CN113156022A - Veterinary drug residue analysis method and platform based on multi-dimensional electronic identity database of veterinary drug compounds in animal-derived food - Google Patents
Veterinary drug residue analysis method and platform based on multi-dimensional electronic identity database of veterinary drug compounds in animal-derived food Download PDFInfo
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Abstract
The invention belongs to the field of food safety analysis and detection, and particularly relates to a veterinary drug residue analysis method and a veterinary drug residue analysis platform based on a multi-dimensional electronic identity database of a veterinary drug compound in animal-derived food. The method of the invention comprises the following steps: (1) establishing a multi-dimensional electronic identity database of a veterinary drug compound standard, comprising: basic information, chromatographic information and mass spectrum information of the veterinary drug compound standard; (2) detecting an animal source food sample to obtain a chromatogram and a mass spectrogram; (3) and comparing the chromatogram and the mass spectrogram with the information in the multi-dimensional electronic identity database to obtain an analysis result of the veterinary drug compound residue in the animal-derived food sample. The technical scheme provided by the invention can carry out non-targeted batch, quick and efficient detection on veterinary drug compound residues in various animal-derived foods. The method has low requirements on experience and professional knowledge level of operators, is beneficial to large-scale use and has high application value.
Description
Technical Field
The invention belongs to the field of food safety analysis and detection, and particularly relates to a veterinary drug residue analysis method and a veterinary drug residue analysis platform based on a multi-dimensional electronic identity database of a veterinary drug compound in animal-derived food.
Background
Animal-derived food is a main source for human beings to intake high-quality protein, fat, essential amino acid and other nutrients, is one of main food raw materials for three meals a day, and has an increasing demand for animal-derived food consumption along with the continuous improvement of the living standard of residents in China. The special effects of the veterinary drug comprise disease prevention and treatment, growth improvement promotion and the like, and the veterinary drug plays an important role in the aspects of protecting the growth of food animals, improving industrial income, guaranteeing the storage quality of animal-derived food and the like. However, the veterinary drug is a 'double-edged sword' with one handle, the contradiction between population growth and food supply can be greatly relieved by reasonable use, and the excessive use not only directly affects the food safety, but also indirectly affects the ecological environment and animal-derived food trade. The veterinary drug residue safety problem is a main problem in animal-derived food safety, and the veterinary drug safety problem in food has been highly valued by government departments and widely concerned by people. Therefore, the detection of veterinary drug residues is a very important task. However, animal-derived food substrates are complex, and the content of various residual veterinary drugs is very small and the polarity difference is large, so that it is difficult to obtain information of various veterinary drug residues by one-time detection. Most of the traditional veterinary drug residue detection methods can only detect a small amount of veterinary drugs with similar basic structures, and the simultaneous detection of a large amount of veterinary drugs with different chemical structures is difficult to realize.
The ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry (UHPLC-Q/Orbitrap) is a method for detecting trace pollutants of pesticides and veterinary drugs in food by combining the high-efficiency separation capability of the ultra-high performance liquid chromatography with the identification and qualitative capability of the electrostatic field orbital trap high resolution mass spectrometry. With the continuous research of the feasibility of the LC-HRMS-based rapid screening detection technology by the broad researchers, the combination of liquid chromatography and high-resolution mass spectrometry is gradually mature.
The UHPLC-Q/Orbitrap has the advantage of high resolution, and is expected to overcome the defect that different veterinary drugs are difficult to detect in one detection. However, due to the complexity of the system, the spectrogram obtained by detecting the food by the UHPLC-Q/Orbitrap is very complex, a large amount of manual analysis work is needed, and the analysis of the spectrogram is very dependent on the related experience and professional knowledge of a spectrum-solving person, so that the wide application of the technology is limited. Therefore, a technology for applying UHPLC-Q/Orbitrap to veterinary drug detection and performing multi-target quick locking and discrimination on veterinary drug residues is still lacked at present.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a veterinary drug residue analysis method and a veterinary drug residue analysis platform based on a multi-dimensional electronic identity database of a veterinary drug compound in animal-derived food, and the aim of quickly identifying the veterinary drug residue based on a detection result of UHPLC-Q/Orbitrap and the multi-dimensional electronic identity database is fulfilled.
