鸡猪排泄物中恩诺沙星和环丙沙星含量HPLC检测方法的建立

Development of HPLC method for determination of enrofloxacin and ciprofloxacin levels in excrements of chicken and swine

建立了测定鸡、猪排泄物中恩诺沙星及环丙沙星含量的高效液相-荧光检测方法.将猪粪和鸡粪尿混合物样品分别用甲醇氨水溶液和醋酸甲醇溶液浸提,猪尿用固相萃取小柱富集净化,流动相为乙腈-三乙胺磷酸溶液(0.02 mol/L),荧光激发波长280 nm,发射波长450 nm.检测结果表明,排泄物样品中恩诺沙星和环丙沙星含量的检测定量限,鸡粪尿混合物为0.010 μg/g,猪尿0.005 μg/mL,猪粪为0.020 μg/g.外标法标准曲线的线性范围鸡粪尿混合物中恩诺沙星为0.010～100.000 μg/g、环丙沙星为0.010～50.000 μg/g,猪尿和猪粪中恩诺沙星和环丙沙星的线性范围分别为0.005～0.500 μg/mL和0.020～1.000 μg/g.恩诺沙星和环丙沙星的回收率分别大于75%和88%.HPLC方法的样品前处理和检测方法简便、快捷,准确性较微生物检测法高.

A HPLC method was developed for determination of enrofloxacin and its main active metabolite ciprofloxacin levels in excrements of chicken and swine. The excreta of chicken and swine were extracted by acetic acid in methanol and ammonia in methanol, respectively. Swine urine was concentrated and cleaned by SPE column. HPLC mobile phase consisted of acetonitrile and phosphate buffer （0.02 mol/L）with triethylamine. Fluorometric excitation wavelength was 280 nm, and emission wavelength was 450 nm. The limit of quantitation of enrofloxacin and ciprofloxacin in excreta of chicken was 0. 010μg/g, in urine was 0. 005μg/mL, and in feces of swine was 0. 020μg/g, respectively. The standard curves were observed to be linear from 0. 010- 100. 000μg/g for enrofloxacin and 0. 010-50. 000μg/g for ciprofloxacin in excreta of chicken, from 0. 005-0. 500μg/mL for both two drugs in pig urine, from 0. 020-1. 000μg/g for both two drugs in pig feces. Recoveries are at least 75% for enrofloxacin, 88% for ciprofloxacin. Coefficients of variation are preferably good for residue analysis.