CN113030300A - Method for determining antibiotics for livestock in farmland environment water - Google Patents
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Abstract
The invention relates to a method for determining veterinary antibiotics in farmland environment water, and belongs to the technical field of environment monitoring. The method for determining the antibiotic for the animals in the farmland environment water comprises the following steps: taking a water sample from a farmland environment, centrifuging, taking supernatant, placing the supernatant into a 60mg/3cc HLB solid-phase extraction column, adding a methanol water solution with the concentration of 10% by volume fraction, leaching, discarding all effluent liquid, performing reduced pressure pumping, and eluting with chromatographic pure methanol to obtain eluent; concentrating the eluent to be dry, dissolving and filtering to obtain a treated sample; and detecting the obtained processed sample by adopting a liquid chromatography-tandem mass spectrometer to obtain the content of the veterinary antibiotic in the farmland environment water. The method for determining the veterinary antibiotics in the farmland environment water can detect atypical veterinary antibiotics, and can detect multiple veterinary antibiotics simultaneously, so that the detection efficiency is greatly improved.
Description
Technical Field
The invention belongs to the technical field of environmental monitoring, and particularly relates to a method for determining veterinary antibiotics in farmland environmental water.
Background
With the increasing development of resistance of pathogenic bacteria to commonly used typical antibiotic drugs, the search for alternative drugs has been accelerated, and some atypical veterinary antibiotic drugs gradually come into the human sight, such as norvancomycin, vancomycin, pirlimycin, clindamycin, tiamulin, valnemulin and tavermectin. However, increasing use of veterinary antibiotics may lead to veterinary antibiotic residues in the agricultural environment. There is therefore a need for the detection of veterinary antibiotics in a farm environment.
At present, detection methods for typical veterinary antibiotics commonly used for quinolones, tetracyclines, penicillins, macrolides and the like are mature, and the detection methods have national standards and industrial standards and also have research papers for simultaneously determining a plurality of veterinary drug residues, but the detection methods have less research on some atypical veterinary antibiotics. In addition, the existing detection methods of the atypical veterinary antibiotics detect single-class veterinary antibiotics, so that the detection efficiency is low, and different processing methods and detection methods are required for multiple detections.
Disclosure of Invention
The invention provides a method for determining veterinary antibiotics in farmland environment water, which can detect atypical veterinary antibiotics and can detect multiple veterinary antibiotics simultaneously, so that the detection efficiency is greatly improved.
The technical scheme for solving the technical problems is as follows: a method for determining veterinary antibiotics in farmland environment water comprises the following steps:
s1, taking a water sample from a farmland environment, centrifuging, taking supernatant, placing the supernatant into a 60mg/3cc HLB solid phase extraction column, adding a methanol water solution with the concentration of 10% by volume fraction for leaching, discarding all effluent liquid, performing reduced pressure draining, and eluting with chromatographic pure methanol to obtain eluent; concentrating the eluent to be dry, dissolving and filtering to obtain a treated sample;
and S2, detecting the processed sample obtained in the step S1 by adopting a liquid chromatography-tandem mass spectrometer, and obtaining the content of the antibiotic for livestock in the farmland environment.
The invention has the beneficial effects that:
(1) by adopting the method, 7 kinds of veterinary antibiotics including norvancomycin, vancomycin, pirlimycin, clindamycin, tiamulin, valnemulin and tylosin can be detected at one time, classification detection is not needed, the detection efficiency is greatly improved, only one-time sample collection is needed, and the method is very convenient;
(2) the method has the advantages of good purification and enrichment effects on 7 veterinary antibiotics, high recovery rate, high detection accuracy, low standard deviation of detection and good reproducibility, and can be widely applied to detection of farmland water environment.
On the basis of the technical scheme, the invention can be further improved as follows.
Further, in step S1, the rotation speed of the centrifugation is 5000-10000r/min, and the time is 5-10 min.
The beneficial effect of adopting the further scheme is that: obtain a clearer sample and is more beneficial to enrichment.
Further, in step S1, the HLB solid-phase extraction column is washed once with chromatographically pure methanol and distilled water, respectively.
The beneficial effect of adopting the further scheme is that: avoiding the influence of impurities.
Further, in step S1, the concentrated solution is dried by placing in a water bath at a temperature of 40-50 ℃ and drying with nitrogen.
