CN108490088A - A kind of detection method of the animal derived product residue of veterinary drug of detection - Google Patents
A kind of detection method of the animal derived product residue of veterinary drug of detection Download PDFInfo
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- CN108490088A CN108490088A CN201810210283.0A CN201810210283A CN108490088A CN 108490088 A CN108490088 A CN 108490088A CN 201810210283 A CN201810210283 A CN 201810210283A CN 108490088 A CN108490088 A CN 108490088A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
The invention belongs to technical field of food safety detection, disclose detection method that is a kind of while quickly detecting animal derived product veterinary drug residue, analyze Extraction solvent, extracting method and purification techniques suitable for sample to be tested, extract component to be measured in purification sample；63 kinds of determinands are detached using liquid chromatogram separation condition, 6 groups of isomers is made to obtain baseline separation；MS/MS detectors, ESI ion sources, schedul MRM patterns, three ions confirmation, inside/outside scalar quantity technology is selected to detect sulfamido, trimethoprim (TMP), quinolones, beta receptor agonist, Tetracyclines, macrolides, quinolines, nitro glyoxaline, cephalo-type, 10 major class of penicillins totally 63 kinds of veterinary drugs simultaneously.Method provided by the invention is applied to the daily detection to 10 63 kinds of veterinary drugs of major class such as sulfamido in animal product, by improving working efficiency, reducing testing cost, quickly answers food-safe accident, generates social benefit and economic benefit.
Description
Technical field
The invention belongs to technical field of food safety detection more particularly to a kind of animal derived product veterinary drug residues of detection
Detection method.
Background technology
Currently, the prior art commonly used in the trade is such：
China is the breeding big country of animal derived food, and according to customs statistics, 2013, Export of Chinese Aquatic Products reached 384
Ten thousand tons, 19,400,000,000 dollars, increase by 4.2% and 7.2% on a year-on-year basis, keeps within continuous 13 years product water outlet amount to rank first place status, aquatic products
The proportion that product export amount accounts for China's agricultural and sideline product total export reaches 22.4%.As aquaculture big country, country goes out successively
Platform policy encourages product water outlet, but aquaculture product because environmental pollution, transmission, germplasm degenerate and cultivation density it is big etc. because
Element, cultivated animals incidence improve year by year, and illegal raiser, retailer are to seek economic interests and Misuse forbidden drugs or limit
It is remained existing despite repeated prohibition with drug behavior, animal product veterinary drug residue control difficulty increases.Antibiotic residue problem makes China animal
Property products export in source baffles, such as the shrimp in the outlet U.S., fish product are due to being detected Furaxone metabolite, chloramphenicol, peacock
The forbidden drugs such as malachite green make the U.S. detain measure automatically to the shrimp product implementation of China's export, seriously affect China's aquatic products
Export to American market.
Residue of veterinary drug refers to animal after having used veterinary drug, accumulates or be stored in zooblast, tissue or intraorganic medicine
Object prototype, virose metabolin or impurity.Mainly there is sulfanilamide (SN) in China in terms of aquatic products and livestock-raising using more veterinary drug
Class, trimethoprim (TMP), quinolones, beta-receptor agonist class, Tetracyclines, macrolide, chloromycetin, glucocorticoid
The compounds such as class, cephalo-type, penicillins, sex hormones and itrofurans.It abuses these drugs and easily causes animal husbandry and fishery product
Serious pollution, and entered in the animal derived foods such as muscle by metabolic pathway, accumulated in human body eventually by food chain,
And these drugs have lead to allergic reaction and make the side effects such as human body develops immunity to drugs, or even the drug having with it is carcinogenic,
The serious side effects such as teratogenesis.But it is driven by economic interests, illegal raiser is illegal to be happened occasionally using the case where forbidden drug, mesh
It is preceding illegal using forbidden drug, abuse antimicrobial and medicated premix etc., it is to cause China's animal derived food veterinary drug residue
It is exceeded, the main reason for causing animal products consumption safety and food trade controversial issue.Therefore, monitoring multiple types veterinary drug is in animal sources
Property product in residual there is positive effect to ensureing food security and food trade, and expand animal derived products export
Effective measures.
Since different types of veterinary drug nature difference is larger, existing wild animal resources are similar generally according to chemical constitution
The corresponding detection method of drug development of the same clan, such as respectively be directed to sulfamido, quinolones, Tetracyclines, beta-receptor agonist
And the inhomogeneities drug such as chloromycetin establishes corresponding detection method, the medicament categories that these analysis methods can be analyzed simultaneously compared with
It is few, current veterinary drug type cannot be met and be continuously increased, the demand that analysis throughput is continuously improved.In modern detection field, no
Generic pesticide multi-residues Simultaneous Detection application reaches its maturity, but the report detected simultaneously in relation to multiple types residue of veterinary drug
Road is less, also without corresponding standard method.
With the fast development of liquid chromatography-tandem mass spectrometry technology and Sample Pretreatment Technique, animal derived complexity food
Analysis is possibly realized while veterinary drug multi-residue determination range in matrix develops into a variety of different classes of veterinary drugs from single classification.
Sample drug residue is analyzed, and the sample pre-treatments occupied time accounts for 95% or more of the bulk analysis time, and
The analysis method of residue of veterinary drug is mostly using extraction, tune acid-base value, extraction, concentration, dissolving, SPE purifications, drying, dissolving at present
Constant volume, upper machine measurement and etc., pre-treatment step is more, cumbersome, not only expends a large amount of reagents, increases testing cost, is easily introduced dry
Object is disturbed, object is caused to lose, and causes testing staff's working efficiency low.Sample pre-treatments be to ensure that testing result it is accurate,
Effectively shorten the committed step of detection cycle.
Therefore, developing the method that multiple types veterinary drug quickly detects simultaneously in the simply animal derived product of pre-treatment has weight
Want Social benefit and economic benefit.
In conclusion problem of the existing technology is：
(1) detection method of the animal derived product residue of veterinary drug of existing detection only detect a kind of residue of veterinary drug and caused by inspection
Survey efficiency is low, detection cycle is long, testing cost is high.
