CN112305091A - UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in Chinese cabbages and soil - Google Patents
UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in Chinese cabbages and soil Download PDFInfo
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Classifications
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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
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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
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Abstract
A UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in Chinese cabbage and soil is characterized in that: the method for pre-treating Chinese cabbage and the soil sample is used for solving the problem of low recovery rate of the soil sample due to easy molecular chelation, a UPLC-MS/MS detection method for detecting streptomycin and dihydrostreptomycin residues in the Chinese cabbage and the soil is established, and a hydrophilic chromatographic column is used for replacing an ion pair reagent to avoid polluting a mass spectrometer. The method can provide detection technical support for monitoring streptomycin in agricultural products and soil.
Description
The technical field is as follows:
the invention belongs to the technical field of agricultural product quality safety detection, relates to an agricultural product and a method for detecting pollutant residues in cultivation soil of the agricultural product, and can meet the technical requirement of pesticide quality safety supervision and spot check.
Background art:
streptomycin and its derivative dihydrostreptomycin belong to aminoglycoside antibiotics, can inhibit the extension of peptide chain and the synthesis of bacterial protein, has effects on tubercle bacillus, gram-negative bacteria and some gram-positive bacteria, has been used in veterinary clinical treatment of bacterial infection before, has been widely used in agricultural production after, can effectively prevent and treat Chinese cabbage soft rot, tomato scab, pepper anthracnose, citrus canker and cucumber bacterial angular leaf spot, etc., but streptomycin residue can cause serious harm to human body, such as anaphylactic shock and hearing loss, etc., and also has potential teratogenic, carcinogenic and mutagenic effects. At present, the production and use of agricultural streptomycin are forbidden in China, so that a method for detecting agricultural products and streptomycin in soil needs to be established urgently.
At present, the reported detection methods of streptomycin and dihydrostreptomycin mainly relate to animal-derived food substrates such as livestock and poultry products, aquatic products, honey and the like, but the reports on the detection of streptomycin and dihydrostreptomycin in agricultural products such as Chinese cabbages and soil are few. The types of the vegetable and soil samples are different from the types of the animal-derived food matrixes, the pretreatment method is not suitable, and the soil is easy to generate molecular chelation with streptomycin due to high ion content. In recent years, a liquid chromatography-tandem mass spectrometry method is mostly adopted for detecting streptomycin and dihydrostreptomycin, and has strong advantages in the aspects of matrix interference resistance, accuracy and qualitative and sensitivity, but due to strong polarity, the detection method is difficult to reserve on a reversed phase chromatographic column, and an ion pair reagent of heptafluoro-n-butyric acid or trifluoroacetic acid needs to be added into a mobile phase. The reagent is a non-volatile ion pair reagent and has a strong ion inhibition effect on mass spectrum negative ion detection.
The method optimizes the sample pretreatment method, utilizes the hydrophilic interaction chromatographic column to replace an ion pair reagent, establishes an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) detection method for streptomycin and dihydrostreptomycin residues in Chinese cabbage and soil, and can provide technical reference for monitoring streptomycin of other agricultural products.
The invention content is as follows:
the invention aims to research a pretreatment method of Chinese cabbage and a soil sample, solve the problem of low recovery rate of the soil sample due to high ion content and easy occurrence of molecular chelation, and establish a UPLC-MS/MS detection method for detecting streptomycin and dihydrostreptomycin residues in the Chinese cabbage and the soil.
The invention aims to realize the following technical scheme, and the UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in Chinese cabbages and soil comprises the following steps:
a. chinese cabbage: weighing 5g of sample to 0.01g, adding 0.02mol/L K into a 50mL polypropylene plastic centrifuge tube2HPO320mL of the solution was vertically shaken on a tissue grinder for 5min and centrifuged at 4000r/min for 5 min. Taking 10mL of supernatant to be purified.
