CN113671013A - Construction method and identification method of portunus trituberculatus origin tracing model - Google Patents
Construction method and identification method of portunus trituberculatus origin tracing model Download PDFInfo
- Publication number
- CN113671013A CN113671013A CN202110946051.3A CN202110946051A CN113671013A CN 113671013 A CN113671013 A CN 113671013A CN 202110946051 A CN202110946051 A CN 202110946051A CN 113671013 A CN113671013 A CN 113671013A
- Authority
- CN
- China
- Prior art keywords
- carbon
- stable isotope
- sample
- portunus trituberculatus
- isotope ratio
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241001533364 Portunus trituberculatus Species 0.000 title claims abstract description 70
- 238000000034 method Methods 0.000 title claims abstract description 40
- 238000010276 construction Methods 0.000 title claims abstract description 18
- CKUAXEQHGKSLHN-UHFFFAOYSA-N [C].[N] Chemical compound [C].[N] CKUAXEQHGKSLHN-UHFFFAOYSA-N 0.000 claims abstract description 53
- 241000238097 Callinectes sapidus Species 0.000 claims abstract description 50
- 229910052788 barium Inorganic materials 0.000 claims abstract description 19
- 229910052785 arsenic Inorganic materials 0.000 claims abstract description 18
- 239000011669 selenium Substances 0.000 claims description 13
- 230000029087 digestion Effects 0.000 claims description 12
- 238000000120 microwave digestion Methods 0.000 claims description 12
- 229910052701 rubidium Inorganic materials 0.000 claims description 12
- 229910052711 selenium Inorganic materials 0.000 claims description 12
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 claims description 11
- 238000004458 analytical method Methods 0.000 claims description 11
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 claims description 11
- DSAJWYNOEDNPEQ-UHFFFAOYSA-N barium atom Chemical compound [Ba] DSAJWYNOEDNPEQ-UHFFFAOYSA-N 0.000 claims description 11
- IGLNJRXAVVLDKE-UHFFFAOYSA-N rubidium atom Chemical compound [Rb] IGLNJRXAVVLDKE-UHFFFAOYSA-N 0.000 claims description 11
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 claims description 10
- 239000003795 chemical substances by application Substances 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 238000000611 regression analysis Methods 0.000 claims description 9
- 210000003205 muscle Anatomy 0.000 claims description 8
- 238000012937 correction Methods 0.000 claims description 7
- 239000007789 gas Substances 0.000 claims description 7
- 230000001419 dependent effect Effects 0.000 claims description 6
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 5
- 229910017604 nitric acid Inorganic materials 0.000 claims description 5
- 238000000889 atomisation Methods 0.000 claims description 4
- 239000012159 carrier gas Substances 0.000 claims description 4
- 238000002485 combustion reaction Methods 0.000 claims description 4
- 239000001307 helium Substances 0.000 claims description 4
- 229910052734 helium Inorganic materials 0.000 claims description 4
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 12
- 229910052748 manganese Inorganic materials 0.000 abstract description 9
- 239000000523 sample Substances 0.000 description 42
- 239000011572 manganese Substances 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 8
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000012795 verification Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000005259 measurement Methods 0.000 description 4
- 229910052757 nitrogen Inorganic materials 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000009616 inductively coupled plasma Methods 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241000406668 Loxodonta cyclotis Species 0.000 description 2
- 241000238122 Paralithodes Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000002790 cross-validation Methods 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 241000243813 Arenicola Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- ATJFFYVFTNAWJD-UHFFFAOYSA-N Tin Chemical compound [Sn] ATJFFYVFTNAWJD-UHFFFAOYSA-N 0.000 description 1
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 229910052793 cadmium Inorganic materials 0.000 description 1
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000012468 concentrated sample Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005485 electric heating Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 229910052712 strontium Inorganic materials 0.000 description 1
- CIOAGBVUUVVLOB-UHFFFAOYSA-N strontium atom Chemical compound [Sr] CIOAGBVUUVVLOB-UHFFFAOYSA-N 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229910052719 titanium Inorganic materials 0.000 description 1
- 239000010936 titanium Substances 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
- G01N27/626—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode using heat to ionise a gas
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
- G01N2001/2866—Grinding or homogeneising
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Electrochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention belongs to the technical field of tracing of aquatic product producing areas, and provides a construction method and an identification method of a tracing model of a portunus trituberculatus producing area. The method starts from the difference of multi-elements and carbon-nitrogen stable isotopes of the organism, only uses the common carbon-nitrogen stable isotope ratio and seven indexes of Mn, As, Se, Rb and Ba to establish a tracing model, has the advantages of simple used instruments, less detection indexes, high detection precision, good reproducibility, simple and convenient method and strong operability, reduces the uncertainty based on commodity labels and subjective judgment, and has the distinguishing rate of the origin of the blue crabs reaching 100 percent.
