum reflects the energy absorption information of hydric group, therefore also just has
It is standby to catch the ability of crops genetic mutation molecule absorption information and determine whether identification transgenosis and not genetically modified reason
By basis.The various groups of ingredient have oneself specific infrared signature absorption peak, can realize some in molecule accordingly
Chemical bond and " fingerprint verification " of functional group.With development and the maturation of spectral technique, this characteristic of infrared spectrum is to turn base
Because the discriminating of agricultural product provides an effective new way.
The content of the invention
In view of the pros and cons present situation of original molecule detection GMOs, of the invention to combine Chemical Measurement using infrared spectrum technology
Method differentiates that to genetically engineered soybean oil the high precision of identification provides effective for the management of efficient Transgene-safty
Detection means.
In order to realize object above, the present invention provides technical scheme below：
A kind of quick nondestructive discrimination method of genetically engineered soybean oil, including step：
S1：Genetically engineered soybean oil and Non-transgenic soybean oil are obtained in 400cm-1～4000cm-1It is infrared in wave-number range
Absorption spectrum information；
S2：Using Savitzky-Golay smooth (SG) pretreatment, principal component (PCA) analysis is carried out；
S3：Spectroscopic data to passing through pretreatment carries out characteristic wavelength selection, and principal component analysis load (PCA is respectively adopted
Loadings) method carries out characteristic wavelength selection；
S4：Modeling collection and forecast set are set up by k-means methods.It is based respectively on original spectrum and the characteristic wave for selecting
It is long, set up PLS-DA, SVM discriminant analysis models.These discriminant analysis models based on different principles, logarithm from different angles
According to carrying out discriminant analysis, and therefrom select suitable discriminant analysis model.
Infrared spectrum is gathered using the Jasco FT/IR-4100 infrared spectrometers of Jasco companies of Japan production in step S1
Information, resolution ratio are 8cm-1, scanning times are 32, and signal to noise ratio S/N is 22000：1.
In step S2, SG is noise, the effective ways of raising signal to noise ratio in a kind of effective removal spectroscopic data.Multinomial
Number of times and smooth points have conclusive impact to its smooth effect.SG of the present invention is smooth flat using 7 points of 2 order polynomial
It is sliding.Specific algorithm is as follows：
In formula, hiFor smoothing factor, H is normalization factor,xkFor the value obtained after wavelength K process；
Modeling collection and forecast set are set up using k-means methods in step S4.Randomly select K cluster center of mass point
(cluster centroids) is μ 1, μ 2 ... ... μ k, repeats procedure below until convergence：
For each sample i, its class that should belong to is calculated
c(i)=argargminj||x(i)-μj||2
For each class j, such barycenter is recalculated
In formula, x(i)For sample i with it is all kinds of in that closest class, μjUs are represented to belonging to same class for barycenter
Center of a sample conjecture；
PLS-DA discriminant analyses model in step S4, replaces chemical score to be analyzed with the integer value for representing classification, according to
Predicting the outcome for obtaining carries out discriminant analysis.In order to judge to the classification of sample, also because predicting the outcome middle predicted value not
Be represent classification integer but actual numerical value, therefore discrimination threshold need to be set.In our current research, it is set as discrimination threshold by 0.5,
I.e. when the absolute value of actual value and the difference of predicted value is more than 0.5, then differentiate mistake, otherwise it is correct to be then considered as differentiation.
SVM discriminant analyses model in step S4, is inferring the class corresponding to arbitrary input x using y=sign (f (x))
Not, output valve only allows to take class label.The present invention SVM modeling in, using RBF (RBF) as kernel function.SVM moulds
The parameter penalty coefficient c Search Ranges of type are 2-8To 28。
Relative to prior art, beneficial effects of the present invention are：After the present invention considers netically modified foods deep processing, its
Foreign protein and foreign gene can occur degraded in various degree, combine chemometrics method based on infrared spectrum technology with this,
Genetically engineered soybean is differentiated.For traditional molecular detecting method, sample preprocessing of the present invention without the need for complexity,
And quick, lossless, high precision.Therefore, it can be said that being with a wide range of applications and value, it is efficient transgenosis peace
Full management is there is provided effective detection means.