Soil: weighing 2g of sample to be accurate to 0.01g, placing the sample in a 50mL polypropylene plastic centrifuge tube, and adding 0.02mol/L Na220mL of EDTA solution, ultrasonic extraction at 40 ℃ for 20min, vertical oscillation on a tissue grinder for 5min, and centrifugation at 4000r/min for 5 min. Taking 10mL of supernatant to be purified.
b. Activating a mixed weak cation exchange (WCX) column by using 3mL of methanol and 3mL of water, loading 10mL of extracting solution, washing by using 3mL of water and 3mL of methanol sequentially, and draining. Eluting with 3mL acetonitrile-water-acetic acid (volume ratio of 20:77:3), collecting eluent, filtering 1.5mL of the eluent with a 0.22 μm organic filter membrane, and detecting.
c. Preparation of standard stock solutions: respectively weighing appropriate amount of streptomycin and dihydrostreptomycin standard substances, dissolving with methanol, preparing standard stock solution with concentration of 100 μ g/mL, storing at 4 deg.C for 6 months.
d. Preparing a mixed standard solution: appropriate amounts of streptomycin and dihydrostreptomycin standard stock solutions are respectively transferred and diluted by methanol to prepare mixed standard solutions with the concentrations of 10 mug/mL and 1 mug/mL. Transferring a proper amount of the mixed standard solution, preparing 0.0083, 0.0167, 0.042, 0.167, 0.417 and 0.833 mu g/mL series of mixed standard working solutions by using a Chinese cabbage blank matrix, and preparing 0.0067, 0.0167, 0.067, 0.167 and 0.333 mu g/mL series of mixed standard working solutions by using a soil blank matrix, and preparing the mixed standard working solutions in situ when the mixed standard working solutions are used.
e. And (3) measuring the mixed standard working solution and the sample to be tested by using an UPLC-MS/MS method, drawing a standard curve by using the concentration as a horizontal coordinate and the peak area of the quantitative ions as a vertical coordinate, and calculating the content of the target substance in the sample to be tested by using the standard curve.
The chromatographic conditions for the UPLC-MS/MS determination are as follows: a chromatographic column: acquityTMCORTECS HILIC column (2.1X 100mm, 1.6 μm); mobile phase: phase A is acetonitrile, phase B is 200mmol/L ammonium formate solution (pH value is adjusted to 3.5 by formic acid); the volume ratio of the two is 50:50 isocratic elution; flow rate: 0.2 mL/min; sample introduction amount: 5 mu L of the solution; column temperature: at 40 ℃.
The mass spectrum conditions are as follows: electrospray positive ion, multiple reaction monitoring and scanning, capillary voltage of 3.0kv, ion source temperature of 150 ℃, desolvation gas temperature of 500 ℃, desolvation gas flow rate of 1000L/hr, cone hole back blowing gas flow rate of 150L/hr, atomization gas pressure of 7bar, retention time and mass spectrum acquisition parameters of 2 compounds are shown in Table 1.
TABLE 1 Mass Spectrometry acquisition parameters for streptomycin and dihydrostreptomycin
According to the characteristics of the Chinese cabbage and the soil sample matrix, the extraction and purification conditions are optimized, and the UPLC-MS/MS method for analyzing the streptomycin and dihydrostreptomycin residues in the Chinese cabbage and the soil is established.
(1) The method is suitable for detecting streptomycin and dihydrostreptomycin residues in Chinese cabbage and soil samples, and no similar report exists at present.
(2) The method of the invention adopts the hydrophilic chromatographic column to replace the ion pair reagent, and can avoid the pollution to the mass spectrometer caused by the use of the ion pair reagent.
(3) The method solves the problem of low recovery rate of streptomycin and dihydrostreptomycin caused by the fact that a soil sample is easy to carry out molecular chelation due to high ionic strength by reducing sample weighing, water bath ultrasound and other methods.
The sensitivity of the method of the invention:
and calculating the detection limit of the method by taking the signal-to-noise ratio as 3, taking the lowest addition concentration as a quantification limit, and meeting the requirement that the signal-to-noise ratio is greater than 10. The detection limit of streptomycin and dihydrostreptomycin in the Chinese cabbage is 0.002mg/kg, and the quantitative limit is 0.01 mg/kg; the detection limit of streptomycin and dihydrostreptomycin in the soil is 0.004mg/kg, and the quantitative limit is 0.02 mg/kg.