Description
Technical Field
The invention relates to the technical field of tracing of aquatic product producing areas, in particular to a construction method and an identification method of a tracing model of a portunus trituberculatus producing area.
Background
Portunus trituberculatus (Portuguetus trituberculatus) is commonly called Portunus trituberculatus, Mariothis gigas and sea crab, belongs to the family of Paralithodes arenicola and genus Paralithodes, and is an important economic crab in China coastal region. China is a big country for catching the blue crabs, and the catching yield is far higher than the breeding yield. With the development of the work of protecting the marine environment, proliferating and releasing the portunus trituberculatus, forbidding the fishing season and the like in China, the resource amount of the portunus trituberculatus is greatly recovered. Origin protection, geographic identification and traditional characteristic protection systems are established for famous portunus trituberculatus in Laizhou, Zhoushan, Xiangshan and the like, and all the measures need to be supported by corresponding analysis technology to be successfully implemented.
The portunus trituberculatus is mainly distributed in yellow Bohai sea and east sea areas, and the price of portunus trituberculatus in different producing areas is greatly different, so that the portunus trituberculatus in a low-value producing area is often used for replacing the portunus trituberculatus in a high-value area in the market.
Therefore, establishing the source tracing technology of the blue crabs has important significance for ensuring the reliability of the source of the blue crabs.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for constructing and identifying a portunus trituberculatus origin tracing model. When the traceability model obtained by the construction method provided by the invention is used for identifying the producing area, the detection indexes are few, the detection precision is high, the method is simple and convenient, and the operability is strong.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for constructing a portunus trituberculatus origin tracing model, which comprises the following steps:
measuring the carbon-nitrogen stable isotope ratio and the element content of a blue crab sample in a known producing area; the elements comprise manganese, arsenic, selenium, rubidium and barium;
and carrying out stepwise regression analysis on the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample to obtain a origin tracing model of blue crab with the carbon-nitrogen stable isotope ratio and the element content as independent variables and the origin as dependent variables.
Preferably, when the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample are measured, the leg muscle of the blue crab sample is used.
Preferably, the determination parameters of the carbon-nitrogen stable isotope ratio comprise: the temperature of the combustion furnace is 900-1020 ℃, the temperature of the column box is 40-70 ℃, and the flow rate of helium is 80-100 mL/min.
Preferably, the process of determining the carbon-nitrogen stable isotope ratio of the blue crab sample further comprises the two-point correction of the isotope standard sample by using USGS40 and USGS41a, wherein the USGS40 comprises delta13C＝-26.39‰，δ15N is-4.52 permillage; the USGS41a includes delta13C＝36.55‰，δ15N＝47.55‰。
Preferably, in the process of determining the carbon-nitrogen stable isotope ratio of the blue crab samples, every 20 blue crab samples are inserted into the isotope standard samples to carry out two-point correction; each blue crab sample was tested in parallel for carbon-nitrogen stable isotope 3 times.
Preferably, the sample for measuring the element content is a digestion solution, and the preparation method of the digestion solution comprises the following steps:
mixing a portunus trituberculatus sample with a digesting agent, and performing microwave digestion to obtain a digesting solution;
the digesting agent comprises concentrated nitric acid; the dosage ratio of the portunus trituberculatus sample to the digesting agent is 0.2 g: (5-10) mL.
Preferably, the microwave digestion procedure comprises: heating to 120 deg.C, and maintaining for 5 min; continuously heating to 150 ℃, and keeping the temperature for 10 min; and continuously heating to 190 ℃ and preserving the heat for 20 min.
Preferably, the determination parameters of the element content include:
radio frequency power: 1200-1500W; plasma gas flow rate: 15L/min; carrier gas flow: 0.4L/min; auxiliary air flow rate: 0.4L/min; temperature of the atomization chamber: 2 ℃; an atomizer: a concentric atomizer.
Preferably, the stepwise regression analysis is performed using SPSS software.
