CN114354823A - Method for detecting chlorothalonil and metabolites thereof in agricultural products - Google Patents
Method for detecting chlorothalonil and metabolites thereof in agricultural products Download PDFInfo
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- 239000005747 Chlorothalonil Substances 0.000 title claims abstract description 86
- CRQQGFGUEAVUIL-UHFFFAOYSA-N chlorothalonil Chemical compound ClC1=C(Cl)C(C#N)=C(Cl)C(C#N)=C1Cl CRQQGFGUEAVUIL-UHFFFAOYSA-N 0.000 title claims abstract description 82
- 238000000034 method Methods 0.000 title claims abstract description 44
- 239000002207 metabolite Substances 0.000 title claims abstract description 42
- 239000000243 solution Substances 0.000 claims abstract description 43
- 239000012224 working solution Substances 0.000 claims abstract description 18
- 239000002253 acid Substances 0.000 claims abstract description 14
- 238000004458 analytical method Methods 0.000 claims abstract description 14
- 238000000605 extraction Methods 0.000 claims description 30
- 238000000746 purification Methods 0.000 claims description 19
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 18
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 15
- MDQKYGOECVSPIW-UHFFFAOYSA-N 4-hydroxychlorothalonil Chemical compound OC1=C(Cl)C(Cl)=C(C#N)C(Cl)=C1C#N MDQKYGOECVSPIW-UHFFFAOYSA-N 0.000 claims description 13
- 238000004445 quantitative analysis Methods 0.000 claims description 13
- MYRTYDVEIRVNKP-UHFFFAOYSA-N 1,2-Divinylbenzene Chemical compound C=CC1=CC=CC=C1C=C MYRTYDVEIRVNKP-UHFFFAOYSA-N 0.000 claims description 12
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 238000005173 quadrupole mass spectroscopy Methods 0.000 claims description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 239000003929 acidic solution Substances 0.000 claims description 10
- 238000004817 gas chromatography Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 239000002904 solvent Substances 0.000 claims description 10
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- 235000019253 formic acid Nutrition 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 9
- 239000000945 filler Substances 0.000 claims description 8
- 238000004811 liquid chromatography Methods 0.000 claims description 8
- 239000002048 multi walled nanotube Substances 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 8
- -1 octadecyl silica gel Chemical compound 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 6
- 239000006229 carbon black Substances 0.000 claims description 6
- 239000000741 silica gel Substances 0.000 claims description 6
- 229910002027 silica gel Inorganic materials 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 4
- 230000014759 maintenance of location Effects 0.000 claims description 4
- 239000012071 phase Substances 0.000 claims description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 3
- RQBBFKINEJYDOB-UHFFFAOYSA-N acetic acid;acetonitrile Chemical compound CC#N.CC(O)=O RQBBFKINEJYDOB-UHFFFAOYSA-N 0.000 claims description 3
- XBJFCYDKBDVADW-UHFFFAOYSA-N acetonitrile;formic acid Chemical compound CC#N.OC=O XBJFCYDKBDVADW-UHFFFAOYSA-N 0.000 claims description 3
- DQFAAIWCCHDCLO-UHFFFAOYSA-N acetonitrile;formic acid;hydrate Chemical compound O.CC#N.OC=O DQFAAIWCCHDCLO-UHFFFAOYSA-N 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 claims description 3
- 238000005571 anion exchange chromatography Methods 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- VBUZIXJFGCVTJL-UHFFFAOYSA-N azanium;methanol;formate Chemical compound [NH4+].OC.[O-]C=O VBUZIXJFGCVTJL-UHFFFAOYSA-N 0.000 claims description 3
- 238000005277 cation exchange chromatography Methods 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 238000012856 packing Methods 0.000 claims description 3
- 239000012521 purified sample Substances 0.000 claims description 3
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims description 3
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 3
- XNZQCYSOYHAYII-UHFFFAOYSA-L disodium;3-carboxy-3-hydroxypentanedioate;hydrate Chemical compound [OH-].[Na+].[Na+].OC(=O)CC(O)(C([O-])=O)CC(O)=O XNZQCYSOYHAYII-UHFFFAOYSA-L 0.000 claims description 2
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 claims description 2
- 230000002000 scavenging effect Effects 0.000 claims description 2
- 239000007790 solid phase Substances 0.000 claims description 2
- 230000000274 adsorptive effect Effects 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 8
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- 238000006243 chemical reaction Methods 0.000 description 10
- 241000220259 Raphanus Species 0.000 description 8
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 8
- 235000012055 fruits and vegetables Nutrition 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 239000012496 blank sample Substances 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 2
- 239000012490 blank solution Substances 0.000 description 2
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- 239000006228 supernatant Substances 0.000 description 2