Description of the drawings
Fig. 1 is identification process figure of the present invention based on the genetically engineered soybean of infrared spectrum.
Fig. 2 is the discriminant analysis result that the present invention is set up based on SVM.
Specific embodiment
With reference to the accompanying drawings and examples, describe in further detail.This specific embodiment is with technical solution of the present invention
Premised under implemented, it should be understood that these modes are only illustrative of the invention and is not intended to limit the scope of the invention.This
As embodiment, the identification of other genetically modified crops processed goods can refer to the embodiment to the bright soybean to turn cry1Ab genes
Method is carried out.As shown in figure 1, quick nondestructive discrimination method is comprised the following steps that in the present embodiment：
1st, the method squeezed using low temperature cold prepares soybean oil:From the soybean of full grains, rinsed using clear water, then powder
It is broken to 20 mesh；By solid-liquid ratio 1：10 plus soybean of the biological enzyme liquid to after crushing in carry out it is swelling；Dry in 90 DEG C of baking ovens；Using
Ke Meite (KOMET) cold oil press of German IBG doors good fortune (IBG Monforts) company carries out physics cold press, the temperature of squeezing of colding pressing
For 40 DEG C, soybean oil is obtained.Totally 240 parts of genetically engineered soybean oil and Non-transgenic soybean oil, every part of 30ml are obtained altogether.
2nd, using 4100 Fourier transform spectrometers of Jasco FTIR (Jsasco Analytical Instruments,
Japan spectral scan is carried out to every part of soybean oil sample), the condition of scanning is：Scanning times 32 times, spectral resolution is 8cm-1,
Spectral region is 4 00～4000cm-1。
3rd, the spectroscopic data obtained using Savitzky-Golay smooth (SG) pretreatment, carries out principal component (PCA) analysis.
SG is noise, the effective ways of raising signal to noise ratio in a kind of effective removal spectroscopic data.The degree of polynomial and
Smooth points have conclusive impact to its smooth effect.SG of the present invention is smooth smooth using 7 points of 2 order polynomial.It is concrete to calculate
Method is as follows：
In formula, hiFor smoothing factor, H is normalization factor,xkFor the value obtained after wavelength K process；
4th, the full spectrum light modal data obtained Jing after SG-7 pretreatments sets up PLS and SVM discriminant analysis moulds respectively as input
Type.In the discriminant analysis model of full spectrum, SVM discrimination model effects are better than PLS discrimination models.SVM modelings collection and forecast set
Rate of accuracy reached to 84.9% and 81.28%.
5th, characteristic wavelength selection is carried out based on principal component analysis load (PCA loadings) method.PCA algorithms are to light
Modal data is obtained during carrying out principal component analysis.PCA algorithms can remove redundancy in data, and be converted into including
The contribution rate (loading) of effect spectral information, while remaining substantial amounts of raw information.When tiring out for the front n principal component for obtaining
When meter contribution rate is more than 85%, then the peak valley of contribution rate under the principal component can be chosen as characteristic wave bands.Obtained using PCA algorithms
Characteristic wave bands include：
6th, PCA loadings methods extract PLS the and SVM models that characteristic wavelength is set up
Based on 21 characteristic wavelengths that PCA loadings methods are extracted, respectively 745cm-1, 917cm-1, 982cm-1,
1156cm-1, 1589cm-1, 2018cm-1, 2147cm-1, 2318cm-1, 2368cm-1, 2451cm-1, 2610cm-1, 2773cm-1,
2781cm-1,2790cm-1, 2867cm-1, 2798cm-1, 3467cm-1, 3614cm-1, 3631cm-1And 3726cm-1。
As shown in Figure 2, modeled using the characteristic wavelength that PCA loadings methods are extracted as input variable, two models
Discriminant analysis effect is all preferable.Preferably, the differentiation accuracy modeling to transgenosis soymilk powder collects the discriminant analysis effect of SVM models
81.73% and 80.54% have been reached with forecast set.The above results explanation fast and effectively can be recognized using the method for the present invention
Transgenosis soymilk powder, has a good application prospect and considerable market value.