The invention also provides a method for identifying the origin of the blue crabs, which comprises the following steps:
determining the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected, wherein the elements comprise manganese, arsenic, selenium, rubidium and barium;
substituting the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected into each origin tracing model obtained by the construction method in the technical scheme to obtain a judgment threshold value;
and dividing the discrimination threshold value by using a Fisher discrimination analysis method to obtain the source tracing and producing area of the portunus trituberculatus to be detected.
The invention provides a method for constructing a portunus trituberculatus origin tracing model, which comprises the following steps: measuring the carbon-nitrogen stable isotope ratio and the element content of a blue crab sample in a known producing area; the elements comprise manganese, arsenic, selenium, rubidium and barium; and carrying out stepwise regression analysis on the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample to obtain a origin tracing model of blue crab with the carbon-nitrogen stable isotope ratio and the element content as independent variables and the origin as dependent variables. The tracing model of the invention establishes the discrimination model based on 7 indexes of the contents of multiple elements (Mn, As, Se, Rb and Ba) and the ratio of stable isotopes of carbon and nitrogen, has the advantages of less detection indexes, high detection precision, good reproducibility, simple method and strong operability, reduces uncertainty based on subjective judgment, and achieves the discrimination rate of the origin of the portunus trituberculatus production area up to 100%.
Drawings
FIG. 1 is a discriminant analysis scattergram of Portunus trituberculatus samples from Zhoushan, Weifang, and Lingyun harbor three producing areas.
Detailed Description
The invention provides a method for constructing a portunus trituberculatus origin tracing model, which comprises the following steps:
measuring the carbon-nitrogen stable isotope ratio and the element content of a blue crab sample in a known producing area; the elements comprise manganese, arsenic, selenium, rubidium and barium;
and carrying out stepwise regression analysis on the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample to obtain a origin tracing model of blue crab with the carbon-nitrogen stable isotope ratio and the element content as independent variables and the origin as dependent variables.
In the present invention, the starting materials used in the present invention are preferably commercially available products unless otherwise specified.
The method is used for measuring the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample in the known producing area.
The production places of the portunus trituberculatus sample are not specifically limited, and the portunus trituberculatus sample is selected according to actual conditions, for example, the origin place protection is established for famous portunus trituberculatus in the state of leizhou, navian and elephant mountain at present, so that the portunus trituberculatus in other places can be used by illegal vendors to pretend, the production places of the portunus trituberculatus are required to be distinguished, and the portunus trituberculatus sample does not belong to the three production places; therefore, in constructing the origin tracing model of the producing area, the portunus trituberculatus samples are preferably from leizhou, navian and elephant mountain.
In the specific embodiment of the invention, the origin tracing model relates to the regions of Zhejiang navian, Shandong Weifang and Jiangsu Lianchun harbor; the portunus trituberculatus sample is from Zhejiang navian, Shandong Weifang and Jiangsu Lianchong.
In the invention, the number of the blue crab samples is preferably more and better, and the more the number of the samples is, the more accurate the tracing model is established. In the embodiment of the invention, the number of the blue crab samples in each producing area is preferably more than or equal to 50, and more preferably 50.
In the present invention, when the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample are measured, the leg muscle of the blue crab sample is preferably used. In the present invention, the leg muscle of the portunus trituberculatus sample is preferably washed, freeze-dried and pulverized before the measurement is performed. The operation of the flushing is not particularly limited in the present invention, as long as the leg muscles can be flushed clean. In the invention, the temperature of the freeze drying is preferably-30 to-50 ℃, and the time is preferably 24 to 48 hours, and more preferably 36 hours. The operation of the pulverization is not particularly limited in the present invention as long as it can pass through a 100-mesh sieve.
In the present invention, the determination of the carbon-nitrogen stable isotope ratio is preferably performed in a stable isotope ratio mass spectrometer. In the present invention, the parameters for measuring the ratio of the carbon-nitrogen stable isotope include: the temperature of the combustion furnace is preferably 900-1020 ℃, and further preferably 960 ℃; the temperature of the column box is preferably 40-70 ℃, and further preferably 70 ℃; the flow rate of the helium gas is preferably 80-100 mL/min, and more preferably 100 mL/min.
In the invention, in the process of determining the carbon-nitrogen stable isotope ratio of the blue crab sample, the method preferably further comprises the step of carrying out two-point correction on the isotope standard sample by using USGS40 and USGS41a, wherein the USGS40 comprises delta13C＝-26.39‰，δ15N is-4.52 permillage; the USGS41a includes delta13C＝36.55‰，δ15N＝47.55‰。
In the invention, in the process of measuring the carbon-nitrogen stable isotope ratio of the blue crab samples, preferably, every 20 blue crab samples are inserted into an isotope standard sample for two-point correction; each blue crab sample is preferably assayed in parallel 3 times for carbon nitrogen stable isotopes.