- 244000291564 Allium cepa Species 0.000 description 1
- 235000002732 Allium cepa var. cepa Nutrition 0.000 description 1
- 241000207199 Citrus Species 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 235000020971 citrus fruits Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 239000012084 conversion product Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
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- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
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- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
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- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000003993 organochlorine pesticide Substances 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000011403 purification operation Methods 0.000 description 1
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- 239000000758 substrate Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- HGPVLOQNBSHYEI-UHFFFAOYSA-J tetrasodium;hydron;2-hydroxypropane-1,2,3-tricarboxylate;trihydrate Chemical compound O.O.O.[Na+].[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O.[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O HGPVLOQNBSHYEI-UHFFFAOYSA-J 0.000 description 1
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Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a method for detecting chlorothalonil and metabolites thereof in agricultural products, and relates to the technical field of instrument analysis. A method for detecting chlorothalonil and metabolites thereof in agricultural products adopts an acid solution as a chlorothalonil metabolism inhibiting solution, so that a solution to be detected and a standard mixed working solution are in a stable concentration state in an instrumental analysis process, and on the basis, an instrumental analysis method is adopted to establish a synchronous full recovery method of chlorothalonil and two main metabolites, realize accurate quantification and truly reflect the residual levels of the chlorothalonil and the metabolites in food.
Description
Technical Field
The invention relates to the technical field of instrument analysis, in particular to a method for detecting chlorothalonil and metabolites thereof in agricultural products.
Background
Chlorothalonil is a broad-spectrum protective bactericide and is widely used for disease control in the production process of planting products such as fruits and vegetables, grains, tea leaves and the like. Abuse or improper use of such drugs can cause chlorothalonil and its metabolites in food, potentially harmful to human and animal health. 4-hydroxychlorothalonil is a well-known metabolite of chlorothalonil. With the rapid advancement of mass spectrometry technology, more and more unknown metabolites were found to be confirmed.
As people do not find that chlorothalonil can be converted into mercaptochlorothalonil in fruit and vegetable products, the national agricultural product quality safety monitoring project only monitors organochlorine pesticide precursors such as chlorothalonil and the like throughout the year, and metabolic conversion products of the chlorothalonil are not concerned, so that the potential risk of pesticide residue is easily underestimated. The inventor finds out through long-term research that: the chlorothalonil can be quickly converted into the mercaptochlorothalonil in fruit and vegetable matrixes such as white radish, onion, citrus and the like, the conversion rate is 10-100%, and the conversion rates of the mercaptochlorothalonil in different fruit and vegetable matrixes are different.
The synchronous full recovery extraction and detection of chlorothalonil, mercaptochlorothalonil and 4-hydroxychlorothalonil are very key for guaranteeing the quality safety of agricultural products. Since mercaptochlorothalonil is a novel metabolite in agricultural product substrates, no research on detection methods of mercaptochlorothalonil has been reported so far. It is worth mentioning that, in the actual sample extraction, purification and detection process, chlorothalonil can be continuously and dynamically converted into mercaptochlorothalonil, and how to inhibit the dynamic conversion of the chlorothalonil matrix standard solution in the instrument analysis process, so that the chlorothalonil matrix standard solution can be used for quantitative analysis? The method is a key and difficult problem to be solved for developing a method for synchronously detecting chlorothalonil and metabolites thereof.
In view of this, the invention is particularly proposed.
Disclosure of Invention
The invention aims to provide a method for detecting chlorothalonil and metabolites thereof in agricultural products, and aims to inhibit the chlorothalonil from being converted into metabolites in the detection process, and further detect the content of the chlorothalonil and the metabolites simultaneously.
