CN111830141A - Method for screening pesticide residues in medicine-food homology food by using pesticide residue mass spectrum database - Google Patents
Method for screening pesticide residues in medicine-food homology food by using pesticide residue mass spectrum database Download PDFInfo
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Abstract
The invention discloses a method for screening pesticide residues in medicinal and edible homology food by a pesticide residue mass spectrum database, which comprises the following steps: step one, establishing an LC-QTOF pesticide mass spectrum database; and step two, establishing a GC-QTOF pesticide mass spectrum database, and step three, carrying out rapid qualitative and quantitative analysis on the sample by adopting the LC-QTOF pesticide mass spectrum database and/or the GC-QTOF pesticide mass spectrum database so as to compare the retention time of the sample to be detected with the retention time of the pesticide compound to judge whether the sample contains pesticide compound residues and quantitatively detect the content of the pesticide compound. The invention simultaneously establishes a primary mass spectrum database and a secondary mass spectrum database of the gas chromatography-quadrupole time-of-flight mass spectrum and the liquid chromatography-quadrupole time-of-flight mass spectrum, and improves the screening accuracy through mutual evidence among the databases and further confirmation of secondary mass spectrum information.
Description
Technical Field
The invention belongs to the field of analytical chemistry, and particularly relates to a method for screening pesticide residues in medicinal and edible homology food by using a pesticide residue mass spectrum database.
Background
The product with homology of medicine and food is a special food with obvious Chinese characteristics, and has a long history in China. However, at present, due to the lack of reliable safety evaluation means, the food and medicine in China does not meet the food safety standard of Europe and America, and the food and medicine are difficult to enter the international market. Up to now, major trade member countries of the european union, japan, usa, korea, etc. have conducted safety evaluation of chemicals in health foods, and established limit regulations and positive lists for illegally adding drugs or pesticide residues. The 'catalog of materials which are food and Chinese medicinal materials according to the tradition' (2018 edition) issued by the national health committee of China includes 105 kinds of medicinal and edible foods, but the detection standards of pesticide residues and illegally added medicines are lacked. Therefore, the database of the spectrum of pesticides and illegal additives in the food with homology of medicine and food has important significance for guaranteeing food safety of people in China and assisting the food with homology of medicine and food to break through international trade barriers.
The early pesticide residue detection method mainly comprises a gas chromatography and a liquid chromatography, the methods mainly aim at the detection of single-class target compounds, the instrument sensitivity can not completely meet the limit requirements of lower and lower residues and undetected residues at home and abroad, and the qualitative capability is poor. Along with the combination of the chromatography and the single-stage or tandem mass spectrometry technology, the sensitivity and the qualitative capability of the detection method are remarkably improved, a gas chromatography-mass spectrometry combined method and a liquid chromatography-tandem mass spectrometry method become mainstream technologies for trace analysis of pesticide residues, and the simultaneous qualitative and quantitative detection of various compounds is realized by selecting an ion monitoring mode and a multi-reaction monitoring mode. However, due to the limitation of the scanning speed of the instrument, the detection flux of the gas chromatography-mass spectrometry combined method and the liquid chromatography-tandem mass spectrometry is limited and is only suitable for detecting the known target object, the method has obvious lag in setting up a limited amount of rules and regulations, and the rapidly-increasing detection requirement cannot be met. With the development of high-resolution mass spectrometry technology, the time-of-flight mass spectrometry is applied to the field of trace analysis by virtue of the advantages of high flux, high resolution, spectrum library retrieval, standard-free analysis and the like. The kit can be matched with functions of high-throughput and full-quality data acquisition, spectral library retrieval and the like to quickly and accurately screen and measure a large number of compounds. Although the technology is adopted to establish food pesticide residue database research in documents, the screening of pesticide compounds in medicinal and edible products or the screening of one type of compounds in various matrixes is not reported. In addition, the pretreatment step of pesticide quick detection at present usually adopts SPE column or QuEChERS step for purification. The SPE method has complicated steps and long purification time, while the adsorbent of the QuEChERS method usually causes adsorption to a target compound and cannot be suitable for all pesticide compounds, so that the method cannot meet the requirements of non-directional and high-throughput screening of cross-classified compounds in various complex matrixes in actual work.
Disclosure of Invention
The invention aims to provide a method for screening pesticide residues in food with homology of medicine and food by using a pesticide residue mass spectrum database.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for screening pesticide residues in medicinal and edible homology food by a pesticide residue mass spectrum database comprises the following steps:
step one, establishing LC-QTOF pesticide mass spectrum database
A. Establishing a primary mass spectrum database of 854 common pesticides for medicinal and edible food: collecting primary map data of pesticide Compound standard products of 854 kinds of medicinal and edible food, obtaining unique retention time of each pesticide Compound, and inputting the retention time into PCDL (Personal Compound Database, PCDL) software for screening pesticide compounds in medicinal and edible food;
B. establishing a secondary mass spectrum database: collecting secondary spectrogram data of the pesticide compound standard substance confirmed by the primary mass spectrum database, and importing the secondary mass spectrum data into a spectrogram editing item in PCDL software for confirming pesticide compounds in food with homology of medicine and food;
step two, establishing a GC-QTOF pesticide mass spectrum database, which comprises the following steps:
A. establishing a primary mass spectrum database of 727 common pesticides for food and medicine homologous food, collecting primary spectrum data of pesticide compound standard products of the 727 common food and medicine homologous food, obtaining unique retention time of each pesticide compound, inputting the retention time into PCDL software, and introducing the primary mass spectrum into a spectrum editing item in the PCDL software for screening the pesticide compounds in the food and medicine homologous food;
B. establishing a secondary mass spectrum database: the primary mass spectrum database confirms the secondary spectrogram data of the pesticide compound standard product; then, importing the secondary mass spectrum data into a spectrogram editing item in PCDL software;
and step three, preparing a sample to be detected, and performing rapid qualitative and quantitative analysis on the sample by adopting the LC-QTOF pesticide spectrum database and the GC-QTOF pesticide spectrum database so as to compare the retention time of the sample to be detected with the retention time of a pesticide compound to judge whether the sample contains pesticide compound residues and quantitatively detect the content of the pesticide compound.