In the present invention, the specific operations of the two-point correction include: calibrating the measured values by using USGS40 and USGS41 a; firstly, zero calibration is carried out, namely, the error between the measured value and a standard value is calculated, the measured value is compensated according to the error, the measured value is raised or lowered to eliminate the system error, and finally, the range calibration is carried out to generate a proportionality coefficient k value to eliminate the measurement error.
In the present invention, the sample for the element content determination is preferably a digestion solution. In the present invention, the method for preparing the digestion solution preferably includes:
and mixing the portunus trituberculatus sample with a digesting agent, and performing microwave digestion to obtain a digesting solution.
In the present invention, the digesting agent is preferably concentrated nitric acid; the mass concentration of the concentrated nitric acid is preferably 65-68%; the dosage ratio of the portunus trituberculatus sample to the digesting agent is preferably 0.2 g: (5-10) mL, more preferably 0.2 g: 10 mL.
In the present invention, the microwave digestion procedure preferably comprises: heating to 120 deg.C, and maintaining for 5 min; continuously heating to 150 ℃, and keeping the temperature for 10 min; and continuously heating to 190 ℃ and preserving the heat for 20 min.
After the microwave digestion, the invention preferably further comprises the steps of performing acid dispelling and volume fixing on the obtained microwave digestion system to obtain a digestion solution. The acid-removing operation is not particularly limited in the present invention, and the acid-removing operation known to those skilled in the art may be used. In the present invention, the constant volume reagent preferably comprises water or digestion tank water wash. In the present invention, the content of the elements in the digestion solution is preferably: 1.55-10.91 mg/kg of manganese; 23.12-37.76 mg/kg of arsenic; 3.18-6.69 mg/kg of selenium; 2.88-6.03 mg/kg of rubidium; 0.47-29.67 mg/kg of barium.
In the present invention, the determination of the element content is preferably performed on an inductively coupled plasma mass spectrometer. In the invention, the determination parameters of the element content comprise: the radio frequency power is preferably 1200-1500W, and more preferably 1500W; the plasma gas flow is preferably 15L/min; the carrier gas flow is preferably 0.4L/min; the auxiliary gas flow is preferably 0.4L/min; the temperature of the atomization chamber is preferably 2 ℃; the atomizer is preferably a concentric atomizer.
In the present invention, the elements include manganese, arsenic, selenium, rubidium, and barium.
After the carbon-nitrogen stable isotope ratio and the element content of the blue crab samples in each known producing area are obtained, the method carries out stepwise regression analysis on the carbon-nitrogen stable isotope ratio and the element content of the blue crab samples to obtain the blue crab producing area tracing model taking the carbon-nitrogen stable isotope ratio and the element content as independent variables and the producing area as dependent variables.
In the present invention, the stepwise regression analysis is preferably performed using SPSS software.
In the embodiment of the invention, when the portunus trituberculatus sample is from Zhoushan, Weichang Weifang and Lianchun hong Kong, the origin tracing models of the producing areas are respectively:
Yzhoushan of Zhejiang province＝-114.258δ13C+29.112δ15N+2.761Mn-0.221As+4.339Se+10.337Rb-0.305Ba-1102.796 (1)；
YShandong Weifang (Weifang Shandong)＝-134.746δ13C+32.635δ15N+5.239Mn-0.505As+6.233Se+10.589Rb-0.319Ba-1513.710 (2)；
YJiangsu Lianyun harbor＝-127.798δ13C+24.958δ15N+3.746Mn-0.583As+5.043Se+18.116Rb-0.082Ba-1316.748 (3)。
The invention also provides a method for identifying the origin of the blue crabs, which comprises the following steps:
determining the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected, wherein the elements comprise manganese, arsenic, selenium, rubidium and barium;
substituting the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected into each origin tracing model obtained by the construction method in the technical scheme to obtain a judgment threshold value;
and dividing the discrimination threshold value by using a Fisher discrimination analysis method to obtain the source tracing and producing area of the portunus trituberculatus to be detected.
The invention determines the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected.
In the present invention, the elements include manganese, arsenic, selenium, rubidium, and barium.
In the invention, the parameters for determining the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected are preferably consistent with the technical scheme, and are not described again.