A veterinary drug residue analysis method based on a multi-dimensional electronic identity database of veterinary drug compounds in animal-derived food comprises the following steps:
(1) establishing a multi-dimensional electronic identity database of a veterinary drug compound standard, the multi-dimensional electronic identity database comprising: basic information, chromatographic information and mass spectrum information of the veterinary drug compound standard;
(2) carrying out UHPLC-Q/Orbitrap detection on the animal source food sample to obtain a chromatogram and a mass spectrum;
(3) and comparing the chromatogram and the mass spectrogram with the information in the multi-dimensional electronic identity database to obtain an analysis result of the veterinary drug compound residue in the animal-derived food sample.
Preferably, the basic information comprises Chinese and English names, CAS numbers, chemical molecular formulas and theoretical precise molecular mass numbers of the compounds;
and/or, the chromatographic information comprises retention time;
and/or the mass spectrum information comprises parent ion mass number, fragment ion mass number, ion abundance ratio and ion addition mode.
Preferably, the multidimensional electronic identity database comprises a first-level accurate mass number fingerprint identification database and a second-level HCD fragment ion reference spectrogram confirmation database;
the first-level accurate mass number fingerprint identification database is formed by sequentially recording basic information, chromatographic information and mass spectrum information of the veterinary drug compound in Trace Finder data acquisition and processing software;
the secondary HCD fragment ion reference spectrogram confirmation library is formed by adding and recording secondary HCD fragment ion mass spectrograms obtained by cracking veterinary drug compounds under the optimal collision energy in a high-energy collision pool in mzVault spectrogram management software, wherein the optimal collision energy is the recorded collision energy value CE (20%, 40%, 60% ev) when 3-5 fragment ions with the largest selective ion abundance ratio appear.
Preferably, the chromatographic information and the mass spectrum information are obtained in step (1), or the chromatogram and the mass spectrum are obtained in step (2), and the following chromatographic conditions are adopted:
the chromatographic column is C18A column; and/or, the mobile phase A is an aqueous solution containing 0.02-0.1% of formic acid and 5-20mmol/L of ammonium acetate, and the mobile phase B is acetonitrile; and/or, the temperature of the chromatographic column is 30-40 ℃; and/or, flow rate: 0.3-0.5 mL/min; and/or, the sample injection amount is 5-15 mu L; and/or, the gradient elution procedure is:
preferably, the chromatographic information and the mass spectrum information are obtained in step (1), or the chromatogram and the mass spectrum are obtained in step (2), and the following Q-Orbitrap high-resolution mass spectrum condition is adopted:
heating an electrospray ion source; scanning and collecting mode: collecting positive and negative ions simultaneously, and performing primary full scanning/data-dependent secondary scanning; and/or, the spray voltage: 3.5kV (+), -3.0kV (-); and/or, ion transfer tube and heater temperatures: 325 ℃ and 450 ℃; and/or, sheath gas and auxiliary gas flow rates: 35arb, 10 arb; and/or, acquisition range: 100 to 1000 m/z; and/or, resolution: full MS 75000, dd-MS217500 (mm); and/or, stemped (n) CE (20%, 40%, 60%); and/or, maximum capacity of ion number in C-Trap: full MS 3X 106、dd-MS2 2×105(ii) a And/or, the maximum injection time in the C-Trap: full MS 100MS, dd-MS2 50ms；
And/or, the tuning correction of the mass axis of the Q-Orbitrap mass spectrum is as follows: injecting the Calmix correction fluid into a mass spectrometer, debugging and observing the characteristic ion peak of the Calmix correction fluid, and automatically correcting by tuning application software when the ion peaks of positive ions and negative ions are stable.
Preferably, in step (3), the criterion for determining the positivity of a veterinary drug compound is that the raw data extracted within the 5ppm mass window satisfies all of the following conditions:
1) retention time of parent ion mass spectrum peakThe error range is +/-15sInternal;
2) the mass number deviation of the parent ions is less than or equal to 5 ppm;
3) the matching number of fragment ions in the secondary mass spectrogram is more than or equal to 2, and the mass number deviation of the fragment ions is less than or equal to 10 ppm.