Linearity of the method of the invention:
and (4) taking the concentration of the standard solution as a horizontal coordinate and the quantitative ion peak area as a vertical coordinate, and drawing a standard curve. The standard curve equation of streptomycin in the Chinese cabbage substrate is as follows: 271550x +722.95, the correlation coefficient is: r is 0.9999; the linear equation of the standard sample of streptomycin in the soil matrix is as follows: 818496x-1130.4, the correlation coefficient is: r is 0.9999. The standard curve equation of dihydrostreptomycin in the Chinese cabbage substrate is as follows: 506935x-1183.1, the correlation coefficient is: r is 0.9998; the standard curve equation of the dihydrostreptomycin in the soil matrix is as follows: 2146801x-6008, the correlation coefficient is: r is 0.9999.
The accuracy and precision of the method of the invention are as follows:
the test of adding and recovering is carried out at three concentration levels of low, medium and high, the adding concentration of the Chinese cabbage is 0.01, 0.1 and 1mg/kg, the adding concentration of the soil is 0.02, 0.1 and 1mg/kg, each concentration is 5 parallel samples, simultaneously, blank samples are used as a reference, extraction, purification and measurement are carried out according to the method, and the average recovery rate and the Relative Standard Deviation (RSD) of the streptomycin and the dihydrostreptomycin are calculated. The results show that the average recovery rate of 2 compounds is between 74% and 91%, the relative standard deviation is between 0.7% and 8.7%, and the requirements of residual analysis are met, and the details are shown in Table 2.
Table 2 add recovery and relative standard deviation (n ═ 5)
Detailed Description
The invention is further illustrated by the following examples, which are illustrative only and are not meant to limit the scope of the invention in any way.
Example 1
The method is characterized in that 72% streptomycin sulfate soluble powder with the dose of 300g a.i/ha is applied for 3 times, and a Chinese cabbage sample collected 21 days after the last application is taken as a sample, the method is adopted for sample pretreatment, and ultra performance liquid chromatography-tandem mass spectrometry is adopted for determination. The preparation method comprises the following steps:
1. instruments and reagents
An American Waters ultra high performance liquid chromatography-tandem mass spectrometer I-Class/Xevo TQ-S triple quadrupole mass spectrometer; a Switzerland Mettler sensitive 0.00001g electronic balance; an ultrasonic extraction instrument; the purity of the pesticide standard product is not lower than 99 percent and is purchased from Dr.E company in Germany; acetonitrile and formic acid are chromatographically pure.
2. Sample pretreatment method
a. Weighing 5g of Chinese cabbage sample to be accurate to 0.01g, adding 0.02mol/L K into a 50mL polypropylene plastic centrifuge tube2HPO320mL of the solution was vertically shaken on a tissue grinder for 5min and centrifuged at 4000r/min for 5 min. Taking 10mL of supernatant to be purified.
b. Activating a mixed weak cation exchange (WCX) column by using 3mL of methanol and 3mL of water, loading 10mL of extracting solution, washing by using 3mL of water and 3mL of methanol sequentially, and draining. Eluting with 3mL acetonitrile-water-acetic acid (volume ratio of 20:77:3), collecting eluent, filtering 1.5mL of the eluent with a 0.22 μm organic filter membrane, and detecting.
3. Preparation of Standard working solutions
Transferring appropriate amount of mixed standard solution, and preparing series of mixed standard working solutions of 0.0083, 0.0167, 0.042, 0.167, 0.417 and 0.833 μ g/mL with Chinese cabbage blank matrix.
4. Measurement method
A chromatographic column: acquityTMCORTECS HILIC column (2.1X 100mm, 1.6 μm); mobile phase: phase A is acetonitrile, phase B is 200mmol/L ammonium formate solution (pH value is adjusted to 3.5 by formic acid); the volume ratio of the two is 50:50 isocratic elution(ii) a Flow rate: 0.2 mL/min; sample introduction amount: 5 mu L of the solution; column temperature: at 40 ℃.
The mass spectrum conditions are as follows: electrospray positive ion, multiple reaction monitoring and scanning, capillary voltage of 3.0kv, ion source temperature of 150 ℃, desolvation gas temperature of 500 ℃, desolvation gas flow rate of 1000L/hr, cone hole back blowing gas flow rate of 150L/hr, atomization gas pressure of 7bar, retention time and mass spectrum acquisition parameters of 2 compounds are shown in Table 1.