After the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected are obtained, the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected are substituted into each origin tracing model obtained by the construction method in the technical scheme, and a judgment threshold value is obtained.
The substitution operation is not particularly limited, and may be performed by a method known to those skilled in the art.
After the judgment threshold value is obtained, the judgment threshold value is divided by using a Fisher judgment analysis method, and the source tracing place of the portunus trituberculatus to be detected is obtained.
The operation of the Fisher discriminant analysis method is not particularly limited in the present invention, and may be performed by a method known to those skilled in the art. In the invention, the discrimination boundary values are divided by using a Fisher discrimination analysis method, and the discrimination boundary values are divided into which type in which boundary value area.
The construction method and the identification method of the portunus trituberculatus origin tracing model provided by the invention are explained in detail with the following embodiments, but the construction method and the identification method are not to be construed as limiting the scope of the invention.
Example 1
1) Sample collection
All portunus trituberculatus crabs are taken from 9 months in 2020, portunus trituberculatus crab samples in navicular are taken from the navicular culture area, portunus trituberculatus crab samples in the garden are taken from the bay of leizhou, portunus trituberculatus crab samples in the continuous hong are taken from the bay of haizhou, the number of samples of each site is 50, and the total number of the portunus trituberculatus crabs is 150.
2) Lyophilized powder of sample
The collected portunus trituberculatus samples are packed in sterile plastic sealing bags during transportation, refrigerated, stored and transported back to a laboratory, leg muscle of the portunus trituberculatus samples are taken, washed by distilled water, freeze-dried for 36 hours, ground, sieved by a 100-mesh sieve and placed in a dryer for storage for later use.
3) Determination of carbon-nitrogen stable isotope ratio
Accurately weighing 0.2mg of sample, putting the sample into a tinfoil cup, and sending the sample into a stable isotope ratio mass spectrometer for detection; the analysis conditions of the stable isotope ratio mass spectrometer include: the temperature of the combustion furnace is 960 deg.C, the temperature of the reduction furnace is 70 deg.C, the flow rate of helium gas is 100mL/min, and the isotope standard sample adopts USGS40 (delta)13C＝-26.39‰，δ15N ═ 4.52 ‰) and USGS41a (δ)13C＝36.55‰，δ15N47.55 ‰) was performed, isotope standards were inserted into every 20 samples during the measurement process for two-point calibration, and the carbon-nitrogen stable isotope ratio of the obtained blue crab sample is shown in table 1.
4) Determination of multiple elements (Mn, As, Se, Rb, Ba)
1. Weighing 0.2g (accurate to 0.005g) of sample, adding 10mL of concentrated nitric acid (mass concentration of 65-68%) into a microwave digestion tank, screwing an outer cover, placing the microwave digestion tank into a microwave digestion instrument, and digesting the sample according to microwave digestion conditions (microwave digestion program is that (1) the temperature is raised to 120 ℃ and is kept for 5min, (2) the temperature is continuously raised to 150 ℃, and is kept for 10min, and (3) the temperature is continuously raised to 190 ℃, and is kept for 20 min). And cooling to room temperature, opening the digestion tank, driving acid on an electric heating plate, washing the digestion tank for 3-4 times by using water, merging washing liquor into a 50mL volumetric flask, metering the volume to the scale by using water, and detecting by using a machine.
The detection instrument is an inductively coupled plasma mass spectrometer, and the instrument conditions comprise: radio frequency power: 1500W; plasma gas flow rate: 15L/min; carrier gas flow: 0.4L/min; auxiliary air flow rate: 0.4L/min; temperature of the atomization chamber: 2 ℃; an atomizer: a concentric atomizer; simultaneously, reagent blanks were made and 3 replicates of each sample were tested.
The contents of Mn, As, Se, Rb and Ba in the obtained blue crab sample are shown in Table 1.