Preferably, the veterinary drug compound is at least one selected from the group consisting of androgens, oestrogens, progestogens, glucocorticoids, zearalanol compounds, beta-receptor agonists, sulphonamide antibiotic drugs, quinolone antibiotics and macrolide antibiotics.
Preferably, the veterinary drug compound is selected from the group consisting of dehydrotestosterone, epitestosterone, fluoxymesterone, methyltestosterone, nandrolone propionate, nandrolone, testosterone propionate, trenbolone, meperirone, nandrolone propionate, stanozolol, norandrostenedione, androstenedione, 17 alpha-methylisotosterone, meperidol, epiandrosterone, 17 beta-hydroxyandrostan-3-one, mesterone, danazol, altrenogest, chlormadinone acetate, levonorgestrel, medroxyprogesterone acetate, norethindrone acetate, 21-α-hydroxyprogesterone, norethindrone, 17-alpha-hydroxyprogesterone, beclomethasone propionate, betamethasone, cortisone acetate, hydrocortisone, methylprednisolone, prednisolone, triamcinolone acetonide, triamcinolone, cortisone, dexamethasone, hydrocortisone acetate, beclomethasone, flumethasone, budesonide, fluocinolone acetonide, fludrocortisone acetate, fluorometholone, clobetasol propionate, aldosterone, prednisone, estriol, estrone, dienestrol, diethylstilbestrol, estriol, estradiol, octylphenol, 4-nonylphenol, bisphenol a, ethinylestradiol, alpha-zearalenol, beta-zearalenol, zearalenone, alpha-zearalanol, beta-zearalanol, clenbuterol, ractopamine, chlorpromazine, pentostatin, oxprenalin, formoterol, clomipramine, clotrimol, clotrimazole, flunisolone, prednisolone, triamcinolone acetonide, flunisolone, and other, flunisolone, and other, Fenoterol, salbutamol, bambuterol, salbutamol, terbutaline, tulobuterol, cimaterol, mabuterol, sulphazopyrazole, sulphanilide, sulphapyrine, sulphacetamide, sulphamethoxamine, sulphaxazole, sulphadimethoxazole, sulphadimethyneAt least one of pyrimidine, sulfamethazine, sulfamethoxazole, sulfamethazine, sulfamethoxypyridazine, sulfamonomethoxine, sulfamquinoxaline, sulfamodiethoxydiazine, sulfachloropyrazine, sulfachlorpyridazine, sulfaguanidine, sulfadiazine, sulfathiazole, sulfamethazine, sulfa nitrobenzene, orbifloxacin, danofloxacin, enrofloxacin, flumequine, fleroxacin, ciprofloxacin, lomefloxacin, nalidixic acid, norfloxacin, pefloxacin, sarafloxacin, difloxacin, sparfloxacin, enoxacin, ofloxacin, erythromycin, kitasamycin, lincomycin, tylosin, tilmicosin, tavermectin, oleandomycin, doramectin, ivermectin, and roxithromycin.
The present invention also provides a computer readable storage medium having stored thereon the above-described multi-dimensional electronic identity database; or have stored thereon a computer program for implementing the above-described method.
The invention also provides a veterinary drug residue analysis platform based on the multi-dimensional electronic identity database of the veterinary drug compounds in the animal-derived food, which comprises the following steps:
the detection module is used for carrying out UHPLC-Q/Orbitrap detection on the animal source food;
a multi-dimensional electronic identity database;
and the analysis module is used for comparing the result of the UHPLC-Q/Orbitrap detection with the multi-dimensional electronic identity database and giving an analysis result of the veterinary drug residue.