The invention is realized by the following steps:
in a first aspect, the present invention provides a method for detecting chlorothalonil and metabolites thereof in agricultural products, comprising:
extracting agricultural products to be detected to obtain a mixed extracting solution containing chlorothalonil and metabolites thereof, wherein the metabolites comprise sulfydryl chlorothalonil and 4-hydroxyl chlorothalonil;
extracting and purifying the mixed extracting solution, and mixing the purified sample solution with an acidic solution to obtain a solution to be detected;
detecting chlorothalonil and metabolites thereof in a liquid to be detected, and carrying out quantitative analysis by using a standard curve;
the standard curve is formed by the following method: preparing a plurality of groups of standard mixed working solutions of chlorothalonil and metabolites thereof with different concentrations, and forming a standard curve for quantitative analysis through instrumental analysis, wherein an acidic solution is added in the preparation process of the standard mixed working solutions.
In an alternative embodiment, the extractive purification is by means of dispersed solid phase extraction or solid phase extractive purification.
In an alternative embodiment, the materials used for the dispersed solid phase extraction include extraction salts and purification packing; preferably, the extraction salt is selected from at least one of anhydrous magnesium sulfate, sodium chloride, trisodium citrate dihydrate and disodium citrate sesquihydrate; preferably, the scavenging filler is selected from at least one of octadecyl silica gel, ethylenediamine-N-propyl silane, carbon black, multi-walled carbon nanotubes, N-vinyl pyrrolidone, and divinyl benzene.
In an alternative embodiment, the chromatographic retention mode used during the solid phase extraction purification is selected from at least one of reverse phase chromatography, anion exchange chromatography and cation exchange chromatography.
In an alternative embodiment, the adsorbent filler used in the solid phase extraction purification process is selected from at least one of octadecyl silica gel, ethylenediamine-N-propyl silane, carbon black, multiwall carbon nanotubes, N-vinyl pyrrolidone, and divinyl benzene.
In an alternative embodiment, the acidic solution is selected from at least one of formic acid, acetic acid, hydrochloric acid, sulfuric acid, and phosphoric acid.
In an alternative embodiment, the volume fraction of the acid in the solution to be detected is controlled to be 0.15-0.25%, and the volume fraction of the acid in the standard mixed working solution is controlled to be 0.15-0.25%.
In an alternative embodiment, the extraction solvent used in the extraction process is selected from any one of acetonitrile, formic acid-acetonitrile solution, acetic acid-acetonitrile solution, and formic acid-water-acetonitrile solution;
preferably, the agricultural product to be detected is homogenized and then mixed with the extraction solvent for extraction.
In an alternative embodiment, the apparatus for quantitative analysis is selected from at least one of gas chromatography, liquid chromatography tandem triple quadrupole mass spectrometry, and gas chromatography tandem triple quadrupole mass spectrometry.
In an optional embodiment, the method comprises the steps of quantitatively analyzing chlorothalonil by using a gas chromatography-tandem triple quadrupole mass spectrometer, and quantitatively analyzing mercaptochlorothalonil and 4-hydroxychlorothalonil by using a liquid chromatography-tandem mass spectrometer;
preferably, when chlorothalonil is detected, gas chromatography is adopted to be connected with triple quadrupole mass spectrometry in series, a chromatographic column is HP-5MS UI, the length of the chromatographic column is 28-32m, the flow rate is 0.8-1.2mL/min, and the injection port temperature is 250-300 ℃; the temperature raising program comprises: firstly heating to 70-90 ℃ and preserving heat for 0.5-2min, then heating to 220 ℃ at the heating rate of 35-45 ℃/min, preserving heat for 4-6min at the temperature of 4-6 ℃/min to 230 ℃ and 250 ℃ at the temperature of 10-15 ℃/min to 310 ℃ and 330 ℃;
preferably, when the mercaptochlorothalonil and the 4-hydroxychlorothalonil are detected, a liquid chromatography tandem mass spectrometer is adopted, a chromatographic column is Agilent XDB C182.1 × 150mm, the diameter of the chromatographic column is 3.5 μm, and the column temperature is as follows: 25 ℃; flow rate: 0.4 mL/min; mobile phase: a is 5mM ammonium formate aqueous solution, B is 5mM ammonium formate methanol; the elution procedure is shown in the following table:
the invention has the following beneficial effects: the acid solution is used as a chlorothalonil metabolism inhibiting solution, so that the solution to be detected and the standard mixed working solution are in a stable concentration state in the instrument analysis process, and on the basis, an instrument analysis method is adopted to establish a directional quantitative analysis method of chlorothalonil, mercaptochlorothalonil and 4-hydroxychlorothalonil.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Aiming at the problem that chlorothalonil is continuously and dynamically converted into mercaptochlorothalonil in the prior art, the inventor can inhibit the dynamic conversion of the chlorothalonil in the instrument analysis process by adding a chlorothalonil conversion inhibitor, and further accurately detect the content of the chlorothalonil and metabolites thereof in a sample.
