CN104931635A - Method and liquid mass database for detecting residual chemicals in animal-derived food - Google Patents
Method and liquid mass database for detecting residual chemicals in animal-derived food Download PDFInfo
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Abstract
The invention relates to a liquid mass database for detecting residual chemicals in animal-derived food and an application method of the liquid mass database. The production method of the liquid mass database includes the steps of firstly, preparing standard solution; secondly, using the extraction purification technology combining fast enzymolysis and fast solid-phase extraction (SPE) to preprocess a to-be-detected sample before analysis; thirdly, performing one-step sample chromatographic analysis; fourthly, building the liquid mass spectrum database.
Description
Technical field
The invention belongs to field of biological detection, in particular to a kind of liquid matter database for detecting left drug in animal-derived food and using method thereof.
Background technology
Animal-derived food accounts for suitable proportion in international agriculture trade, regulation limitations for animal-derived food drug residue is more and more stricter, for " positive list " system that Japan implements in May, 06, it has formulated concrete limit standard to 236 kinds of veterinary drugs and feed addictive, and field of food safety at home, from early stage " chloromycetin, nitrofuran " " malachite green finally, clenbuterol hydrochloride ", the food safety affair caused because of medicament residue takes place frequently and highlights China's food safety Regulation hierarchy of control technical support scarce capacity, rapid screening method is remained in the urgent need to setting up multiple types.At present, single class multi-residue analysis has been developed into by single retention analysis for animal-derived food drug residue detection technique and method, since 03 year, the multi-residue determination standard of dividing with substance classes circle occupies more than 70% of retention analysis country and industry standard.Same time, mass spectrophotometry has become the technical way of multi-residue determination, and the technical indicator rule being source with European Union 2002/657/EC regulation is also by being admitted in the world and accepting.Along with the development of retention analysis technology, multiple types retention analysis has advantage in the many-side such as quantity of information, detection efficiency and becomes research and development focus, and mass spectroscopy device field is as Q-TOF, the tandem mass spectrometers such as Q-Trap practical, the spectrum storehouse compare of analysis based on senior mass-spectrometric technique gradually maturation also for multiple types remain express-analysis provide technology may.But, be no matter in method exploitation, checking and standardization or in regulation is perfect etc., detect that multiple types is residual all exists more difficulty simultaneously:
(1) variety classes veterinary drug physico-chemical property difference is very large, polarity coverage is wide, in different substrates, residual form differs, and indivedual veterinary drug derivatization of still needing can effectively detect, and is therefore difficult to realize effective breakthrough on general pre-treating method and the analysis of chromatogram single injected sampling;
(2) regulation requires the limitation of different veterinary drug (forbidden drugs and limit the use of medicine) and uses regulation to differ, and multiple types residue detection proof rule is not yet perfect;
(3) bare substrate that method exploitation needs is difficult to obtain, mixed standard solution preparation difficulty;
(4) analysis ability of instrument.In structure liquid mass spectral database, although there is commercialization to compose storehouse as AB company legal medical expert database of poisons (1250 kinds of target analytes), basic veterinary drug database (139 kinds of target analytes); The small-molecule drug database of Freiburg university hospital, but existence applies limitation as follows:
1) MS/MS data are only had, without chromatographic information characteristics;
2) without online information feature;
3) without unified pre-treating method and the coupling of instrumental analysis phase;
4) extraction standard data are lacked.
Therefore, under existence conditions, be difficult to the effectively multiple types realized based on liquid mass spectral database technology and remain quick Screening analysis.
Summary of the invention
In view of domestic and international technological difficulties such as sample universal pre-treatment, instrument express-analysis and screening technique checking existed in animal-derived food multiple types left drug is analyzed at present.The present invention remains for the purpose of quick selective mechanisms system to set up animal-derived food medium or high risk medicine multiple types, extract by adopting multi-solvent system piecewise combination, set up sample high flux versatility pretreatment technology, complex optimum chromatographic realizes multiobjective analysis thing wide polarity scope liquid chromatography and effectively retains and be separated, and by building the liquid chromatography-mass spectrography/mass spectrometric data storehouse containing chromatography-mass spectroscopy multidimensional information and the research and development carrying out the gordian technique contents such as screening technique checking according to laws and regulations requirement and detection practice, break through multiple types retention analysis technical barrier, set up one and remain versatility pretreatment technology with multiple types, quick separating system and liquid mass spectral database are the open express-analysis platform of technical characteristic and effective examination detection technique system.