TABLE 1 determination of elemental content and carbon-nitrogen stable isotope ratio (mean + -SD) of Portunus trituberculatus in different areas
SPSS software is used for carrying out one-factor variance analysis on the measurement results in the table 1, and the different producing areas delta13The average values of C values are ranked in Zhoushan>Linyuankang>Weifang, delta15Respectively Weifang's respective N-value average sorting>Zhoushan (a Chinese character of 'Zhoushan')>In hong Kong, the difference in carbon and nitrogen stable isotopes (P) between the three producing areas is significant<0.001); meanwhile, the content of 5 elements in leg muscles of portunus trituberculatus crabs in three producing areas has different characteristics, Mn and Se in the portunus trituberculatus crabs in the Weifang are obviously higher than those in the Zhoushan and the Negrong, while the As content in the Zhoushan producing area is high, the Rb and Ba content in the Negrong is highest, especially the Ba content is As high As 29.67mg/kg, and the Weifang (P) are obviously higher than those in the Zhoushan and the Weifang (P)<0.001)。
Performing stepwise regression analysis on the element content of the portunus trituberculatus sample in combination with the carbon-nitrogen stable isotope ratio by using SPSS 22.0, and respectively establishing Fisher's origin tracing models (discriminant models) of 3 origins, which are as follows:
Yzhoushan of Zhejiang province＝-114.258δ13C+29.112δ15N+2.761Mn-0.221As+4.339Se+10.337Rb-0.305Ba-1102.796 (1)；
YShandong Weifang (Weifang Shandong)＝-134.746δ13C+32.635δ15N+5.239Mn-0.505As+6.233Se+10.589Rb-0.319Ba-1513.710 (2)；
YJiangsu Lianyun harbor＝-127.798δ13C+24.958δ15N+3.746Mn-0.583As+5.043Se+18.116Rb-0.082Ba-1316.748 (3)；
Meanwhile, discriminant analysis scatter diagrams of portunus trituberculatus samples of three producing areas, namely Zhoushan, Weifang and Lingyun harbor, are obtained, and are shown in figure 1.
Directly predicting and verifying 150 blue crab samples, namely initial verification; and performing prediction verification on 150 blue crab samples by adopting a leave-one-out cross verification method, which is called cross verification. The initial validation and cross-validation results obtained are shown in table 2.
TABLE 2 origin classification results and percentage of correct classification of observed values for Portunus trituberculatus samples
As can be seen from table 2: the discrimination accuracy of the back-up test is 100%, which shows that the seven indexes can effectively realize the discrimination of the producing areas of the portunus trituberculatus, the Weifang and the Never-cloud-harbor portunus trituberculatus.
Comparative example 1
Sodium, magnesium, aluminum, potassium, calcium, titanium, manganese, iron, cobalt, copper, zinc, arsenic, selenium, rubidium, strontium, silver, cadmium and barium in the digestion solution were measured by the method of example 1; according to the indexes given in the table 3 as independent variables and the producing area as dependent variables, a producing area tracing model is established for 150 blue crab samples by using the SPSS, and the producing areas of the 150 blue crab samples are subjected to initial verification and cross verification by using the obtained producing area tracing model, and the results are shown in the table 3.
TABLE 3 prediction accuracy of the origin tracing model and the origin tracing model established by different indexes on the concentrated samples
As can be seen from table 3: when the carbon-nitrogen stable isotope ratio and the element content are respectively and solely used for independent judgment, partial misjudgment can occur in the judgment result, and after the carbon-nitrogen stable isotope ratio and the element content are jointly judged, the judgment accuracy is greatly improved and reaches 100%, and the judgment effect is good. In addition, in order to further screen effective indexes and exclude elements with high relevance, five elements of Mn, As, Se, Rb and Ba and delta are further screened13C、δ15N is combined for judgment, and the accuracy can reach 100%; verifying remaining elements simultaneously with delta13C、δ15And (3) the N combined judgment result shows that misjudgment occurs during cross validation, the judgment result accuracy is slightly lower than that of the combined judgment of the five elements and the carbon and nitrogen stable isotope, and the five elements of Mn, As, Se, Rb, Ba and delta obtained by screening are shown13C、δ15The N-joint discrimination can meet the requirements of less detection indexes and high detection precision, and the discrimination effect is optimal.
As can be seen from the above embodiments: the fingerprint characteristics of the element and the carbon-nitrogen stable isotope in leg tissues of the blue crabs from different producing areas show obvious regional difference, and the delta is measured by respectively utilizing a stable Isotope Ratio Mass Spectrometer (IRMS) and an inductively coupled plasma mass spectrometer (ICP-MS)13C、δ15N, Mn, As, Se, Rb and Ba, and the element difference in leg muscle tissues of the portunus trituberculatus samples from different production places is judged by using a mathematical statistical analysis method to obtain a tracing model of stable isotope fingerprint characteristics of the portunus trituberculatus production places, and the origin places are distinguished by using the tracing model, so that the information of the portunus trituberculatus production places can be accurately and fairly identified.