(2) since different types of veterinary drug nature difference is larger, existing wild animal resources are generally according to chemical constitution
Similar drug of the same clan develops corresponding method, such as using GB/T 21313-2007, GB/T 22286-2008, SN/T
Beta-receptor agonist class residue of veterinary drug in the property product of 1924-2007 analyzing animals source, using GB/T 20366-2006, GB/T
Quinolones residue of veterinary drug in 20751-2006, GB/T 21312-2007 analyzing animals source property product, using GB/T 20759-
2006, Sulfonamides remain in No. 1077 GB/T 21316-2007, Ministry of Agriculture bulletin -1-2008 analyzing animals source property products,
Using Tetracyclines residue of veterinary drug in GB/T 21317-2007, GB/T 22990-2008 analyzing animals source property product, using GB/
Chloromycetin residue of veterinary drug in T 20756-2006 analysis aquatic products, using GB/T19857-2005, GB/T 20361-2006 points
Analyse triphenylmenthane series bactericidal agent, cephalosporin analog antibiotic analyzed using GB/T 20755-2006, using GB/T 21314-2007,
GB/T 22960-2008, SN/T 1988-2007, SN/T 1988-2007 analyze penicillin antibiotics, using GB/T
22957-2008, No. 1031 bulletin -2-2008 of the Ministry of Agriculture, SN/T 2222-2008 analyze glucocorticoids medicine, using GB/T
23406-2009, SN/T 1928-2007 analyze nitro glyoxaline deinsectization medicine, using No. 2086 bulletin -5-2014 analyses of the Ministry of Agriculture
Quinolines, male sex hormone drug is analyzed using SN/T 2677-2010, and polypeptide is analyzed using SN/T 2748-2010
Veterinary drug analyzes macrolide antibiotics using SN/T 1777.2-2007.It is only agriculture in standard method prevailing for the time being in force
Bulletin-the 1-2008 of portion 1077《17 kinds of sulfamidos and the measurement liquid of 15 kinds of quinolones medicament relict amounts in animal derived food
Phase chromatography-tandem mass spectrometry》Sulfamido and quinolones veterinary drug can be analyzed simultaneously, and in addition to this, there is presently no can the same time-division
Analyse the standard detecting method of multiple types residue of veterinary drug.
The Detecting Pesticide field in modern food, the method application that different classes of pesticide residue detects simultaneously is
Maturation, but in related food multiple types residue of veterinary drug simultaneously detect not only without standard method and document report it is also less, with
The fast development of liquid chromatography-tandem mass spectrometry detection technique and Sample Pretreatment Technique, it is animal derived complexity food substrate in
While veterinary drug multi-residue determination range develops into a variety of different classes of veterinary drugs from single classification quantification and qualification become can
Energy.The scholars such as domestic Guo De China, bear Chunlan also study veterinary drug multi-residue determination.
Guo Dehua etc. establishes Solid Phase Extraction-high performance liquid chromatography-tandem mass method while measuring 6 in animal derived food
The detection method of 76 kinds of residues of veterinary drug of class, sample is using acetonitrile and contains Mg+2Citrate buffer solution extracts, with slow after removal organic phase
Fliud flushing weight is molten, and polymer and cation exchange solid-phase extraction column series connection purification, and liquid chromatography-tandem mass spectrometry measures beta-receptor simultaneously
Agonist, triphenylmethane, nitro glyoxaline, Benzodiazepines, sulfamido and benzimidazole 76 kinds of veterinary drugs of totally 6 class.The party
Method fails to detect the common veterinary drugs of aquacultures such as quinolones, Tetracyclines and macrolides, while the method is net using twin columns
Change method has the deficiencies such as testing cost is high, operation difficulty is big, time for sample pretreatment is long, and testing staff's efficiency is low.
Bear Chunlan etc. establishes high performance liquid chromatography-tandem mass while measuring quinolones in aquatic products, macrolide
Class, sulfamido, trimethoprim (TMP), woods can amine, nitro glyoxaline, quinolines and polypeptide class 38 kinds of residues of veterinary drug of totally 8 class, examination
Sample is extracted with 1% acetic acid acetonitrile solution, and after chilled centrifugation and n-hexane purification, HPLC-MS/MS is measured.The method
Pre-treatment purifying step only carries out ungrease treatment, can not effectively remove and interfere larger phospholipid impurity in sample, can not be effective
Generated matrix effect when HPLC-MS/MS detections is eliminated or reduced, there is larger impact to measurement result.
Cao Yafei etc. establishes one kind and quickly measuring macrolides, sulfamido, quinolones and Tetracyclines in cheese
Na is being added in the analysis method of totally 50 kinds of residues of veterinary drug, sample2After edta buffer liquid and ceramics proton, in 5% acetate acetonitrile
Under the conditions of carry out mechanical shaking extraction, sodium chloride and anhydrous sodium sulfate are for saltouing, after the purification of C18 column adsorbents, for liquid chromatogram-
Tandem mass spectrum measures.The method only measures four classes, 50 kinds of residues of veterinary drug in cheese sample, and detection substrate range is single and detects object only
Limit four class veterinary drugs；Using absorption type Solid Phase Extraction column purification sample extracting solution, operation difficulty is larger, and pillar is easy to block up and causes to repeat
Property is poor.
Solve the difficulty and meaning of above-mentioned technical problem：
Using the technology of the present invention route exploitation establish " 10 major class, 63 kinds of Residual Veterinary Medicines are quickly surveyed in animal derived food
Determine method and pass through formula SPE purifications-high performance liquid chromatography-tandem mass method ", can be applied to amino sulfanilamide (SN) in animal derived food,
Sulphaguanidine, sulphadiazine, sulfapryidine, sulphathiazole, sulfamethazine, sulfacetamide, 5-methoxysulfadiazine, sulfanilamide (SN)
The different pyrimidine of diformazan, daimeton, sulfamethoxy-pyridazine, sulfamethoxazole, sulfamethazole, sulfabenzamide,
Sulfachlorpyridazine, sulfaquinoxaline, fanasil, N'-phenylsulfanilamide pyrene azoles, sulfanilamide (SN) Sulfafurazole, bensulfa
Pyrimidine, ayerlucil, the pyridine of kynix benzyl, flumequine, oxolinic acid, Norfloxacin, Enoxacin, Ciprofloxacin, training fluorine
Sha Xing, Lomefloxacin, Dan Nuosha stars, Enrofloxacin, Ofloxacin, marbofloxacin, fleraxacin, sarafloxacin, Si Baisha stars,
Orbifloxacin, Difloxacin, acidum nalidixicum, salbutamol, Terbutaline, Cimaterol, Ke Lunsailuo, Ractopamine, benzene second
Hydramine A, horse-ride step spy sieve, Clenbuterol, ampicillin, benzyl penicillin, Ceftiofur, carbadox, quinoline sigh ketone, quinoline match more, Fourth Ring
Element, terramycin, lincomycin, tylosin, spiramvcin, Tilmicosin, clindamycin, metronidazole, Dimetridazole, Lip river nitre reach
The rapid screening of 63 kinds of veterinary drug projects of azoles measures.This method has high sensitivity, selectivity strong, accuracy, repeatability and reproduction
Property meets《Good Laboratory controls specification Physico-chemical tests》With《Retention analysis controls guide》Requirement；This method is realized a variety of
Class is remained more while being analyzed, and primary measure can monitor 10 major class, 63 kinds of veterinary drug projects simultaneously, more can eat agricultural production by full-scope safeguards
Product safety, quickly answers food-safe accident, assists enterprise's breaks through foreign tradeing mutual compensation, promotes the disengaging of agricultural product
Mouth trade, has larger social benefit；This method is used through formula SPE purification techniques, and sample pre-treatments are simply, conveniently, soon
Victory, realize multiple types multiple medicine it is residual while detect, can substantially reduced intensity of workers, testing agency can pass through save examination
Agent spending reduces testing cost, improves working efficiency, shortens detection cycle, generates bigger economic benefit.Therefore, this method can expire
The job requirement quick to testing laboratory, accurate, efficient, reliable of foot society, has stronger dissemination.
