摘要
利用免疫亲和色谱净化技术建立了可同时检测动物肝脏组织中10种喹诺酮类药物(麻保沙星、环丙沙星、诺氟沙星、单诺沙星、洛美沙星、恩诺沙星、沙拉沙星、二氟沙星、恶喹酸和氟甲喹)的高效液相色谱检测方法。对利用喹诺酮抗体制备的免疫亲和色谱柱的性能、操作条件进行了考察和优化。抗体的偶联量为5g/L,其对10种喹诺酮药物的柱容量为3.75～6.67μmol/Lgel(1425～2135mg/Lgel),选用V(甲醇):V(PBS)=7:3作为洗脱溶液,连续使用12次后,QNs的柱容量仍能达到初始柱容量的38%～45%;IAC柱重复使用20次后,药物的回收率与样品的净化效果无明显变化。动物肝脏组织样品用PBS溶液提取,IAC柱净化,HPLC-FLD检测。方法的线性范围为0.15～200μg/L,相关系数大于0.9989,检出限为0.05～0.15μg/kg;10种喹诺酮类药物在动物肝脏的平均回收率为74.7%～94.8%,相对标准偏差为3.9%～12.1%。
An immunoaffinity chromatographic column （IAC） was developed for the analysis of 10 quinolones （marbofloxacin, norfloxacin, ciprofloxacin, lomefloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine） in animal liver tissues by HPLC-FLD. The performance of the IAC and the operating strategies were optimized. The efficient elution of quinolones captured by IAC was achieved with 4 mL of 70% methanol in PBS. The dynamic column capacities of 13 QNs were from 3.75 μmol/L gel to 6.67 μmol/L gel （1425 -2135μg/L gel）. The column capacity was about 35% -48% of the original capacity after 12 cycles of use at the optimized procedure. After 20 cycles, the recoveries and purified capacity were not significant different using the IAC. The samples were extracted with PBS solution, cleaned up with IAC and then determined by HPLC-FLD. The linear range was 0.15 -200 μg/L with correlation coefficients more than 0. 9989. The limits of detection were 0.05 - 0. 15 μg/kg. The mean recoveries for each analyte in chicken and pig muscles ranged from 74.7% to 94.8% with relative standard deviation from 3.9% to 12.1% at 1.0 - 100 g/kg fortification levels.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2009年第3期335-340,共6页
Chinese Journal of Analytical Chemistry
基金
国家杰出青年科学基金(No.30325032)
“十一五”国家科技支撑计划(No.2006BAK02A08)资助项目
关键词
喹诺酮类药物
免疫亲和色谱
肝脏组织
高效液相色谱法
多残留
Quinolones, immunoaffinity chromatography, liver tissues,high performance liquid chromatography, multi-residue
