CN115372492A - Method for detecting residual quantity of thiophosphoryl methyl by LC-MS/MS and application thereof - Google Patents
Method for detecting residual quantity of thiophosphoryl methyl by LC-MS/MS and application thereof Download PDFInfo
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- CN115372492A CN115372492A CN202111540392.7A CN202111540392A CN115372492A CN 115372492 A CN115372492 A CN 115372492A CN 202111540392 A CN202111540392 A CN 202111540392A CN 115372492 A CN115372492 A CN 115372492A
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- VEZYCISMGXNHOV-UHFFFAOYSA-N [dimethyl-[(trimethylsilylamino)silylamino]silyl]methane Chemical group C[Si](C)(C)N[SiH2]N[Si](C)(C)C VEZYCISMGXNHOV-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Library & Information Science (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention discloses a method for detecting the residual quantity of thiophosphoryl methyl by LC-MS/MS, which comprises the following steps: the method comprises the steps of sample extraction, sample purification, sample determination and result analysis, and improves the purification effect on analytes, reduces the interference of other components on the thiophosphoryl methyl and improves the detection accuracy by adopting the specific polystyrene filler solid-phase extraction column. And the octadecyl silicagel column blocked by heptamethyltrisiloxane is used as a liquid chromatographic column, so that the combination stability of an analysis sample and a stationary phase is improved, and the reproducibility and precision of experimental data are improved. Meanwhile, by setting the high-low mass resolution of a four-stage rod in the LC-MS/MS, the linear distribution of analyte linear measurement is improved, the interference of false positive signals is reduced, and the detection accuracy is improved. The method is particularly suitable for trace detection of the thiophosphoryl methyl in the traditional Chinese medicinal materials, and is particularly suitable for pesticide residue analysis of the thiophosphoryl methyl in the salvia miltiorrhiza bunge and the gallnuts.
Description
Technical Field
The invention relates to a method for detecting the residual quantity of thiophosphoryl methyl by LC-MS/MS, relates to G01N, and particularly relates to a method for testing or analyzing a material by means of determining the chemical or physical properties of the material.
Background
Disclosure of Invention
In order to improve the detection sensitivity of trace amount of methylthiocyclophosphate in traditional Chinese medicinal materials and reduce the influence of interferents, the first aspect of the invention provides a method for detecting the residual amount of methylthiocyclophosphate by LC-MS/MS, which comprises the following steps:
(1) Sample extraction: sieving the ground sample, adding sodium chloride, mixing, adding an organic solvent, homogenizing, centrifuging, and taking the supernatant;
(2) Sample purification: passing the supernatant obtained in the step 1 through a solid phase extraction column to obtain an effluent liquid;
(3) And (3) sample determination: performing LC-MS/MS detection on the effluent liquid obtained in the step 2, setting parameters and performing detection; (4) analysis of results: and analyzing the spectrogram, and comparing with a standard curve to obtain the residual quantity of the thiophosphoryl methyl.
As a preferred embodiment, the sample extraction in step 1 is: the weight ratio of the sample to the sodium chloride was 5:1, the rotation speed of the homogenate treatment is 13000-15000r/min, the time is 2min, and the rotation speed of the centrifugation is 5000-6000r/min; then adding organic solvent into the centrifuged precipitate again, homogenizing at 13000-15000r/min for 2min at 5000-6000r/min, mixing the two supernatants, concentrating under reduced pressure to 3-5ml, and diluting with organic solvent to 10ml.
As a preferred embodiment, the sample in step 1 is a 50-mesh undersize.
As a preferred embodiment, the organic solvent in step 1 is selected from one or a combination of acetone, benzene, ethanol, acetonitrile, isopropanol, acetic acid, toluene and styrene.
As a preferred embodiment, the organic solvent in step 1 is acetonitrile.
As a preferred embodiment, the solid phase extraction column in step 2 is selected from one of a reverse phase solid phase extraction column and an ion exchange solid phase extraction column, and the reverse phase solid phase extraction column is preferably selected from one of an octadecyl bonded silica gel solid phase extraction column, a graphitized carbon black filler solid phase extraction column, a polystyrene filler solid phase extraction column, and a propionylcyanane bonded silica gel solid phase extraction column.
