CN105424675A - Ruminant animal-origin feedstuff identification method based on lipid Raman spectrums - Google Patents
Ruminant animal-origin feedstuff identification method based on lipid Raman spectrums Download PDFInfo
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Abstract
The invention provides a ruminant animal-origin feedstuff identification method based on lipid Raman spectrums. The ruminant animal-origin feedstuff identification method includes the steps that animal-origin feedstuff with known origins and of different species are collected; lipid samples are extracted; Raman spectrum information data of the lipid samples are collected, and the spectrum range is 400-3600 cm<-1>; a distinguishing and analyzing model of the ruminant animal-origin feedstuff is built; the built distinguishing and analyzing model is evaluated; lipid samples of the animal-origin feedstuff to be detected are extracted, and the Raman spectrum information data are collected and input into the distinguishing and analyzing model for species identification. The Raman spectrums of the lipid samples are directly collected, spectrum information and species origin information are correlated, the ruminant animal-origin feedstuff can be identified and analyzed fast and effectively, the analyzing requirement for species identification of the ruminant animal-origin feedstuff of feed quality safety supervision in China is met, and it can be ensured that while 'bovine spongiform encephalopathy' is efficiently prevented, and sustainable development and cyclic utilization of the feed industry are effectively implemented.
Description
Technical field
The invention belongs to the discriminating field of animalsderived feedstuffs, particularly a kind of discriminatory analysis method utilizing Raman spectrum to detect animalsderived feedstuffs raw material.
Background technology
Rabid ox disease (BSE) is a kind of zoonosis, and its cause of disease prion can be propagated by food chain.Meat meal tankage is the animal-based protein feed that slaughtering animal Hou Fei edible tissue makes after pulverizing and pyroprocessing, has the features such as rich in protein, is once widely used in feed industry field.There are some researches show that rabid ox disease causes owing to using the meat meal tankage of the animals such as ill ox, sheep to be added on feeding of ruminant animals in feed.In order to take precautions against the propagation of rabid ox disease, countries in the world are formulated relevant law, regulation limitations one after another or are prohibitted the use the animalsderived feedstuffs such as meat meal tankage.Meat meal tankage is the protein feeds source that a kind of nutritive value is higher, often adds the protein deficient for supplementary grain trough in feed to.Because domestic protein feeds production capacity is not enough, under the ordering about of interests, the behavior of the Misuse animalsderived feedstuffs such as the contraband of import and violated interpolation remains incessant after repeated prohibition, and this is very disruptive market order not only, the interests of infringement consumer, also result in very large rabid ox disease hidden danger.Visible, control the reasonable employment of meat meal tankage, forbid the propagation of carrying prion meat meal tankage, be ensure that feed safety, animal husbandry are sustainablely sent a letter, the important means of food security and human health.Therefore higher requirement is proposed to the authentication technique of different genera especially ruminant source feed.On the one hand, whether mutual feeding animalsderived feedstuffs need to be refined between different genera; On the other hand, need discrimination method fast, to adapt to the needs of extensive Feed Manufacturing.
At present, the animalsderived feedstuffs detection technique that in international coverage, investigation and application is more mainly contains: the microscopic analysis based on histological characteristic, the enzyme linked immunosorbent assay analysis method based on protein analysis, the polymerase chain reaction method based on DNA analysis and the near infrared spectrum based on organo-functional group is analyzed detect analytical technology.The animalsderived feedstuffs examination criteria method wherein detected for official's arbitration is mainly microscopic analysis and polymerase chain reaction method, and current China mainly adopts polymerase chain reaction method.Although these two kinds of methods above-mentioned can carry out the discriminatory analysis of different genera animal derived materials, all there is limitation in various degree, the species discrimination that single use is difficult to meet efficiently and accurately analyzes requirement.Microscopic method cannot carry out the detection of ruminant kind, and polymerase chain reaction method can be carried out ruminant species discrimination but self be existed a lot of not enough in problems such as responsive to temperature, false positive height.Existing near-infrared spectrum technique all detects based on raw material, belongs to method for quick but accuracy of detection is all relatively on the low side, and has certain limitation for the discriminatory analysis of kind equally.
Summary of the invention
In order to solve the problems of the technologies described above, the invention provides a kind of ruminant source feed raw material discrimination method of the Raman spectrum based on fat characteristics.
Animalsderived feedstuffs raw material of the present invention comprises the protein feeds such as terrestrial animal meat meal tankage, terrestrial animal digested tankage, terrestrial animal bone meal and fish meal.On Vehicles Collected from Market, animalsderived feedstuffs mostly is single kind animal and makes, and the present invention is also that the animalsderived feedstuffs raw material made for multiple different single kind animal carries out discriminatory analysis.