Invention content
In view of the problems of the existing technology, the present invention provides 10 major class, 63 kinds of beasts in a kind of animal derived product of detection
The remaining detection method of medicine.Detection sensitivity of the present invention is high, quantitative accurate, qualitative reliable, overcomes mesh former standard method only
A kind of residue of veterinary drug of detection and caused by the deficiencies of detection efficiency is low, detection cycle is long, testing cost is high.
The invention is realized in this way a kind of detection method of the animal derived product residue of veterinary drug of detection, is：
(1) suitable Extraction solvent and extracting method are analyzed, the object that property in sample to be tested differs all is extracted,
Determinand is set to be transferred in extracting solution from tissue；
(2) it selects, suitable for purification techniques, to purify sample extracting solution using by formula PRiME HLB solid phase extraction columns, lead to
The purifying step is crossed, matrix effect can be effectively eliminated or mitigate, but is not caused damages again to object, while being purified by formula
Technology purifies scheme compared to hold mode, can effectively overcome palpus activation, balance, upper prop, elution and the elution of hold mode purification techniques
Extracting solution need only directly be crossed column, collect efflux, quick, time saving and favorable reproducibility easy to operate by equal tedious steps.
(3) suitable chromatographic condition is selected, the optimization to chromatographic column, column temperature, mobile phase and gradient condition, separation 63 are passed through
Kind determinand, makes parent ion and the identical isomer of daughter ion reach baseline separation, while when the reservation of 63 kinds of determinands
Between distribute it is reasonable, so that mass spectrum has each pair of ion enough number of scan points；
(4) series connection quadrupole rod detector, ESI ion sources, scheduled MRM patterns, three ions confirmation, inside/outside are selected
Scalar quantity technology detects sulfamido, trimethoprim (TMP), quinolones, beta-receptor agonist, Tetracyclines, macrolide simultaneously
Class, quinolines, nitro glyoxaline, cephalo-type, 10 major class of penicillins totally 63 kinds of veterinary drugs.
Further, the detection method of the animal derived product residue of veterinary drug of detection includes：
(1) optimize Mass Spectrometry Conditions, sample introduction pattern pumped using wriggling needle, the Mass Spectrometry Conditions of each object are optimized,
Optimize the impact energy of each pair of ion, remove cluster voltage, lens voltage Mass Spectrometry Conditions, forms MRM ion pairs；Select sensitivity highest
Fragment ion establishes the triple quadrupole bar MRM of 10 major class, 63 kinds of veterinary drug projects as quota ion, second a height of qualitative ion
Ion pair.Optimize atomization gas, spray voltage, auxiliary gas, heating temperature source parameters.
(2) optimize liquid chromatogram separation condition, select mass spectral analysis column, adjust the liquid phases such as mobile phase, flow velocity and column temperature point
From condition.Through research, select waters Attantis T3 chromatographic columns as analytical column, 0.1% aqueous formic acid and methanol+second
Nitrile (50：50(v：V), contain 0.1% formic acid) it is tested component as eluent gradient elution separation, 35 DEG C of column temperature is used as liquid chromatogram
Separation condition, under the conditions of this liquid phase separation, 63 kinds of object separation are preferably, peak shape is sharp, retention time distribution is reasonable, retain
Time terminates from 3min beginning 23min, and the setting of gradient makes 6 groups of isomers in measured object that can obtain baseline separation, special
It is not that 5-methoxysulfadiazine, daimeton and kynix three kinds of isomers of this group obtain baseline point
From.The retention time for determining 63 kinds of objects establishes scheduled MRM ion pairs；Establish liquid chromatography-tandem mass spectrometry detection
The method of 10 major class totally 63 kinds of veterinary drugs；The standard curve for establishing 63 kinds of drugs determines minimum detection limit and the range of linearity；
(3) select ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization influence extraction efficiency because
Element.By comparing the extraction efficiency of 10 kinds of Extraction solvents, project uses 1% formic acid Acidifying acetonitrile-Na2EDTA is with homogeneous extraction side
Formula extracts determinand in sample.The sample extraction method, a pre-treatment can extract 10 major class, 63 kinds of veterinary drugs in sample, effectively
Overcome current variety classes detection of veterinary drugs in food need to using various criterion method and caused by inefficiency, detection cycle length etc.
It is insufficient.
(4) suitable thickening-purification technology technical finesse sample extracting solution is selected, matrix effect is mitigated or eliminated.Through comparing,
Method is used by the way that after the purification of formula PRiME HLB solid phase extraction columns, which is suitble to the veterinary drug that most of property differs,
It will not cause damages to object, and fat and phosphatide impurity can be effectively removed, the results showed that 99% phosphatide etc. can be removed
Nonpolar chaff interferent.After purification techniques processing, baseline noise is can obviously reduce, signal-to-noise ratio is improved, matrix is mitigated or eliminated
Effect.
(5) inside authentication is carried out, selects representational sample low in detection lower bound, twice of detection lower bound and ten times of measurement
Three levels of limit are added recycling verification accuracy；Select positive or negative sample addition recycling verification precision.Through
Verification, this method have following technical indicator.