The present invention is directed to 12 kinds of Limited Doses at the following excessive risk left drug of 1.0 μ g/kg (category-A residuals) as detecting target, this group medicine is respectively,
(1) steroid hormone compounds: testosterone (T), prednisolone (PNSL), betamethasone (BTS), dexamethasone (DTS);
(2) nitroimidazoles medicine and metabolic product thereof: flagyl (MNZ), cough up nitre and to rattle away azoles (RNZ), Dimetridazole (DMZ);
(3) Beta-receptor stimulating agent class material: Clenbuterol (CLB), Tulobuterol (TUL), Cimaterol (CIM);
(4) dye class material: crystal violet (CV) and leuco crystal violet (LCV);
First the present invention relates to a kind of construction method detecting the rapid screening liquid mass spectral database that animal-derived food medium or high risk medicine multiple types remains, and described animal-derived food is animal muscle based food, dairy products, specifically comprises the steps,
(1) preparation of standard working solution;
Steroid hormone compounds, nitroimidazoles medicine and metabolic product thereof, Beta-receptor stimulating agent class material: adopt its hybrid standard stock solution (0.01g/L) of acetonitrile classification preparation;
The materials such as dye class adopt its hybrid standard stock solution (0.01g/L) of methyl alcohol classification preparation;
Mixing and standard working solution (0.1mg/L), pipetting testosterone, prednisolone, betamethasone, dexamethasone, metronidazole respectively, to cough up rattle away azoles, Dimetridazole, Clenbuterol, Tulobuterol, Cimaterol, crystal violet and leuco crystal violet Standard Stock solutions of nitre appropriate, is the mixed standard solution of 0.1mg/L by acetontrile concentration;
(2) analyze pre-treatment, adopt the extraction and cleaning technology of rapid enzymolysis (release Conjugate polyamines)+quick Solid-Phase Extraction (SPE) to carry out sample to be tested and analyze pre-treatment work, concrete extraction and cleaning process is as follows:
1) extract: take homogeneous samples 5g, be placed in 50mL teflon centrifuge tube, add ammonium acetate buffer (0.2mol/L, pH 5.2) 15mL, β-glucosiduronic acid/sulfatase 50 μ L, vortex mixes, 50 DEG C of water-bath vibration 2h, let cool to room temperature, in 0 DEG C, the centrifugal 5min of 15000rpm, transfer supernatant is in another clean centrifuge tube, pH to 9.0 is regulated by NaOH solution (5mol/L), add ethyl acetate 20mL again, sodium chloride 6g, vibration vortex 5min, in 0 DEG C, the centrifugal 5min of 15000rpm, get upper organic phase 35 DEG C and revolve steaming near dry, nitrogen dries up, residue uses 5mL 0.1mol/L dissolve with hydrochloric acid solution, ultrasonic 1min, wait until column purification,
2) purify: MCX post (Waters Oasis MCX SPE pillar) is installed on (this device is documented in utility model patent CN201020014620.X) on solid-phase extraction device, use methyl alcohol 3mL successively, water 3mL, 3mL 0.1mol/L hydrochloric acid solution activates.Extraction solution is carried on solid-phase extraction column, flows out under gravity, use 3mL water successively, 3mL methyl alcohol and 5mL normal hexane drip washing pillar, drain 2min, add 6mL eluent (ethyl acetate 50mL, methyl alcohol 45mL, ammoniacal liquor 5mL, mixing) wash-out.At eluent 35 DEG C, nitrogen stream dries up, sample dissolution liquid is used (to measure 0.1% formic acid acetonitrile (V/V) solution 5mL, mix with ammonium formate (5mmol/L)-formic acid (0.1%)-aqueous solution 95mL) 0.5mL dissolved residue, ultrasonic 1min, crosses 0.22 μM of miillpore filter; Described solid-phase extraction device integrate reagent rack, Nitrogen evaporator function and on-stream pressure is relatively uniform, it is made up of reagent rack, operator's console, 8 glass storehouses, 8 pear shape bottles and 1 automatic getter device, can analyze the quick SPE process of thing by realize target.
(3) disposable sample introduction stratographic analysis
Use chromatographic column is Kinetex C18 post, and employing acetonitrile is organic phase, and formic acid is as ionization reinforcing agent, and formates optimizes agent as peak shape, using the combination of aqueous phase/acid acetonitrile as the flow visualizing optimized;
Actual conditions is: chromatographic column: Kinetex C18,2.6 μm, 2.1mm × 100mm i.d.;
Flow velocity: 0.2mL/min;
Sample size: 10 μ L;
Column temperature: 30 DEG C;
Gradient elution program: (A:0.1% formic acid-acetonitrile solution; B: ammonium formate (5mmol/L)-formic acid (0.1%)-aqueous solution)
0～2min：5％A；
～8min：20％A；
～15min：95％A；
～16min：100％A；
～19min：100％A；
20min：5％A；
(4) liquid mass spectral database is built
Adopt the senior drainage pattern of level Four bar/ion trap tandem mass spectrometry: presetting many reaction detection (sMRM)-Information Dependent gathers (IDA)-enhancer ion scan (EPI);
Mass spectrometry parameters deterministic process is as follows: using initial flow according to target to analyze thing classification dilution mixed mark stock solution to concentration is mutually 0.2mg/L, then uses constant current syringe pump to inject mass ion source with the flow velocity of 5 μ L/min and carries out parameter optimization,
Use the parent ion of the scan pattern determination target analytes such as Q1MS, Q1Multiple Ions, Product Ion, MRM respectively, daughter ion, and use Ramp function optimization and determine the chemical parameters such as depolymerization voltage (DP), collision cell entrance potential (EP), collision energy (CE), collision cell exit potential (CXP);
Mass Spectrometry Conditions (API 4000 and API 4000Q-TRAP):
A) ion gun: electric spray ion source;
B) scan mode: positive ion scans;
C) detection mode: sMRM-IDA-EPI
D) electron spray voltage: 5500V;
E) atomization gas pressure: 40psi;
F) gas curtain atmospheric pressure: 30psi;
G) assisted gas pressure: 45psi;
H) ion source temperature: 475 DEG C;
I) sMRM optimum configurations: MRM detection window is set to 60s, object is set to 1.4s sweep time;
J) IDA rule: response lag: 3000cps; Dynamic background is deducted; The strongest ion is chosen as 1 to 3;
K) enhancer ion scan (EPI) optimum configurations: quality of scanning number scope is 70 ~ 1000Da; Sweep velocity is 10000Da/s; Scanning accumulative frequency is 1; Collision energy is 35eV; Expansion collision energy is 15eV;
By the mass spectral results for 12 kinds of category-A materials, build liquid mass spectral database.
What use in the structure of database builds the Analyst1.5 that library software is AB SCIEX company, and the standard of building storehouse is set as according to mentality of designing:
(1) provide each target analytes use this research to set liquid phase chromatogram condition system under RT value;
(2) provide each target analytes related chemistry information (as title, chemical formula, molecular weight, No. CAS, compounds category, ID, molecular structural formula etc.);
(3) the EPI spectrogram according to the described collection of 3.2.2 item under each target analytes at least 5 different conditions is provided, namely CE is 20eV, 35eV, 50eV and CE 4 EPI spectrograms when be 35eV, CES being 15eV, in addition, one will at least also be had CE is 35eV, CES under chromatographic condition EPI spectrogram when being 15eV;
(4) provide the EPI spectrogram of target analytes in different substrates as far as possible.
First 3 to build library standard be the condition that must meet, and the 4th article is the alternatives according to actual conditions.
The chromatographic selected according to above condition and Mass Spectrometry Conditions achieve how residual efficient of 115 kinds of veterinary drugs comprising 12 kinds of high residue category-A materials of the present invention and are separated.Although there is individual compound retention time close, it extracts ion current collection of illustrative plates all can realize mass spectrum separation, can carry out quantitative test, in addition, EPI composes a storehouse target analytes patch information feature, and representation compound is structure " fingerprint " information qualitatively, can be accurately qualitative to target analytes.The thus obtained liquid mass spectral database comprising the liquid matter information of described category-A residuals, can realize remaining rapid screening to described animal-derived food medium or high risk medicine multiple types.