In a preferred embodiment, the solid phase extraction column in step 2 is a polystyrene packing solid phase extraction column.
In a preferred embodiment, the stationary phase of the polystyrene filler solid phase extraction column is polystyrene-divinylbenzene.
As a preferred embodiment, the polystyrene packed solid phase extraction column has a specification of 200mg,6mL.
During the experiments the applicant found that mixing the sample with sodium chloride 5:1, the polarity of the acetonitrile can be increased, the polarity of the formed extractant is closer to that of the thiophosphoryl methyl, thereby being beneficial to extracting the thiophosphoryl methyl, and the applicant further finds that the extract is purified by adopting a polystyrene filler solid phase extraction column, the stationary phase polystyrene-divinylbenzene contains hydrophilic groups and lipophilic groups in specific proportion, a hydrophobic divinylbenzene structure retains a non-polar compound, a hydrophilic polyvinylpyrrolidone retains a polar compound, the throughput of the whole solid phase extraction column is adjusted by adjusting the divinylbenzene structure and the polyvinylpyrrolidone structure, the thiophosphoryl methyl is allowed to pass to the maximum extent, but the interference is prevented from passing, the polar range of the passing substance is narrowed, the purity of the extract is improved from the extraction source, the interference concentration is reduced, and the higher accuracy is realized.
The applicant also finds that the interference is large when the methyl sulfur cyclo-phosphate is extracted because the salvia miltiorrhiza contains more volatile oil components, and the solid phase chromatographic column with the specification of 200mg,6mL can be adopted to increase the absorption of the volatile oil components, so that the influence of the interference is further reduced.
As a preferred embodiment, said step 3 is detected by LC-MS/MS, wherein LC is selected from at least one of adsorption chromatography column, distribution chromatography column, ion exchange chromatography column, gel chromatography column.
In a preferred embodiment, the adsorption chromatographic column is at least one selected from the group consisting of a silica gel chromatographic column, an alumina chromatographic column, a polystyrene-divinyl chromatographic column and a polymethacrylate chromatographic column.
In a preferred embodiment, the adsorption column is a silica gel column.
As a preferred embodiment, the silica gel column is selected from one of an octadecylsilane bonded silica gel column, an octylsilane bonded silica gel column, and a cyclohexylsilane bonded silica gel column.
As a preferred embodiment, the octadecylsilane bonded silica gel chromatography column is a heptamethyltrisiloxane-terminated octadecylsilane bonded silica gel chromatography column.
In a preferred embodiment, the size of the stationary phase of the octadecylsilane chemically bonded silica chromatographic column is 1.7-2.3 μm.
As a preferred embodiment, the particle size of the stationary phase of the octadecylsilane chemically bonded silica chromatographic column is 1.8 μm.
The applicant finds that the liquid chromatography-triple quadrupole tandem mass spectrometry is used for analysis and detection, so that a sample enters the liquid chromatography firstly, the octadecyl bonded silica gel with the particle size of 1.8 mu m has a good retention effect on polar substances and metabolites, the analyte can enter the pore structure of the particles more easily, the balanced retention performance is provided for polar and hydrophobic molecules, the analyte is prevented from directly flowing out without being combined with resin, and the detection precision is reduced. And the stationary phase in the liquid chromatographic column is octadecyl bonded silica gel blocked by heptamethyltrisiloxane, the bonded silica gel blocked by heptamethyltrisiloxane has good adsorption stability with analytes, still has reaction activity after multiple tests, avoids larger deviation generated in each test, ensures the repeatability of detection, and ensures that the methylthiocyclo-phosphorus has higher detection precision. And because the pesticide residue in the traditional Chinese medicine is low and the retention time of similar components is approximate, the end capping is carried out by heptamethyltrisiloxane, so that the tailing of the thiophosphoryl methyl can be reduced, a peak with good peak shape is formed, and the detection precision is improved.