The beneficial effect of adopting the further scheme is that: is beneficial to the volatilization of the methanol.
Further, in step S1, the solvent used for the dissolution is an aqueous methanol solution having a concentration of 10% by volume fraction.
The beneficial effect of adopting the further scheme is that: the method is beneficial to reducing the solvent effect and the dissolution of impurities during the measurement by an instrument.
Further, in step S1, the filtration is performed using a PTFE filter with a pore size of 0.22 μm.
The beneficial effect of adopting the further scheme is that: is favorable for reducing the adsorption of the filter membrane to the compound and improving the recovery rate.
Further, in step S2, the liquid chromatography-tandem mass spectrometry detects the chromatographic conditions as: an ACQUITY UPLC BEH C18 chromatographic column with an inner diameter of 2.1mm, a length of 100mm and a particle size of 1.7 μm; the flow rate of the mobile phase is 0.3mL/min, and the column temperature of the chromatographic column is 40 ℃; the mobile phase was methanol and 0.1% volume fraction aqueous formic acid, sampled at 2 μ L.
The beneficial effect of adopting the further scheme is that: the 7 antibiotics can achieve better chromatographic separation in a shorter time, and have higher sensitivity and less interference; the detection efficiency is improved.
Further, in step S2, the mass spectrometry conditions detected by the liquid chromatography-tandem mass spectrometry are: the ion source temperature is 150 ℃, and the capillary tube voltage is 3.0 kV; the desolventizing gas is nitrogen, the temperature of the desolventizing gas is 300 ℃, the airflow rate of the desolventizing gas is 800L/h, the collision gas is argon, and the airflow rate of the collision gas is 0.15 mL/min.
The beneficial effect of adopting the further scheme is that: is beneficial to the qualitative and quantitative detection of 7 antibiotics.
Drawings
FIG. 1 is an extracted ion chromatogram of pirlimycin in Experimental example 2 of the present invention;
FIG. 2 is an extracted ion chromatogram of vancomycin in Experimental example 2 of the present invention;
FIG. 3 is an extracted ion chromatogram of norvancomycin in Experimental example 2 of the present invention;
FIG. 4 is an extracted ion chromatogram of taenifamycin in Experimental example 2 of the present invention;
FIG. 5 is an extracted ion chromatogram of valnemulin in Experimental example 2 of the present invention;
FIG. 6 is an extracted ion chromatogram of tiamulin in Experimental example 2 of the present invention;
FIG. 7 is an extracted ion chromatogram of clindamycin in Experimental example 2 of the present invention;
FIG. 8 is a bar graph showing the recovery of 7 antibiotics using two solid phase extraction columns in Experimental example 3 of the present invention.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.
Example 1
The embodiment provides a method for determining veterinary antibiotics in farmland environment water, which comprises the following steps:
s1, taking a water sample from a farmland environment, shaking uniformly, putting 10mL into a 15mL centrifugal tube, firstly centrifuging for 5min at the rotation speed of 5000r/min, taking out the centrifugal tube, taking 60mg/3cc of an HLB solid-phase extraction column, sequentially adding 3mL of methanol and 3mL of ultrapure water, respectively cleaning once to realize pretreatment, after a cleaning solution completely flows out, adding 8mL of centrifuged supernatant into the HLB solid-phase extraction column, adding 3mL of 10% volume fraction methanol aqueous solution to elute the HLB solid-phase extraction column, discarding all effluent liquid to obtain a solid-phase extraction column to be eluted, decompressing and draining the solid-phase extraction column to be eluted, eluting the solid-phase extraction column to be eluted by using 3mL of methanol, collecting effluent liquid by using a glass test tube to obtain eluent, placing the eluent at the water bath temperature of 40 ℃, drying by using nitrogen to obtain solid residues, the solid residue was dissolved in 1mL of a 10% volume fraction aqueous methanol solution and filtered through a 0.22 μm PTFE filter to obtain a treated sample.
And S2, detecting the processed sample obtained in the step S1 by adopting a liquid chromatography-tandem mass spectrometer, and obtaining the content of the veterinary antibiotics in the farmland environment water.