Under above-mentioned chromatogram and Mass Spectrometry Conditions, except 12 kinds of category-A materials, category-B 103 kinds the total ion current figure of totally 115 kinds of target analytes, typical Selective ion mode flow graph, each target analytes extract ion flow graph and when selecting CES to expand the database spectrogram of (CE 35eV, CES 15eV) as Fig. 1 ~ Fig. 4.
The invention still further relates to by said method build obtain for detecting the residual rapid screening liquid mass spectral database of animal-derived food medium or high risk medicine multiple types, described animal-derived food is animal muscle based food, dairy products;
Described excessive risk medicine be Limited Doses at the following excessive risk left drug of 1.0 μ g/kg, be specially:
(a) steroid hormone compounds: testosterone (T), prednisolone (PNSL), betamethasone (BTS), dexamethasone (DTS);
(b) nitroimidazoles medicine and metabolic product thereof: flagyl (MNZ), cough up nitre and to rattle away azoles (RNZ), Dimetridazole (DMZ);
(c) Beta-receptor stimulating agent class material: Clenbuterol (CLB), Tulobuterol (TUL), Cimaterol (CIM);
(d) dye class material: crystal violet (CV) and leuco crystal violet (LCV).
The invention still further relates to described detection animal-derived food medium or high risk medicine multiple types and remain rapid screening liquid mass spectral database in the application detecting the medicament residue in animal-derived food.
The invention still further relates to and use described liquid mass spectral database to carry out method that is qualitative or quantitative test to the excessive risk medicament residue in target animal derived food, described method comprises the steps,
(1) preparation standard solution,
(2) analyze pre-treatment, adopt the extraction and cleaning technology of rapid enzymolysis (release Conjugate polyamines)+quick Solid-Phase Extraction (SPE) to carry out sample to be tested and analyze pre-treatment work,
(3) chromatogram and the analysis of liquid matter are carried out to sample to be tested, obtain liquid mass spectrogram, contrast described liquid mass spectral database, carry out qualitative or quantitative test detection.
Described qualitative analysis adopts library searching to carry out, and qualitative criteria is as follows:
(1) in sample, compound extracts retention time and the standard solution of ion current or adds target analytes in sample and extract compared with the retention time of ion current, and amplitude of variation is no more than 5%.
(2) parent ion/daughter ion (transmission ion pair) of target analytes must occur simultaneously, and the signal to noise ratio (S/N ratio) (S/N) >=3 of transmission ion pair.
(3) in sample, compd E PI spectrogram is compared with close concentration level (same concentration numbers magnitude) standard solution in spectrum storehouse or extraction standard solution E PI spectrogram, and spectrogram mates purity (Purity value) >=60.
The quantivative approach of described quantitative test is as follows:
Use external standard one-point method quantitative, calculate the content of target analytes by formula (1).
In formula (1):
Target analytes content in X-sample, μ g/kg;
The concentration of target analytes in c-sample solution, μ g/L;
The constant volume of V-sample solution, mL;
The quality of m-sample, g;
The R-recovery, %.
Accompanying drawing explanation
The total ion current figure of Fig. 1 .A class material, category-B material totally 115 kinds of target analytes.
The typical Selective ion mode flow graph of Fig. 2 .A class material, category-B material totally 115 kinds of target analytes.
The target analytes of Fig. 3 .A class material extracts ion flow graph.
The target analytes spectrum database data figure of Fig. 4 .A class material.
Embodiment
The preparation of embodiment 1. standard working solution
Steroid hormone compounds, nitroimidazoles medicine and metabolic product thereof, Beta-receptor stimulating agent class material: adopt its hybrid standard stock solution (0.01g/L) of acetonitrile classification preparation, the equal Absorbable organic halogens of standard of physical storing solution 12 months.
The materials such as dye class adopt its hybrid standard stock solution (0.01g/L) of methyl alcohol classification preparation, Absorbable organic halogens 3 months.
Mixing and standard working solution (0.1mg/L), pipette testosterone, prednisolone, betamethasone, dexamethasone, metronidazole respectively, to cough up rattle away azoles, Dimetridazole, Clenbuterol, Tulobuterol, Cimaterol, crystal violet and leuco crystal violet Standard Stock solutions of nitre appropriate, be the mixed standard solution of 0.1mg/L by acetontrile concentration, this solution-20 DEG C of Absorbable organic halogens 1 month.
Embodiment 2. analyzes pre-treating method
12 kinds of target compounds (A group material) are under the jurisdiction of 4 classes of compounds such as steroid hormone class, nitro glyoxaline, beta-receptor activator class and dye class respectively, it performs limitation claimed range at 0.05 μ g/kg ~ 0.8 μ g/kg, is one group of medicine that limitation requirement is the strictest in the world at present.Therefore, the step that purification is concentrated is considered when pretreatment process designs, to meet strict limitation requirement.For the classes of compounds involved by A group material, the extraction and cleaning technology of the Solid-Phase Extraction (SPE) of research and design rapid enzymolysis (release Conjugate polyamines)+fast, fast and effeciently can complete the pretreatment process of 12 kinds of ultra-low limit quantity of material.Concrete extraction and cleaning process is as follows:
(1) extract: take homogeneous samples 5g (being accurate to 0.01g), be placed in 50mL teflon centrifuge tube (as interpolation sample need be made, the A group material hybrid standard working solution of debita spissitudo is added in this step, and place 30min in dark place), add ammonium acetate buffer (0.2mol/L, pH 5.2) 15mL, β-glucosiduronic acid/sulfatase 50 μ L, vortex mixes, 50 DEG C of water-bath vibration 2h, let cool to room temperature, in 0 DEG C, the centrifugal 5min of 15000rpm, transfer supernatant is in another clean centrifuge tube, pH to 9.0 is regulated by NaOH solution (5mol/L), add ethyl acetate 20mL again, sodium chloride 6g, vibration vortex 5min, in 0 DEG C, the centrifugal 5min of 15000rpm, get upper organic phase 35 DEG C and revolve steaming near dry, nitrogen dries up, residue uses 5mL 0.1mol/L dissolve with hydrochloric acid solution, ultrasonic 1min, wait until column purification.
