高效液相色谱法同时测定尿液中溴敌隆和溴鼠灵的方法研究

Simultaneous determination of bromadiolone and brodifacoum in human urine samples by reverse-phase high-performance liquid chromatography

目的:建立反相高效液相色谱法同时快速检测尿液中的溴敌隆、溴鼠灵的分析方法。方法:尿样经乙酸乙酯提取过滤后直接进样,以甲醇-1%甲酸(90:10,V/V)为流动相,流速1.0 ml/min,经ODS-C18色谱柱(150 mm×4.6 mmi.d,5.0μm)40℃分离,在波长265 nm处检测。结果:分离测定过程在10 min之内完成,保留时间分别为溴敌隆3.770 min、溴鼠灵8.110 min。溴敌隆、溴鼠灵线性范围均为0.02～50.0μg/ml,最低检出量为0.16 ng。低、中、高3个浓度水平的样品加标平均回收率为86.0%～109.0%,保留时间与峰面积的日内和日间相对标准偏差分别<1%和<5%。结论:本方法简便、快速、稳定、准确,实用性强,满足中毒应急检测及临床治疗的需要。

Objective：To develop a RP - HPLC method for the simultaneous determination of two rodenticides of bromadiolone and brodifacoum in human urine samples. Methods ：The objects of analysis were extracted from urine samples by ethyl acetate and were injected into an ODS- C18 column （150 mm×4. 6 mm i. d,5 μm） after being filtered. The elution was performed with the mobile phase comprising of methanol -1% formic acid （90：10, V/V）. The flow rate was 1.0 ml/min, the temperature of column was 40℃, and the dectecion wavelength was 265 nm for both. Results：The seperation and detection was completed within 10 minutes. The retention time was 3. 770 min and 8.110 min for bromadiolone and brodifacoum, respectively. There were two linear ranges, 0. 02 - 1.0μg/ml and 0. 5 - 50. 0μg/ml for both of bromadiolone and brodifacoum. The recoveries from urine samples spiked at low, middle and higher levels ranged from 86.0% to 109.0% , and the precision of retention times and areas of intra - day and inter-day expressing RSD was less than 1% and 5% ,respectively. Conclution：The method developed is simple, fast, stable and available. It can meet the requirements for simultaneous determination of two rodenticides for poisoning patients in clinic.