5. Calculation of results
And (3) measuring the mixed standard solution and the sample to be tested with the series of concentrations by using an UPLC-MS/MS method, drawing a standard curve by using the concentration as a horizontal coordinate and the peak area of the quantitative ions as a vertical coordinate, and calculating the content of the target pesticide in the sample to be tested. The calculation results are shown in Table 3.
Calculating the formula:
wherein:
c-residual amount of target (mg/kg);
C0-concentration of target in sample solution (μ g/mL) calculated from standard curve;
V1-volume of extraction solvent (mL);
V2-volume of extract used for purification (mL);
V3-final volumetric volume (mL);
m-weight sample (g).
6. Measurement results
The method is used for detecting by taking a Chinese cabbage sample which is collected 21 days after the last application as a sample, wherein the application is carried out 3 times at a dose of 300g a.i/ha of 72% streptomycin sulfate soluble powder, the application interval is 7 days, and the last application is carried out. The results show that streptomycin and dihydrostreptomycin residues are detected in the Chinese cabbage sample, and the specific data are shown in Table 3.
Table 3 actually measured streptomycin and dihydrostreptomycin residue in Chinese cabbage sample
Claims (3)
1. A UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in Chinese cabbage and soil is characterized in that: the method for pre-treating Chinese cabbage and the soil sample is used for solving the problem of low recovery rate of the soil sample due to easy molecular chelation, a UPLC-MS/MS detection method for detecting streptomycin and dihydrostreptomycin residues in the Chinese cabbage and the soil is established, and a hydrophilic chromatographic column is used for replacing an ion pair reagent to avoid polluting a mass spectrometer. The method can provide detection technical support for monitoring streptomycin in agricultural products and soil.
The method comprises the following specific steps:
a. chinese cabbage: weighing 5g of sample to 0.01g, adding 0.02mol/L K into a 50mL polypropylene plastic centrifuge tube2HPO320mL of the solution was vertically shaken on a tissue grinder for 5min and centrifuged at 4000r/min for 5 min. Taking 10mL of supernatant to be purified.
Soil: weighing 2g of sample to be accurate to 0.01g, placing the sample in a 50mL polypropylene plastic centrifuge tube, and adding 0.02mol/L Na220mL of EDTA solution, ultrasonic extraction at 40 ℃ for 20min, vertical oscillation on a tissue grinder for 5min, and centrifugation at 4000r/min for 5 min. Taking 10mL of supernatant to be purified.
b. Activating a mixed weak cation exchange (WCX) column by using 3mL of methanol and 3mL of water, loading 10mL of extracting solution, washing by using 3mL of water and 3mL of methanol sequentially, and draining. Eluting with 3mL acetonitrile-water-acetic acid (volume ratio of 20:77:3), collecting eluent, filtering 1.5mL of the eluent with a 0.22 μm organic filter membrane, and detecting.
c. Preparation of standard stock solutions: respectively weighing appropriate amount of streptomycin and dihydrostreptomycin standard substances, dissolving with methanol, preparing standard stock solution with concentration of 100 μ g/mL, storing at 4 deg.C for 6 months.
d. Preparing a mixed standard solution: appropriate amounts of streptomycin and dihydrostreptomycin standard stock solutions are respectively transferred and diluted by methanol to prepare mixed standard solutions with the concentrations of 10 mug/mL and 1 mug/mL. Transferring a proper amount of the mixed standard solution, preparing 0.0083, 0.0167, 0.042, 0.167, 0.417 and 0.833 mu g/mL series of mixed standard working solutions by using a Chinese cabbage blank matrix, and preparing 0.0067, 0.0167, 0.067, 0.167 and 0.333 mu g/mL series of mixed standard working solutions by using a soil blank matrix, and preparing the mixed standard working solutions in situ when the mixed standard working solutions are used.
e. And (3) measuring the mixed standard working solution and the sample to be tested by using an UPLC-MS/MS method, drawing a standard curve by using the concentration as a horizontal coordinate and the peak area of the quantitative ions as a vertical coordinate, and calculating the content of the target substance in the sample to be tested by using the standard curve.