(2) purify: (Waters Oasis MCX SPE pillar, to MCX post this type pillar filling adsorption agent add-SO on the basis of HLB pillar NVP-DVB multipolymer
3h ion exchanging function base.) be installed on solid-phase extraction device (this device is documented in utility model patent CN201020014620.X), use methyl alcohol 3mL successively, water 3mL, 3mL 0.1mol/L hydrochloric acid solution activates.Extraction solution is carried on solid-phase extraction column, flows out under gravity, use 3mL water successively, 3mL methyl alcohol and 5mL normal hexane drip washing pillar, drain 2min, add 6mL eluent (ethyl acetate 50mL, methyl alcohol 45mL, ammoniacal liquor 5mL, mixing) wash-out.At eluent 35 DEG C, nitrogen stream dries up, sample dissolution liquid is used (to measure 0.1% formic acid acetonitrile (V/V) solution 5mL, mix with ammonium formate (5mmol/L)-formic acid (0.1%)-aqueous solution 95mL) 0.5mL dissolved residue, ultrasonic 1min, cross 0.22 μM of miillpore filter, liquid chromatography tandom mass spectrometry determination.
The step of extraction and cleaning process is described as follows: testosterone in A group material, the materials such as Tulobuterol adhere to hormone and beta-receptor activator class separately, literature survey shows this kind of normal to exist in conjunction with state in animal tissue, especially this phenol type of Tulobuterol medicine, therefore β-glucosiduronic acid/sulfatase enzymolysis residual tissue conjugates need be added, to discharge free state medicine, add the pH value environment that damping fluid one is to provide enzymolysis needs, also be use as Extraction solvent in addition, this step compare existing method have employed improve hydrolysis temperature (by now methodical 36 DEG C be promoted to 50 DEG C) the mode processes shrink that (16h) react that makes originally to need to spend the night be 2h, greatly accelerate enzymolysis process, crystal violet and Recessive Crystal Violet prove also to be extracted in this step through test, freezing ultracentrifugal object is adopted to be precipitation fat and albumen, promote that nitro glyoxaline compound and beta-receptor activator are by the distribution of aqueous phase to organic phase by extract alkalization and the object that adds sodium chloride.Purifying step is owing to relating to the purification of multiple types material, and SPE post have selected the reversed material of versatility.Research compares hydrophilic-lipophilic balance (HLB) post and common C18 post, all fails to realize good effect.Final research adopts Waters Oasis MCX SPE pillar, and this type pillar filling adsorption agent adds-SO3H ion exchanging function base on the basis of HLB pillar NVP-DVB multipolymer.Therefore this SPE post possesses anti-phase and mixing and absorption function that is ion-exchange, and when low ph value, energy adsorb acidic, alkalescence and neutral compound, has stronger detergent power compared with HLB and C18 post.The design of elution program is reported with reference to pertinent literature and in the step of drip washing, adds the step of normal hexane drip washing, disturb with better place to go lipid, elute soln have employed the mixed solution of ethyl acetate (50%)-methyl alcohol (45%)-ammonia (5%) water, through test, 6mL can complete the wash-out to target analytes.
In addition, in the process using SPE post to detect, because existing SPE system not easily realizes fast operating, seminar have developed a kind of novel solid-phase extraction device, this device can integrate reagent rack, Nitrogen evaporator function and on-stream pressure is relatively uniform, it is made up of reagent rack, operator's console, 8 glass storehouses, 8 pear shape bottles and 1 automatic getter device, can analyze the quick SPE process of thing by realize target.
To sum up, although A group material pre-treating method adopts classical solvent extraction/Solid phase extraction pre-treatment flow process, but in the design of rapid enzymolysis and the quick SPE purification process of versatility, all consider that multiple types is residual to be extracted and flux process factor, compare existing method and there is fast and convenient advantage, and in a pretreatment process, complete effective extraction of ultra-low limit quantity of material in 4 classes 12, meet the requirement of fast Screening analysis.
The disposable sample introduction chromatographic analysis system research of embodiment 3.
1. the selection of chromatographic column
The liquid chromatography of multiobjective analysis thing is separated to be considered to use ultrafine particle (particle diameter < 2 μm) chromatographic column at present usually, but this research adopts conventional H PLC system, it is withstand voltage, and the upper limit is 400Bar, therefore cannot use Ultra Performance Liquid Chromatography post.For realizing the separation object of design, consider that the particle size range of chromatographic column controls between 2 μm ~ 3 μm by system pressure restriction, chromatogram column length scope is 100mm ~ 150mm; In addition, because classes of compounds is more, polarity range spans is comparatively large, just have selected the C18 chromatographic column being suitable for being separated wide polarity scope in chromatographic column type.By above-mentioned restriction, this research have selected the one in every compounds according to polarity (selecting with reference to LogD value), has 4 kinds of materials represent examination 4 type liquid-phase chromatographic column as analysis thing altogether.Screening conditions and the results are shown in Table 2.
The selectivity experimental design of table 2 chromatographic column and compartment analysis result
As seen from the above table, compare other chromatographic columns, Kinetex C18 post degree of separation and sensitivity all better.Although this chromatographic column particle diameter is minimum, its post back pressure is still in the withstand voltage scope of conventional liquid phase.And filler particles have employed advanced core-shell structure copolymer technology, compared with the complete Bio-sil post of tradition, can obviously improve degree of separation and sensitivity.Determine and use this type chromatographic column to be preferable separate post.
2. the selection of flow visualizing and the determination of other chromatographic conditions
Mass spectrophotometry is commonly used to mobile phase (water, formic acid-aqueous solution, acetic acid-aqueous solution, methyl alcohol, acetonitrile, acidified methanol, acidifying acetonitrile, Ammonium formate buffer, ammonium acetate buffer etc.) and formed use 12 kinds of hybrid standard working fluids and screens.On the basis considering the factors such as degree of separation, sensitivity and analysis time, finally determine to adopt acetonitrile to be organic phase, formic acid is as ionization reinforcing agent, and formates optimizes agent as peak shape, using the combination of aqueous phase/acid acetonitrile as the flow visualizing optimized.