According to the invention, before the preparation of the sample to be detected in the third step, the method further comprises a sample pretreatment step, wherein the sample pretreatment step comprises the following steps: weighing a sample in a centrifuge tube, adding excessive acetonitrile and a QuEChERS extraction bag, wherein the QuEChERS extraction bag contains anhydrous magnesium sulfate and sodium acetate in a mass ratio of 4-6: 1-1.5, and performing vortex oscillation; centrifuging; placing a SinChERS small column into a centrifugal tube filled with an extracting solution, wherein the SinChERS small column is filled with anhydrous magnesium sulfate, 300i graphitized carbon black, C18 and PSA, and the mass ratios of the anhydrous magnesium sulfate, 200-300 i graphitized carbon black, C18 and PSA to a sample are respectively 10-15%, 1-2% and 0.5-1%; then, 500. mu.L of the supernatant was aspirated and the volume was adjusted to 1mL with acetonitrile, and the supernatant was filtered through a nylon filter and subjected to LC-QTOF measurement.
Preferably, the sample pretreatment step is: weighing a sample in a centrifuge tube, adding excessive acetonitrile and a QuEChERS extraction bag, wherein the QuEChERS extraction bag contains anhydrous magnesium sulfate and sodium acetate in a mass ratio of 4:1, and performing vortex oscillation; centrifuging; placing a SinChERS small column in a centrifugal tube filled with an extracting solution, wherein the SinChERS small column is filled with anhydrous magnesium sulfate, 300i graphitized carbon black, C18 and PSA, and the mass ratios of the anhydrous magnesium sulfate, the 300i graphitized carbon black, the C18 and the PSA to the sample are respectively 12%, 2% and 0.5%; then, 500. mu.L of the supernatant was aspirated and the volume was adjusted to 1mL with acetonitrile, and the supernatant was passed through a 0.22 μm nylon filter and subjected to LC-QTOF measurement. According to the invention, the first-stage mass spectrum database of the first step is established by the following steps: inputting the information of the pesticide compound into an edited compound information item in PCDL software; then, respectively preparing the collected pesticide compound standard substance into a standard stock solution and a standard intermediate solution, adjusting chromatographic conditions and mass spectrum conditions, and acquiring primary spectrogram data of a working solution of the standard substance in an MS mode to obtain information such as retention time, parent ions, mass deviation, ionization form and the like of a target object; finally, the retention time of each pesticide is input in PCDL software;
the information of the pesticide compound includes name, CAS number, molecular formula, precise molecular weight, etc.;
the second-level mass spectrum database in the first step is established by the following steps: under the condition of confirmation by primary mass spectrum data, according to the chromatographic conditions and mass spectrum conditions of a primary mass spectrum database, giving different collision energy to target ions to a standard working solution in a Targeted MS/MS mode, and collecting secondary spectrum data; then, the secondary mass spectrometry data is imported under a spectrogram editing entry in PCDL software.
Further: preparing a standard stock solution in the first step: accurately weighing each pesticide compound standard substance in a volumetric flask respectively, dissolving with a small amount of methanol or n-hexane, and then preparing to a constant volume to obtain a 1.0mg/mL standard stock solution; preparing a standard intermediate solution in the first step: a certain amount of pesticide solution is sucked, diluted by methanol or acetonitrile and prepared into standard intermediate solution with the concentration of 50 mu g/mL in groups.
According to the invention, the chromatographic conditions and mass spectrometry conditions of step one are respectively:
(1) chromatographic conditions
Mobile phase: a: 5mmol/L aqueous ammonium acetate and 0.1% formic acid, B: acetonitrile; flow rate: 0.4 mL/min; sample introduction amount: 5 mu L of the solution;
(2) conditions of Mass Spectrometry
An Agilent 6540 liquid chromatogram/four-level rod-flight time mass spectrometer adopts an ESI source, the capillary voltage is 5500V, the declustering voltage is 150V, the taper hole voltage is 65V, and the eight-level rod radio frequency voltage is 750V; the collision gas, the atomization gas and the drying gas are all nitrogen; the temperature of the sheath gas is 350 ℃, and the flow rate of the sheath gas is 11L/min; the temperature of the drying gas is 280 ℃, and the flow rate of the drying gas is 9L/min.
According to the invention: the second step of establishing a primary mass spectrum database of 727 common pesticides for medicinal and edible food comprises the following steps: firstly, inputting information such as compound name, CAS number, molecular formula, accurate molecular weight and the like of pesticide into an edited compound information item in PCDL software; then, respectively preparing the collected standard substance of the library building compound into a standard stock solution and a working solution, adjusting chromatographic conditions and mass spectrum conditions, and acquiring primary spectrogram data of the working solution of the standard substance in an MS mode to obtain information such as retention time, parent ions, mass deviation, ionization forms and the like of the target substance; finally, inputting the retention time of each pesticide in the PCDL software, and importing the primary mass spectrogram into a spectrogram editing item in the PCDL software;
the second-level mass spectrum database establishing step in the second step is as follows: firstly, under the condition of confirmation by primary mass spectrum data, according to the chromatographic condition and mass spectrum condition of a primary mass spectrum database, giving different collision energy to target ions to a standard working solution in a Targeted MS/MS mode, and collecting secondary spectrogram data; and then, importing the secondary mass spectrum information into a spectrogram editing item in PCDL software.
According to the invention, the chromatographic conditions and mass spectrometry conditions of step two are respectively:
(1) chromatographic conditions
Sample inlet temperature: 280 ℃; and (3) sample introduction mode: pulse non-shunting sample introduction; sample introduction amount: 2 mu L of the solution; carrier gas: helium gas; flow rate: locking by taking the retention time of the chlorpyrifos-methyl as 18.00 min; transmission line temperature: 280 ℃;
(2) conditions of Mass Spectrometry
An ion source: an EI source; solvent retardation: 5 min; current intensity: 2 mA. Temperature rising procedure: 60 deg.C (keeping for 1min), 5 deg.C/min to 280 deg.C, and keeping for 5 min.
Further, the mass spectrum acquisition conditions of the step two are as follows: when the mass spectrum acquisition mode is the MS mode, the mass spectrum acquisition range is 100-1100 m/z; the collection speed is 2 spectra/s; when the mass spectrum acquisition mode is a Target-MS/MS mode, the mass spectrum acquisition range is 100 and 1100 m/z; the collection speed is 2 spectra/s; MS response absolute broadness 200; the MS/MS response is absolutely broad 10.
The invention has the beneficial effects that:
1. the invention covers 1066 kinds of pesticides in the medicinal and edible food which may be introduced in related links of raw material production, product processing, storage, sale and the like of medicinal and edible products at home and abroad, and the compound types cover various limit regulations at home and abroad. The detection limit reaches 10 mug/kg, and the pesticide residue screening requirement of various countries in the world is met.
2. Aiming at different types of pesticides, the adsorbent formula of QuEChERS and SinChERS small columns is improved, the recovery rate of acid pesticides and plane six-membered ring structure pesticides is improved, and the applicability of the method is improved.