Claims (6)
1. the quick nondestructive discrimination method of a kind of genetically engineered soybean oil, it is characterised in that including step：
(1) ir data of genetically engineered soybean oil and Non-transgenic soybean oil is gathered using infrared spectrometer；
(2) to described ir data using the smooth pretreatments of Savitzky-Golay, and carry out principal component analysis；
(3) for passing through the ir data for pre-processing, characteristic wavelength is extracted using the method for PCA-loading；
(4) set up the discriminant analysis model of infrared spectrum and characteristic wavelength respectively, including PLS-DA discriminant analyses model and/or
SVM discriminant analysis models；
(5) for transgenosis soymilk powder to be measured and non-transgenic sample, step (1)-step (4) is passed sequentially through, by sample to be tested
Infrared spectrum, estimate classification through described discriminant analysis model.
2. the quick nondestructive discrimination method of genetically engineered soybean according to claim 1 oil, it is characterised in that in step (1),
Infrared spectrum information is gathered using Jasco FT/IR-4100 infrared spectrometers, resolution ratio is 8cm-1, scanning times are 32, letter
It is 22000 to make an uproar than S/N：1.
3. the quick nondestructive discrimination method of genetically engineered soybean according to claim 1 oil, it is characterised in that Savitzky-
The smooth pretreatments of Golay are smoothed for 7 points using 2 order polynomial, and specific algorithm is as follows：
In formula, hiFor smoothing factor, H is normalization factor,xkFor the value obtained after wavelength K process.
4. the quick nondestructive discrimination method of genetically engineered soybean according to claim 1 oil, it is characterised in that set up and differentiate point
Before analysis model, modeling collection and forecast set are set up using k-means methods；It is μ 1, μ 2 ... ... μ to randomly select K cluster center of mass point
K, repeats procedure below until convergence：
For each sample i, its class that should belong to is calculated
c(i)=argargminj||x(i)-μj||2
For each class j, such barycenter is recalculated
In formula, x(i)For sample i with it is all kinds of in that closest class, μjUs are represented to belonging to of a sort sample for barycenter
The conjecture at center.
5. the quick nondestructive discrimination method of genetically engineered soybean according to claim 1 oil, it is characterised in that the feature of extraction
Wavelength is respectively 745cm-1, 917cm-1, 982cm-1, 1156cm-1, 1589cm-1, 2018cm-1, 2147cm-1, 2318cm-1,
2368cm-1, 2451cm-1, 2610cm-1, 2773cm-1, 2781cm-1, 2790cm-1, 2867cm-1, 2798cm-1, 3467cm-1,
3614cm-1, 3631cm-1And 3726cm-1。
6. the quick nondestructive discrimination method of genetically engineered soybean according to claim 1 oil, it is characterised in that adopt low temperature cold
The method prepare transgenosis soybean oil of squeezing and Non-transgenic soybean oil：From the soybean of full grains, rinsed using clear water, then
It is crushed to 20 mesh；By solid-liquid ratio 1：10 plus soybean of the biological enzyme liquid to after crushing in carry out it is swelling；Dry in 90 DEG C of baking ovens；Adopt
Physics cold press is carried out with cold oil press, the temperature of squeezing of colding pressing is 40 DEG C, obtains soybean oil.
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| CN114113035A (en) * | 2021-11-18 | 2022-03-01 | 北京理工大学 | Transgenic soybean oil identification method |
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| CN114113035A (en) * | 2021-11-18 | 2022-03-01 | 北京理工大学 | Transgenic soybean oil identification method |
| CN114113035B (en) * | 2021-11-18 | 2024-02-02 | 北京理工大学 | Identification method of transgenic soybean oil |
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