Be optimized liquid chromatography parameters such as sample size, column temperature and flow velocitys, main considerations comprises chromatographic resolution, sensitivity, reappearance and matrix effect etc.By using 12 kinds of object mixed standard solutions at the enterprising circumstances in which people get things ready for a trip spectrum separation test of screening target concentration levels (0.5 μ g/kg) and determining optimal value.
On the basis optimizing correlation parameter and more repeatedly test findings, the actual conditions of the disposable liquid phase chromatographic resolution system originally determined is: chromatographic column: Kinetex C18,2.6 μm, 2.1mm × 100mm i.d.; Flow velocity: 0.2mL; Sample size: 10 μ L; Column temperature: 30 DEG C.Gradient elution program: (A:0.1% formic acid-acetonitrile solution; B: ammonium formate (5mmol/L)-formic acid (0.1%)-aqueous solution) 0-2min:5%A; 8min:20%A; 15min:95%A; 16-19min:100%A; 20-35min:5%A.
Embodiment 4. liquid mass spectral database builds
1. the foundation of mass spectrum acquisition method and liquid mass spectral database build
(1) foundation of mass spectrum acquisition method
This research have employed the senior drainage pattern of level Four bar/ion trap tandem mass spectrometry: presetting many reaction detection (sMRM)-Information Dependent gathers (IDA)-enhancer ion scan (EPI).The foundation of this drainage pattern, first target analytes retention time in chromatographic will be determined, next sets up multiple-reaction monitoring (MRM) mass spectrometry method, and then setting triggers the condition (i.e. IDA setting) that EPI gathers, and is finally the condition setting determining that EPI gathers.
For setting up corresponding mass spectrum acquisition method, first test setting is carried out to the mass spectrometry parameters of 12 kinds of target analytes.Research adopts first Classified optimization chemical parameters (comprising parent ion, daughter ion and depolymerization voltage, collision energy etc.), then chooses representation compound optimization source parameters (comprising atomization gas pressure, ion source temperature etc.).
1. chemical parameters optimization is determined.About the basis of design chromatographic resolution result of RT parameter.Mass spectrometry parameters deterministic process is as follows: using initial flow according to target to analyze thing classification dilution mixed mark stock solution to concentration is mutually 0.2mg/L, but note in same classification, having the target analytes of identical molecular weight will prepare separately (because studied use instrument is standard resolution, cannot isomers be distinguished), then use constant current syringe pump to inject mass ion source with the flow velocity of 5 μ L/min and carry out parameter optimization.Use Q1 MS respectively, Q1 Multiple Ions, Product Ion, the parent ion of the scan pattern determination target analytes such as MRM, daughter ion (at least choose 2 for subsequent use to transmission ion by often kind of target analytes, the collection ion of a pair optimum ion as sMRM is chosen according to the disturbed condition in the signal to noise ratio (S/N ratio) of response and bare substrate) in follow-up test, and use Ramp function optimization and determine depolymerization voltage (DP), collision cell entrance potential (EP), collision energy (CE), the chemical parameters such as collision cell exit potential (CXP), 12 kinds of materials compounds parameter optimizations the results are shown in Table 3.
Table 3 12 kinds of target analytes optimize Mass Spectrometry Conditions and reference retention time
Note: 1. in this table, listed ion pair is the optimization ion pair determined after follow-up test examination matrix effect and signal to noise ratio (S/N ratio), in the parameter of this unlisted ion pair for subsequent use; 2. target analytes retention time (RT) sets in window (15s) to change at RT and all belongs to normal.
2. the optimization of source parameters is determined.Adopt Flow Injection Analysis (FIA) optimization, because instrument adopts representation compound to be optimized to this research of Limited Number system optimizing ion pair, representation compound selection principle responds relatively low target analytes when being and adopting chemical parameters to optimize, through test, source parameters optimum results is as follows: gas curtain atmospheric pressure: 30psi; Spray voltage: 5500V; Ion source temperature: 475 DEG C; Atomization gas pressure: 40psi; Assisted gas pressure: 45psi.
3. IDA condition setting.In the setting of IDA condition, the most important thing is the mensuration of response lag, screening target concentration levels (0.025 μ g/kg) is analyzed extraction standard (using the standard solution of " bare substrate " extract preparation), with 1/2 of minimum response substance responds intensity for degree setting response lag.It should be noted that, this threshold value has matrix dependence, different matrix threshold values is different, under normal circumstances, threshold value setting principle is ensure that all responses meeting examination aimed concn all can trigger EPI and gather as far as possible, improves threshold value to reduce data acquisition amount (being easy to analyze) simultaneously as far as possible.Investigate two kinds of animal sources matrix (animal muscle based food, dairy products) in this research, be set as 3000cps with minimum.
Secondly, also to select instrument dynamic background deduction function in IDA setting kind, effectively can reduce the generation of invalid data.
4. the setting of EPI parameter.
A) EPI acquisition quality number scope: in this research, the mass number scope of 12 kinds of target analyte molecule quasi-molecular ions is 140Da ~ 940Da, fragment ion masses number scope 80Da ~ 880Da, therefore EPI acquisition quality number range set is at 70Da ~ 1000Da;
B) EPI acquisition scans speed: this research use equipment is AB SCIEX 5500Q-Trap mass spectrometer, EPI gathers total 1000Da/s, 10000Da/s and 20000Da/s 3 grades of speed, for ensureing the quality of data, 10000Da/s is selected to be EPI acquisition scans speed, so both take into account the analysis acquisition rate of 12 kinds of target analytes, in turn ensure that spectrogram quality;
C) EPI gathers the setting of DP value and EP value: for ensureing spectrum database data quality, carry out segmentation statistics to 12 kinds of target analytes DP values and EP value, the median getting section is at high proportion final setting value.