2. The UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in Chinese cabbage and soil as claimed in claim 1, wherein the UPLC-MS/MS method is 0.02mol/L K2HPO3The preparation method of the solution comprises the following steps: 4.56g of dipotassium phosphate is weighed, dissolved by adding 1000mL of ultrapure water, and the pH value is adjusted to 7.2-7.4 by hydrochloric acid. Wherein 0.02mol/L of Na2The preparation method of the EDTA solution comprises the following steps: 3.72g of Na were weighed2EDTA, dissolved in 500mL of ultrapure water, 0.5mL of phosphoric acid, and the pH is adjusted to 7.2 to 7.4 with 10mol/L sodium hydroxide solution.
3. The UPLC-MS/MS method for analyzing streptomycin and dihydrostreptomycin residues in chinese cabbage and soil as claimed in claim 1, wherein the chromatographic conditions are as follows: a CORTECS HILIC chromatographic column, wherein a mobile phase consists of an organic phase and a water phase, the organic phase is acetonitrile, the water phase is 200mmol/L ammonium formate solution, and the pH value of the ammonium formate solution is adjusted to 3.5 by formic acid; isocratic elution is adopted, the volume ratio of acetonitrile to ammonium formate solution is 50:50, the flow rate is 0.2mL/min, the column temperature is 40 ℃, and the sample injection volume is 5 mu L. The mass spectrometry conditions used were: electrospray positive ion ionization, multi-reaction monitoring and scanning, capillary voltage of 3.0kv, ion source temperature of 150 ℃, desolvation gas temperature of 500 ℃, desolvation gas flow rate of 1000L/hr, cone hole back blowing gas flow rate of 150L/hr, atomization gas pressure of 7bar, cone hole voltage of 10-90V, and collision energy of 10-90V.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548412A (en) * | 2016-03-07 | 2016-05-04 | 山东出入境检验检疫局检验检疫技术中心 | Method for measuring residual quantities of five aminoglycoside drugs in food simultaneously |
| MX2016011285A (en) * | 2016-08-31 | 2018-02-27 | Prontius Laboratorios S A De C V | Method for the confirmation and quantification of streptomycin in honey using the analytical technique of liquid chromatography coupled to mass spectrometry. |
| CN108519456A (en) * | 2018-03-26 | 2018-09-11 | 烟台杰科检测服务有限公司 | It is a kind of to analyze a variety of remaining methods of aminoglycoside compound in agriculture beast product simultaneously |
| CN108872425A (en) * | 2018-06-19 | 2018-11-23 | 山东省食品药品检验研究院 | The method that hydrophilic Interaction Chromatography-tandem mass spectrometry quickly measures honey streptomycin and dihydrostreptomycin |
-
2019
- 2019-07-31 CN CN201910701048.8A patent/CN112305091A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548412A (en) * | 2016-03-07 | 2016-05-04 | 山东出入境检验检疫局检验检疫技术中心 | Method for measuring residual quantities of five aminoglycoside drugs in food simultaneously |
| MX2016011285A (en) * | 2016-08-31 | 2018-02-27 | Prontius Laboratorios S A De C V | Method for the confirmation and quantification of streptomycin in honey using the analytical technique of liquid chromatography coupled to mass spectrometry. |
| CN108519456A (en) * | 2018-03-26 | 2018-09-11 | 烟台杰科检测服务有限公司 | It is a kind of to analyze a variety of remaining methods of aminoglycoside compound in agriculture beast product simultaneously |
| CN108872425A (en) * | 2018-06-19 | 2018-11-23 | 山东省食品药品检验研究院 | The method that hydrophilic Interaction Chromatography-tandem mass spectrometry quickly measures honey streptomycin and dihydrostreptomycin |
Non-Patent Citations (4)
| Title |
|---|
| MAŁGORZATA GBYLIK-SIKORSKA 等: "Occurrence of veterinary antibiotics and chemotherapeutics in fresh water, sediment, and fish of the rivers and lakes in Poland", 《BULL VET INST PULAWY》 * |
| 何强 等: "奶粉中链霉素与双氢链霉素残留量的超高效液相色谱-串联质谱法测定", 《分析测试学报》 * |
| 吴映璇 等: "亲水作用色谱-质谱法同时检测鱼肉中链霉素和双氢链霉素残留", 《食品安全质量检测学报》 * |
| 李敏青 等: "固相萃取/高效液相色谱-串联质谱法测定植物性食品中链霉素与双氢链霉素", 《分析测试学报》 * |
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