Wherein, the chromatographic conditions detected by the liquid chromatography-tandem mass spectrometry method are an acquisition UPLC BEH C18 chromatographic column with the inner diameter of 2.1mm, the length of 100mm and the particle size of 1.7 μm. The flow rate of the mobile phase was 0.3mL/min and the column temperature of the column was 40 ℃. The sample volume is 2 mu L; the mobile phase A is methanol, the mobile phase B is 0.1 percent of formic acid aqueous solution by volume fraction, and the gradient elution procedure is as follows: 0-2.0 min, 90-80% of mobile phase B; 2.0-3.5 mim, 80% -50% mobile phase B; 3.5-5.0 min, 50-40% of mobile phase B; 5.0-6.5 min, 40-20% of mobile phase B; 6.5-7.0 min, 20-10% of mobile phase B, 7.0-7.1 min, 10-90% of mobile phase B, 7.1-8.5 min and 90-90% of mobile phase B.
Wherein, the mass spectrum conditions detected by the liquid chromatogram-tandem mass spectrometry are as follows: electrospray ion source (ESI), positive ion scanning mode, Multiple Reaction Monitoring (MRM) mode. The ion source temperature in mass spectrometric detection was 150 ℃ and the capillary voltage was 3.0 kV. The desolventizing gas in mass spectrometry is nitrogen, the temperature of the desolventizing gas is 300 ℃, the airflow rate of the desolventizing gas is 800L/h, the collision gas is argon, and the airflow rate of the collision gas is 0.15 mL/min.
Example 2
S1, taking a water sample from a farmland environment, shaking uniformly, putting 10mL into a 15mL centrifugal tube, firstly centrifuging at a rotation speed of 7000r/min for 10min, taking out the centrifugal tube, taking 60mg/3cc of an HLB solid-phase extraction column, sequentially adding 3mL of methanol and 3mL of ultrapure water, respectively cleaning once to realize pretreatment, after a cleaning solution completely flows out, adding 8mL of centrifuged supernatant into the HLB solid-phase extraction column, adding 3mL of 10% volume fraction methanol aqueous solution to elute the HLB solid-phase extraction column, discarding all effluent liquid to obtain a solid-phase extraction column to be eluted, decompressing and draining the solid-phase extraction column to be eluted, sampling 3mL of methanol to elute the solid-phase extraction column to be eluted, collecting effluent liquid by using a glass test tube to obtain eluent, placing the eluent at a water bath temperature of 45 ℃, drying by using nitrogen to obtain solid residues, the solid residue was dissolved in 1mL of a 10% volume fraction aqueous methanol solution and filtered through a 0.22 μm PTFE filter to obtain a treated sample.
And S2, detecting the processed sample obtained in the step S1 by adopting a liquid chromatography-tandem mass spectrometer, and obtaining the content of the veterinary antibiotics in the farmland environment water.
Wherein, the chromatographic conditions detected by the liquid chromatography-tandem mass spectrometry method are an acquisition UPLC BEH C18 chromatographic column with the inner diameter of 2.1mm, the length of 100mm and the particle size of 1.7 μm. The flow rate of the mobile phase was 0.3mL/min and the column temperature of the column was 40 ℃. The sample volume is 2 mu L; the mobile phase A is methanol, the mobile phase B is 0.1 percent of formic acid aqueous solution by volume fraction, and the gradient elution procedure is as follows: 0-2.0 min, 90-80% of mobile phase B; 2.0-3.5 mim, 80% -50% mobile phase B; 3.5-5.0 min, 50-40% of mobile phase B; 5.0-6.5 min, 40-20% of mobile phase B; 6.5-7.0 min, 20-10% of mobile phase B, 7.0-7.1 min, 10-90% of mobile phase B, 7.1-8.5 min and 90-90% of mobile phase B.
Wherein, the mass spectrum conditions detected by the liquid chromatogram-tandem mass spectrometry are as follows: electrospray ion source (ESI), positive ion scanning mode, Multiple Reaction Monitoring (MRM) mode. The ion source temperature in mass spectrometric detection was 150 ℃ and the capillary voltage was 3.0 kV. The desolventizing gas in mass spectrometry is nitrogen, the temperature of the desolventizing gas is 300 ℃, the airflow rate of the desolventizing gas is 800L/h, the collision gas is argon, and the airflow rate of the collision gas is 0.15 mL/min.