By analysis, target compound DP value strengthens in 60V ~ 100V section ratio, and getting median 80V is the DP setting value that EPI gathers.Same, target group compd E P value, at 9V ~ 11V section large percentage, gets the EP setting value that median 10V gathers as EPI.After determining DP and EP setting value, DP value and EP value EPI are gathered, confirm that the DP value of setting and the EPI data acquisition quality of EP value to this compounds have no significant effect;
D) EPI gathers the setting of collision energy (CE) value, and with reference to table 3, the CE value of general common compounds is at 20eV ~ 30eV, but individual compound is if the CE value of crystal violet is when 45 ~ 60eV, just has the crumb data that quality is higher.For this reason, first fixation of C E value is 35, and resetting expansion CE value (CES) is that EPI spectrogram quality are examined or check in 3 kinds of combinations such as 5,10,15.Finally choosing CE is 35eV, CES is the CE setting that 15eV (being equivalent to the progressive mean of 3 spectrograms when CE is respectively 20eV, 35eV, 50eV) gathers as EPI, this setting value can take into account high, medium and low collision energy section preferably, can obtain high-quality data spectrogram.
By above method, establish the senior acquisition method of mass spectrum (sMRM-IDA-EPI) of 12 kinds of target analytes, this pattern can be utilized to carry out examination collection (using the collection ion pair of sMRM), then EPI collection is carried out to the object meeting examination rule (concentration-response exceedes screening aimed concn), obtain detailed ion information to carry out qualitative analysis.
(2) liquid mass spectral database builds
1. online EPI spectral data gathers.After using 12 kinds of object hybrid standard working fluids to use mobile phase dilution, compound concentration is for screening the mixed standard solution of target concentration levels, upper machine analysis gathers online EPI data in sMRM-IDA-EPI mode, use Analyst1.5 software building spectrum storehouse, and improve target analysis information (as Chinese and English title, chemical formula, No. CAS, chemical structural drawing etc.).
2. off-line EPI spectral data gathers.The standard reserving solution of classification preparation is used (in similar, to have isomers, need independent sample introduction) use mobile phase to be diluted to 0.2mg/L, direct mass spectrum sample introduction, off-line EPI data are gathered with reference to online EPI condition, in addition gather 3 CE level (basic, normal, high) EPI spectrograms separately again, can adopt according to compound actual nature the collision energy optimized.By above-mentioned image data typing spectrum storehouse.
3. liquid mass spectral database judges service regeulations.Library searching is one of the most effective qualitative tool, but there is no any regulation and technical stipulation at present in the matched rule formulation of liquid mass spectral database retrieval.This research determines the decision rule of retention time and signal to noise ratio (S/N ratio) according to European Union and U.S.'s relevant regulations; The determination of the EPI spectrogram comparison matching tolerance factor adopts 10 " bare substrate " (each 5 of muscle, milk sample) to prepare extraction standard, the standard EPI collection of illustrative plates composed in storehouse with EPI uses Analyst 1.5 software to carry out the matching analysis, obtain Reinheitszahl (purity), obtain mean value and standard deviation.Through comparison, all object Purity mean value deducts standard deviation and is all greater than 60, for controlling false negative rate to greatest extent, originally determines purity value 60 for the matching tolerance factor.To sum up, the library searching rule determined is:
A) in sample, compound extracts retention time and the standard solution of ion current or adds target analytes in sample and extract compared with the retention time of ion current, and amplitude of variation is no more than 5%;
B) in table 3, the parent ion/daughter ion (transmission ion pair) of listed target analytes must occur simultaneously, and the signal to noise ratio (S/N ratio) (S/N) >=3 of transmission ion pair;
C) spectrogram coupling purity (Purity value) or the matching tolerance factor >=60.
Spectrum storehouse comparison qualitative function is powerful, and the information provided because of it is more.Specify according to European Union 2002/657/EC, this research drainage pattern can obtain confirmation counts >=5.5 (the strictest forbidding veterinary drug of European Union only require confirmation count >=4).If therefore library searching coupling, can carry out fast qualitative confirmation to target analytes.
Claims (10)
1. detect a construction method for the rapid screening liquid mass spectral database that animal-derived food medium or high risk medicine multiple types remains, specifically comprise the steps,
(1) preparation of standard working solution;
(2) analyze pre-treatment, adopt the extraction and cleaning technology of rapid enzymolysis release Conjugate polyamines+quick Solid-Phase Extraction (SPE) to carry out sample to be tested and analyze pre-treatment work;
(3) disposable sample introduction stratographic analysis;
(4) liquid mass spectral database is built;
Described animal-derived food is animal muscle based food, dairy products,
Described excessive risk medicine be Limited Doses at the following excessive risk left drug of 1.0 μ g/kg (A group medicine), be specially:
(a) steroid hormone compounds: testosterone (T), prednisolone (PNSL), betamethasone (BTS), dexamethasone (DTS);
(b) nitroimidazoles medicine and metabolic product thereof: flagyl (MNZ), cough up nitre and to rattle away azoles (RNZ), Dimetridazole (DMZ);
(c) Beta-receptor stimulating agent class material: Clenbuterol (CLB), Tulobuterol (TUL), Cimaterol (CIM);
(d) dye class material: crystal violet (CV) and leuco crystal violet (LCV).
2. method according to claim 1, is characterized in that, the preparation method of the standard working solution described in step (1) is:
Steroid hormone compounds, nitroimidazoles medicine and metabolic product thereof, Beta-receptor stimulating agent class material: adopt its hybrid standard stock solution (0.01g/L) of acetonitrile classification preparation;
The materials such as dye class adopt its hybrid standard stock solution (0.01g/L) of methyl alcohol classification preparation;
Mixing and standard working solution (0.1mg/L), pipetting testosterone, prednisolone, betamethasone, dexamethasone, metronidazole respectively, to cough up rattle away azoles, Dimetridazole, Clenbuterol, Tulobuterol, Cimaterol, crystal violet and leuco crystal violet Standard Stock solutions of nitre appropriate, is the mixed standard solution of 0.1mg/L by acetontrile concentration.