The technical scheme of the invention has the following beneficial effects:
(1) the invention provides a construction method and a detection platform of a multi-dimensional electronic identity database of veterinary drug compounds in animal-derived food based on UHPLC-Q/Orbitrap, wherein the multi-dimensional electronic identity database of 128 veterinary drug compounds is established based on UHPLC-Q/Orbitrap technology on the basis of obtaining chromatographic and mass spectrum information, and the electronic identity replaces a real standard substance, so that automatic, multi-target, high-precision and high-efficiency non-target veterinary drug residue detection can be realized. The method has important significance for realizing multi-target quick locking and discrimination confirmation of hundreds of trace veterinary drug residues in animal-derived food. The method of the invention is simpler and more efficient when being used for batch processing of various veterinary drug compounds, has lower requirements on knowledge and experience level of operators, and is beneficial to wide application of the technology of the invention.
(2) The invention relates to a high-flux detection method based on a multi-dimensional electronic identity database and a detection platform of a veterinary drug compound, which replaces the traditional targeted qualitative analysis method based on real standard reference, and realizes the cross-over development of a high-flux non-targeted detection technology from the targeted detection of a known analyte to an unknown analyte of veterinary drug residues. The method has the advantages of shortening sample analysis time, improving sample detection efficiency, saving resources, reducing pollution and meeting the requirements of sustainable development and environmental friendliness.
(3) In the preferred scheme, the liquid chromatography condition and the high-resolution mass spectrum parameter are optimized, so that the optimal instrument condition is obtained, and a rapid, efficient, stable and reliable instrument analysis method is established.
(4) In the preferred scheme, the database (MS) is identified by first-level accurate mass number fingerprint identification through searching and comparing the acquired information of the veterinary drug compound to be detected and the multidimensional electronic identity information in the database according to the retention time, accurate mass number, ion abundance ratio, collision energy, second-level HCD fragment ion graph and other information of the target compound1) Screening and identifying the medium-color mass spectrum fingerprint information, and matching with a secondary HCD fragment ion reference spectrogram verification library (MS)2) The medium characteristic spectrogram confirms, and the accuracy of the accurate and qualitative screening result of the medicine is ensured.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Drawings
Fig. 1 is a flow chart of the construction of a multi-dimensional electronic identity database and a detection platform for veterinary drug compounds.
Fig. 2 is a multi-dimensional electronic identity database model of veterinary drug residual chemical pollutants.
Detailed Description
It should be noted that, in the embodiment, the algorithm of the steps of data acquisition, transmission, storage, processing, etc. which are not specifically described, as well as the hardware structure, circuit connection, etc. which are not specifically described, can be implemented by the contents disclosed in the prior art.
The following examples were prepared using the following laboratory instruments: vanqish UHPLC/Q active Plus ultra high performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry system (HP-1510, containing 65 μm PDMS/DVB extraction head, Thermo Fisher company, USA); Milli-Q ultrapure water system (Millipore, USA); electronic balance model ME203 (Mettler Toledo, Switzerland), Waters Acquity BEH C18Column (2.1 mm. times.100 mm, 1.7 μm, Waters, USA), Q-exact Orbitrap Special Mass axis calibration solution (Thermo Scientific, USA).
Example 1 creation of a multidimensional electronic identity database
As shown in fig. 1 and 2, the multidimensional electronic identity database of veterinary compounds comprises basic information, chromatographic information and mass spectrometric information of veterinary compound standards. The basic information comprises Chinese and English names of the compounds, CAS numbers, chemical molecular formulas and theoretical accurate molecular mass numbers; the chromatographic information comprises retention time; the mass spectrum information comprises a parent ion mass number, a fragment ion mass number, an ion abundance ratio and an ion addition mode.
The multi-dimensional electronic identity database comprises a first-level accurate mass number fingerprint identification database (MS)1) And secondary HCD fragment ion reference spectrum validation library (MS)2). The first-class accurate mass number fingerprint identification database is formed by sequentially recording the basic information, chromatographic information and mass spectrum information of the veterinary drug compound in Trace Finder4.1 data acquisition and processing software. The secondary HCD fragment ion reference spectrogram confirmation library is the most important veterinary drug compound in the high-energy collision poolAdding a spectrogram confirmation library formed by recording a secondary HCD fragment ion mass spectrogram obtained by cracking under the optimal collision energy into mzVault spectrogram management software, wherein the optimal collision energy is a collision energy value recorded when 3-5 fragment ions with the maximum abundance ratio of selected ions appear, and Stepped (N) CE (20%, 40%, 60%).