3. The invention simultaneously establishes a primary mass spectrum database and a secondary mass spectrum database of the gas chromatography-quadrupole time-of-flight mass spectrum and the liquid chromatography-quadrupole time-of-flight mass spectrum, and improves the accuracy by mutual evidence among the databases and further confirmation of secondary mass spectrum information.
4. The method can complete the detection, the qualification and the confirmation of 27 classes of organophosphorus insecticides, organochlorine insecticides and the like within 1 hour, and totally 1066 pesticides (shown in table 1) can be detected, and compared with the traditional detection method, the method can greatly improve the working efficiency.
5. The GC-QTOF MS and LC-QTOF MS established by the invention have the following detection technical indexes: scanning range 50-1100 m/z: the accurate mass measured can reach 0.0001 m/z: the quality precision can be controlled within 5 ppm: the collection speed was 2 spectra/s.
Drawings
FIG. 1A is a superimposed diagram of LC-QTOF extracted ion chromatography of representative pesticides in sea buckthorn.
FIG. 1B shows the screening results of LC-QTOF for representative pesticides in Hippophae rhamnoides.
FIG. 2A is a superimposed view of LC-QTOF extraction ion chromatography of representative pesticides in radix Puerariae.
FIG. 2B shows the screening result of LC-QTOF for representative pesticide in radix Puerariae.
FIG. 3A is a superimposed graph of LC-QTOF extracted ion chromatography of representative pesticides in lonicera confusa.
FIG. 3B shows the screening results of LC-QTOF for representative pesticide in Lonicera confusa.
FIG. 4 is a LC-QTOF primary mass spectrometry total ion flow chart (TIC) of 2,4-D in a sample of Hippophae rhamnoides.
FIG. 5A is an LC-QTOF primary Extracted Ion Chromatogram (EIC) of 2,4-D in a sample of Hippophae rhamnoides.
FIG. 5B shows the LC-QTOF primary mass spectrum and PCDL matching results of 2,4-D in the sample of Hippophae rhamnoides.
FIG. 6 is a LC-QTOF secondary mass spectrum total ion flow chart (TIC) of a sample of Hippophae rhamnoides.
FIG. 7A is a LC-QTOF secondary Extracted Ion Chromatogram (EIC) of 2,4-D in a sample of Hippophae rhamnoides.
FIG. 7B is a mirror image comparison of the LC-QTOF secondary mass spectrum and PCDL match of 2,4-D in a sample of Hippophae rhamnoides.
FIG. 8A is a GC-QTOF extracted ion chromatography overlay of a representative pesticide in Hippophae rhamnoides.
FIG. 8B shows the result of GC-QTOF screening of a representative pesticide in Hippophae rhamnoides.
FIG. 9A is a GC-QTOF extraction ion chromatography overlay of a representative pesticide in kudzu.
FIG. 9B shows the result of GC-QTOF screening for representative pesticides in kudzu roots.
FIG. 10A is a GC-QTOF extracted ion chromatography overlay of a representative pesticide in Lonicera confusa.
FIG. 10B shows the result of GC-QTOF screening of representative pesticides in Lonicera confusa.
FIG. 11 is a GC-QTOF primary mass spectrometry total ion flow chart (TIC) of hexachlorobenzene in a flos lonicerae sample.
FIG. 12A is a GC-QTOF primary Extraction Ion Chromatogram (EIC) of hexachlorobenzene in a flos lonicerae sample.
FIG. 12B is a mirror image comparison of GC-QTOF primary mass spectrum and PCDL of hexachlorobenzene in lonicera confusa samples.
FIG. 13 is a GC-QTOF secondary mass spectrum total ion flow chart (TIC) of the lonicera confusa.
FIG. 14A is a GC-QTOF secondary Extraction Ion Chromatogram (EIC) of hexachlorobenzene in a flos lonicerae sample.
FIG. 14B is a mirror image comparison of GC-QTOF secondary mass spectrum and PCDL match of hexachlorobenzene in a flos lonicerae sample.
FIG. 15A is a graph comparing the effect of PSA content in SinChERS columns on recovery of weakly acidic pesticides from Hippophae rhamnoides.
FIG. 15B is a graph comparing the effect of PSA content in SinChERS cartridge on recovery of weakly acidic pesticides from kudzu.
FIG. 15C is a graph comparing the effect of PSA content in SinChERS columella on recovery of weakly acidic pesticides from Lonicera confusa.
Fig. 16A is a graph comparing the effect of the particle size of graphitized carbon black in sinchrs pillars on the recovery rate of planar-structure pesticides in hippophae rhamnoides.
FIG. 16B is a graph comparing the effect of the particle size of graphitized carbon black in the SinChERS cartridge on the recovery rate of planar structure pesticides in kudzu roots.
FIG. 16C is a graph comparing the effect of the graphitized carbon black particle size in the SinChERS cartridge on the recovery rate of the plane structure pesticide in flos lonicerae.
Detailed Description
The present invention will be further described with reference to the following examples. It should be understood that the following examples are illustrative only and are not intended to limit the scope of the present invention.
The invention aims at 105 kinds of medicinal and edible food which are recorded according to the catalog of materials which are both food and traditional Chinese medicinal materials (2018 edition), combs relevant links of raw material production, product processing, storage, sale and the like, combines the hazardous substance residues specified for the products in main economic body aiming laws and regulations of China, European Union, Japan and the like, finds out pesticide residues which are possibly introduced in different links, the food is purified by an improved QuEChERS combined SinChERS small column purification method, liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF MS) and gas chromatography quadrupole time-of-flight tandem mass spectrometry (GC-QTOF MS) detection equipment are adopted, a mass spectrum database (shown in table 1) of 1066 pesticide residues in medicinal and edible food is established by utilizing the high separation capacity of chromatography and the high resolution of modern mass spectrometry, and the aim of rapidly screening unknown pesticides is fulfilled by the mass spectrum database. The invention can solve the problem of difficult high-flux screening and rapid detection of harmful components in medicinal and edible products, and realizes the detection, judgment and control of quality safety of medicinal and edible products at import and export, thereby promoting import and export trade of medicinal and edible products in China. (it should be understood that since 1066 kinds of pesticides shown in table 1 are common pesticides that may be introduced into food with homology of medicine and food, there is no one list, and only representative pesticides of each kind of pesticides are listed).
TABLE 1 chemical information of 1066 pesticides of the present invention
Example 1
This example selects 52 representative pesticides covering all types in table 1 from 1066 pesticide databases established for sorting and analysis to establish a Q-TOF mass spectrum database (see table 2).