1. under conditions selected, 63 kinds of object separation are preferable, peak shape is sharp, and retention time distribution is reasonable, retention time
Terminate from 3min beginning 23min, the setting of gradient makes 6 groups of isomers in measured object that can obtain baseline separation；
2. 17 kinds of fluoroquinolones, 22 kinds of sulfa drugs, a kind of trimethoprim (TMP), 3 kinds of quinolines, 5 kinds big
Cyclic lactone class drug, ampicillin, benzyl penicillin, the minimum quantitative limit of Ceftiofur are 10 μ g/kg, and tetracycline and terramycin are most
Light basis weight is limited to 50 μ g/kg, and 8 kinds of beta-receptor agonists and 3 kinds of the minimum of nitroimidazole are quantitatively limited to 0.5 μ g/kg；Detection limit is full
Foot country relevant laws and regulations are to these residue of veterinary drug limitation requirements.
3. the linear regression coeffficient of 6 concentration points of 63 kinds of veterinary drug projects is 0.9928 or more；
4. using shrimp, fish and pig muscle as background sample, three concentration level mark-on reclaims experiments, indoor recovery of standard addition exists
Between 52.8%~145%, the coefficient of variation of 6 measurement is between 4.6%~28.3%；Using pork as background sample, four
Eight measurement results of measuring unit, between room recovery of standard addition between 55.2%~140%, between room precision 6.1%~
Between 26.2%.
5. being compared to blank sample and blank addition standard results, the region that target analytes occur does not have signal dry
It disturbs, each object retention time is stablized, method high specificity；
6. choosing Enrofloxacin, sulfamethoxazole and tetracycline project, compare this law and the difference of national standard method, t
Value is respectively 1.907,1.94,0.963, respectively less than t0.05,14=2.145, illustrate method that project is worked out and standard method without
Notable difference；Selection standard substance (proficiency testing remaining sample) carries out two to Enrofloxacin, sulphadiazine and sulfamethoxazole
Kind method compares, and the relative differences of two kinds of assay methods are less than 20%, meet the requirement of research project plan charter；
7. matrix effect evaluation result shows that matrix effect does not influence the critical index of the screening technique.
(6) verified carrying out room to the detection method established.With four units, eight testing result accuracy in computation indexs
The rate of recovery and the reproducibility index coefficient of variation.The rate of recovery between 55.2%~140.0%, 8 coefficient of variation 6.1%~
Between 26.2%.
Another object of the present invention is to provide a kind of using shrimp as the detection of the animal derived product residue of veterinary drug of detection of object
Method, including：
Optimize Mass Spectrometry Conditions, determine the parent ion of more than 50 kinds of veterinary drug, select two pairs of daughter ions, optimizes the collision of each pair of ion
Cluster voltage, lens voltage Mass Spectrometry Conditions can, be removed, MRM ion pairs are formed；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature
Spend source parameters；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations items such as mobile phase, flow velocity and column temperature
Part makes parent ion and the identical isomerism physical efficiency of daughter ion reach and efficiently separates；Establish liquid chromatography-tandem mass spectrometry detection 9
The method of major class totally 50 kinds of veterinary drugs；The standard curve for establishing 50 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；It establishes animal derived
The sample-pretreating method of 9 50 kinds of detection of veterinary drugs in food of major class in product；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement
Three levels are added recycling verification accuracy；Select positive or negative sample addition recycling verification precision.
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
Another object of the present invention is to provide a kind of using bullfrog leg as the animal derived product residue of veterinary drug of detection of object
Detection method, including：
Optimize Mass Spectrometry Conditions, determine the parent ion of more than 50 kinds of veterinary drug, select two pairs of daughter ions, optimizes the collision of each pair of ion
Cluster voltage, lens voltage Mass Spectrometry Conditions can, be removed, MRM ion pairs are formed；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature
Spend source parameters；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations items such as mobile phase, flow velocity and column temperature
Part makes parent ion and the identical isomerism physical efficiency of daughter ion reach and efficiently separates；Establish liquid chromatography-tandem mass spectrometry detection 9
The method of major class totally 50 kinds of veterinary drugs；The standard curve for establishing 50 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；It establishes animal derived
The sample-pretreating method of 9 50 kinds of detection of veterinary drugs in food of major class in product；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement
Three levels are added recycling verification accuracy；Select positive or negative sample addition recycling verification precision.
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
Another object of the present invention is to provide a kind of using the flesh of fish as the inspection of the animal derived product residue of veterinary drug of detection of object
Survey method, including：
Optimize Mass Spectrometry Conditions, determine the parent ion of more than 50 kinds of veterinary drug, select two pairs of daughter ions, optimizes the collision of each pair of ion
Cluster voltage, lens voltage Mass Spectrometry Conditions can, be removed, MRM ion pairs are formed；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature
Spend source parameters；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations items such as mobile phase, flow velocity and column temperature
Part makes parent ion and the identical isomerism physical efficiency of daughter ion reach and efficiently separates；Establish liquid chromatography-tandem mass spectrometry detection 9
The method of major class totally 50 kinds of veterinary drugs；The standard curve for establishing 50 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；It establishes animal derived
The sample-pretreating method of 9 50 kinds of detection of veterinary drugs in food of major class in product；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement
Three levels are added recycling verification accuracy；Select positive or negative sample addition recycling verification precision.
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
Another object of the present invention is to provide a kind of using pig muscle as the animal derived product residue of veterinary drug of detection of object
Detection method, including：
Optimize Mass Spectrometry Conditions, determine the parent ion of more than 50 kinds of veterinary drug, select two pairs of daughter ions, optimizes the collision of each pair of ion
Cluster voltage, lens voltage Mass Spectrometry Conditions can, be removed, MRM ion pairs are formed；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature
Spend source parameters；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations items such as mobile phase, flow velocity and column temperature
Part makes parent ion and the identical isomerism physical efficiency of daughter ion reach and efficiently separates；Establish liquid chromatography-tandem mass spectrometry detection 9
The method of major class totally 50 kinds of veterinary drugs；The standard curve for establishing 50 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；It establishes animal derived
The sample-pretreating method of 9 50 kinds of detection of veterinary drugs in food of major class in product；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement
Three levels are added recycling verification accuracy；Select positive or negative sample addition recycling verification precision.
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
In conclusion advantages of the present invention and good effect are：
The present invention 10 major class 63 such as sulfamido suitable for the animal derived products such as shrimp, bullfrog leg, the flesh of fish and animal muscle
Kind of detection of veterinary drugs in food, the sensitivity of this method meet relevant laws and regulations and the regulation of trading country, and standard curve is linear, precision
(RSD), sample recovery of standard addition, accuracy meet GB/T27404-2008 annex F《The confirmation of detection method》Requirement.This hair
The method of bright offer is applied to the daily detection to 10 63 kinds of veterinary drugs of major class such as sulfamido in animal product, generates social benefit
And economic benefit.