The chromatographic conditions for collecting chromatographic information based on UHPLC-Q/Orbitrap are as follows:
a chromatographic column: waters Acquity BEH C18(2.1 mm. times.100 mm, 1.7 μm). Temperature of the column: at 40 ℃. Flow rate: 0.5 mL/min. Mobile phase a (aqueous phase) was ammonium acetate (20mmol/L) in water with 0.1% formic acid and mobile phase B (organic phase) was acetonitrile. Sample introduction amount: 10 μ L. Gradient elution procedure:
table 1 liquid chromatography mobile phase gradient elution procedure
The mass spectrum conditions for collecting chromatographic information based on UHPLC-Q/Orbitrap are as follows:
heating an electrospray ion source; scanning and collecting mode: simultaneous positive and negative ion acquisition, first-order Full scan/data-dependent second-order scan (Full MS/dd-MS)2) (Top N); spraying voltage: 3.5kV (+), -3.0kV (-); ion transfer tube and heater temperatures: 325 ℃ and 450 ℃; sheath gas and auxiliary gas (N)2) Flow rate: 35arb, 10 arb; the collection range is as follows: 100 to 1000 m/z; resolution ratio: 75000(Full MS), 17500 (dd-MS)2) (ii) a Stepped (N) CE (20%, 40%, 60%); maximum capacity of ion number in C-Trap (AGC target): 3X 106(Full MS)、2×105(dd-MS2) (ii) a Maximum injection time in C-Trap: 100MS (full MS), 50MS (dd-MS)2)。
And (3) mass axis tuning correction of the Q/Orbitrap mass spectrum, wherein the mass axis tuning correction is as follows: firstly, a special adaptive injector is picked up to absorb a proper amount of Calmix correction fluid, the Calmix correction fluid is directionally and slowly injected into a mass spectrometer, characteristic ion peaks of the injection fluid are debugged and observed, and when the ion peaks of positive ions and negative ions are stable, automatic correction is carried out by using tuning application software Q active 2.1. The characteristic ion peak of the injection is debugged and observed when the positive ion is from the low Caffeine (Caffeine) mass-to-charge ratio to the middle MRFA (poly tyrosine short peptide), and gradually rises to the high mass-to-charge ratio until the ion peak appears and ends with 22 all the time; meanwhile, the negative ions are from low K12 (sodium dodecyl sulfate) to medium S T (sodium taurocholate) and then to high mass-to-charge ratio until the ion peak appears and ends with 80; the ion peaks of the positive and negative ions are stable at this time.
In this example, a multidimensional electronic identity database of 128 veterinary compounds was created according to the above method, and the specific information of the 128 veterinary compounds is shown in the following table:
TABLE 2 UHPLC-Q/Orbitrap128 veterinary drug Compound List
Example 2 high throughput detection technique for UHPLC-Q/Orbitrap of 128 veterinary drugs in livestock and poultry meat
1. Preparation of samples to be tested
Collecting 24 batches of pork, beef, mutton and chicken in a province city in southwest, selecting edible parts in samples, preparing samples according to GB/T30891-2014, and filling the samples into clean containersThe container is sealed and marked. Accurately weighing 2.5g (to the percentile) of the sample to be detected after the uniform mixing treatment, transferring the sample to a 50mL PP centrifuge tube, and slowly adding 10mL of acetonitrile (80%) -formic acid (0.2%) aqueous solution. Standing for 10min, adding ceramic homogenous protons (2 particles), rapidly and violently shaking and carrying out vortex oscillation for 30min (2000r/min) to fully disperse the sample. Centrifuging the sample in a high speed centrifuge at 9500r/min for 5min, collecting supernatant 5mL, and passing through 6cc 200mg of the supernatant at 1 drop/s
PRIME HLB solid phase extraction column. Collecting the effluent liquid of the latter half section and accurately measuring 2mL, N2Blow-dry, fix volume with 1mL initial mobile phase, filter with 0.22 μm Teflon filter membrane, and test with UHPLC-Q/Orbitrap HRMS.