Chemical information for representative pesticides in Table 252
Inputting the collected information of the names, CAS numbers, molecular formulas, precise molecular weights and the like of the pesticides shown in the table 2 into an edited Compound information item in Personal Compound Database (PCDL) software; then, preparing the collected standard substance of the library-building compound into a standard stock solution and a working solution respectively, adjusting chromatographic conditions and mass spectrum conditions, and acquiring primary spectrogram data of the working solution of the standard substance in an MS mode by using the following instruments, equipment and specific methods:
instruments and equipment: model 1290 high performance liquid chromatograph-model 6545 quadrupole-time-of-flight mass spectrometer, equipped with MassHunter chromatography workstation and PCDL software, Agilent, usa; 7890 type A gas chromatograph-7200 quadrupole-time-of-flight mass spectrometer, Agilent, USA; sensory balance, Mettler Toledo, usa.
1. Preparation of standard stock solution, standard intermediate solution and standard working solution: accurately weighing each pesticide standard substance in a volumetric flask respectively, dissolving with a small amount of methanol or n-hexane, and then preparing into 1.0mg/mL standard stock solution by constant volume. According to the requirement, a certain amount of pesticide solution is sucked at the time of use, diluted by methanol or acetonitrile and prepared into a standard intermediate solution with the concentration of 50 mug/mL in groups. A certain amount of pesticide solution is absorbed according to the requirement, and the pesticide solution is diluted by a blank medicinal and edible food substrate to prepare a standard working solution with proper concentration.
2. Liquid chromatography conditions and mass spectrometry conditions:
(1) chromatographic conditions
Mobile phase: A. 5mmol/L ammonium acetate water solution + 0.1% formic acid, B, 0.1% formic acid-methanol;
flow rate: 0.4 mL/min;
sample introduction amount: 5 mu L of the solution;
elution gradient: 5% A (0min), 5% A (1min), 60% A (6min), 100% A (16min), 100% A (20.0min), 5% A (20.1min), 5% A (25 min).
(2) Mass spectrum conditions: an Agilent 6540 liquid chromatogram/four-level rod-flight time mass spectrometer adopts an ESI source, the capillary voltage is 5500V, the declustering voltage is 150V, the taper hole voltage is 65V, and the eight-level rod radio frequency voltage is 750V; the collision gas, the atomization gas and the drying gas are all nitrogen; the temperature of the sheath gas is 350 ℃, and the flow rate of the sheath gas is 11L/min; the temperature of the drying gas is 280 ℃, and the flow rate of the drying gas is 9L/min.
An ion source: an EI source;
and (3) monitoring mode: a positive ion mode, a negative ion mode;
current intensity: 2 mA.
When the mass spectrum acquisition mode is the MS mode, the mass spectrum acquisition range is 100-1100 m/z; the collection speed is 2 spectra/s;
when the mass spectrum acquisition mode is a Target-MS/MS mode, the mass spectrum acquisition range is 100 and 1100 m/z; the collection speed is 2 spectra/s; MS response absolute broadness 200; the MS/MS response is absolutely broad 10.
3. Establishing and optimizing an LC-QTOF mass spectrum database (liquid chromatogram quadrupole time-of-flight tandem mass spectrum database): is divided into a primary mass spectrum database and a secondary mass spectrum database
Establishing a primary mass spectrum database: first, the collected information of the compound name, CAS number, structural formula, precise molecular weight, etc. is entered under the edit compound information entry in the PCDL software. And then, preparing the collected standard library compound according to the solution preparation method in the step 1, and respectively collecting gas phase and liquid phase primary mass spectrum data in an MS mode according to the chromatographic condition and the mass spectrum condition in the step 2. Finally, judging whether the mass number of the primary ions is consistent with the mass number of the parent ions, and if so, finishing building a primary mass spectrum database; if the information is not consistent, the information needs to be retrieved again, whether the information collection is correct or not is confirmed, and whether the standard solution is correct or not is confirmed until the information is consistent (see table 3). Then, the retention time of each pesticide is input into PCDL software, and the primary mass spectrogram is introduced into the spectrogram editing item in the PCDL software.
Table 353 primary mass spectrum parent ion information for representative pesticides
Remarking: the "\\" in Table 3 indicates that GC or LC cannot be used for walking.
The secondary mass spectrum database is established as follows: firstly, under the condition of primary mass spectrum data confirmation, according to the chromatographic condition and mass spectrum condition in the step 2, giving 10-50V of collision energy to the parent ions to the standard working solution in a Targeted MS/MS mode to obtain a secondary mass spectrum, subtracting a background mass spectrum from the obtained secondary mass spectrum, and introducing the secondary mass spectrum into a spectrum editing item in PCDL software. And finally, completing the establishment of the whole colorant compound mass spectrum database after the second-stage mass spectrum data of all the compounds are recorded.
4. Gas chromatography conditions and mass spectrometry conditions:
(1) chromatographic conditions
Sample inlet temperature: 280 ℃; and (3) sample introduction mode: pulse non-shunting sample introduction; sample introduction amount: 2 mu L of the solution; carrier gas: helium gas; flow rate: locking by taking the retention time of the chlorpyrifos-methyl as 18.00 min; transmission line temperature: 280 ℃;
(2) conditions of Mass Spectrometry
An ion source: an EI source; solvent retardation: 5 min; current intensity: 2 mA; temperature rising procedure: 60 deg.C (keeping for 1min), 5 deg.C/min to 280 deg.C, and keeping for 5 min.
When the mass spectrum acquisition mode is the MS mode, the mass spectrum acquisition range is 100-1100 m/z; the collection speed is 2 spectra/s; when the mass spectrum acquisition mode is a Target-MS/MS mode, the mass spectrum acquisition range is 100 and 1100 m/z; the collection speed is 2 spectra/s; MS response absolute broadness 200; the MS/MS response is absolutely broad 10.
5. Establishing and optimizing a GC-QTOF mass spectrum database (gas chromatography quadrupole time-of-flight tandem mass spectrum database): is divided into a primary mass spectrum database and a secondary mass spectrum database
Establishing a primary mass spectrum database: first, the collected information of the compound name, CAS number, structural formula, precise molecular weight, etc. is entered under the edit compound information entry in the PCDL software. And then, preparing the collected standard library compound according to the solution preparation method in the step 1, and respectively collecting gas phase and liquid phase primary mass spectrum data in an MS mode according to the chromatographic condition and the mass spectrum condition in the step 4. Finally, judging whether the mass number of the primary ions is consistent with the mass number of the parent ions, and if so, finishing building a primary mass spectrum database; if the two types of pesticides are not consistent, the two types of pesticides need to be retrieved again, whether the information collection is correct or not and whether the standard solution is correct or not are confirmed until the information is consistent (see table 2), the retention time of each pesticide is input into the PCDL software, and the primary mass spectrogram is introduced into the PCDL software under spectrogram editing items.
