高效液相色谱法测定羊奶中五种氟喹诺酮类药物残留量的研究

Study on Detection Method Quinolones Residues in Sheep milk by HPLC

关键词:  高效液相色谱法  氟喹诺酮类药物  残留  羊奶

Key words:  HPLC  quinolones  residues  sheep milk

摘要:

建立一种可同时检测羊奶中五种氟喹诺酮类药物（环丙沙星、达氟沙星、恩诺沙星、沙拉沙星、二氟沙星）残留的高效液相色谱—荧光检测法。羊奶样品经乙腈提取，正己烷去脂肪，C18柱净化。以0.05 mol/L磷酸三乙胺溶液和乙腈溶液为流动相，流速为0.8 mL/min，激发波长280 nm，发射波长450 nm。结果表明环丙沙星、恩诺沙星、沙拉沙星、二氟沙星在10-200 ng/mL，达氟沙星在2-40 ng/mL呈良好的线性关系，R2均大于0.999。环丙沙星、恩诺沙星、沙拉沙星、二氟沙星的最低定量限为30 μg/kg，达氟沙星的最低定量限为6 μg/kg。环丙沙星、恩诺沙星、沙拉沙星、二氟沙星在30~200 μg/kg的浓度添加水平上，达氟沙星在6~40 μg/kg的浓度添加水平上其回收率均在70~100 %，批内、批间的相对标准偏差小于20 %。该方法简单快速、灵敏度高、重复性好，适用于羊奶中五种氟喹诺酮类药物残留检测。

Abstract:

A method for determination of quinolones residues in sheep milk with HPLC by FLD detection was established. Ciprofloxacin、Danofloxacin 、Enrofloxacin、Sarafloxacin and difloxacin were extracted from sheep milk with acetonitrile and degreased in hexane, dissolved with pH7.0 phosphate solution after evaporation, then the solution was purified with C18 column. The mobile phase were 0.05 mol/L triethylammonium phosphate solution and acetonitrile, the flow rate was 0.8 mL/min .Fluorescence detection was carried out at excitation and emission wavelengths of 280 nm and 450 nm . A good linearity of the calibration curve was obtained with the range of 10-200 ng/mL for Ciprofloxacin、Enrofloxacin、Sarafloxacin and difloxacin, and 2-40 ng/mL for Danofloxacin（R2>0.999）. The limit of quantification of Ciprofloxacin、Enrofloxacin、Sarafloxacin and difloxacin was 30 μg/kg, and Danofloxacin was 6 μg/kg. The average recoveries of quinolones residues from spiked sheep milk at the three concentrations of 30、100、200 μg/kg（Ciprofloxacin、Enrofloxacin、Sarafloxacin and difloxacin）and 6、20、40 μg/kg（Danofloxacin）ranged from 70-100 %(n=5) respectively. In day-between days batch variation coefficients were both less than 20 %.This method can be applied for the determination of quinolones residues in sheep milk.