2. UHPLC-Q/Orbitrap HRMS operating conditions
The compounds were separated by ultra high performance liquid chromatography system, UHPLC conditions: a chromatographic column: waters Acquity BEH C18(2.1 mm. times.100 mm, 1.7 μm). Temperature of the column: at 25 ℃. Flow rate: 0.5 mL/min. Mobile phase a (aqueous phase) was ammonium acetate (20mmol/L) in water with 0.1% formic acid and mobile phase B (organic phase) was acetonitrile. Gradient elution procedure: 0-0.5 min, 5% B; 0.5-15 min, 98% B; 15-20 min, 98% B; 20-20.1 min, 5% B; 20.1-25 min, 5% B. Sample introduction amount: 10 μ L.
Q/Orbitrap high resolution mass spectrometry conditions: heating an electrospray ion source; scanning and collecting mode: simultaneous positive and negative ion acquisition, first-order Full scan/data-dependent second-order scan (Full MS/dd-MS)2) (Top N); spraying voltage: 3.5kV (+), -3.0kV (-); ion transfer tube and heater temperatures: 325 ℃ and 450 ℃; sheath gas and auxiliary gas (N)2) Flow rate: 35arb, 10 arb; the collection range is as follows: 100 to 1000 m/z; resolution ratio: 75000(Full MS), 17500 (dd-MS)2) (ii) a Stepped (N) CE (20%, 40%, 60%); maximum capacity of ion number in C-Trap (AGC target): 3X 106(Full MS)、2×105(dd-MS2) (ii) a Maximum injection time in C-Trap: 100MS (full MS), 50MS (dd-MS)2)。
3. Data processing and analysis
Using the first-class accurate mass number fingerprinting database (MS) established in example 1 in TraceFinder 4.1 analysis software1) Carrying out retrieval matching analysis on sample data acquired by UHPLC-Q/Orbitrap, carrying out comparative qualitative identification (according to set screening standard) on retention time, accurate parent ion mass number, fragment ion mass number and other related parameters in Database by utilizing automatic deconvolution function of mass spectrum software, and aiming at suspicious compound utilization and secondary HCD fragment ion reference spectrogram validation library (MS)2) Further confirmation analysis.
4. The detection result
After the detection is finished, counting the information of the veterinary drug residues contained in the sample solution, detecting 17 veterinary drug residues in the livestock and poultry meat detection, and detecting the veterinary drug residues for 59 times in total, wherein the sample is 59 batches (17 pork and chicken, 14 beef and 11 mutton). The specific results are shown in Table 3.
TABLE 3 detection results of veterinary drug residues in livestock and poultry meat samples
Example 3: technical example for detecting UHPLC-Q/Orbitrap high flux of 128 veterinary drugs in fish
1. 9 batches of turbot, mandarin fish, snakehead and grass carp are collected in a certain province city in the southwest region, and the preparation of a sample to be detected, the UHPLC-Q/Orbitrap HRMS operation conditions and the data processing and analyzing process refer to example 2.
2. The detection result
After the detection is finished, the information of the veterinary drug residues contained in the sample solution is counted, 13 veterinary drug residues are detected in the fish meat detection, and the veterinary drug residues are detected 22 times in total, wherein 18 times of samples are related (7 batches of turbot, 5 batches of mandarin fish, 4 batches of grass carp and 2 batches of snakehead). The specific results are shown in Table 4.
TABLE 4 detection results of veterinary drug residues in fish samples
Example 4: high-throughput detection technology for 128 veterinary drugs UHPLC-Q/Orbitrap in pork liver and chicken liver
1. 8 batches of pork liver and chicken liver are collected in a certain province city in the southwest region, and the preparation of a sample to be detected, the operation conditions of UHPLC-Q/Orbitrap HRMS and the data processing and analyzing process refer to example 2.
2. The detection result
After the detection is finished, the information of the veterinary drug residues contained in the sample solution is counted, 7 veterinary drug residues are detected by the detection of the pig liver and the chicken liver, and the frequency of the detected veterinary drug residues is 12 times in total, which relates to 8 batches of samples (4 pig livers and 3 chicken livers). The specific results are shown in Table 5.
