高效液相色谱-荧光检测法同时分析鸡粪中六种氟喹诺酮类抗生素

Simultaneous Determination of Six Fluoroquinolones in Chicken Manures by High Performance Liquid Chromatography with Fluorescence Detection

通过对样品预处理过程优化、提取剂与洗脱剂的筛选和检测色谱条件优化,建立了基于固相萃取-高效液相色谱荧光检测法分析鸡粪中6种氟喹诺酮类(FQs)的检测方法。该方法选用Mcllvaine-Na2EDTA缓冲液∶乙腈(1∶1,V/V)(pH=2)作为提取剂,超声提取时间15 min,经HLB固相柱净化,选用甲醇为洗脱剂,浓缩后用HPLC/FLD检测,采用乙腈/0.7%磷酸作为流动相。FQs在0.01～1.0 mg·kg-1浓度范围内呈现良好线性关系,R2为0.996 9～0.999 9。加标(浓度水平0.01、0.5、1.0 mg·kg-1干重)回收率达76.7%～106.7%,相对标准偏差0.7%～14.4%(n=4)。方法检出限为0.002～0.022 mg·kg-1,方法定量限为0.006 8～0.074 mg·kg-1。应用该方法对北京地区5个养鸡场内鸡粪样品中6种FQs进行分析,检出浓度为ND～1.13 mg·kg-1。

An analytical method for simultaneous determination of six fluoroquinolones（FQs ） in chicken manures based on solid-phase extraction and high performance liquid chromatography with a fluorescence detection （HPLC/FLD） was developed in the current study. Samples were ultrasonically extracted in Mcllvaine-Na2EDTA buffer/acetonitrile（ 1：1 ）（pH=2） for 15 min. The extract was cleaned by a HLB solid-phase extraction column, and then eluted with methanol solvent. The fluoroquinolones in the extract were determined by HPLC/FLD with acetonitrile/0.7% phosphate as mobile phase. In a concentration range of 0.01-1.0 mg· kg-1, R2 of calibration was greater than 0.99. The average percent recoveries ranged from 76.7% to 106.7% for six compounds in chicken manure samples at three different spiked levels （0.1, 0.5 mg· kg-1 and 1.0 mg· kg-1）. The detection limits （LOD） were from 0.002 mg·kg-1 to 0.022 mg·kg-1, and limit of quantification （LOQ） from 0.006 8 mg· kg-1 to 0.074 mg· kg-1. This method was used to analyze six FQs in ehicken manures collected from 5 chicken farms in Beijing. All six FQs were detected in most samples, with concentrations ranging from LOD to 1.13 mg· kg-1.