Example 3
S1, taking a water sample from a farmland environment, shaking uniformly, putting 10mL into a 15mL centrifugal tube, firstly centrifuging for 8min at a rotation speed of 10000r/min, taking out the centrifugal tube, taking 60mg/3cc of an HLB solid-phase extraction column, sequentially adding 3mL of methanol and 3mL of ultrapure water, respectively cleaning once to realize pretreatment, after a cleaning solution completely flows out, adding 8mL of centrifuged supernatant into the HLB solid-phase extraction column, adding 3mL of 10% volume fraction methanol aqueous solution to elute the HLB solid-phase extraction column, discarding all effluent liquid to obtain a solid-phase extraction column to be eluted, decompressing and draining the solid-phase extraction column to be eluted, sampling 3mL of methanol to elute the solid-phase extraction column to be eluted, collecting effluent liquid by using a glass test tube to obtain eluent, placing the eluent at a water bath temperature of 50 ℃, drying by using nitrogen to obtain solid residues, the solid residue was dissolved in 1mL of a 10% volume fraction aqueous methanol solution and filtered through a 0.22 μm PTFE filter to obtain a treated sample.
And S2, detecting the processed sample obtained in the step S1 by adopting a liquid chromatography-tandem mass spectrometer, and obtaining the content of the veterinary antibiotics in the farmland environment water.
Wherein, the chromatographic conditions detected by the liquid chromatography-tandem mass spectrometry method are an acquisition UPLC BEH C18 chromatographic column with the inner diameter of 2.1mm, the length of 100mm and the particle size of 1.7 μm. The flow rate of the mobile phase was 0.3mL/min and the column temperature of the column was 40 ℃. The sample volume is 2 mu L; the mobile phase A is methanol, the mobile phase B is 0.1 percent of formic acid aqueous solution by volume fraction, and the gradient elution procedure is as follows: 0-2.0 min, 90-80% of mobile phase B; 2.0-3.5 mim, 80% -50% mobile phase B; 3.5-5.0 min, 50-40% of mobile phase B; 5.0-6.5 min, 40-20% of mobile phase B; 6.5-7.0 min, 20-10% of mobile phase B, 7.0-7.1 min, 10-90% of mobile phase B, 7.1-8.5 min and 90-90% of mobile phase B. The addition of certain acids to the mobile phase improves the peak shape and retention of the compounds, and facilitates the separation of the mixture.
Wherein, the mass spectrum conditions detected by the liquid chromatogram-tandem mass spectrometry are as follows: electrospray ion source (ESI), positive ion scanning mode, Multiple Reaction Monitoring (MRM) mode. The ion source temperature in mass spectrometric detection was 150 ℃ and the capillary voltage was 3.0 kV. The desolventizing gas in mass spectrometry is nitrogen, the temperature of the desolventizing gas is 300 ℃, the airflow rate of the desolventizing gas is 800L/h, the collision gas is argon, and the airflow rate of the collision gas is 0.15 mL/min.
Experimental example 1
ESI scanning and mass spectrum condition optimization are carried out on the antibiotics needing to be detected, so that mass spectrum parameters of the antibiotics are determined. The specific antibiotic is norvancomycin, vancomycin, pirlimycin, clindamycin, tiamulin, valnemulin and tiamulin. Antibiotic specific mass spectral parameters are as in table 1.
Table 17 MRM mass spectral parameters of antibiotics
Is a quantitative ion
Experimental example 2
7 antibiotics of norvancomycin, vancomycin, pirlimycin, clindamycin, tiamulin, valnemulin and tylosin are added into a water sample to be detected by a high performance liquid chromatography-tandem mass spectrometry method, and a formic acid aqueous solution with methanol-0.1% volume fraction is taken as a mobile phase to carry out gradient elution, so that chromatographic conditions are determined to ensure that the corresponding 7 antibiotics can be separated and detected. Wherein the addition amount of 7 antibiotics is 1 ng/mL. As shown in fig. 1-7.