The embodiment of the invention provides a method for detecting chlorothalonil and metabolites thereof in agricultural products, which comprises the following steps:
s1, extraction
Extracting agricultural products to be detected to obtain mixed extracting solution containing chlorothalonil and metabolites thereof, wherein the metabolites comprise sulfydryl chlorothalonil and 4-hydroxyl chlorothalonil. In the actual operation process, the agricultural products to be detected are crushed and then mixed with the extraction solvent for extraction by adopting a solvent extraction method, so as to obtain a mixed extracting solution containing chlorothalonil and metabolites thereof.
In some embodiments, the extraction solvent employed in the extraction process is selected from any one of acetonitrile, formic acid-acetonitrile solution, acetic acid-acetonitrile solution, and formic acid-water-acetonitrile solution; the extraction solvents can effectively extract chlorothalonil, sulfydryl chlorothalonil and 4-hydroxyl chlorothalonil, and the extraction rate is ideal.
In a preferred embodiment, the agricultural product to be detected is homogenized homogeneously and then extracted by mixing with an extraction solvent. The homogenizing and homogenizing can be processed by adopting a common homogenizer and a homogenizer, and the homogenizing and homogenizing is mixed with the extraction solvent to obviously improve the extraction efficiency.
S2 preparation of solution to be detected
And extracting and purifying the mixed extracting solution, and mixing the purified sample solution with an acidic solution to obtain a solution to be detected. The extracted effective components are extracted by extraction and purification and then are mixed with an acidic solution to obtain the liquid to be detected of the sample, and the acidic solution plays a role in inhibiting the chlorothalonil from being converted into metabolites, so that the detection can be effectively carried out.
In some embodiments, the extraction and purification is performed by dispersive solid-phase extraction or solid-phase extraction and purification, both of which are suitable for the method provided in the embodiments of the present invention, and can effectively extract chlorothalonil and metabolites thereof. Both dispersed solid phase extraction and solid phase extraction are existing extraction methods.
In some embodiments, materials employed for dispersed solid phase extraction include extraction salts and purification packing; the extract salt is selected from anhydrous magnesium sulfate (MgSO)4) Sodium chloride (NaCl), trisodium citrate dihydrate (Na)3Cit·2H2O) and citric acid disodium salt sesquihydrate (Na)2Cit·1.5H2At least one of O); the purifying filler is at least one selected from octadecyl silica gel (C18), ethylenediamine-N-Propyl Silane (PSA), carbon black (GCB), multi-wall carbon nanotubes (MWCNTs), N-vinyl pyrrolidone and divinyl benzene. The extracted salts are suitable for extracting chlorothalonil and metabolites thereof, impurities can be effectively removed through the purification filler, and the detection accuracy is improved.
In some embodiments, the chromatographic retention mode used in the solid phase extraction purification process is selected from at least one of reverse phase chromatography, anion exchange chromatography, and cation exchange chromatography. The adsorption filler used in the solid phase extraction purification process is at least one selected from octadecyl silica gel (C18), ethylenediamine-N-Propyl Silane (PSA), carbon black (GCB), multi-walled carbon nanotubes (MWCNTs), N-vinyl pyrrolidone and divinylbenzene. Through screening adsorption filler, can reach better extraction purifying effect, promote the accuracy that detects.
