| Fluoroquinolones（FQs） and tetracyclines（TCs） have been widely used to prevent andremedy animal diseases in animal husbandry because of their wide-spectrum and strongantibiotic activity. But unreasonable or abuse adding medicated feed additives in feeds willlead to the drugs remaining in animal derived food and environment, which will harm thehuman health through food chain. Most of the antibiotic can’t completely absorbed by thebody, about60percent of the antibiotic in the form of original shape or metabolite enter theenvironment through animal’s manure and urine, then polluted the groundwater, surface-waterand soil.The two types of antibiotics in animal manure can’t completely be degradated in thesoil, but can be enriched in the soil.Be absorbed in the plant’s roots through the way of passivediffusion and active transport, and then were enriched in plants’ root,stalk and so on.Theplant-food which enriched much antibiotics entered the food chain, affecting food security.Atlast antibiotics eventually be absorbed by the human,then produce toxic effects.Thus, it’snecessary to establish a method for fluoroquinolones and tetracyclines detection.With enrofloxacin, pefloxacin, sarafloxacin, danofloxain, tetracycline and chlortetracy asdetecting objects, the pretreatment method and detection technology of suckling manureswere studied. The method for simultaneous extraction of FQs and TCs in Manure was studied,and then established the assay methods with fluoroquinolones and tetracyclines respectivelyby high performance capillary electrophoresis and high-performance liquid chromatography.Finally, the method has been successfully used in the detection of six antibiotics around thecity of Hefei.The specific experimental results are as follows:1The Study on Simultaneous Extraction of Fluoroquinolones （FQs）and Tetracyclines （TCs） in ManureThe experiment compared the extraction effects of fluoroquinolones （FQs） andtetracyclines （TCs） in manure by using different extraction, and then the method of usingmixed extractions of EDTA―McIlvaine and Mg（NO3）2―NH3·H2O was determined. Theexperiment explored the proportion of two extractions. The result showed that when thevolume ratio of EDTA―McIlvaine and Mg （NO3）2―NH3·H2O was3:1, the extraction ratiosof the six antibiotics were83.6%92.1%.On the based of simple extraction, the study used a method for purifying the sample bysolid-phase extraction （SPE）: the experiment investigated the type, amount of drench lotionand elution agents, identified the SPE method by distilled water as drench lotion and methanol as eluent; the recovery rate was85.2%88.5%.2The method of separating fluoroquinolones and tetracyclines byhigh-performance capillary electrophoresis（HPCE）was builtA method the simultaneous separation of fluoroquinolone（sFQs）and tetracycline（sTCs）by high performance capillary electrophoresis（HPCE） was developed. The effects of severalfactors such as the concentration and pH of turning buffer,the organic added to theelectrolyte,the voltage and the temperature were investigated.The optimal conditions were：the running butter consisted of50mmol/L NaB4O7―200mmol/L H3BO3，pH8.41，separationvoltage at18kV,the UV deception wavelength was270nm.The results showed that the sixremedies can be well separated within12min.Each had a good linear relationship（R2=0.99860.9993）.The RSD was0.25%0.40%and7.3%9.8%.3The method of separating fluoroquinolones and tetracyclines byhigh-performance liquid chromatography（HPLC）was builtThe method for determining respectively four kinds of fluoroquinolones and two kinds oftetracyclines drugs by HPLC was built. The effect of some important factors such asdetection wavelength，column temperature，composition，ratio and flow rate of the mobilephase were investigated The optimal chromatographic conditions were: the UV detectionwavelength was270nm，the column temperature was35℃，the flow rate was1ml/min，themobile phase was consisted of acetonitrile and0.1%acetic acid. The gradient elution was:0min:19/81,5min:40/60,6min:85/15,7min:85/15,8min:19/81,18min:19/81. The resultsshowed that the six remedies can be well separated within18.0min. Each had a good linearrelationship（r=0.99900.9999）.The RSD of migration time and peak area separately was0.07%0.39%and1.98%3.12%.The method for simultaneously extracting the six antibiotics with EDTA―McIlvaine andMg（N03）2―NH3·H2O was built.The method for determinating the4kinds of FQs and2kindsof TCs drugs by high performance capillary electrophoresis has the following advantages:simple operation, chromatographic columns without pollution, fast analysis, low runningcosts and so on; the method by high performance liquid chromatography determinate4kindsof FQs and2kinds of TCs drugs has the following advantages: good reproducibility,sensitivity high. The sample preparation method and apparatus for detecting technology bythis experiment is successfully used in poultry and animal feces veterinary drug residuedetection in surrounding areas in Hefei city in Anhui Province. |
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