In a preferred embodiment, in the LC-MS/MS, the ion source of the triple quadrupole tandem mass spectrometer is an electrospray ion source, and the scanning mode is: positive ion scan mode, capillary voltage 1.26Kv, capillary temperature 149 ℃, ion collision energy 30V, low mass resolution 505-515, high mass resolution 1890-1990.
In a preferred embodiment, the low mass resolution of the first quadrupole is 511, the high mass resolution is 1895, the low mass resolution of the second quadrupole is 507, and the high mass resolution is 1989.
In the experimental process, the applicant finds that the retention time of the thiophosphoryl methyl separated from the liquid chromatogram and a plurality of interferents is 34-40s, the separation effect between peaks is poor, and the interference is large when the thiophosphoryl methyl is subjected to quantitative analysis. The possible reasons for guessing are: the low mass resolution of the two quadrupole rods is favorable for judging main fragments, so that the noise of other interferents can be reduced, the signal of low-content methylthiocyclo-phosphate is amplified, the accurate determination of trace amount is realized, more mass spectrum fragments can be obtained through high mass resolution, the peak position and the peak height of the target determination substance methylthiocyclo-phosphate are more accurately judged, and qualitative and quantitative analysis is realized. Meanwhile, the interference of false positive peaks possibly occurring can be further confirmed through the high-quality resolution of the second quadrupole rod, and the interference of false positive signals is avoided.
In a preferred embodiment, the LC-MS/MS is performed at a column temperature of 40 ℃ and a system pressure of 2000-7000psi.
In a preferred embodiment, the LC-MS/MS has a column temperature of 40 deg.C, a minimum system pressure of 2676.6psi and a maximum system pressure of 6298.9psi.
As a preferred embodiment, in the LC-MS/MS, the mobile phase of the LC is selected from at least one of formic acid-acetonitrile, water-ethanol, acetic acid-acetonitrile.
As a preferred embodiment, in the LC-MS/MS, the mobile phase of LC is water-acetonitrile, and the volume ratio of water to acetonitrile at the beginning of elution is 7: and 3, the volume ratio of water to acetonitrile at the end of elution is 0:10, the elution time is 14min.
The applicant finds that the thiophosphoryl methyl is oily liquid in the experimental process, in the detection of pesticide residues of the traditional Chinese medicines, because a lot of traditional Chinese medicines contain a large amount of volatile oil components and have similar properties with the thiophosphoryl methyl, the thiophosphoryl methyl is difficult to separate, and the applicant finds that the thiophosphoryl methyl is easy to denaturize in high-temperature and illumination environments and has low stability in the experiment, so that the thiophosphoryl methyl needs to be rapidly detected in a low-temperature environment, and the detection accuracy is prevented from being influenced by the denaturation of the thiophosphoryl methyl. The applicant finds that gradient elution is carried out by using a mixed solution of water and acetonitrile, the column temperature is 40 ℃, the minimum system pressure is 2676.6psi, the maximum system pressure is 6298.9psi, the outflow time of an analyte can be shortened, and an octadecyl silicagel column blocked by heptamethyltrisiloxane has stronger pressure resistance, so that the octadecyl silicagel column still can keep higher separation efficiency under high-pressure conveying. And the volume ratio of mobile phase water to acetonitrile is from 7: 10, the interfering substance can be kept in the column as much as possible, and the thiophosphoryl methyl flows out in the column rapidly, so that tailing is reduced.
The second aspect of the invention provides application of a method for detecting the residual quantity of thiophosphoryl methyl by LC-MS/MS, which is applied to the detection of pesticide residues in traditional Chinese medicinal materials.
As a preferred embodiment, the Chinese medicinal material is selected from one of salvia miltiorrhiza, panax notoginseng, american ginseng, gallnut and liquorice.
Compared with the prior art, the invention has the following beneficial effects:
(1) According to the method for detecting the residual quantity of the thiophosphoryl methyl by using the LC-MS/MS, the specific polystyrene filler solid-phase extraction column is adopted, so that the purification effect on an analyte is improved, the interference of other components on the thiophosphoryl methyl is reduced, and the detection accuracy is improved.