Further, the acid solution is at least one selected from formic acid, acetic acid, hydrochloric acid, sulfuric acid and phosphoric acid, and the acids are all suitable for the analysis method in the embodiment of the invention, so that the conversion of chlorothalonil can be effectively inhibited, and the detection can be smoothly carried out. Specifically, the volume fraction of the acid in the liquid to be detected is controlled to 0.15 to 0.25%, and the volume fraction of the acid in the liquid to be detected is preferably controlled within the above range, in order to more effectively suppress the conversion of chlorothalonil, and the volume fraction of the acid in the liquid to be detected may be 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, or the like, or may be any value between the above adjacent volume fractions.
S3 preparation of standard mixed working solution
Preparing a plurality of groups of standard mixed working solutions of chlorothalonil and metabolites thereof with different concentrations so as to form a standard curve for quantitative analysis through instrumental analysis, wherein an acidic solution is added in the preparation process of the standard mixed working solutions; the acid content of the standard mixed working solution is 0.15-0.25%. Acid is also added into the standard mixed working solution to inhibit the conversion of chlorothalonil in the standard mixed working solution, a more accurate standard curve is obtained through detection and analysis, and the detection accuracy is improved.
In some embodiments, the standard mixed working solution may be prepared on the basis of a blank matrix extracting solution in the preparation process, if the substance to be detected is white radish, white radish which does not substantially contain chlorothalonil is extracted to obtain a blank matrix, the blank matrix is mixed with an acid solution to obtain an acidic blank matrix, the acidic blank matrix is mixed with the standard mixed solution of chlorothalonil and metabolites, and a plurality of groups of standard mixed working solutions with different concentrations are prepared to form a standard curve.
S4, quantitative analysis
And (3) quantitatively analyzing the chlorothalonil and the metabolites thereof in the liquid to be detected by adopting common analytical instruments such as a chromatograph, a mass spectrometer and the like.
In some embodiments, the apparatus for quantitative analysis is selected from at least one of gas chromatography, liquid chromatography tandem triple quadrupole mass spectrometry, and gas chromatography tandem triple quadrupole mass spectrometry. The above instruments can accurately detect chlorothalonil and metabolites thereof.
In a preferred embodiment, in order to improve the detection accuracy, the gas chromatography-tandem triple quadrupole mass spectrometry is adopted to perform quantitative analysis on the chlorothalonil, and the liquid chromatography-tandem mass spectrometer is adopted to perform quantitative analysis on the mercaptochlorothalonil and the 4-hydroxychlorothalonil.
Specifically, when chlorothalonil is detected, gas chromatography is adopted to be connected with triple quadrupole mass spectrometry in series, a chromatographic column is HP-5MS UI, the length of the chromatographic column is 28-32m, the flow rate is 0.8-1.2mL/min, and the injection port temperature is 250-300 ℃; the temperature raising program comprises: firstly heating to 70-90 ℃ and preserving heat for 0.5-2min, then heating to 220 ℃ at the heating rate of 35-45 ℃/min, preserving heat for 4-6min at the temperature of 4-6 ℃/min to 230 ℃ and 250 ℃ at the temperature of 10-15 ℃/min to 310 ℃ and 330 ℃;
specifically, when detecting mercaptochlorothalonil and 4-hydroxychlorothalonil, a liquid chromatography tandem mass spectrometer is adopted, a chromatographic column is Agilent XDB C182.1 multiplied by 150mm, the column temperature is 3.5 mu m: 25 ℃; flow rate: 0.4 mL/min; mobile phase: a is 5mM ammonium formate aqueous solution, B is 5mM ammonium formate methanol; the elution procedure is shown in table 1:
table 1 mobile phase gradient elution procedure
Test examples
In order to illustrate that the detection analysis method in the embodiment of the invention can accurately detect chlorothalonil and metabolites thereof in agricultural products, the inventor provides the following tests:
taking white radish without chlorothalonil and metabolites thereof as a sample, introducing the chlorothalonil, and testing the content of the chlorothalonil and the metabolites thereof in the final sample. The method comprises the following specific steps:
pretreatment of QuEChERS samples
Weighing 10g of homogenized white radish sample (accurate to 0.01g) into a 50mL plastic centrifuge tube, and adding a certain concentration level of chlorothalonil standard solution. 15mL of acetonitrile was added, extraction package P-QuEChERS-EN 1101 (content: 4g MgSO. RTM4，1g NaCl，1g Na3Cit·2H2O，0.5g Na2Cit·1.5H2O), after 1min of vigorous shaking, 4200r/min of the suspension is centrifuged for 5 min.