3. method according to claim 1, is characterized in that, the analysis pre-treating method described in step (2) is as follows,
1) extract:
Take homogeneous samples 5g, be placed in 50mL teflon centrifuge tube, add ammonium acetate buffer (0.2mol/L, pH 5.2) 15mL, β-glucosiduronic acid/sulfatase 50 μ L, vortex mixes, 50 DEG C of water-bath vibration 2h, let cool to room temperature, in 0 DEG C, the centrifugal 5min of 15000rpm, transfer supernatant is in another clean centrifuge tube, pH to 9.0 is regulated by NaOH solution (5mol/L), add ethyl acetate 20mL again, sodium chloride 6g, vibration vortex 5min, in 0 DEG C, the centrifugal 5min of 15000rpm, get upper organic phase 35 DEG C and revolve steaming near dry, nitrogen dries up, residue uses 5mL0.1mol/L dissolve with hydrochloric acid solution, ultrasonic 1min, wait until column purification,
2) purify:
MCX post (Waters Oasis MCX SPE pillar) is installed on solid-phase extraction device, uses methyl alcohol 3mL successively, and water 3mL, 3mL 0.1mol/L hydrochloric acid solution activates.Extraction solution is carried on solid-phase extraction column, flows out under gravity, use 3mL water successively, 3mL methyl alcohol and 5mL normal hexane drip washing pillar, drain 2min, add 6mL eluent (ethyl acetate 50mL, methyl alcohol 45mL, ammoniacal liquor 5mL, mixing) wash-out.At eluent 35 DEG C, nitrogen stream dries up, sample dissolution liquid is used (to measure 0.1% formic acid acetonitrile (V/V) solution 5mL, mix with ammonium formate (5mmol/L)-formic acid (0.1%)-aqueous solution 95mL) 0.5mL dissolved residue, ultrasonic 1min, crosses 0.22 μM of miillpore filter;
Described solid-phase extraction device integrates the function of reagent rack, Nitrogen evaporator, and on-stream pressure is relatively uniform, it is made up of reagent rack, operator's console, 5 ~ 10 glass storehouses, 5 ~ 10 pear shape bottles and 1 ~ 2 automatic getter device, can analyze the quick SPE process of thing by realize target.
4. method according to claim 1, is characterized in that, the disposable sample introduction chromatogram analysis method described in step (3) is as follows,
Use chromatographic column is Kinetex C18 post, and employing acetonitrile is organic phase, and formic acid is as ionization reinforcing agent, and formates optimizes agent as peak shape, using the combination of aqueous phase/acid acetonitrile as the flow visualizing optimized;
Actual conditions is: chromatographic column: Kinetex C18,2.6 μm, 2.1mm × 100mm i.d.;
Flow velocity: 0.2mL;
Sample size: 10 μ L;
Column temperature: 30 DEG C;
Gradient elution program is as following table:
A eluent is: 0.1% formic acid-acetonitrile solution;
B eluent is: ammonium formate (5mmol/L)-formic acid (0.1%)-aqueous solution).
5. method according to claim 1, is characterized in that, step (4) described structure liquid mass spectral database method is:
Adopt the senior drainage pattern of level Four bar/ion trap tandem mass spectrometry: presetting many reaction detection (sMRM)-Information Dependent gathers (IDA)-enhancer ion scan (EPI);
Mass Spectrometry Conditions (API 4000 and API 4000Q-TRAP):
Ion gun: electric spray ion source;
Scan mode: positive ion scans;
Detection mode: sMRM-IDA-EPI
Electron spray voltage: 5500V;
Atomization gas pressure: 40psi;
Gas curtain atmospheric pressure: 30psi;
Assisted gas pressure: 45psi;
Ion source temperature: 475 DEG C;
SMRM optimum configurations: MRM detection window is set to 60s, object is set to 1.4s sweep time;
IDA rule: response lag: 3000cps; Dynamic background is deducted; The strongest ion is chosen as 1 to 3;
Enhancer ion scan (EPI) optimum configurations: quality of scanning number scope is 70 ~ 1000Da; Sweep velocity is 10000Da/s; Scanning accumulative frequency is 1; Collision energy is 35eV; Expansion collision energy is 15eV;
By the mass spectral results for 12 kinds of excessive risk medicines, build liquid mass spectral database.
6. method according to claim 5, is characterized in that, what use in the structure of database builds the Analyst1.5 that library software is AB SCIEX company, and the standard of building storehouse is set as according to mentality of designing:
Step (1) provide each target analytes use this research to set liquid phase chromatogram condition system under RT value;
Step (2) provide each target analytes related chemistry information (as title, chemical formula, molecular weight, No. CAS, compounds category, ID, molecular structural formula etc.);
The EPI spectrogram that step (3) gathers under providing each target analytes at least 5 different conditions, namely CE is 20eV, 35eV, 50eV and CE 4 EPI spectrograms when be 35eV, CES being 15eV, in addition, one will at least also be had CE is 35eV, CES under chromatographic condition EPI spectrogram when being 15eV;
Or, optionally increase the EPI spectrogram that step (4) provides target analytes in different substrates as far as possible.
7. the rapid screening liquid mass spectral database that the detection animal-derived food medium or high risk medicine multiple types prepared by the arbitrary described method of claim 1-6 is residual, described animal-derived food is animal muscle based food, dairy products;
Described excessive risk medicine be Limited Doses at the following excessive risk left drug of 1.0 μ g/kg, be specially:
(a) steroid hormone compounds: testosterone (T), prednisolone (PNSL), betamethasone (BTS), dexamethasone (DTS);
(b) nitroimidazoles medicine and metabolic product thereof: flagyl (MNZ), cough up nitre and to rattle away azoles (RNZ), Dimetridazole (DMZ);
(c) Beta-receptor stimulating agent class material: Clenbuterol (CLB), Tulobuterol (TUL), Cimaterol (CIM);
(d) dye class material: crystal violet (CV) and leuco crystal violet (LCV).
8. rapid screening liquid mass spectral database according to claim 7 is detecting the application of the medicament residue in animal-derived food.
9. use the rapid screening liquid mass spectral database described in claim 7 to carry out method that is qualitative or quantitative test to the excessive risk medicament residue in target animal derived food, described method comprises the steps,
(1) preparation standard solution,
(2) analyze pre-treatment, adopt the extraction and cleaning technology of rapid enzymolysis (release Conjugate polyamines)+quick Solid-Phase Extraction (SPE) to carry out sample to be tested and analyze pre-treatment work,
(3) chromatogram and the analysis of liquid matter are carried out to sample to be tested, obtain liquid mass spectrogram, contrast described liquid mass spectral database, carry out qualitative or quantitative test detection.
