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A detection method for enrofloxacin residues in earthworms
Received:September 20, 2015  
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KeyWord:enrofloxacin;earthworm;HPLC;recovery rate;filter paper exposure
Author NameAffiliationE-mail
DING Li-ling Department of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China  
LU Xiao-xu Department of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China  
LI Yin-sheng Department of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China yinshengli@sjtu.edu.cn 
QIU Jiang-ping Department of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China  
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Abstract:
      An assay method using high performance liquid chromatography(HPLC) with fluorescence detection was established to quantify enrofloxacin residues in earthworms. The extraction solution comprised 0.1 mol·L-1 phosphoric acid(pH12)+acetonitrile(1+3,V/V). The initial extracts were defatted, evaporated, and re-dissolved in mobile phase consisting of 0.025 mol·L-1 phosphoric acid/triethylamine buffer(pH3.0)+ acetonitrile(83+17,V/V). The fluorescence detector excitation wavelength was 280 nm, and the emission wavelength 450 nm. The linear range of the standard curve was 5~500 ng·mL-1. The recoveries of 3 concentration groups were between 81% and 88% with an average of 84.5% and the coefficient of variation was less than 10%. The detection limit of enrofloxacin was 2 ng·g-1. Earthworms were exposed to filter papers soaked in enrofloxacin with concentrations of 0, 0.16, 1.6, 16μg·g-1 and 32μg·cm-2, and 4 replicates in each case. After 48 hours of exposure, the average enrofloxacin concentration in earthworms was 0, 1.53, 16.81, 113.32μg·g-1 and 120.83μg·g-1, respectively. These results show that the absorption of enrofloxacin by earthworms increased linearly up to 1.6μg·cm-2 exposure concentration, and then asymptotically to a maximum of approximately 120μg enrofloxacin·g-1 earthworm tissue.