(2) According to the method for detecting the residual quantity of the thiophosphoryl methyl by LC-MS/MS, the octadecyl silica gel column capped by heptamethyltrisiloxane is used as a liquid chromatographic column, so that the combination stability of an analysis sample and a stationary phase is improved, and the reproducibility and precision of experimental data are improved.
(3) According to the method for detecting the residual quantity of the thiophosphoryl methyl by LC-MS/MS, the linear distribution of analyte linear measurement is improved by setting the high-low mass resolution of a four-level rod in the LC-MS/MS, the interference of false positive signals is reduced, and the detection accuracy is improved.
(4) According to the method for detecting the residual quantity of the thiophosphoryl methyl by LC-MS/MS, acetonitrile is used as an eluent during solid-phase extraction, water-acetonitrile is used as a mobile phase in liquid chromatogram, and the analytes are subjected to gradient elution, so that the period of analysis and detection is shortened, the separation capacity of the thiophosphoryl methyl is improved, and the tailing of peaks is reduced.
(5) The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl has low detection limit, can be used for trace detection of the thiophosphoryl methyl in the traditional Chinese medicinal materials, and is particularly suitable for pesticide residue analysis of the thiophosphoryl methyl in the salvia miltiorrhiza and the gallnuts.
Detailed Description
The present invention will be specifically described below by way of examples. It should be noted that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention, and that the insubstantial modifications and adaptations of the present invention by those skilled in the art based on the above disclosure are still within the scope of the present invention.
In addition, the starting materials used are all commercially available, unless otherwise specified.
Example 1
A method for detecting the residual quantity of thiophosphoryl methyl by LC-MS/MS comprises the following steps:
(1) Sample extraction: sieving ground Saviae Miltiorrhizae radix, adding sodium chloride, mixing, adding acetonitrile, homogenizing, centrifuging, and collecting supernatant;
(2) Sample purification: passing the supernatant obtained in the step 1 through a solid phase extraction column to obtain an effluent liquid;
(3) And (3) sample determination: performing LC-MS/MS detection on the effluent liquid obtained in the step 2, setting parameters and performing detection;
(4) And (4) analyzing results: and analyzing the spectrogram, and comparing with a standard curve to obtain the residual quantity of the thiophosphoryl methyl.
As a preferred embodiment, the sample extraction in step 1 is: adding 5g of salvia miltiorrhiza and 1g of sodium chloride into 50mL of acetonitrile, homogenizing at the rotation speed of 15000r/min for 2min, and centrifuging at the rotation speed of 5500r/min; and then adding 50mL of acetonitrile into the centrifuged precipitate again, homogenizing at the rotation speed of 15000r/min for 2min and at the rotation speed of 5000r/min, combining the two supernatants, concentrating under reduced pressure to 5mL, and diluting with an organic solvent to 10mL.
The sample in the step 1 is a 50-mesh screen underflow.
The solid phase extraction column in the step 2 is a polystyrene filler solid phase extraction column, the stationary phase is polystyrene-divinylbenzene, the specification is 200mg,6mL, and the solid phase extraction column is purchased from Shanghai Anpu.
In the step 3LC-MS/MS, the liquid chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, the stationary phase is heptamethyltrisiloxane end-capped octadecylsilane chemically bonded silica, the particle size of the stationary phase is 1.8 mu m, and the model is ACQUITY
HSS T3, available from Watts corporation.
In the LC-MS/MS, an ion source of a triple quadrupole tandem mass spectrometer is an electrospray ion source, and the scanning mode is as follows: positive ion scan mode, capillary voltage 1.26Kv, capillary temperature 149 ℃, ion collision energy 30V, low mass resolution of 511 for the first quadrupole, high mass resolution of 1895, low mass resolution of 507 for the second quadrupole, high mass resolution of 1989. The column temperature of the liquid chromatography is 40 ℃, the minimum system pressure is 2676.6psi, the maximum system pressure is 6298.9psi, the mobile phase of the liquid chromatography is water-acetonitrile, and the volume ratio of water to acetonitrile at the beginning of elution is 7: and 3, the volume ratio of water to acetonitrile at the end of elution is 0:10, elution time 14min.
