浊点萃取-高效液相色谱法同时测定人尿中的4种抗凝血鼠药  

Simultaneous Determination of Four Anticoagulant Rodenticides in Human Urine Using High Performance Liquid Chromatography-Cloud Point Extraction

作　　者：孟庆玉[1] 黎源倩[1] 邹晓莉[1] 郑波[1] 

机构地区：[1]四川大学华西公共卫生学院卫生检测教研室

出　　处：《分析化学》2008年第6期760-764,共5页Chinese Journal of Analytical Chemistry

基　　金：郑州市科技发展计划资助项目(No.200619-58)

摘　　要：建立了基于浊点萃取同时测定人尿中杀鼠灵、杀鼠醚、溴敌隆和溴鼠灵的反相高效液相色谱法。以非离子表面活性剂TritonX-114为萃取剂,考察了色谱测定条件,优化了浊点萃取参数。5mL尿样中加入0.030gNaCl,30μL甲酸和5%(w/v)TritonX-114400μL,50℃萃取15min,4种目标物的萃取效率能达到85%以上。在选择的色谱条件下,以V(甲醇)∶V(1%甲酸)=90∶10为流动相,待测组分经反相C18色谱柱分离,于306nm波长处用二极管阵列检测器检测,10min可完成分析。4种鼠药中杀鼠醚在0.01～5.0mg/L,其余3种在0.02～5.0mg/L范围内呈良好的线性关系(r≥0.997);尿样的标准加入回收率为71.2%～114.5%;相对标准偏差小于7.0%。对于5mL尿液,富集相用50μL甲醇稀释,萃取浓缩因子可达到28倍。定量检出浓度分别为:杀鼠灵0.016mg/L,杀鼠醚0.001mg/L,溴敌隆0.005mg/L,溴鼠灵0.003mg/L。本法简便快速、灵敏可靠、绿色环保,能满足鼠药中毒应急检测的要求。A new method was developed for the simultaneous determination of four rodenticides including warfarin, coumatetralyl, bromadiolone and brodifacoum in human urine samples by reverse-phase high performance liquid chromatography after clound point extraction（CPE）by Triton X-114. The chromatographic conditions and extraction parameters were optimized. Under the optimum CPE conditions： 0. 030 g NaCl, 30 μL formic acid and 5% （w/V） Triton X-114 400 μL for 5 mL unine sample with the extraction temperature of 50℃ and 15 min of equilibration time, the extraction efficiencies were more than 85%. Then the extracts were seperated and eluted by an ODS-C18column（ 150 mm×4.6 mm i. d. , 5 8zm） and the mobile phase comprising of methanol-1% formic acid （90：10, V/V） at 1.0 mL/min under 40 ℃. The dectecion was done at wavelength of 306 nm and completed within 10 minutes. The linear ranges of four rodenticides were in the range of 0.02 -5.0 mg/L except for coumatetralyl 0.01 -5.0 mg/L with the correlation coefficients more than 0. 997. The recoveries of the four rodenticides from spiked urine samples ranged from 71.2% to 114.5% and the relative standard deviations were less than 7.0%. The concentration factor（Fc） could be changed with the final diluted volume of surfactant-rich phase, and the theory concentration factor could reach to 28 times when 50 μL methanol was added after CPE of 5 mL sample. The limits of quantification were 0. 016 mg/L, 0.001 mg/L, 0. 005 mg/L, 0. 003 mg/L for warfarin, coumatetralyl, bromadiolone and brodifacoum, respectively. The proposed method is simple, rapid, sensitive, accurate and friendly to environment. It can meet the requirements for simultaneous determination of four rodenticides for poisoning patients.

关 键 词：浊点萃取 高效液相色谱法 非离子表面活性剂 抗凝血杀鼠剂 尿样 

分 类 号：R446.1[医药卫生—诊断学]