Study On Detection And Analysis Method Of Aminoglycoside Antibiotics In Antibiotic Residue And Environmental Samples

With the current advancement of biotechnology,the production of antibiotics has been achieved.Antibiotics play an important role in the medical and livestock,aquaculture and other field,due to the selective action of antibiotics,it will cause the induction of bacterial resistance,which will gradually invalidate the sterilization treatment and ultimately damage the public body.Health,increase the economic burden of society.Antibiotic residues are now a non-negligible problem in the field of environmental safety.In this paper,gentamicin was used as an example to study the sample pretreatment techniques and detection methods for gentamicin residue analysis in slag,soil and plant samples,and to establish samples in slag,soil and plant matrix.Pretreatment method,solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry?SPE-UPLC-MS/MS?analysis method and solid phase extraction-ultra-high performance liquid chromatography-evaporative light scattering?SPE-UPLC-ELSD?Detection analysis method.In this paper,sample pretreatment methods for gentamicin slag,soil and plants were established.The sample preparation includes two processes of extraction and purification.In the extraction process of the sample,the extractant type,extractant concentration,auxiliary extraction method,extraction time,extraction times and sample matrix dosage of the residue,soil and plant samples were optimized to select gentamicin recovery.The best extraction conditions are:5%of the best extractant for the residue sample(trichloroacetic acid?including 2 mmol/L EDTA?:acetonitrile=8:2;the best extractant for plant samples It is 5%trichloroacetic acid?containing 1mmol/L EDTA?;the best extractant for soil samples is 0.5 mol/L Na₂CO₃,which is extracted by ultrasonic extraction.The best extraction time is 20 min and extracted twice.The sample base is used in an amount of 10 ml/g.In the process of purifying the extract?in which the pH of the soil extract is adjusted to 4?,the solid phase extraction purification method is used for the solid phase extraction cartridge type,the eluent type and concentration,the eluent pH,and the washing.The conditions of the type of liquid removal and the volume of the eluent were optimized.The gentamicin recovery rate was measured.It is found that the MCX column is used in the solid phase extraction column,and the eluent was 5 mL of 0.1 mol/L pH 8.The recovery of gentamicin was highest when the phosphate buffer solution of was eluted with 10 mL of ammonia in a volume ratio of 1:9.The MCX solid phase extraction cartridge is eluted.A method for the detection of gentamicin residues was established based on ultra performance liquid chromatography-tandem mass spectrometry?UPLC-MS/MS?.Qualitative and quantitative analysis of each C component of gentamicin was carried out by selecting the type of column,the choice of mobile phase species,the adjustment of flow ratio,and the selection and optimization of mass spectrometry parameters such as cone voltage and capillary voltage.The results showed that the recoveries of gentamicin in the residue,soil and plants are all about 80%.The intraday precision was in line with the requirements.The linear range of gentamicin was 36³065?g/kg.The LOD of the method is 1.16-6.86?g/L,the LOQ is 3.86-22.86?g/L,and the linearity is good in the linear range,and the R2 values are all above 0.99.In addition,this paper establishes a method for the detection of gentamicin residues based on ultra performance liquid chromatography-evaporative light scattering?UPLC-ELSD?.Qualitative quantification of each C component of gentamicin by selection of column type,selection of mobile phase species,flow ratio,column temperature adjustment,evaporation temperature,selection and optimization of detector parameters such as nebulizer temperature Riddle.The gentamicin has a good linear range from 1.58 to 278?g/mL,LOD is 1.185-2.385?g/mL,and LOQ is 3.658-7.362?g/mL.Based on the UPLC-ELSD method for detecting gentamicin,a UPLC-ELSD residue detection method for streptomycin and spectinomycin is established.Streptomycin and spectinomycin are qualitatively and quantitatively detected by gradient elution by adjusting the chromatographic conditions and detector conditions.The linear range of streptomycin is 10-1000?g/mL,the linear equation is y=0.9138x+1.412,the LOD of the method is 0.312?g/mL,the LOQ is 1.25?g/mL,and the linear range of spectinomycin is 10.-1000?g/mL,the linear equation is y=0.9239x+1.0283,the LOD of the method is 1.304?g/mL,the LOQ is 4.35?g/mL;the R2 values are all above 0.99.