TABLE 5 detection results of veterinary drug residues in pork liver and chicken liver samples
According to the embodiments, the technical scheme provided by the invention can carry out non-targeted batch, rapid and efficient detection on veterinary drug compound residues in various animal-derived foods. The method has low requirements on experience and professional knowledge level of operators, is beneficial to large-scale use and has high application value.
Claims (10)
1. A veterinary drug residue analysis method based on a multi-dimensional electronic identity database of veterinary drug compounds in animal-derived food is characterized by comprising the following steps:
(1) establishing a multi-dimensional electronic identity database of a veterinary drug compound standard, the multi-dimensional electronic identity database comprising: basic information, chromatographic information and mass spectrum information of the veterinary drug compound standard;
(2) carrying out UHPLC-Q/Orbitrap detection on the animal source food sample to obtain a chromatogram and a mass spectrum;
(3) and comparing the chromatogram and the mass spectrogram with the information in the multi-dimensional electronic identity database to obtain an analysis result of the veterinary drug compound residue in the animal-derived food sample.
2. The analytical method of claim 1, wherein: the basic information comprises Chinese and English names of the compounds, CAS numbers, chemical molecular formulas and theoretical accurate molecular mass numbers;
and/or, the chromatographic information comprises retention time;
and/or the mass spectrum information comprises parent ion mass number, fragment ion mass number, ion abundance ratio and ion addition mode.
3. The analytical method according to claim 1 or 2, wherein: the multidimensional electronic identity database comprises a first-level accurate mass number fingerprint identification database and a second-level HCD fragment ion reference spectrogram confirmation database;
the first-level accurate mass number fingerprint identification database is formed by sequentially recording basic information, chromatographic information and mass spectrum information of the veterinary drug compound in Trace Finder data acquisition and processing software;
the secondary HCD fragment ion reference spectrogram confirmation library is formed by adding and recording secondary HCD fragment ion mass spectrograms obtained by cracking veterinary drug compounds under the optimal collision energy in a high-energy collision pool in mzVault spectrogram management software, wherein the optimal collision energy is the recorded collision energy value CE (20%, 40%, 60% ev) when 3-5 fragment ions with the largest selective ion abundance ratio appear.
4. The analytical method of claim 1, wherein: the chromatographic information and the mass spectrum information are obtained in the step (1), or the chromatographic conditions are adopted when the chromatogram and the mass spectrum are obtained in the step (2):
the chromatographic column is C18A column; and/or, the mobile phase A is an aqueous solution containing 0.02-0.1% of formic acid and 5-20mmol/L of ammonium acetate, and the mobile phase B is acetonitrile; and/or, the temperature of the chromatographic column is 30-40 ℃; and/or, flow rate: 0.3-0.5 mL/min; and/or, the sample injection amount is 5-15 mu L; and/or, gradient washThe removing procedure is as follows:
5. the analytical method of claim 1, wherein: the chromatographic information and the mass spectrum information are obtained in the step (1), or the chromatogram and the mass spectrum are obtained in the step (2), the following Q-Orbitrap high-resolution mass spectrum condition is adopted:
heating an electrospray ion source; scanning and collecting mode: collecting positive and negative ions simultaneously, and performing primary full scanning/data-dependent secondary scanning; and/or, the spray voltage: 3.5kV (+), -3.0kV (-); and/or, ion transfer tube and heater temperatures: 325 ℃ and 450 ℃; and/or, sheath gas and auxiliary gas flow rates: 35arb, 10 arb; and/or, acquisition range: 100 to 1000 m/z; and/or, resolution: full MS 75000, dd-MS217500 (mm); and/or, stemped (n) CE (20%, 40%, 60%); and/or, maximum capacity of ion number in C-Trap: full MS 3X 106、dd-MS2 2×105(ii) a And/or, the maximum injection time in the C-Trap: full MS 100MS, dd-MS2 50ms；
And/or, the tuning correction of the mass axis of the Q-Orbitrap mass spectrum is as follows: injecting the Calmix correction fluid into a mass spectrometer, debugging and observing the characteristic ion peak of the Calmix correction fluid, and automatically correcting by tuning application software when the ion peaks of positive ions and negative ions are stable.