The secondary mass spectrum database is established as follows: firstly, under the condition of primary mass spectrum data confirmation, giving 10-50V collision energy to the parent ion according to the chromatographic condition and mass spectrum condition in the step 4 to the standard working solution in a Targeted MS/MS mode to obtain a secondary mass spectrum, subtracting a background mass spectrum from the obtained secondary mass spectrum, and introducing the secondary mass spectrum into a spectrum editing item in PCDL software. And finally, completing the establishment of the whole colorant compound mass spectrum database after the second-stage mass spectrum data of all the compounds are recorded.
In the library building process, the structure information of each compound is analyzed in advance, and the mass spectrum ion source mode is analyzed, so that the library building efficiency is improved. When a certain compound has strong response in the positive and negative ion modes, mass spectrum information in the positive and negative modes can be recorded simultaneously.
Example 2 screening and confirmation of LC-QTOF for pesticides in Hippophae rhamnoides, Pueraria lobata and Lonicera confusa
In this embodiment, the LC-QTOF mass spectrum database provided in embodiment 1 is used to screen and confirm pesticide residues in a representative large number of medicinal and edible food products, namely, sea buckthorn, radix puerariae, and lonicera confusa, so as to verify the feasibility of the LC-QTOF mass spectrum database for detecting pesticide residues in medicinal and edible food products.
The instrument comprises the following steps: model 1290 high performance liquid chromatograph-model 6540 quadrupole-time-of-flight mass spectrometer, equipped with MassHunter chromatography workstation and PCDL software, Agilent, usa; a centrifuge: 4500r/min, Eppendorf; a touch oscillator, Vision VORTEX MIXER, KMC-1300V; sensory balance, Mettler Toledo, usa.
Materials: commercial samples of hippophae rhamnoides, pueraria lobata and lonicera hypoglauca (determined to be free of the 52 representative pesticides of example 1) were added to three samples at a concentration of 10 μ g/kg of the 52 representative pesticides, simulating positive samples.
The implementation steps are as follows:
1. sample preparation:
weigh 10.0g of sample into a 50mL centrifuge tube. 10mL acetonitrile, QuEChERS extract pack (6g anhydrous magnesium sulfate, 1.5g sodium acetate) was added and vortexed for 1 min. Centrifuging at 4000r/min for 5 min. Placing SinChERS small column (1200mg anhydrous magnesium sulfate, 200mg 300i graphitized carbon black, 200mg C18, 50mg PSA) in centrifugal tube containing extractive solution, and slowly pressing down to scale to purify the extractive solution. And sucking 500 mu L of supernatant liquid, metering to 1mL by acetonitrile, passing through a 0.22 mu m nylon filter membrane, and performing LC-QTOF measurement.
2. Liquid chromatography conditions and mass spectrometry conditions:
(1) chromatographic conditions
Mobile phase: mobile phase: a: 5mmol/L aqueous ammonium acetate + 0.1% formic acid, B: 0.1% formic acid-methanol; flow rate: 0.4 mL/min; sample introduction amount: 5 μ L. Elution gradient: 5% A (0min), 5% A (1min), 60% A (6min), 100% A (16min), 100% A (20.0min), 5% A (20.1min), 5% A (25 min).
(2) Conditions of Mass Spectrometry
The Agilent 6540 liquid chromatogram/four-stage rod-flight time mass spectrometer adopts a Dual ESI source, the capillary voltage is 4000V, the de-clustering voltage is 125V, the taper hole voltage is 65V, and the eight-stage rod radio frequency voltage is 750V; the collision gas, the atomization gas and the drying gas are all nitrogen; the temperature of the sheath gas is 350 ℃, and the flow rate of the sheath gas is 11L/min; the temperature of the drying gas is 280 ℃, and the flow rate of the drying gas is 9L/min.
An ion source: an EI source; and (3) monitoring mode: a positive ion mode, a negative ion mode; current intensity: 2 mA.
When the mass spectrum acquisition mode is the MS mode, the mass spectrum acquisition range is 100-1100 m/z; the collection speed is 2 spectra/s; when the mass spectrum acquisition mode is a Target-MS/MS mode, the mass spectrum acquisition range is 100 and 1100 m/z; the collection speed is 2 spectra/s; MS response absolute broadness 200; the MS/MS response is absolutely broad 10.
3. And (3) qualitative analysis:
the qualitative identification of the unknown substances is realized by searching and matching with the LC-QTOF mass spectrogram library described in the embodiment 1, the extracted ion chromatography overlay graphs of 52 pesticides in sea buckthorn, radix puerariae and lonicera confusa are shown in figures 1A, 2A and 3A, and the matching results are shown in figures 1B, 2B and 3B.
The qualitative steps are illustrated by 2,4-D in Hippophae rhamnoides. Firstly, taking the sample pretreatment liquid in the step 1, performing on-machine determination according to the instrument conditions in the step 2, acquiring primary mass spectrum spectrogram data in an MS mode to obtain a TIC (see figure 4), deconvoluting the TIC, and performing retrieval, matching and scoring on the accurate mass number obtained by deducting the background of the obtained Extracted Ion Chromatogram (EIC) (see figure 5A) and a compound in PCDL software and a mass spectrum spectrogram library to find that the matching degree of an unknown substance and 2,4-D in the spectrogram library is high (see figure 5B). And then, applying different collision energy to the target parent ions in a Targeted MS/MS mode to obtain a secondary mass spectrum total ion flow graph (see fig. 6), deconvoluting the TIC graph, extracting mass spectrograms from the obtained Extracted Ion Chromatogram (EIC) (see fig. 7A), and performing secondary mass spectrum data retrieval and matching in MassHunter software to find that the suspected compound has higher matching degree with the 2,4-D secondary mass spectrum data in the mass spectrum library (see fig. 7B). Thus, the unknown substance was confirmed to be 2, 4-D.
4) Quantitative analysis: respectively sucking standard stock solutions with different volumes, respectively preparing standard working solutions with the concentrations of 0ng/mL, 1ng/mL, 2ng/mL, 5ng/mL, 10ng/mL and 20ng/mL by using blank sea buckthorn, kudzu root and lonicera confusa matrix solutions, and injecting samples according to the sequence of the standard working solutions, reagent blanks, sample blanks, addition recovery, sample samples and the standard solutions. The concentration of the target was plotted on the abscissa and the peak area of the target was plotted on the ordinate, and the standard curve was used for quantification (see table 4).