The technical scheme realizing the object of the invention is:
Based on a discrimination method for the Raman spectrum ruminant source feed of fat characteristics, comprise the following steps:
1) collect the different genera animalsderived feedstuffs raw material in known source, grind and be prepared as feed sample; Described animalsderived feedstuffs raw material comprises terrestrial animal meat meal tankage, terrestrial animal digested tankage, terrestrial animal bone meal and fish meal;
2) soxhlet extraction is utilized to extract lipid samples from feed sample;
3) gather lipid samples Raman spectral information data with FTRaman spectrometer, the spectral range of collection is 3600 to 400cm
-1;
4) according to step 3) in the Raman spectral information data of lipid samples that gather, set up ruminant source feed raw material discriminatory analysis model;
5) evaluation procedure 4) the discriminatory analysis model set up;
6) extract tested animal source feed lipid samples, and gather Raman spectral information data, input step 4) in the discriminatory analysis model set up carry out species discrimination analysis.
Wherein, described step 1) in, terrestrial animal is selected from pig, ox, Yang Heji, represents mammal, ruminant and bird respectively; Cyclone mill pulverizing is carried out to the different genera animalsderived feedstuffs in collected known source, then crosses 1.0mm sieve.
Feedstuff crosses 1.0mm sieve after pulverizing, to carry out surname extraction.
Preferably, described step 3) in the resolution of Raman spectrum be 4cm
-1, scanning times is 60 times.
Wherein, described step 3) in principal component analysis (PCA) is carried out to Fourier's Raman spectral information data of lipid samples.
Wherein, described step 4) in, lipid Raman spectral information is associated with the Species origin information of sample, adopts the Return Law and leaving-one method validation-cross, set up ruminant source feed raw material partial least squares discriminant analysis model.
Wherein, step 5) in, adopt discrimination Sensitivity and reject rate Specificity two indices to evaluate discrimination model, Sensitivity and Specificity is more close to 1, and discrimination model precision is higher;
Sensitivity＝PA/(PA+ND)(1)
Specificity＝NA/(PD+NA)(2)
In formula: PA is for differentiating positive number, and ND is for differentiating false negative sample number, and NA is for differentiating negative sample number, and PD is for differentiating false positive sample number.Such as, suppose that positive is the pig source sample in experiment, then negative sample is the non-pig source sample in experiment, and false negative is pig source sample for actual and be judged as non-pig source sample, and false positive is non-pig source sample for actual and be judged as pig source sample.
Particularly, when described discrimination Sensitivity and reject rate Specificity is 0.85-1.0, described discriminatory analysis model is effective.
Wherein, step 6) in extract the lipid samples of tested animal source feed material sample with soxhlet extraction, gather lipid samples Raman spectral information basic data with attenuated total reflectance, input step 4) in the discriminatory analysis model set up carry out the discriminatory analysis of ruminant source feed raw material.
Further, step 6) in extract the lipid samples of tested animal source feed material sample with soxhlet extraction, lipid samples Raman spectral information basic data is gathered with FTRaman spectrometer, input discriminatory analysis model, differentiates that tested animal source feed material sample belongs to fish, ruminant, bird or mammal.
Because present feed market only has cattle and sheep bone meal to be used to feedstuff, identify feed and whether derive from cattle and sheep, just effectively can judge the security of feed.
Beneficial effect of the present invention is:
1) the present invention adopts soxhlet extraction to carry out lipid samples extraction, can the pretreatment sample of conventional, easy acquisition different genera animalsderived feedstuffs raw material.
2) the present invention carries out the process of lipid samples Raman spectral information basic data by Fourier Transform Technique, intuitively and significantly can embody different genera animalsderived feedstuffs starting lipid composition difference characteristic.
3) the present invention utilizes the Return Law and leaving-one method validation-cross to set up different genera animalsderived feedstuffs partial least squares discriminant analysis model, and evaluated by discrimination Sensitivity and reject rate Specificity two indices, while effectively ensureing discrimination model stability and the discriminatory analysis ability of display model directly perceived.
4) the present invention directly gathers the Raman spectrum of lipid samples, spectral information is associated with Species origin information, quick, effective discriminatory analysis of ruminant source feed raw material can be realized, thus meet the analysis requirement of China's feeding quality security control for ruminant source feed species discrimination, can ensure while efficient strick precaution " rabid ox disease ", effectively implement the sustainable development of feed industry and recycle.
Accompanying drawing explanation
Fig. 1 is the different genera animalsderived feedstuffs principal component analysis (PCA) figure based on the Raman spectrum of lipid samples in embodiment 1.
Fig. 2 is PLS-DA Model checking result in embodiment.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, the amendment do the inventive method, step or condition or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that technological means used in embodiment is well known to those skilled in the art.