6. The analytical method of claim 1, wherein: in step (3), the criterion for determining that a certain veterinary drug compound is positive is that the raw data extracted within the 5ppm mass window satisfies all of the following conditions:
1) the retention time error range of the mass spectrum peak of the parent ion is within +/-15 s;
2) the mass number deviation of the parent ions is less than or equal to 5 ppm;
3) the matching number of fragment ions in the secondary mass spectrogram is more than or equal to 2, and the mass number deviation of the fragment ions is less than or equal to 10 ppm.
7. The analytical method of claim 1, wherein: the veterinary drug compound is at least one of androgen, estrogen, progestogen, glucocorticoid, zearalanol compound, beta-receptor stimulant, sulfonamide antibiotic drug, quinolone antibiotic or macrolide antibiotic.
8. The analytical method of claim 7, wherein: the veterinary drug compound is selected from dehydrotestosterone, epitestosterone, fluoxymesterone, methyltestosterone, nandrolone propionate, nandrolone, testosterone propionate, trenbolone, meperiandrone, nandrolone phenylpropionate, stanozolol, norandrostenedione, androstenedione, 17 alpha-methylisotosterone, meiandrol, epiandrosterone, 17 beta-hydroxyandrostan-3-one, mesterone, danazol, altrenogest, chlormadinone acetate, levonorgestrel, medroxyprogesterone acetate, megestrol acetate, norelgestromin acetate, norethisterone acetate, 21-alpha-hydroxyprogesterone, norethindrone, 17-alpha-hydroxyprogesterone, beclomethasone propionate, betamethasone, cortisone acetate, hydrocortisone, methylprednisolone, prednisolone, triamcinolone, cortisone, dexamethasone, hydrocortisone acetate, hydrocortisone, medroxyprogesterone acetate, norgestrel, norgestimate, medroxyprogesterone acetate, medroxyprogesterone, medrysone, and, Beclomethasone, flumethasone, budesonide, fluocinonide, fludrocortisone acetate, fluorometholone, clobetasol propionate, aldosterone, prednisone, estriol, estrone, dienestrol, diethylstilbestrol, estradiol, octylphenol, 4-nonylphenol, bisphenol A, ethinylestradiol, alpha-zearalenol, beta-zearalenol, zearalenone, alpha-zearalanol, beta-zearalanol, clenbuterol, ractopamine, chlorpromazine, penbuterol, metaproterenol, formoterol, fenoterol, secobronol, bambuterol, salbutamol, terbutaline, tuobuterol, cimaterol, mabuterol, pyrazolsulfaphenamide, sulfapyridine, sulfacetamide, sulfamethoxydiazine, sulfamethoxazole, sulfamethazine dimethyidipine, sulfamethazine, At least one of sulfamethoxazole, sulfamethazine, sulfamethoxypyridazine, sulfamonomethoxine, sulfamoxine, sulfamodiethoxypyrimidine, sulfamchloropyrazine, sulfamamidine, sulfadiazine, sulfathiazole, sulfamethazine, sulfa nitrobenzene, orbifloxacin, danofloxacin, enrofloxacin, flumequine, fleroxacin, ciprofloxacin, lomefloxacin, nalidixic acid, norfloxacin, fleroxacin, sarafloxacin, difloxacin, sparfloxacin, enoxacin, ofloxacin, erythromycin, kitasamycin, lincomycin, tylosin, tilmicosin, tavermectin, oleandomycin, doramectin, ivermectin, and roxithromycin.
9. A computer-readable storage medium characterized by: a multi-dimensional electronic identity database as claimed in any one of claims 1 to 8 stored thereon;
or having stored thereon a computer program for implementing the method of any one of claims 1-8.
10. A veterinary drug residue analysis platform based on a multi-dimensional electronic identity database of veterinary drug compounds in animal-derived food, comprising:
the detection module is used for carrying out UHPLC-Q/Orbitrap detection on the animal source food;
the multi-dimensional electronic identity database of any one of claims 1-8;
and the analysis module is used for comparing the result of the UHPLC-Q/Orbitrap detection with the multi-dimensional electronic identity database and giving an analysis result of the veterinary drug residue.
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