1mL of supernatant was placed in a plastic centrifuge tube and QuEChERS purification pack (contents: 150mg MgSO 2) was added425mg PSA, 5mg GCB). Mixing by vortex for 1min, centrifuging at 4200r/min for 5min, repeating the above operation for 2 times, accurately transferring 1mL supernatant, addingAdd 10. mu.L of formic acid solution (20%, v/v), vortex for 10s and mix, filter 0.22 μm into the sample vial for assay.
2. Preparation of Standard solutions
When the matrix standard solution is prepared, the blank sample extracting solution needs to be subjected to parallel repeated purification operation for a certain number of times in advance to obtain a sufficient amount of blank solution (the blank solution refers to a sample obtained after extracting and extracting the white radish which does not contain chlorothalonil) for preparing the matrix standard solution. 10.00mL of the purified blank sample solution was removed, and 20. mu.L of formic acid was added to prepare a blank matrix solution containing 0.2% formic acid. Accurately sucking a certain amount of mixed standard solution of chlorothalonil, mercaptochlorothalonil and hydroxychlorothalonil, gradually diluting the mixed standard solution into mixed standard matrix working solution with the mass concentration of 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L and 0.2mg/L by using blank sample matrix solution containing 0.2% formic acid, and using the mixed standard working solution for quantitative determination of chlorothalonil and metabolites.
Note: the mass concentration of the diluted solution refers to the mass concentration of formic acid.
3. Measurement of
(1) Determination of chlorothalonil by gas chromatography-tandem mass spectrometer
The instrument conditions were as follows:
a chromatographic column: HP-5MS UI 30m × 0.25mm,0.25 μm; flow rate: 1.0 mL/min; sample inlet temperature: 280 ℃; temperature rising procedure: 80 deg.C (1min), 40 deg.C/min to 200 deg.C, 5 deg.C/min to 240 deg.C, 12 deg.C/min to 320 deg.C (5 min); sample introduction amount: 1 mu L of the solution; and (3) sample introduction mode: no shunt sampling; electron bombardment source: 70 eV; transmission line temperature: 280 ℃; ion source temperature: 280 ℃; solvent retardation: 3 min; monitoring multiple reactions: all ion pairs to be detected are detected in time intervals according to the peak appearance sequence. The retention time, quantitative ion pair, qualitative ion pair and collision voltage of chlorothalonil are shown in table 2:
TABLE 2 test parameters of the gas chromatography-tandem mass spectrometry method
(2) Method for measuring hydroxyl chlorothalonil and sulfydryl chlorothalonil by adopting liquid chromatography tandem mass spectrometer
The contents of the hydroxychlorothalonil and the mercaptochlorothalonil were determined by a liquid chromatography tandem mass spectrometer, and the specific test parameters are shown in table 3:
TABLE 3 liquid chromatography-tandem mass spectrometry method test parameters
4. The result of the detection
The results of the tests on chlorothalonil, mercaptochlorothalonil and hydroxychlorothalonil are shown in table 4.
Table 4 test results of white radish added samples
The accuracy and precision of the detection method are verified by carrying out a chlorothalonil addition recovery experiment, and detection results show that most of the added chlorothalonil standard solution of the white radish is converted into mercaptochlorothalonil in the pretreatment process, the overall recovery rate of the chlorothalonil and metabolites thereof is 98.2%, and the Relative Standard Deviation (RSD) is 1.2%. The detection limit of chlorothalonil, hydroxychlorothalonil and mercaptochlorothalonil is 10 mug/kg.
According to the test example, the detection method provided by the embodiment of the invention can accurately detect the content of chlorothalonil and metabolites thereof, and is a feasible technical scheme.
Comparative test example 1
This test example 2 differs from test example 1 only in that: formic acid is not added in the preparation processes of the detection solution and the standard solution.