The method starts from the difference of multi-element and carbon-nitrogen stable isotope of the organism, only uses seven indexes of common carbon-nitrogen stable isotope ratio, Mn content, As content, Se content, Rb content and Ba content to establish a discrimination model, has simple used instruments, few detection indexes, high detection precision, good reproducibility, simple and convenient method and strong operability, reduces the uncertainty based on commodity labels and subjective judgment, and has the discrimination rate of the origin of the portunus trituberculatus production area reaching 100 percent.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A method for constructing a portunus trituberculatus origin tracing model comprises the following steps:
measuring the carbon-nitrogen stable isotope ratio and the element content of a blue crab sample in a known producing area; the elements comprise manganese, arsenic, selenium, rubidium and barium;
and carrying out stepwise regression analysis on the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample to obtain a origin tracing model of blue crab with the carbon-nitrogen stable isotope ratio and the element content as independent variables and the origin as dependent variables.
2. The construction method according to claim 1, wherein leg muscles of the blue crab sample are used for measuring the carbon-nitrogen stable isotope ratio and the element content of the blue crab sample.
3. The construction method according to claim 1 or 2, wherein the determination parameters of the carbon-nitrogen stable isotope ratio include: the temperature of the combustion furnace is 900-1020 ℃, the temperature of the column box is 40-70 ℃, and the flow rate of helium is 80-100 mL/min.
4. The construction method of claim 3, wherein the process of determining the carbon-nitrogen stable isotope ratio of the blue crab sample further comprises two-point correction of the isotope standard sample by using USGS40 and USGS41a, wherein the USGS40 comprises delta13C＝-26.39‰，δ15N is-4.52 permillage; the USGS41a includes delta13C＝36.55‰，δ15N＝47.55‰。
5. The construction method according to claim 1 or 4, characterized in that in the process of determining the carbon-nitrogen stable isotope ratio of the blue crab samples, every 20 blue crab samples are inserted into an isotope standard sample for two-point correction; each blue crab sample was tested in parallel for carbon-nitrogen stable isotope 3 times.
6. The construction method according to claim 1 or 2, wherein the sample for element content determination is a digestion solution, and the preparation method of the digestion solution comprises:
mixing a portunus trituberculatus sample with a digesting agent, and performing microwave digestion to obtain a digesting solution;
the digesting agent comprises concentrated nitric acid; the dosage ratio of the portunus trituberculatus sample to the digesting agent is 0.2 g: (5-10) mL.
7. The construction method according to claim 6, wherein the microwave digestion procedure comprises: heating to 120 deg.C, and maintaining for 5 min; continuously heating to 150 ℃, and keeping the temperature for 10 min; and continuously heating to 190 ℃ and preserving the heat for 20 min.
8. The construction method according to claim 6, wherein the parameters for measuring the content of the elements comprise:
radio frequency power: 1200-1500W; plasma gas flow rate: 15L/min; carrier gas flow: 0.4L/min; auxiliary air flow rate: 0.4L/min; temperature of the atomization chamber: 2 ℃; an atomizer: a concentric atomizer.
9. The method of establishing of claim 1, wherein the stepwise regression analysis is performed using SPSS software.
10. The method for identifying the origin of the blue crabs is characterized by comprising the following steps:
determining the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected, wherein the elements comprise manganese, arsenic, selenium, rubidium and barium;
substituting the carbon-nitrogen stable isotope ratio and the element content of the portunus trituberculatus to be detected into each origin tracing model obtained by the construction method according to any one of claims 1 to 9 to obtain a judgment threshold value;
and dividing the discrimination threshold value by using a Fisher discrimination analysis method to obtain the source tracing and producing area of the portunus trituberculatus to be detected.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110946051.3A CN113671013A (en) | 2021-08-17 | 2021-08-17 | Construction method and identification method of portunus trituberculatus origin tracing model |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110946051.3A CN113671013A (en) | 2021-08-17 | 2021-08-17 | Construction method and identification method of portunus trituberculatus origin tracing model |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113671013A true CN113671013A (en) | 2021-11-19 |
Family
ID=78543421