Example 2
A method for detecting the residual quantity of thiophosphoryl methyl by LC-MS/MS comprises the following steps:
(1) Sample extraction: sieving pulverized Galla chinensis, adding sodium chloride, mixing, adding acetonitrile, homogenizing, centrifuging, and collecting supernatant;
(2) Sample purification: passing the supernatant obtained in the step 1 through a solid phase extraction column to obtain an effluent liquid;
(3) And (3) sample determination: performing LC-MS/MS detection on the effluent liquid obtained in the step 2, setting parameters and performing detection;
(4) And (4) analyzing results: and analyzing the spectrogram, and comparing with a standard curve to obtain the residual quantity of the thiophosphoryl methyl.
As a preferred embodiment, the sample extraction in step 1 is: 5g of gallnut and 1g of sodium chloride, adding 50mL of acetonitrile, homogenizing at 13000r/min for 2min, and centrifuging at 6000r/min; and then adding 50mL of acetonitrile into the centrifuged precipitate again, homogenizing at 13000r/min for 2min, and 5500r/min, combining the two supernatants, concentrating under reduced pressure to 3mL, and diluting to 10mL by using an organic solvent.
The sample in the step 1 is the undersize material of a 50-mesh sieve.
The solid phase extraction column in the step 2 is a polystyrene filler solid phase extraction column, and the stationary phase is polystyrene-divinylbenzene with the specification of 200mg,6mL and purchased from Shanghai Anpu.
In the step 3LC-MS/MS, the liquid chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, the stationary phase is heptamethyltrisiloxane end-capped octadecylsilane chemically bonded silica, the particle size of the stationary phase is 1.8 mu m, and the model is ACQUITY
HSST 3, available from Watts corporation.
In the LC-MS/MS, an ion source of a triple quadrupole tandem mass spectrometer is an electrospray ion source, and the scanning mode is as follows: positive ion scan mode, capillary voltage 1.26Kv, capillary temperature 149 ℃, ion collision energy 30V, low mass resolution of 511 for the first quadrupole, high mass resolution of 1895, low mass resolution of 507 for the second quadrupole, and high mass resolution of 1989. The column temperature of the liquid chromatography is 40 ℃, the minimum system pressure is 2676.6psi, the maximum system pressure is 6298.9psi, the mobile phase of the liquid chromatography is water-acetonitrile, and the volume ratio of water to acetonitrile at the beginning of elution is 7: and 3, the volume ratio of water to acetonitrile at the end of elution is 0:10, elution time 14min.
Example 3
The steps of the method for detecting the residual quantity of the thiophosphoryl methyl by LC-MS/MS are the same as those of the example 1, and the difference is that the solid-phase extraction column in the step 2 is a graphitized carbon black filler solid-phase extraction column.
Example 4
The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl and the application of the method are the same as the example 1, and the difference is that in the step 3LC-MS/MS, a liquid phase chromatographic column is an octadecylsilane chemically bonded silica chromatographic column, and a stationary phase is hexamethyltrisilazane end-capped octadecylsilane chemically bonded silica.
Example 5
The steps of a method for detecting the residual quantity of methylthiocyclophosphate by LC-MS/MS are the same as those of example 1, and the difference is that the mobile phase of liquid chromatogram is formic acid-acetonitrile.
Performance testing
1. Linear value, detection limit: preparing a standard solution: taking the salvia miltiorrhiza extract which does not contain the thiophosphoryl methyl through detection, preparing the salvia miltiorrhiza extract into a blank solution, preparing a series of standard solutions with different concentrations by the thiophosphoryl methyl and the blank solution within the range of 5-500 mu g/L, drawing a standard curve by adopting the detection methods of the embodiments 1,3,4 and 5, and calculating the linear values and the lowest detection limit corresponding to different embodiments.
In the same test method, the gallnut extract which is detected to be free of thiophosphoryl methyl is used as a blank solution, a standard curve is drawn by adopting the detection method of the example 2, and the corresponding linear value and the lowest detection limit are calculated.
The sample concentration at detection limit D =3 times the signal-to-noise ratio; the linear value is a calculation method of the linear value in the prior art.