The results show that: the chlorothalonil matrix standard solution is in a dynamic conversion state, and an effective chlorothalonil standard working curve cannot be obtained, so that the quantitative detection of the chlorothalonil in a sample cannot be carried out.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for detecting chlorothalonil and metabolites thereof in agricultural products, comprising:
extracting agricultural products to be detected to obtain a mixed extracting solution containing chlorothalonil and metabolites thereof, wherein the metabolites comprise sulfydryl chlorothalonil and 4-hydroxyl chlorothalonil;
extracting and purifying the mixed extracting solution, and mixing the purified sample solution with an acidic solution to obtain a solution to be detected;
detecting chlorothalonil and metabolites thereof in the liquid to be detected, and carrying out quantitative analysis by using a standard curve;
the standard curve is formed by the following method: preparing a plurality of groups of standard mixed working solutions of chlorothalonil and metabolites thereof with different concentrations so as to form a standard curve for quantitative analysis through instrumental analysis, wherein the acidic solution is added in the preparation process of the standard mixed working solutions.
2. The method of claim 1, wherein the extractive purification is by dispersed solid phase extraction or solid phase extraction purification.
3. The method of claim 2, wherein the materials used for the dispersed solid phase extraction include extraction salts and purification packing;
preferably, the extraction salt is selected from at least one of anhydrous magnesium sulfate, sodium chloride, trisodium citrate dihydrate and disodium citrate sesquihydrate;
preferably, the scavenging filler is selected from at least one of octadecyl silica gel, ethylenediamine-N-propyl silane, carbon black, multi-walled carbon nanotubes, N-vinyl pyrrolidone, and divinyl benzene.
4. The method of claim 2, wherein the chromatographic retention mode used in the solid phase extraction purification process is selected from at least one of reverse phase chromatography, anion exchange chromatography and cation exchange chromatography.
5. The method of claim 4, wherein the adsorptive filler used in the solid phase extractive purification process is selected from at least one of octadecyl silica gel, ethylenediamine-N-propyl silane, carbon black, multiwall carbon nanotubes, N-vinyl pyrrolidone, and divinylbenzene.
6. The method of claim 1, wherein the acidic solution is selected from at least one of formic acid, acetic acid, hydrochloric acid, sulfuric acid, and phosphoric acid.
7. The method according to claim 6, wherein the volume fraction of the acid in the liquid to be detected is controlled to be 0.15 to 0.25%, and the volume fraction of the acid in the standard mixed working solution is controlled to be 0.15 to 0.25%.
8. The method according to claim 1, wherein the extraction solvent used in the extraction process is selected from any one of acetonitrile, formic acid-acetonitrile solution, acetic acid-acetonitrile solution, and formic acid-water-acetonitrile solution;
preferably, the agricultural product to be detected is homogenized and then mixed with the extraction solvent for extraction.
9. The method of claim 1, wherein the quantitative analysis is performed by an instrument selected from the group consisting of gas chromatography, liquid chromatography tandem triple quadrupole mass spectrometry, and gas chromatography tandem triple quadrupole mass spectrometry.
10. The method of claim 9, wherein the chlorothalonil is quantitatively analyzed by gas chromatography tandem triple quadrupole mass spectrometry, and the mercaptochlorothalonil and the 4-hydroxychlorothalonil are quantitatively analyzed by liquid chromatography tandem mass spectrometry;
preferably, when chlorothalonil is detected, gas chromatography is adopted to be connected with triple quadrupole mass spectrometry in series, a chromatographic column is HP-5MS UI, the length of the chromatographic column is 28-32m, the flow rate is 0.8-1.2mL/min, and the injection port temperature is 250-300 ℃; the temperature raising program comprises: firstly heating to 70-90 ℃ and preserving heat for 0.5-2min, then heating to 220 ℃ at the heating rate of 35-45 ℃/min, preserving heat for 4-6min at the temperature of 4-6 ℃/min to 230 ℃ and 250 ℃ at the temperature of 10-15 ℃/min to 310 ℃ and 330 ℃;