From fig. 1-7, it can be seen that the 7 antibiotics can achieve better chromatographic separation in a shorter time, with higher sensitivity and less interference. During chromatographic qualitative detection of each antibiotic, the antibiotic is divided into two pairs of ions, wherein fig. 1 shows an extracted ion chromatogram of pirlimycin, and the parent ion/daughter ion 1(411.16/112.06) and the parent ion/daughter ion 2(411.16/363.13) are shown. FIG. 2 shows an extracted ion chromatogram of vancomycin, wherein parent ion/daughter ion 1(724.93/144.06) and parent ion/daughter ion 2(724.93/100.06) are shown. FIG. 3 shows an extracted ion chromatogram of norvancomycin, wherein parent ion/daughter ion 1(717.93/144.06) and parent ion/daughter ion 2 (717.93/99.99). FIG. 4 is an extracted ion chromatogram of tylosin, parent/daughter ion 1(1042.45/109.00), parent/daughter ion 2 (1042.45/174.07). FIG. 5 shows the extracted ion chromatogram of valnemulin, parent ion/daughter ion 1(565.39/263.06), and parent ion/daughter ion 2 (565.39/163.86). Fig. 6 shows an extracted ion chromatogram of tiamulin, parent ion/daughter ion 1(494.33/192.00), and parent ion/daughter ion 2 (494.33/119.00). FIG. 7 is an extracted ion chromatogram of clindamycin, parent ion/daughter ion 1(424.99/126.06), and parent ion/daughter ion 2 (424.99/377.09).
Experimental example 3
7 antibiotics of norvancomycin, vancomycin, pirlimycin, clindamycin, tiamulin, valnemulin and tylosin are respectively added into a water sample, purified by 2 solid phase extraction columns of C18(200mg/3cc) and Oasis HLB (60mg/3cc), and the recovery rates of the 7 antibiotics are compared according to the method of example 1, as shown in FIG. 8.
As can be seen from FIG. 8, the recovery rates of norvancomycin and vancomycin were low when the C18(200mg/3cc) solid phase extraction column was used, whereas the recovery rates of 7 antibiotics were 70% or more when the Oasis HLB (60mg/3cc) solid phase extraction column was used. Therefore, the method has high detection accuracy on the 7 antibiotics, and the qualitative and quantitative analysis of the sample matrix on the target compound is basically free from interference when the detection is carried out under the selected instrument condition.
Experimental example 4
7 antibiotics of norvancomycin, vancomycin, pirlimycin, clindamycin, tiamulin, valnemulin and tavermectin are prepared into 5 mixed standard working solutions with different concentrations by using a blank sample solution, so that the concentrations of norvancomycin and vancomycin in the 5 mixed standard working solutions are respectively 2, 4, 20, 100 and 200ng/mL, and the concentrations of pirlimycin, clindamycin, tiamulin, valnemulin and tavermectin are respectively 0.2, 0.4, 2, 10 and 20ng/mL, detection is carried out according to the method of the embodiment 1, a linear regression equation is fitted, and specific results are shown in Table 2.
TABLE 27 regression equation, correlation coefficient (r), limit of detection (LOD) and limit of quantitation (LOQ) for antibiotics
As can be seen from the data in Table 2, the linear relation between the norvancomycin and the vancomycin is good in the range of 2-200ng/mL, and the correlation coefficients are all greater than 0.999; within the range of 0.2-20ng/mL, the linear relation of pirlimycin, clindamycin, tiamulin, valnemulin and tiamulin is good, and the correlation coefficient is more than 0.999. The detection limit of the method for obtaining the norvancomycin and the vancomycin through a standard addition recovery experiment is 0.2ng/mL, and the quantification limit is 0.4 ng/mL; the detection limits of the methods of pirlimycin, clindamycin, tiamulin, valnemulin and tylosin all reach 0.02ng/mL, and the quantification limits all reach 0.04 ng/mL.
Experimental example 5
Mixed standard working solutions with different concentration levels are added into a blank sample of the environmental water, so that the content of norvancomycin and vancomycin in the environmental water sample is 0.4ng/mL, 2ng/mL and 20ng/mL, the content of pirlimycin, clindamycin, tiamulin, valnemulin and tavermectin is 0.04ng/mL, 0.2ng/mL and 2ng/mL, 6 parallel repetitions are set for each addition concentration, blank control is simultaneously carried out, the method of example 1 is carried out, and the average recovery rate and the Relative Standard Deviation (RSD) are calculated, and the results are shown in Table 3.