5) And (3) comparing results: and (3) sequentially detecting the standard working solution (containing 5 concentration points and zero point), the reagent blank, the sample blank, the addition recovery, the sample and the standard solution in the step 4) by using Agilent 1290-6410 liquid chromatography-tandem quadrupole mass spectrometry, and drawing a standard curve for quantification by using the concentration of the target as a horizontal coordinate and the peak area of the target as a vertical coordinate. The results of the assay of 52 representative compounds in samples of hippophae rhamnoides, pueraria lobata and lonicera confusa are shown in table 4. According to the results, the deviation between the LC-QTOF quantitative result and the LC-QQQ (liquid chromatogram-triple tandem quadrupole liquid chromatograph-mass spectrometer) quantitative result is less than 10%.
TABLE 4 quantitative results of LC-QTOF and LC-QQQ of Pueraria lobata, Hippophae rhamnoides and Lonicera confusa
And (4) conclusion: the LC-QTOF mass spectrum database can be used for qualitatively and quantitatively detecting the pesticide residue in food with homology of medicine and food.
Example 3 GC-QTOF confirmation of representative pesticides in Hippophae rhamnoides, Pueraria lobata and Lonicera confusa
In this embodiment, the GC-QTOF mass spectrum database provided in embodiment 1 is used to screen and confirm pesticide residues in a representative large number of medicinal and edible food products, namely sea buckthorn, radix puerariae, and lonicera confusa, so as to verify the feasibility of the GC-QTOF mass spectrum database for detecting pesticide residues in medicinal and edible food products.
The instrument comprises the following steps: model 1290 high performance liquid chromatograph-model 6540 quadrupole-time-of-flight mass spectrometer, equipped with MassHunter chromatography workstation and PCDL software, Agilent, usa; a centrifuge: 4500r/min, Eppendorf; a touch oscillator, Vision VORTEX MIXER, KMC-1300V; sensory balance, Mettler Toledo, usa.
Materials: commercial samples of kudzu root, kudzu root and lonicera confusa were tested to be free of the 52 representative pesticides of example 1, and 52 representative pesticides at a concentration of 10 μ g/kg were added to the three samples to simulate positive samples.
The implementation steps are as follows:
1. sample preparation:
weigh 10.0g of sample into a 50mL centrifuge tube. 10mL acetonitrile, QuEChERS extract pack (6g anhydrous magnesium sulfate, 1.5g sodium acetate) was added and vortexed for 1 min. Centrifuging at 4000r/min for 5 min. Placing SinChERS small column (1200mg anhydrous magnesium sulfate, 200mg 300i graphitized carbon black, 200mg C18, 50mg PSA) in centrifugal tube containing extractive solution, and slowly pressing down to scale to purify the extractive solution. And sucking 500 mu L of supernatant liquid, metering to 1mL by acetonitrile, filtering by a 0.22 mu m nylon filter membrane, and performing GC-QTOF determination.
2. Gas chromatography conditions and mass spectrometry conditions:
(1) chromatographic conditions
Sample inlet temperature: 280 ℃; and (3) sample introduction mode: pulse non-shunting sample introduction; sample introduction amount: 2 mu L of the solution; carrier gas: helium gas; flow rate: locking by taking the retention time of the chlorpyrifos-methyl as 18.00 min; transmission line temperature: 280 ℃;
(2) the mass spectrum conditions are as follows: an ion source: an EI source; solvent retardation: 5 min; current intensity: 2 mA. Temperature rising procedure: 60 deg.C (keeping for 1min), 5 deg.C/min to 280 deg.C, and keeping for 5 min.
When the mass spectrum acquisition mode is the MS mode, the mass spectrum acquisition range is 100-1100 m/z; the collection speed is 2 spectra/s; when the mass spectrum acquisition mode is a Target-MS/MS mode, the mass spectrum acquisition range is 100 and 1100 m/z; the collection speed is 2 spectra/s; MS response absolute broadness 200; the MS/MS response is absolutely broad 10.
3. And (3) qualitative analysis: the qualitative identification of the unknown substances is realized by searching and matching with the GC-QTOF mass spectrum library, the extracted ion chromatography overlay graphs of 52 pesticides in sea buckthorn, kudzu vine root and lonicera confusa are shown in figures 8A, 9A and 10A, and the matching results are shown in figures 8B, 9B and 10B.
The qualitative procedure is illustrated by using hexachlorobenzene in lonicera confusa as an example. Firstly, taking the sample pretreatment liquid in the step 1, performing on-machine determination according to the instrument conditions in the step 2, acquiring primary mass spectrum spectrogram data in an MS mode to obtain a TIC (see figure 11), deconvoluting the TIC, extracting a mass spectrogram of an obtained Extracted Ion Chromatogram (EIC) (see figure 12A), searching and matching the mass spectrogram with a background subtracted, performing primary mass spectrum spectrogram data in MassHunter software, and finding that the matching degree of an unknown substance and hexachlorobenzene in a spectrogram library is high (see figure 12B). Therefore, the unknown substance was confirmed to be suspected hexachlorobenzene. And then, applying different collision energy to the target parent ions in a Targeted MS/MS mode to obtain a secondary mass spectrum total ion flow graph (see fig. 13), deconvoluting the TIC graph, performing secondary mass spectrum spectrogram data retrieval and matching on the obtained Extracted Ion Chromatogram (EIC) (see fig. 14A) in MassHunter software, and finding that the suspected compound has higher matching degree with the hexachlorobenzene secondary mass spectrum data in the mass spectrum library (see fig. 14B).
4. Quantitative analysis: respectively sucking standard stock solutions with different volumes, preparing standard working solutions with the concentrations of 0ng/mL, 5ng/mL, 10ng/mL, 20ng/mL, 50ng/mL and 100ng/mL by using blank radix puerariae matrixes, and injecting samples according to the sequence of the standard working solutions (comprising 5 concentration points and zero points), reagent blanks, sample blanks, addition and recovery, sample samples and the standard solutions. 366.9444, 212.9489, 366.9444 and 254.9714 were selected as quantitative and qualitative ion pairs, respectively, and a standard curve was plotted with the concentration of the target substance as abscissa and the peak area of the target as ordinate for quantitative determination (see table 5).
5. And (3) comparing results: and (4) sequentially detecting the standard working solution, the reagent blank, the sample blank, the addition recovery, the sample and the standard solution in the step (4) by using Agilent 7890-7000 gas chromatography-tandem quadrupole mass spectrometry, and drawing a standard curve for quantification by using the concentration of the target as a horizontal ordinate and the peak area of the target as a vertical ordinate. The results of the assays for representative compounds in samples of hippophae rhamnoides, pueraria lobata and lonicera confusa are shown in table 5. According to the results, the deviation between the LC-QTOF quantitative result and the LC-QQQ quantitative result is less than 10%.