preferably, when the mercaptochlorothalonil and the 4-hydroxychlorothalonil are detected, a liquid chromatography tandem mass spectrometer is adopted, a chromatographic column is Agilent XDB C182.1 × 150mm, the diameter of the chromatographic column is 3.5 μm, and the column temperature is as follows: 25 ℃; flow rate: 0.4 mL/min; mobile phase: a is 5mM ammonium formate aqueous solution, B is 5mM ammonium formate methanol; the elution procedure is shown in the following table:
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114349672A (en) * | 2022-01-07 | 2022-04-15 | 中国农业科学院农业质量标准与检测技术研究所 | Sulfydryl chlorothalonil and synthesis method, purification method and application thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4069342A (en) * | 1973-12-12 | 1978-01-17 | Diamond Shamrock Corporation | Method for regulating rests using alkylthiotrichlorodicyanobenzenes |
| WO1994023018A1 (en) * | 1993-03-29 | 1994-10-13 | Isk Biosciences Corporation | Immunoassay for tetrachloroisophthalonitrile (chlorothalonil®), its derivatives and breakdown products |
| JP2009294161A (en) * | 2008-06-09 | 2009-12-17 | Miura Co Ltd | Analysis method of residual agricultural chemical |
| WO2015124662A1 (en) * | 2014-02-19 | 2015-08-27 | Basf Se | Aqueous co-formulation of metalaxyl |
| US20190248714A1 (en) * | 2018-02-11 | 2019-08-15 | Gary David McKnight | Non-Aqueous Organo Liquid Delivery Systems containing dispersed Organo Polycarboxylate Functionalities that improves efficiencies and properties of nitrogen sources |
| CN111257492A (en) * | 2020-02-28 | 2020-06-09 | 上海瑞鑫科技仪器有限公司 | Multifunctional food environment analyzer |
| CN113109471A (en) * | 2021-04-12 | 2021-07-13 | 诺安实力可商品检验（青岛）有限公司 | LCMSMS (liquid crystal display module medium) method for rapidly and simultaneously detecting chlorothalonil and metabolites thereof in vegetables and fruits |
| CN114349672A (en) * | 2022-01-07 | 2022-04-15 | 中国农业科学院农业质量标准与检测技术研究所 | Sulfydryl chlorothalonil and synthesis method, purification method and application thereof |
-
2022
- 2022-01-07 CN CN202210016233.5A patent/CN114354823B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4069342A (en) * | 1973-12-12 | 1978-01-17 | Diamond Shamrock Corporation | Method for regulating rests using alkylthiotrichlorodicyanobenzenes |
| WO1994023018A1 (en) * | 1993-03-29 | 1994-10-13 | Isk Biosciences Corporation | Immunoassay for tetrachloroisophthalonitrile (chlorothalonil®), its derivatives and breakdown products |
| JP2009294161A (en) * | 2008-06-09 | 2009-12-17 | Miura Co Ltd | Analysis method of residual agricultural chemical |
| WO2015124662A1 (en) * | 2014-02-19 | 2015-08-27 | Basf Se | Aqueous co-formulation of metalaxyl |
| US20190248714A1 (en) * | 2018-02-11 | 2019-08-15 | Gary David McKnight | Non-Aqueous Organo Liquid Delivery Systems containing dispersed Organo Polycarboxylate Functionalities that improves efficiencies and properties of nitrogen sources |
| CN111257492A (en) * | 2020-02-28 | 2020-06-09 | 上海瑞鑫科技仪器有限公司 | Multifunctional food environment analyzer |
| CN113109471A (en) * | 2021-04-12 | 2021-07-13 | 诺安实力可商品检验（青岛）有限公司 | LCMSMS (liquid crystal display module medium) method for rapidly and simultaneously detecting chlorothalonil and metabolites thereof in vegetables and fruits |
| CN114349672A (en) * | 2022-01-07 | 2022-04-15 | 中国农业科学院农业质量标准与检测技术研究所 | Sulfydryl chlorothalonil and synthesis method, purification method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| LIWEI TANG 等: "Biodegradation of chlorothalonil by Enterobacter cloacae TUAH-1", INTERNATIONAL BIODETERIORATION & BIODEGRADATION, vol. 121, pages 122 - 130 * |
| 李凌璞 等: "QuEChERS/气相色谱-质谱/质谱法测定蔬菜中58种高风险农药残留量", 常州大学学报（自然科学版）, vol. 31, no. 3, pages 40 - 48 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114349672A (en) * | 2022-01-07 | 2022-04-15 | 中国农业科学院农业质量标准与检测技术研究所 | Sulfydryl chlorothalonil and synthesis method, purification method and application thereof |
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