10. method according to claim 9, is characterized in that,
Described qualitative analysis adopts library searching to carry out, and qualitative criteria is:
(1) in sample, compound extracts retention time and the standard solution of ion current or adds target analytes in sample and extract compared with the retention time of ion current, and amplitude of variation is no more than 5%;
(2) parent ion/daughter ion (transmission ion pair) of target analytes must occur simultaneously, and the signal to noise ratio (S/N ratio) (S/N) >=3 of transmission ion pair;
(3) in sample, compd E PI spectrogram is compared with close concentration level (same concentration numbers magnitude) standard solution in spectrum storehouse or extraction standard solution E PI spectrogram, and spectrogram mates purity (Purity value) >=60;
The method of described quantitative test is as follows:
Use external standard one-point method quantitative, calculate the content of target analytes by formula (1),
Formula (1):
Wherein:
Target analytes content in X-sample, μ g/kg;
The concentration of target analytes in c-sample solution, μ g/L;
The constant volume of V-sample solution, mL;
The quality of m-sample, g;
The R-recovery, %.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105784870A (en) * | 2016-04-15 | 2016-07-20 | 广西壮族自治区梧州食品药品检验所 | Method for online solid-phase extraction of clenbuterol |
| CN106841428A (en) * | 2017-01-04 | 2017-06-13 | 华中农业大学 | A kind of discrimination method of organic liquid milk |
| CN107664671A (en) * | 2017-08-18 | 2018-02-06 | 广东省药品检验所（广东省药品质量研究所、广东省口岸药品检验所） | A kind of left drug detection method of animal-derived food |
| CN108267525A (en) * | 2016-12-30 | 2018-07-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | The detection method of glucocorticoid in dairy products |
| CN109324128A (en) * | 2018-10-22 | 2019-02-12 | 嘉兴迈维代谢生物科技有限公司 | A kind of metabolism group detects general quality-control product and its quality control method |
| CN108037192B (en) * | 2017-10-18 | 2020-02-11 | 华中农业大学 | Liquid chromatography-tandem mass spectrometry method capable of simultaneously detecting five types of medicines in animal edible product and feed |
| CN112362783A (en) * | 2020-11-10 | 2021-02-12 | 江苏省淡水水产研究所 | High-flux and rapid screening method for potential pollutants in non-medicines for fishing |
| CN112858493A (en) * | 2019-11-28 | 2021-05-28 | 内蒙古蒙牛乳业（集团）股份有限公司 | Method for detecting beta-agonists in milk |
| CN115078610A (en) * | 2022-07-22 | 2022-09-20 | 郑州大学第一附属医院 | Method for detecting content of hydrophobic drugs in body fluid |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063791A (en) * | 2012-12-06 | 2013-04-24 | 国家烟草质量监督检验中心 | Method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine |
| CN104297406A (en) * | 2014-10-21 | 2015-01-21 | 中国农业大学 | Method for broad spectrum identification of beta-receptor stimulant medicines |
-
2015
- 2015-06-23 CN CN201510349005.XA patent/CN104931635B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103063791A (en) * | 2012-12-06 | 2013-04-24 | 国家烟草质量监督检验中心 | Method of simultaneously determining contents of 1-OHP ,3-OHB[a]P and 3-OHB[a]A in urine |
| CN104297406A (en) * | 2014-10-21 | 2015-01-21 | 中国农业大学 | Method for broad spectrum identification of beta-receptor stimulant medicines |
Non-Patent Citations (8)
| Title |
|---|
| BRIAN KINSELLA 等: "Current trends in sample preparation for growth promoter and veterinary drug residue analysis", 《JOURNAL OF CHROMATOGRAPHY A》, vol. 1216, 9 September 2009 (2009-09-09), pages 7977 - 8015, XP 026704138, DOI: doi:10.1016/j.chroma.2009.09.005 * |
| M.D. MARAZUELA 等: "A review of novel strategies of sample preparation for the determination of antibacterial residues in foodstuffs using liquid chromatography-based analytical methods", 《ANALYTICA CHIMICA ACTA》, no. 645, 3 May 2009 (2009-05-03), pages 5 - 17 * |
| STANISLAVA G. DMITRIENKO 等: "Recent advances in sample preparation techniques and methods of sulfonamides detection – A review", 《ANALYTICA CHIMICA ACTA》, no. 850, 17 August 2014 (2014-08-17), pages 6 - 25, XP 029079262, DOI: doi:10.1016/j.aca.2014.08.023 * |
| 张毅 等: "分散固相萃取净化与液相色谱／串联质谱法测定牛奶中８类禁用药物残留", 《分析化学研究报告》, vol. 40, no. 5, 31 May 2012 (2012-05-31), pages 724 - 729 * |
| 张鸿伟 等: "液相色谱-四极杆/离子阱质谱同时确证和测定肌肉中16种同化甾体激素残留", 《色谱》, vol. 30, no. 10, 31 October 2012 (2012-10-31), pages 991 - 1001 * |
| 林黎明 等: "液相色谱/串联质谱线性组合法测定动物组织中硝基呋喃代谢产物", 《分析化学研究报告》, vol. 33, no. 8, 31 August 2005 (2005-08-31), pages 1081 - 1086 * |
| 王凤美 等: "超高效液相色谱-串联质谱测定动物源性食品和尿液中4种β-受体激动剂残留", 《分析化学》, vol. 36, no. 12, 31 December 2008 (2008-12-31), pages 1629 - 1635 * |
| 田宏哲 等: "农产品中50余种农药LC-MS/MS质谱数据库的建立及应用", 《食品科学》, vol. 31, no. 4, 28 February 2010 (2010-02-28), pages 218 - 222 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105784870A (en) * | 2016-04-15 | 2016-07-20 | 广西壮族自治区梧州食品药品检验所 | Method for online solid-phase extraction of clenbuterol |
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| CN108037192B (en) * | 2017-10-18 | 2020-02-11 | 华中农业大学 | Liquid chromatography-tandem mass spectrometry method capable of simultaneously detecting five types of medicines in animal edible product and feed |
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