The present invention uses newest sample purification pretreatment technology and Liquid Chromatography-Tandem Mass Spectrometry detection technique, once
Sulfamido in sample, trimethoprim (TMP), quinolones, beta-receptor agonist, Tetracyclines, nitroimidazole are analyzed in pre-treatment simultaneously
Class, macrolides, quinolines, penicillins, 10 major class of cephalo-type totally 63 kinds of residues of veterinary drug.Through quality inspection research and development management information system
The websites such as system, middle National IP Network, National Science and Technology Book Document Center, the general information of dimension, free and unfettered visitor bar bar are inquired, and are not found using this
Invention Sample Pretreatment Technique, Liquid Chromatography-Tandem Mass Spectrometry detect 63 kinds of veterinary drugs of this 10 major class in animal derived product simultaneously
Report.
The present invention is known in looking into new result：
《China Science and Technology Achievements Database》、《National science and technology achievement library》、《Chinese patent》、《Chinese Journals data
Library》、《China Dissertation Database》、《China academia meeting paper database》Deng.By liquid phase * chromatographies * series connection * mass spectrum * beasts
Medicine * (residual+inspection * survey) * PRiME*HLB, liquid phase * chromatographies * series connection * mass spectrum * animal sources * (sulfanilamide (SN)+quinolone+trimethoprim (TMP)+
Quinolines+tetracycline+nitroimidazole+macrolides+β * receptor stimulating agents+penicillins+cephalo-type) retrieval mode carry out
Retrieval.In the range of search of above-mentioned document and database, using " 10 major class in animal derived food of the above-mentioned technology of this project
63 kinds of Residual Veterinary Medicines rapid assay methods pass through formula SPE purifications-high performance liquid chromatography-tandem mass method ", the country has no identical
Research Literature and achievement report.
Description of the drawings
Fig. 1 is the detection method flow chart of the animal derived product residue of veterinary drug of detection provided in an embodiment of the present invention.
Specific implementation mode
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
The present invention analyzes suitable Extraction solvent, carries using animal products such as shrimp, bullfrog leg, the flesh of fish, pig muscles as object
Component to be measured in method and purification techniques extraction and cleaning sample is taken, interference is measured to improve extraction efficiency to greatest extent and eliminate；
63 kinds of determinands are detached using suitable liquid chromatogram separation condition, isomerism physical efficiency is made to reach baseline separation；
Select MS/MS detectors, ESI ion sources, scheduled MRM patterns, three ions confirmation, inside/outside scalar quantity skill
Art detect simultaneously sulfamido, trimethoprim (TMP), quinolones, beta-receptor agonist, Tetracyclines, macrolides, quinolines,
Nitro glyoxaline, cephalo-type, 10 major class of penicillins totally 63 kinds of veterinary drugs.
The detection method of the animal derived product residue of veterinary drug of detection provided in an embodiment of the present invention,
The present invention is analysis object with shrimp, bullfrog leg, the flesh of fish, pig muscle etc., selects suitable Extraction solvent and extraction side
Formula extracts more than 50 kinds of veterinary drug in sample, and extraction efficiency (rate of recovery) is made to meet the requirements；Suitable purification techniques is analyzed, as far as possible
(reduction) matrix effect is eliminated, the interference to measurement is reduced, to improve detection sensitivity as far as possible；Analyze the liquid phase of 63 kinds of veterinary drugs
Chromatographic separation condition makes the isomerism physical efficiency in this 63 kinds of veterinary drugs reach baseline separation；It is detected using quadrupole rod tandem mass spectrum
Device, multiple-reaction monitoring pattern, three ions confirmation, internal standard or quantified by external standard method technology detect more than 50 kinds of residue of veterinary drug simultaneously, finally
It establishes liquid chromatography-tandem mass spectrometry while detecting sulfamido, trimethoprim (TMP), quinolones, beta-receptor in animal derived product
Agonist, Tetracyclines, macrolides, quinolines, nitro glyoxaline, cephalo-type, 10 major class of penicillins totally 63 kinds of veterinary drugs
Remaining method makes this method reach high sensitivity, quantitative accurate, qualitative reliable, and this method is applied to routine testing work
Make.
As shown in Figure 1, the detection method of the animal derived product residue of veterinary drug of detection provided in an embodiment of the present invention, including：
S101：Optimize Mass Spectrometry Conditions, determine the parent ion of 63 kinds of veterinary drugs, selects the maximum two pairs of daughter ions of sensitivity, it is excellent
Change the impact energy of each pair of ion, remove the Mass Spectrometry Conditions such as cluster voltage, lens voltage, forms MRM ion pairs.Optimize atomization gas, spraying
Voltage, auxiliary gas, heating temperature plasma source parameter.
S102：Optimize liquid chromatogram separation condition, select suitable mass spectral analysis column, adjusts mobile phase, flow velocity and column temperature
Etc. liquid phase separations condition, so that parent ion and the identical isomerism physical efficiency of daughter ion is reached and efficiently separate.Establish liquid chromatogram-string
Join the instrumental method of Mass Spectrometer Method 10 major class totally 63 kinds of veterinary drugs.The standard curve for establishing 63 kinds of drugs, linearly meets the requirements, and determines
Minimum detection limit and the range of linearity.
S103：Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence extraction efficiency
Factor makes the rate of recovery of various components to be measured meet the technical requirements.
S104：Suitable thickening-purification technology technical finesse sample extracting solution is selected, to effectively remove the impurity such as fat and phosphatide,
So that this method is improved detection sensitivity to greatest extent, and measurement interference can be reduced.Establish 10 major class 63 in animal derived product
The sample-pretreating method of kind detection of veterinary drugs in food.
S105：Inside authentication is carried out to detection method, selects representational sample in detection lower bound, twice of detection lower bound
And three levels such as ten times of lower limit of measurement are added recycling verification accuracy；Select positive or negative sample addition recycling
The precision (no less than 6 times) of verification method；Selected section detection project compares this law with corresponding national standard method
Compared with.
S106：Qualified third party laboratory is selected, is verified carrying out room to the detection method established.
S107：It is for statistical analysis to detection data.
With reference to good effect, the invention will be further described.