2. Recovery, relative standard deviation: two salvia miltiorrhiza samples containing the thiophosphoryl methyl are extracted by the method of the embodiment 1,3,4 and 5, one part is added with the thiophosphoryl methyl standard solution of 10 mu g/L, and two parts are detected by the method of the embodiment 1,3,4 and 5, are horizontally repeated for 6 times, and the average recovery rate and the precision are measured.
Two samples of Galla chinensis containing thiophosphoryl methyl were extracted as described in example 2, and 10. Mu.g/L of thiophosphoryl methyl standard solution was added to one sample, and the two samples were tested as described in example 2, and the level was repeated 5 times to determine the average recovery and precision.
Recovery = (measured with standard solution-measured without standard solution)/10 μ g/L100%
Precision is standard deviation, which is a prior art method of calculating standard deviation.
The examples were tested according to the above criteria and the results are shown in table 1.
TABLE 1
| Linear value | Detection limit D [ mu ] g/kg | Percent recovery% | Precision RSD/%) | |
| Example 1 | 0.9966 | 7 | 108 | 8.9 |
| Example 2 | 0.9954 | 10 | 103 | 9.5 |
| Example 3 | 0.9885 | 12 | 65 | 11.2 |
| Example 4 | 0.9881 | 11 | 68 | 10.3 |
| Example 5 | 0.9752 | 13 | 66 | 10.9 |
Claims (10)
1. A method for detecting the residual quantity of thiophosphoryl methyl by LC-MS/MS is characterized by comprising the following steps:
(1) Sample extraction: sieving the ground sample, adding sodium chloride, mixing, adding an organic solvent, homogenizing, centrifuging, and taking the supernatant;
(2) Sample purification: passing the supernatant obtained in the step 1 through a solid phase extraction column to obtain an effluent liquid;
(3) And (3) sample determination: performing LC-MS/MS detection on the effluent liquid obtained in the step 2, setting parameters and performing detection;
(4) And (4) analyzing results: and analyzing the spectrogram, and comparing with a standard curve to obtain the residual quantity of the thiophosphoryl methyl.
2. The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to claim 1, wherein the sample in the step 1 is a 50-mesh screen underflow.
3. The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to claim 1, wherein the organic solvent in the step 1 is one or a combination of more of acetone, benzene, ethanol, acetonitrile, isopropanol, acetic acid, toluene and styrene.
4. The LC-MS/MS method for detecting the residual quantity of thiophosphoryl methyl according to claim 1, wherein the solid phase extraction column in the step 2 is selected from one of a reverse phase solid phase extraction column or an ion exchange solid phase extraction column, and preferably the reverse phase solid phase extraction column is selected from one of an octadecyl bonded silica gel solid phase extraction column, a graphitized carbon black filler solid phase extraction column, a polystyrene filler solid phase extraction column and a propanecyanoalkane bonded silica gel solid phase extraction column.
5. The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to the claim 1, wherein the step 3 is performed by LC-MS/MS, and LC is selected from at least one of an adsorption chromatographic column, a distribution chromatographic column, an ion exchange chromatographic column and a gel chromatographic column.
6. The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to claim 5, wherein the adsorption chromatographic column is at least one selected from a silica gel chromatographic column, an alumina chromatographic column, a polystyrene-divinyl chromatographic column and a polymethacrylate chromatographic column.
7. The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to claim 1, wherein in the LC-MS/MS, the column temperature of LC is 40 ℃, and the system pressure is 2000-7000psi.
8. The LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to claim 1, wherein in the LC-MS/MS, the mobile phase of the LC is selected from at least one of formic acid-acetonitrile, water-ethanol and acetic acid-acetonitrile.
9. The application of the LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to any one of claims 1 to 8 is characterized in that the method is applied to the detection of pesticide residues of Chinese medicinal materials.
10. The application of the LC-MS/MS method for detecting the residual quantity of the thiophosphoryl methyl according to claim 9, wherein the Chinese medicinal material is one selected from the group consisting of salvia miltiorrhiza, panax notoginseng, american ginseng, gallnut and liquorice.
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