Embodiment 1
1) sample collection and preparation is studied
Research sample is the animalsderived feedstuffs of 70 parts of known animal species, comprise and collect product from home and overseas protein feed enterprise and self-control sample two parts through relevant quality testing department, sample covers terrestrial animal digested tankage, bone meal, meat meal tankage and fish meal etc., wherein 21, ruminant source sample (11, ox source, 9, sheep source), 32, non-ruminant animal source sample (11, pig source, 13, chicken source) and 26, source of fish sample.
All research samples adopt Cyclone mill to pulverize, and then cross 1.0mm sieve.
2) lipid samples extracts
Soxhlet extraction is adopted to utilize full-automatic Milko-Tester (SoxtecTM2050, FOSS company of Denmark) to extract above-mentioned research sample lipid samples.
3) lipid samples Raman spectral information data
BrukerOpticsFT-RamanVERTEX70v (German Bruker company) is adopted to carry out the collection of lipid samples Raman spectral information basic data.Wherein spectra collection scope is 3600 to 400cm
-1, resolution is 4cm
-1, scanning times is 60 times.
4) discriminatory analysis model Establishment and evaluation
Adopt Matlab software (R2012b, Mathworks company of the U.S.), first principal component analysis (PCA) (PCA) is carried out to the animalsderived feedstuffs lipid Raman spectral information data in different genera source.Principal component analysis (PCA) is the algorithm routine that Matlab runs, try to achieve after bringing data into and comprise the maximum number of principal components certificate of spectral information amount, representative large principal ingredient is chosen according to differentiation degree, obtain principal component scores figure (Fig. 1, transverse axis is the first principal component that reflection spectral information is maximum, and the longitudinal axis is Second principal component).
Fig. 1 shows: first and second major component accounts for 22.48% and 7.80% of total variation respectively, and in research sample, the sample of ruminant source and non-ruminant animal source and the source of fish is in the group be separated from each other respectively, has good degree of separation.
According to above-mentioned gathered lipid samples Raman spectral information basic data, lipid Raman spectral information is associated with the Species origin information of sample, adopt the Return Law and leaving-one method validation-cross, set up ruminant source feed raw material partial least squares discriminant analysis (PLS-DA) model.
PLS-DA Model checking the results are shown in Figure 2.In Fig. 2, a dot-and-dash line of top represents discriminant line (discrimY).Discrimination Sensitivity and the reject rate Specificity of model the results are shown in Table 1:
Table 1 ruminant source feed raw material discriminatory analysis model result
Result shows, and experiment sample discrimination Sensitivity and the reject rate Specificity in ruminant source are respectively 0.96 and 1.00, show higher discriminatory analysis model accuracy.
5) discriminatory analysis modelling verification
Select 10 experiment samples to be measured, wherein 4, ruminant source sample (2, ox source, 2, sheep source), 4, non-ruminant animal source sample (2, pig source, 2, chicken source) and 2, source of fish sample, set up discriminatory analysis model is verified.
The result shows, the species discrimination analysis result of 4 ruminant source samples is all correct, and discrimination Sensitivity and the reject rate Specificity of sample are 1.00.
Comparative example
Its method and following analytical approach are compared analysis by the present embodiment.
Adopt soxhlet extraction to utilize full-automatic Milko-Tester (SoxtecTM2050, FOSS company of Denmark) to extract research sample lipid samples, measured the content of fatty acid of sample by gas chromatograph (GC-2014C, Japanese Shimadzu Corporation).Concrete for fatty acid content information is associated with sample type source-information, sets up ruminant source feed partial least squares discriminant analysis (PLS-DA) model.
Test findings: table 2 is the discriminatory analysis model result of the inventive method and vapor-phase chromatography.Result shows, based on the inventive method of lipid Raman spectral information, be more or less the same with the discriminatory analysis result based on content of fatty acid information, but save sample content of fatty acid and measure this time and effort consuming, spend step that is high and that have strict operative technique to require to mensuration personnel.Therefore, it is fast that the inventive method has finding speed compared with vapor-phase chromatography, do not expend chemical reagent, easy and simple to handle grade for remarkable advantage.
The comparative analysis of table 2 ruminant source feed raw material discriminatory analysis model
Above embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various modification that the common engineering technical personnel in this area make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determine.