(1), establish for the first time at home《The quick side of measurement of 10 major class, 63 kinds of Residual Veterinary Medicines in animal derived food
Method passes through formula SPE purifications-high performance liquid chromatography-tandem mass method》, the method can detect simultaneously sulfamido in animal derived food,
Trimethoprim (TMP), quinolones, beta-receptor agonist, Tetracyclines, nitro glyoxaline, macrolides, quinolines, penicillin
Class, 10 major class of cephalo-type totally 63 kinds of residues of veterinary drug, the method achieve animal derived product high throughputs, low cost, multiple types, more
Analyzed while veterinary drug, realize the simplification that low no solutions of chlorine is used and operated, be can detect simultaneously at present veterinary drug type compared with
More methods；
(2), this method just can extract 10 major class in sample using suitable Extraction solvent, extracting method, a pre-treatment
63 kinds of veterinary drugs, effectively overcome variety classes detection of veterinary drugs in food need to carry out sample pre-treatments respectively using various criterion method and
Caused by work repetition, inefficiency, long detection cycle the deficiencies of.
(3), this method is used through formula PRiME HLB Solid Phase Extraction column purification sample extracting solutions, which is suitble to big
The veterinary drug that some properties differ can remove the nonpolar chaff interferent such as 99% phosphatide, while the purification techniques greatly improves sample
The analysis throughput of product can effectively eliminate measurement interference, reduce or eliminate matrix effect, improve detection sensitivity, overcome at present
The variety classes veterinary drug of standard method defined need to using different purification techniques and caused by detection efficiency is low, testing cost is high etc.
It is insufficient；
(4), this method detection range is wide, covers the animal derived production of the aquatic products and pig muscle etc. such as shrimp, bullfrog leg, flesh of fish
Product, detection project is more, can once detect 10 major class, 63 kinds of residues of veterinary drug simultaneously, realizes that more residuals, entry monitor simultaneously, can have
Effect improves testing staff's working efficiency, reduces testing cost, shortens detection cycle, and speed passenger flow speed, safeguards Agricultural Products Trade,
External tradeing mutual compensation is successfully managed, ensures the health for the crowd of eating.
With reference to final goal, the invention will be further described.
The present invention finally establishes the detection for quickly detecting ten 63 kinds of residues of veterinary drug of major class in animal derived product simultaneously
Method, this method must reach following technical indicator：
1., under conditions selected, the separation of 63 kinds of objects is preferable, and peak shape is sharp, and retention time distribution is reasonable, retention time
Terminate from 3min beginning 23min, the setting of gradient makes 6 groups of isomers in measured object that can obtain baseline separation；
2., this method amino sulfanilamide (SN), sulphaguanidine, sulphadiazine, sulfanilamide (SN) suitable for shrimp, bullfrog leg, the flesh of fish and pig muscle
First between pyridine, sulphathiazole, sulfamethazine, sulfacetamide, 5-methoxysulfadiazine, domian, sulfanilamide (SN)
Oxygen pyrimidine, sulfamethoxy-pyridazine, sulfamethoxazole, sulfamethazole, sulfabenzamide, sulfachlorpyridazine, sulfanilamide (SN) quinoline are disliked
Quinoline, fanasil, N'-phenylsulfanilamide pyrene azoles, sulfanilamide (SN) Sulfafurazole, madribon, ayerlucil,
The pyridine of kynix benzyl, flumequine, oxolinic acid, Norfloxacin, Enoxacin, Ciprofloxacin, pefloxacin, Lomefloxacin, red promise
Sha Xing, Enrofloxacin, Ofloxacin, marbofloxacin, fleraxacin, sarafloxacin, Si Baisha stars, Orbifloxacin, Difloxacin,
Acidum nalidixicum, salbutamol, Terbutaline, Cimaterol, Ke Lunsailuo, Ractopamine, phenolethanolamine A, horse-ride step spy sieve, Ke Lun
Special sieve, ampicillin, benzyl penicillin, Ceftiofur, carbadox, quinoline sigh ketone, quinoline match more, tetracycline, terramycin, lincomycin, Thailand
Happy rhzomorph, spiramvcin, Tilmicosin, clindamycin, metronidazole, Dimetridazole, Lip river nitre reach the detection of 63 kinds of veterinary drug projects of azoles；
3., 17 kinds of fluoroquinolones, 22 kinds of sulfa drugs, a kind of trimethoprim (TMP), 3 kinds of quinolines, 5 kinds
Macrocyclolactone lactone kind medicine, ampicillin, benzyl penicillin, the minimum quantitative limit of Ceftiofur are 10 μ g/kg, tetracycline and terramycin
Minimum to be quantitatively limited to 50 μ g/kg, 8 kinds of beta-receptor agonists and 3 kinds of the minimum of nitroimidazole are quantitatively limited to 0.5 μ g/kg；Detection limit
Meet domestic relevant laws and regulations to these residue of veterinary drug limitation requirements.
4., the linear regression coeffficients of 6 concentration points of 63 kinds of veterinary drug projects is 0.9928 or more；
5., using shrimp, fish and pig muscle as background sample, the experiment of three concentration level mark-on reclaims, indoor recovery of standard addition
Between 52.8%~145%, the coefficient of variation of 6 measurement is between 4.6%~28.3%；Using pork as background sample, four
Family's eight measurement results of measuring unit, between room recovery of standard addition between 55.2%~140%, between room precision 6.1%~
Between 26.2%.
6., to blank sample and blank addition standard results compare, target analytes occur region there is no signal
Interference, each object retention time are stablized, method high specificity；
7., choose Enrofloxacin, sulfamethoxazole and tetracycline project, compare this law and the difference of national standard method, t
Value is respectively 1.907,1.94,0.963, respectively less than t0.05,14=2.145, illustrate method that project is worked out and standard method without
Notable difference；Selection standard substance (proficiency testing remaining sample) carries out two to Enrofloxacin, sulphadiazine and sulfamethoxazole
Kind method compares, and the relative differences of two kinds of assay methods are less than 20%, meet the requirement of research project plan charter；
8., matrix effect evaluation result show that matrix effect does not influence the critical index of the screening technique.
2, after method of the invention is established, it is applied to the detection to this 10 major class, 63 kinds of residue of veterinary drug in animal derived product
Work detects while reaching multiple types drug residual, to improve testing staff's working efficiency, reduce testing cost, shortening
Detection cycle reaches the effect that synergy subtracts consumption, to generate apparent Social benefit and economic benefit.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement etc., should all be included in the protection scope of the present invention made by within refreshing and principle.