TABLE 5 quantitative results of GC-QTOF and GC-QQQ of Pueraria lobata, Hippophae rhamnoides and Lonicera confusa
And (4) conclusion: the GC-QTOF mass spectrum database can be used for qualitatively and quantitatively detecting the pesticide residue in food with homology of medicine and food.
Example 4
The instrument comprises the following steps: the same as example 1 and example 2.
Materials: commercial samples of hippophae rhamnoides, pueraria lobata and lonicera hypoglauca were determined to be free of 52 representative pesticides.
The method comprises the following steps: samples of Hippophae rhamnoides, Pueraria lobata and Lonicera confusa not containing 52 representative agricultural chemicals were measured and quantified by adding 8 representative acidic agricultural chemicals (added concentration: 10. mu.g/kg) according to the methods of example 1 and example 2. In addition, PSA was increased from 50mg to 150mg in the SinChERS column, and other components were unchanged in the SinChERS column, and the remaining steps were determined and quantified as in example 2. The recovery rates were calculated and the results are shown in tables 6A, 6B, 6C, FIGS. 15A, 15B and 15C.
The results show that when the dosage of PSA is reduced from 150mg to 50mg, the average recovery rate of 8 acidic pesticides in sea buckthorn, kudzu root and lonicera confusa can be improved to more than 70%, and the detection requirement of No. 2386 bulletin of pesticide ministry is met.
And (4) conclusion: the PSA content in the SinChERS small column in the sample pretreatment step is 50mg, and the recovery rate of the sample is good, so that the SinChERS filler formula provided by the invention can greatly improve the recovery rate of weakly acidic pesticides and enhance the applicability of the method to weakly acidic pesticides.
When the content of PSA in the SinChERS small column in the sample pretreatment step is 0.5% -1% of the sample content, the recovery rate of the sample is gradually reduced, and when the content of PSA is 0.5% of the sample content, the recovery rate of the sample is good, so that when the content of PSA in the SinChERS filler formula provided by the invention is 0.5% -1% of the sample content, the recovery rate of weakly acidic pesticides can be greatly improved, and the applicability of the method to weakly acidic pesticides is enhanced.
Table 6A average recovery (%) of representative acid pesticides from hippophae rhamnoides at different PSA levels in SinChERS columns (n ═ 6)
Table 6B average recovery (%) of representative acid pesticides from puerariae radix with different PSA levels in SinChERS cartridge (n ═ 6)
Table 6C average recovery (%) of representative acid pesticides from lonicera confusa at different PSA levels in SinChERS cartridge (n ═ 6)
Example 5
The instrument comprises the following steps: the same as example 1 and example 2.
Materials: commercial samples of hippophae rhamnoides, pueraria lobata and lonicera hypoglauca were determined to be free of 52 representative pesticides.
The method comprises the following steps:
samples of Hippophae rhamnoides, Pueraria lobata and Lonicera confusa not containing 52 representative pesticides were measured and quantified by adding 7 representative plane-structured pesticides (added concentration: 10. mu.g/kg) according to the methods of example 1 and example 2. The grain size of GCB in the SinChERS packing was reduced to 300i, and other components in the SinChERS column were not changed, and the measurement and quantification were performed in the same manner as in examples 1 and 2. The recovery rates were calculated and the results are shown in tables 7A, 7B, 7C, FIGS. 16A, 16B and 16C.
Table 7A average recovery (%) of graphitized carbon blacks of different particle sizes in SinChERS cartridge for representative planar structure pesticides in hippophae rhamnoides (n ═ 6)
TABLE 7 average recovery (%) of graphitized carbon blacks of different particle sizes in SinChERS cartridge for representative planar structure pesticides in kudzu root (n ═ 6)
TABLE 7 average recovery (%) of graphitized carbon blacks of different particle sizes in SinChERS cartridge for representative plane structure pesticides in Lonicera confusa (n ═ 6)
From the results, it is found that when the particle size of the graphitized carbon black is increased from 100i to 300i, the recovery rate of hexachlorobenzene can be improved to 75% or more, and the detection requirement of the ministry of agricultural chemicals publication No. 2386 is satisfied.
And (4) conclusion: when the particle size of the graphitized carbon black in the SinChERS small column in the sample pretreatment step is 200i-300i, the recovery rate of the sample is gradually increased, and when the particle size of the graphitized carbon black is 300i, the recovery rate of the sample is good, so that when the particle size of the graphitized carbon black in the SinChERS filler formula provided by the invention is 200i-300i, the accuracy of the plane structure pesticide can be greatly improved, and the applicability to the plane structure pesticide is enhanced.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are given by way of illustration of the principles of the present invention, and that various changes and modifications may be made without departing from the spirit and scope of the invention as defined by the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (8)
1. A method for screening pesticide residues in medicinal and edible homology food by a pesticide residue mass spectrum database is characterized by comprising the following steps:
step one, establishing LC-QTOF pesticide mass spectrum database
A. Establishing a primary mass spectrum database of 854 common pesticides for medicinal and edible food: collecting primary map data of 854 pesticide compound standard products of medicinal and edible food to obtain unique retention time of each pesticide compound, and inputting the retention time into PCDL software for screening the pesticide compounds in the medicinal and edible food;
B. establishing a secondary mass spectrum database: collecting secondary spectrogram data of the pesticide compound standard substance confirmed by the primary mass spectrum database, and importing the secondary mass spectrum data into a spectrogram editing item in PCDL software for confirming pesticide compounds in food with homology of medicine and food;
step two, establishing a GC-QTOF pesticide mass spectrum database, which comprises the following steps:
A. establishing a primary mass spectrum database of 727 common pesticides for food and medicine homologous food, collecting primary spectrum data of pesticide compound standard products of the 727 common food and medicine homologous food, obtaining unique retention time of each pesticide compound, inputting the retention time into PCDL software, and introducing the primary mass spectrum into a spectrum editing item in the PCDL software for screening the pesticide compounds in the food and medicine homologous food;
B. establishing a secondary mass spectrum database: the primary mass spectrum database confirms the secondary spectrogram data of the pesticide compound standard product; then, importing the secondary mass spectrum data into a spectrogram editing item in PCDL software;
step three, preparation of a sample to be detected: and rapidly, qualitatively and quantitatively analyzing the sample by using the LC-QTOF pesticide mass spectrum database and the GC-QTOF pesticide mass spectrum database to detect the retention time of the sample to be detected, and comparing the retention time with the retention time of the pesticide compound to judge whether the sample contains pesticide compound residues and quantitatively detect the content of the pesticide compound.