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110946051.3A Pending CN113671013A (en) | 2021-08-17 | 2021-08-17 | Construction method and identification method of portunus trituberculatus origin tracing model |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113671013A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109283240A (en) * | 2018-11-06 | 2019-01-29 | 中国农业科学院农产品加工研究所 | Identify the method in the maca place of production |
| CN110646496A (en) * | 2018-06-26 | 2020-01-03 | 中国科学技术大学 | Method for identifying hairy crab producing area by strontium isotope |
| CN110687189A (en) * | 2019-10-08 | 2020-01-14 | 浙江省检验检疫科学技术研究院 | Method for distinguishing milk producing area tracing and identifying based on multiple elements and multiple indexes |
| CN111024802A (en) * | 2019-12-26 | 2020-04-17 | 中国水产科学研究院黄海水产研究所 | Stichopus japonicus origin tracing model construction and identification method based on lead-Pb stable isotope |
-
2021
- 2021-08-17 CN CN202110946051.3A patent/CN113671013A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110646496A (en) * | 2018-06-26 | 2020-01-03 | 中国科学技术大学 | Method for identifying hairy crab producing area by strontium isotope |
| CN109283240A (en) * | 2018-11-06 | 2019-01-29 | 中国农业科学院农产品加工研究所 | Identify the method in the maca place of production |
| CN110687189A (en) * | 2019-10-08 | 2020-01-14 | 浙江省检验检疫科学技术研究院 | Method for distinguishing milk producing area tracing and identifying based on multiple elements and multiple indexes |
| CN111024802A (en) * | 2019-12-26 | 2020-04-17 | 中国水产科学研究院黄海水产研究所 | Stichopus japonicus origin tracing model construction and identification method based on lead-Pb stable isotope |
Non-Patent Citations (2)
| Title |
|---|
| 张政权等: "矿物元素结合稳定同位素的中华绒螯蟹产地溯源", 《食品科学》, vol. 41, no. 24, pages 125 - 130 * |
| 胡桂仙;邵圣枝;张永志;朱加虹;赵首萍;袁玉伟;: "杨梅中稳定同位素和多元素特征在其产地溯源中的应用", 核农学报, no. 12 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108982692B (en) | Method for distinguishing milk powder producing area by element analysis-stable isotope mass spectrum | |
| Huang et al. | Qualitative discrimination of Chinese dianhong black tea grades based on a handheld spectroscopy system coupled with chemometrics | |
| Baffi et al. | Food traceability using the 87 Sr/86 Sr isotopic ratio mass spectrometry | |
| CN111735886B (en) | Pretreatment method of thermally processed food and detection method of acrylamide | |
| CN111024802B (en) | Stichopus japonicus origin tracing model construction and identification method based on lead-Pb stable isotope | |
| Bai et al. | Multi-element analysis of three Codonopsis Radix varieties in China and its correlation analysis with environmental factors | |
| Bai et al. | Elemental analysis of wild Eriocheir sinensis: Determining the geographic origin and human health risk assessment | |
| CN111024665A (en) | Method for measuring content of organic selenium in selenium-enriched rice or product prepared from selenium-enriched rice | |
| CN102899409B (en) | PCR identification kit for Thunberg fritillary bulb and identification method | |
| CN109239758B (en) | Method for detecting and analyzing gamma energy spectrum nuclides in biological sample | |
| CN105738516B (en) | A kind of method that mark characterized by biogenic amine differentiates bee venom species | |
| CN113671013A (en) | Construction method and identification method of portunus trituberculatus origin tracing model | |
| Taylor et al. | Atomic spectrometry update: review of advances in the analysis of clinical and biological materials, foods and beverages | |
| CN107884301B (en) | Method for identifying fermentation year of thick broad-bean sauce | |
| CN108593829A (en) | A kind of Stable Isotopic Analysis method for influence factor of tracing to the source the milk place of production | |
| CN102759563A (en) | Method for determining chrominum content in edible gelatine | |
| CN107421845A (en) | A kind of quick analysis and identification Weihai sea cucumber, Russian red ginseng and the method for eggplant ginseng | |
| CN110596079A (en) | Method for effectively distinguishing sturgeons in different producing areas | |
| CN106074679B (en) | A kind of root of kudzu vine particle and its Chinese medicine preparation | |
| CN105136899B (en) | A kind of method for differentiating different manufacturers, different geographical and producing donkey-hide gelatin | |
| CN109900532B (en) | Pretreatment method of lycium barbarum during determination of stable isotope of dried lycium barbarum | |
| CN105651705A (en) | Method for nondestructively detecting content of counterfeit ingredients in cordyceps sinensis powder/tablets | |
| CN112649276A (en) | Method for digesting geological sample by using ammonium bifluoride or ammonium fluoride | |
| CN111610073A (en) | Preparation method of heavy metal-containing Yunnan pseudo-ginseng standard substance | |
| KR102348622B1 (en) | A method of provide data for checking pork in Korea |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211119 |