Claims (6)
1. a kind of detection method of the animal derived product residue of veterinary drug of detection, which is characterized in that the animal derived product of detection
The detection method of residue of veterinary drug is：
Analysis extracts in purification sample to be measured group suitable for Extraction solvent, extracting method and the purification techniques of detected sample
Point；
63 kinds of determinands are detached using liquid chromatogram separation condition, make isomer baseline separation；
Select MS/MS detectors, ESI ion sources, schedul MRM patterns, three ions confirmation, inside/outside scalar quantity technology simultaneously
Detect sulfamido, trimethoprim (TMP), quinolones, beta-receptor agonist, Tetracyclines, macrolides, quinolines, nitro miaow
Azole, cephalo-type, 10 major class of penicillins totally 63 kinds of veterinary drugs.
2. detecting the detection method of animal derived product residue of veterinary drug as described in claim 1, which is characterized in that the detection
The detection method of animal derived product residue of veterinary drug includes：
Optimize Mass Spectrometry Conditions, determine the parent ion of 63 kinds of veterinary drugs, select two pairs of daughter ions, optimizes the impact energy of each pair of ion, goes
Cluster voltage, lens voltage Mass Spectrometry Conditions form MRM ion pairs；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature ion
Source parameter；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations conditions such as mobile phase, flow velocity and column temperature,
Make parent ion and the identical isomerism physical efficiency of daughter ion reach to efficiently separate；It is big to establish liquid chromatography-tandem mass spectrometry detection 10
The method of class totally 63 kinds of veterinary drugs；The standard curve for establishing 63 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；Establish animal derived product
In 10 63 kinds of detection of veterinary drugs in food of major class sample-pretreating method；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement three
Level is added recycling verification accuracy；Select positive or negative sample addition recycling verification precision；
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
3. a kind of using shrimp as the detection method of the animal derived product residue of veterinary drug of detection of object, which is characterized in that described with shrimp
Include for the detection method for detecting animal derived product residue of veterinary drug of object：
Optimize Mass Spectrometry Conditions, determine the parent ion of 63 kinds of veterinary drugs, select two pairs of daughter ions, optimizes the impact energy of each pair of ion, goes
Cluster voltage, lens voltage Mass Spectrometry Conditions form MRM ion pairs；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature ion
Source parameter；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations conditions such as mobile phase, flow velocity and column temperature,
Make parent ion and the identical isomerism physical efficiency of daughter ion reach to efficiently separate；It is big to establish liquid chromatography-tandem mass spectrometry detection 10
The method of class totally 63 kinds of veterinary drugs；The standard curve for establishing 63 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；Establish animal derived product
In 10 63 kinds of detection of veterinary drugs in food of major class sample-pretreating method；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement three
Level is added recycling verification accuracy；Select positive or negative sample addition recycling verification precision；
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
4. a kind of using bullfrog leg as the detection method of the animal derived product residue of veterinary drug of detection of object, which is characterized in that described
Include by the detection method of the animal derived product residue of veterinary drug of detection of object of bullfrog leg：
Optimize Mass Spectrometry Conditions, determine the parent ion of 63 kinds of veterinary drugs, select two pairs of daughter ions, optimizes the impact energy of each pair of ion, goes
Cluster voltage, lens voltage Mass Spectrometry Conditions form MRM ion pairs；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature ion
Source parameter；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations conditions such as mobile phase, flow velocity and column temperature,
Make parent ion and the identical isomerism physical efficiency of daughter ion reach to efficiently separate；It is big to establish liquid chromatography-tandem mass spectrometry detection 10
The method of class totally 50 kinds of veterinary drugs；The standard curve for establishing 63 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；Establish animal derived product
In 10 63 kinds of detection of veterinary drugs in food of major class sample-pretreating method；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement three
Level is added recycling verification accuracy；Select positive or negative sample addition recycling verification precision；
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
5. a kind of using the flesh of fish as the detection method of the animal derived product residue of veterinary drug of detection of object, which is characterized in that it is described with
The flesh of fish is that the detection method of the animal derived product residue of veterinary drug of detection of object includes：
Optimize Mass Spectrometry Conditions, determine the parent ion of 63 kinds of veterinary drugs, select two pairs of daughter ions, optimizes the impact energy of each pair of ion, goes
Cluster voltage, lens voltage Mass Spectrometry Conditions form MRM ion pairs；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature ion
Source parameter；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations conditions such as mobile phase, flow velocity and column temperature,
Make parent ion and the identical isomerism physical efficiency of daughter ion reach to efficiently separate；It is big to establish liquid chromatography-tandem mass spectrometry detection 10
The method of class totally 63 kinds of veterinary drugs；The standard curve for establishing 63 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；Establish animal derived product
In 10 63 kinds of detection of veterinary drugs in food of major class sample-pretreating method；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement three
Level is added recycling verification accuracy；Select positive or negative sample addition recycling verification precision；
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
6. a kind of using pig muscle as the detection method of the animal derived product residue of veterinary drug of detection of object, which is characterized in that described
Include by the detection method of the animal derived product residue of veterinary drug of detection of object of pig muscle：
Optimize Mass Spectrometry Conditions, determine the parent ion of 63 kinds of veterinary drugs, select two pairs of daughter ions, optimizes the impact energy of each pair of ion, goes
Cluster voltage, lens voltage Mass Spectrometry Conditions form MRM ion pairs；Optimize atomization gas, spray voltage, auxiliary gas, heating temperature ion
Source parameter；
Optimize liquid chromatogram separation condition, select mass spectral analysis column, adjusts the liquid phase separations conditions such as mobile phase, flow velocity and column temperature,
Make parent ion and the identical isomerism physical efficiency of daughter ion reach to efficiently separate；It is big to establish liquid chromatography-tandem mass spectrometry detection 10
The method of class totally 63 kinds of veterinary drugs；The standard curve for establishing 63 kinds of drugs determines minimum detection limit and the range of linearity；
Ingredient to be measured in suitable Extraction solvent, extracting method extraction sample, optimization is selected to influence the factor of extraction efficiency；
Select suitable thickening-purification technology technical finesse sample extracting solution, removal fat and phosphatide impurity；Establish animal derived product
In 10 63 kinds of detection of veterinary drugs in food of major class sample-pretreating method；
Inside authentication is carried out, selects representational sample in detection lower bound, twice of detection lower bound and ten times of lower limit of measurement three
Level is added recycling verification accuracy；Select positive or negative sample addition recycling verification precision；
It is verified carrying out room to the detection method established；
It is for statistical analysis to detection data.