2. The method as claimed in claim 1, wherein before the sample to be tested in the third step is prepared, a sample pre-treatment step is further included, and the sample pre-treatment step is as follows: weighing a sample in a centrifuge tube, adding excessive acetonitrile and a QuEChERS extraction bag, wherein the QuEChERS extraction bag contains anhydrous magnesium sulfate and sodium acetate in a mass ratio of 4-6: 1-1.5, and performing vortex oscillation; centrifuging; placing a SinChERS small column into a centrifugal tube filled with an extracting solution, wherein the SinChERS small column is filled with anhydrous magnesium sulfate, 300i graphitized carbon black, C18 and PSA, and the mass ratios of the anhydrous magnesium sulfate, 200-300 i graphitized carbon black, C18 and PSA to a sample are respectively 10-15%, 1-2% and 0.5-1%; then, 500. mu.L of the supernatant was aspirated and the volume was adjusted to 1mL with acetonitrile, and the supernatant was filtered through a nylon filter and subjected to LC-QTOF measurement.
3. The method as claimed in claim 2, wherein before the sample to be tested in the third step is prepared, the method further comprises a sample pre-treatment step, wherein the sample pre-treatment step comprises: weighing a sample in a centrifuge tube, adding excessive acetonitrile and a QuEChERS extraction bag, wherein the QuEChERS extraction bag contains anhydrous magnesium sulfate and sodium acetate in a mass ratio of 4:1, and performing vortex oscillation; centrifuging; placing a SinChERS small column in a centrifugal tube filled with an extracting solution, wherein the SinChERS small column is filled with anhydrous magnesium sulfate, 300i graphitized carbon black, C18 and PSA, and the mass ratios of the anhydrous magnesium sulfate, the 300i graphitized carbon black, the C18 and the PSA to the sample are respectively 12%, 2% and 0.5%; then, 500. mu.L of the supernatant was aspirated and the volume was adjusted to 1mL with acetonitrile, and the supernatant was passed through a 0.22 μm nylon filter and subjected to LC-QTOF measurement.
4. The method of claim 1, wherein the first-stage mass spectrum database of the first step is created by: inputting the information of the pesticide compound into an edited compound information item in PCDL software; then, respectively preparing the collected pesticide compound standard substance into a standard stock solution and a standard intermediate solution, adjusting chromatographic conditions and mass spectrum conditions, and acquiring primary spectrogram data of a working solution of the standard substance in an MS mode to obtain information such as retention time, parent ions, mass deviation, ionization form and the like of a target object; finally, the retention time of each pesticide is input in PCDL software;
the information of the pesticide compound includes name, CAS number, molecular formula, precise molecular weight, etc.;
the second-level mass spectrum database in the first step is established by the following steps: under the condition of confirmation by primary mass spectrum data, according to the chromatographic conditions and mass spectrum conditions of a primary mass spectrum database, giving different collision energy to target ions to a standard working solution in a Targeted MS/MS mode, and collecting secondary spectrogram data; then, the secondary mass spectrometry data is imported under a spectrogram editing entry in PCDL software.
5. The method of claim 4, wherein the chromatographic conditions and mass spectrometry conditions of step one are:
(1) chromatographic conditions are as follows: mobile phase: a: 5mmol/L aqueous ammonium acetate and 0.1% formic acid, B: acetonitrile; flow rate: 0.4 mL/min; sample introduction amount: 5 mu L of the solution;
(2) mass spectrum conditions: an Agilent 6540 liquid chromatogram/four-level rod-flight time mass spectrometer adopts an ESI source, the capillary voltage is 5500V, the declustering voltage is 150V, the taper hole voltage is 65V, and the eight-level rod radio frequency voltage is 750V; the collision gas, the atomization gas and the drying gas are all nitrogen; the temperature of the sheath gas is 350 ℃, and the flow rate of the sheath gas is 11L/min; the temperature of the drying gas is 280 ℃, and the flow rate of the drying gas is 9L/min.
6. The method of claim 1, wherein the step two of establishing the primary mass spectrum database of 727 common pesticides for food and drug homologous food comprises the following steps: firstly, inputting information such as compound name, CAS number, molecular formula, accurate molecular weight and the like of pesticide into an edited compound information item in PCDL software; then, respectively preparing the collected standard substance of the library building compound into a standard stock solution and a working solution, adjusting chromatographic conditions and mass spectrum conditions, and acquiring primary spectrogram data of the working solution of the standard substance in an MS mode to obtain information such as retention time, parent ions, mass deviation, ionization forms and the like of the target substance; finally, inputting the retention time of each pesticide in the PCDL software, and importing the primary mass spectrogram into a spectrogram editing item in the PCDL software;
the second-level mass spectrum database establishing step in the second step is as follows: firstly, under the condition of confirming with primary mass spectrum data, according to the chromatographic condition and mass spectrum condition of a primary mass spectrum database, giving different collision energy to target ions to a standard working solution in a Targeted MS/MS mode, and collecting secondary spectrum data; and then, importing the secondary mass spectrum information into a spectrogram editing item in PCDL software.
7. The method of claim 1, wherein the chromatographic conditions and mass spectrometry conditions of step two are: (1) chromatographic conditions are as follows: sample inlet temperature: 280 ℃; and (3) sample introduction mode: pulse non-shunting sample introduction; sample introduction amount: 2 mu L of the solution; carrier gas: helium gas; flow rate: locking by taking the retention time of the chlorpyrifos-methyl as 18.00 min; transmission line temperature: 280 ℃;
(2) mass spectrum conditions: an ion source: an EI source; solvent retardation: 5 min; current intensity: 2 mA; temperature rising procedure: 60 deg.C (keeping for 1min, 5 deg.C/min to 280 deg.C, keeping for 5 min).
8. The method of claim 7, wherein the mass spectrometry acquisition conditions of step two are: when the mass spectrum acquisition mode is the MS mode, the mass spectrum acquisition range is 100-1100 m/z; the collection speed is 2 spectra/s; when the mass spectrum acquisition mode is a Target-MS/MS mode, the mass spectrum acquisition range is 100 and 1100 m/z; the collection speed is 2 spectra/s; MS response absolute broadness 200; the MS/MS response is absolutely broad 10.
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| CN114487224A (en) * | 2022-01-20 | 2022-05-13 | 中国农业科学院农业质量标准与检测技术研究所 | Method for detecting acidic pesticide in fruits and vegetables |
| CN114487224B (en) * | 2022-01-20 | 2022-11-08 | 中国农业科学院农业质量标准与检测技术研究所 | Method for detecting acidic pesticide in fruits and vegetables |
| CN115144457A (en) * | 2022-06-27 | 2022-10-04 | 中验科学仪器(福建)有限公司 | Portable mass spectrum analyzer, analysis method and terminal |
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