Claims (9)
1., based on a discrimination method for the ruminant source feed raw material of lipid Raman spectrum, comprise the following steps:
1) collect the different genera animalsderived feedstuffs raw material in known source, grind and be prepared as feed sample; Described animalsderived feedstuffs raw material comprises terrestrial animal meat meal tankage, terrestrial animal digested tankage, terrestrial animal bone meal and fish meal;
2) soxhlet extraction is utilized to extract lipid samples from feed sample;
3) gather lipid samples Raman spectral information data with FTRaman spectrometer, the spectral range of collection is 3600 to 400cm
-1;
4) according to step 3) in the Raman spectral information data of lipid samples that gather, set up ruminant source feed raw material discriminatory analysis model;
5) evaluation procedure 4) the discriminatory analysis model set up;
6) extract tested animal source feed lipid samples, and gather Raman spectral information data, input step 4) in the discriminatory analysis model set up carry out species discrimination analysis.
2. discrimination method according to claim 1, is characterized in that, described step 1) in, terrestrial animal is selected from pig, ox, Yang Heji, represents mammal, ruminant and bird respectively; Cyclone mill pulverizing is carried out to the different genera animalsderived feedstuffs in collected known source, then crosses 1.0mm sieve.
3. discrimination method according to claim 1, is characterized in that, described step 3) in the resolution of Raman spectrum be 4cm
-1, scanning times is 60 times.
4. discrimination method according to claim 1, is characterized in that, described step 3) in principal component analysis (PCA) is carried out to the Fourier Raman spectral information data of lipid samples.
5. discrimination method according to claim 1, it is characterized in that, described step 4) in, lipid Raman spectral information is associated with the Species origin information of sample, adopt the Return Law and leaving-one method validation-cross, set up ruminant source feed raw material partial least squares discriminant analysis model.
6. according to the arbitrary described discrimination method of Claims 1 to 5, it is characterized in that, described step 5) in, discrimination Sensitivity and reject rate Specificity two indices is adopted to evaluate discrimination model, Sensitivity and Specificity is more close to 1, and discrimination model precision is higher;
Sensitivity＝PA/(PA+ND)(1)
Specificity＝NA/(PD+NA)(2)
In formula: PA is for differentiating positive number, and ND is for differentiating false negative sample number, and NA is for differentiating negative sample number, and PD is for differentiating false positive sample number.
7. discrimination method according to claim 6, is characterized in that, when described discrimination Sensitivity and reject rate Specificity is 0.85-1.0, described discriminatory analysis model is effective.
8. discrimination method according to claim 1, it is characterized in that, step 6) in extract the lipid samples of tested animal source feed material sample with soxhlet extraction, gather lipid samples Raman spectral information basic data with attenuated total reflectance, input step 4) in the discriminatory analysis model set up carry out the discriminatory analysis of ruminant source feed raw material.
9. discrimination method according to claim 8, it is characterized in that, step 6) in extract the lipid samples of tested animal source feed material sample with soxhlet extraction, lipid samples Raman spectral information basic data is gathered with FTRaman spectrometer, input discriminatory analysis model, differentiates that tested animal source feed material sample belongs to fish, ruminant, bird or mammal.
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Cited By (7)
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| CN106990096A (en) * | 2017-03-27 | 2017-07-28 | 中国农业大学 | Meat meal tankage kind detection method and system based on LIBS |
| CN108801965A (en) * | 2018-05-04 | 2018-11-13 | 中国农业大学 | Meat meal tankage kind detection method and system based on genomic DNA infrared spectrum |
| CN110631967A (en) * | 2019-10-16 | 2019-12-31 | 复旦大学 | Raman spectrum-based atmospheric black carbon aerosol source analysis method |
| CN110672582A (en) * | 2019-10-08 | 2020-01-10 | 浙江大学 | Raman characteristic spectrum peak extraction method based on improved principal component analysis |
| CN111141719A (en) * | 2019-12-27 | 2020-05-12 | 梧州市食品药品检验所 | Rapid nondestructive identification method for anti-AIDS medicine |
| CN111401794A (en) * | 2020-04-24 | 2020-07-10 | 江苏傲农生物科技有限公司 | Feed quality control method based on near infrared spectrum |
| CN114819712A (en) * | 2022-05-17 | 2022-07-29 | 江苏邦鼎科技有限公司 | Pet feed recommendation method and system based on artificial intelligence |
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| CN110631967A (en) * | 2019-10-16 | 2019-12-31 | 复旦大学 | Raman spectrum-based atmospheric black carbon aerosol source analysis method |
| CN111141719A (en) * | 2019-12-27 | 2020-05-12 | 梧州市食品药品检验所 | Rapid nondestructive identification method for anti-AIDS medicine |
| CN111401794A (en) * | 2020-04-24 | 2020-07-10 | 江苏傲农生物科技有限公司 | Feed quality control method based on near infrared spectrum |
| CN114819712A (en) * | 2022-05-17 | 2022-07-29 | 江苏邦鼎科技有限公司 | Pet feed recommendation method and system based on artificial intelligence |
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Application publication date: 20160323 |