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108828110A (en) * | 2018-09-11 | 2018-11-16 | 广东出入境检验检疫局检验检疫技术中心 | The measuring method of benzimidazoles residues residual quantity in one breeder fat |
| CN109061018A (en) * | 2018-10-19 | 2018-12-21 | 山东省新世纪检测认证中心有限公司 | A kind of pre-treating method preventing high protein content matrix gelation |
| CN109115920A (en) * | 2018-11-02 | 2019-01-01 | 中国水产科学研究院东海水产研究所 | The method for detecting 18 kinds of beta-receptor agonists in aquatic products simultaneously using Liquid Chromatography-Tandem Mass Spectrometry |
| CN109298109A (en) * | 2018-12-07 | 2019-02-01 | 昌邑市检验检测中心 | In meat product a variety of beasts it is residual and meanwhile detection method |
| CN109406680A (en) * | 2018-12-19 | 2019-03-01 | 山东省食品药品检验研究院 | The method of 10 kinds of quinolone antibiotics in ultra performance liquid chromatography-tandem mass spectrometry measurement bean sprouts |
| CN112162052A (en) * | 2020-11-06 | 2021-01-01 | 深圳市格物正源质量标准系统有限公司 | Method for determining multiple residues of veterinary drugs in aquatic products |
| CN112526054A (en) * | 2020-12-31 | 2021-03-19 | 镇江华大检测有限公司 | Method for measuring residual quantity of tetracycline veterinary drugs in animal-derived food |
| CN113866335A (en) * | 2021-09-23 | 2021-12-31 | 南京市产品质量监督检验院 | Method for determining 81 veterinary drug residues in animal tissues by using tandem liquid chromatography-mass spectrometry and application |
| CN115469029A (en) * | 2022-09-14 | 2022-12-13 | 中国人民解放军军事科学院军事医学研究院 | Detection method for various veterinary drug residues in animal muscle and application thereof |
| CN118191186A (en) * | 2024-05-16 | 2024-06-14 | 西南林业大学 | Extraction method and application of sulfonamide antibiotics in water body |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106596773A (en) * | 2016-12-16 | 2017-04-26 | 深圳天祥质量技术服务有限公司 | Method for simultaneous and rapidly screening and testing multiple veterinary drug residues in animal-derived food |
| CN107228912A (en) * | 2017-05-26 | 2017-10-03 | 河南省兽药饲料监察所 | The screening method of 79 kinds of antibacterials in a kind of animal food |
| CN107576744A (en) * | 2017-09-29 | 2018-01-12 | 沈阳出入境检验检疫局检验检疫综合技术中心 | A kind of method for detecting animal derived food veterinary drug residue |
| CN107607645A (en) * | 2017-09-13 | 2018-01-19 | 山东出入境检验检疫局检验检疫技术中心 | Method that is a kind of while detecting multiple types residue of veterinary drug in the flesh of fish |
-
2018
- 2018-03-14 CN CN201810210283.0A patent/CN108490088A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106596773A (en) * | 2016-12-16 | 2017-04-26 | 深圳天祥质量技术服务有限公司 | Method for simultaneous and rapidly screening and testing multiple veterinary drug residues in animal-derived food |
| CN107228912A (en) * | 2017-05-26 | 2017-10-03 | 河南省兽药饲料监察所 | The screening method of 79 kinds of antibacterials in a kind of animal food |
| CN107607645A (en) * | 2017-09-13 | 2018-01-19 | 山东出入境检验检疫局检验检疫技术中心 | Method that is a kind of while detecting multiple types residue of veterinary drug in the flesh of fish |
| CN107576744A (en) * | 2017-09-29 | 2018-01-12 | 沈阳出入境检验检疫局检验检疫综合技术中心 | A kind of method for detecting animal derived food veterinary drug residue |
Non-Patent Citations (5)
| Title |
|---|
| KUN WANG等: "A Simple and Fast Extraction Method for the Determination of Multiclass Antibiotics in Eggs Using LC-MS/MS", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| SHERRI B. TURNIPSEED等: "Wide-Scope Screening Method for Multiclass Veterinary Drug Residues in Fish, Shrimp, and Eel Using Liquid Chromatography−Quadrupole High-Resolution Mass Spectrometry", 《JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY》 * |
| 孙雷等: "猪肉中92种兽药残留的UPLC-Qtrap高通量筛查和定量方法的研究", 《中国兽药杂志》 * |
| 沃特世公司: "《Oasis PRiME HLB食品应用文集》", 30 September 2015 * |
| 郝杰等: "固相萃取-超高效液相色谱-串联质谱法同时测定动物源性食品中多种兽药残留", 《食品科学》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108828110A (en) * | 2018-09-11 | 2018-11-16 | 广东出入境检验检疫局检验检疫技术中心 | The measuring method of benzimidazoles residues residual quantity in one breeder fat |
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| CN109115920A (en) * | 2018-11-02 | 2019-01-01 | 中国水产科学研究院东海水产研究所 | The method for detecting 18 kinds of beta-receptor agonists in aquatic products simultaneously using Liquid Chromatography-Tandem Mass Spectrometry |
| CN109298109A (en) * | 2018-12-07 | 2019-02-01 | 昌邑市检验检测中心 | In meat product a variety of beasts it is residual and meanwhile detection method |
| CN109406680A (en) * | 2018-12-19 | 2019-03-01 | 山东省食品药品检验研究院 | The method of 10 kinds of quinolone antibiotics in ultra performance liquid chromatography-tandem mass spectrometry measurement bean sprouts |
| CN112162052A (en) * | 2020-11-06 | 2021-01-01 | 深圳市格物正源质量标准系统有限公司 | Method for determining multiple residues of veterinary drugs in aquatic products |
| CN112526054A (en) * | 2020-12-31 | 2021-03-19 | 镇江华大检测有限公司 | Method for measuring residual quantity of tetracycline veterinary drugs in animal-derived food |
| CN113866335A (en) * | 2021-09-23 | 2021-12-31 | 南京市产品质量监督检验院 | Method for determining 81 veterinary drug residues in animal tissues by using tandem liquid chromatography-mass spectrometry and application |
| CN115469029A (en) * | 2022-09-14 | 2022-12-13 | 中国人民解放军军事科学院军事医学研究院 | Detection method for various veterinary drug residues in animal muscle and application thereof |
| CN118191186A (en) * | 2024-05-16 | 2024-06-14 | 西南林业大学 | Extraction method and application of sulfonamide antibiotics in water body |
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