TABLE 37 recovery of antibiotics in biogas slurry with standard addition and RSD results (n ═ 6)
As can be seen from Table 3, the recovery rates of the 7 antibiotics in the environmental water sample are 73.7% -96.7%, and the relative standard deviation is 1.8% -6.1%, which indicates that the method has high accuracy and good reproducibility, and is suitable for the determination of the 7 atypical antibiotics in the environmental water.
Experimental example 6
In practice, the method of embodiment 1 is applied to detect 25 environmental water samples, wherein tiamulin is detected in 11 environmental water samples, and the detected value range is 0.30-32 μ g/L, and norvancomycin, vancomycin, pirlimycin, clindamycin, valnemulin and tavermectin are not detected in the 25 environmental water samples.
At present, no method for simultaneously determining the 7 antibiotics in the water environment exists, generally, one class of antibiotics such as pirlimycin and clindamycin (lincomycin), or tiamulin and valnemulin (diterpenoid antibiotic), or norvancomycin and vancomycin (glycopeptide antibiotic) is simultaneously determined, and the determined samples are mostly livestock and poultry products and aquatic products. The method has the advantages of advancement, innovation and operability, and is suitable for simultaneously measuring 7 atypical antibiotics in the water environment.
It is to be noted that "comprising" in the present invention means that it may include other components in addition to the components described, and the "comprising" may be replaced with "being" or "consisting of … …" in a closed manner.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (8)
1. A method for determining veterinary antibiotics in farmland environment water is characterized by comprising the following steps:
s1, taking a water sample from a farmland environment, centrifuging, taking supernatant, placing the supernatant into a 60mg/3cc HLB solid phase extraction column, adding a methanol water solution with the concentration of 10% by volume fraction for leaching, discarding all effluent liquid, performing reduced pressure draining, and eluting with chromatographic pure methanol to obtain eluent; blowing the eluent by nitrogen, dissolving and filtering to obtain a treated sample;
and S2, detecting the processed sample obtained in the step S1 by adopting a liquid chromatography-tandem mass spectrometer, and obtaining the content of the veterinary antibiotics in the farmland environment water.
2. The method for determining antibiotics for animals in farmland environment water as claimed in claim 1, wherein in step S1, the rotation speed of the centrifugation is 5000-10000r/min, and the time is 5-10 min.
3. The method for determining antibiotic for animals in agricultural environmental water according to claim 1, wherein the HLB solid phase extraction column is previously activated and equilibrated with chromatographically pure methanol and ultrapure water in step S1.
4. The method for determining antibiotic for animals in farmland environment water according to claim 1, wherein in step S1, the concentration to dryness is carried out at a bath temperature of 40-50 ℃ and dried by nitrogen.
5. The method for determining veterinary antibiotics in agricultural and horticultural environments of water according to claim 1, wherein the solvent used for dissolution in step S1 is a 10% volume fraction methanol aqueous solution.
6. The method for determining antibiotic for animals in agricultural and horticultural environment water as claimed in claim 1, wherein in step S1, the filtration is performed with a PTFE filter membrane having a pore size of 0.22 μm.
7. The method for determining veterinary antibiotics in farmland environment water according to any one of claims 1-6, characterized in that in step S2, the liquid chromatography-tandem mass spectrometry method detects the chromatographic conditions as follows: an ACQUITY UPLC BEH C18 chromatographic column with an inner diameter of 2.1mm, a length of 100mm and a particle size of 1.7 μm; the flow rate is 0.3mL/min, and the column temperature is 40 ℃; the mobile phase was methanol and 0.1% volume fraction aqueous formic acid, sampled at 2 μ L.
8. The method for determining veterinary antibiotics in farmland environment water according to any one of claims 1-6, characterized in that in step S2, the mass spectrometric conditions detected by the liquid chromatography-tandem mass spectrometry are: the ion source temperature is 150 ℃, and the capillary tube voltage is 3.0 kV; the desolventizing gas is nitrogen, the temperature of the desolventizing gas is 300 ℃, the airflow rate of the desolventizing gas is 800L/h, the collision gas is argon, and the airflow rate of the collision gas is 0.15 mL/min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118243830A (en) * | 2024-05-30 | 2024-06-25 | 成都汇标检测技术有限公司 | Method for detecting agricultural antibiotics in grains and vegetables |
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