摘要
目的建立尿样中毒死蜱及其代谢物3,5,6-三氯-2-吡啶醇(TCPyr)的高效液相色谱分析方法。方法采用乙酸乙酯-正己烷混合溶剂提取,Inertsil ODS-3反相分离柱,以乙腈-甲醇-水-0.05%三氟乙酸为流动相梯度洗脱,毒死蜱和TCPyr的紫外检测波长分别为290 nm与298 nm,并利用二极管阵列检测器对毒死蜱和TCPyr的紫外图谱进行鉴定。结果毒死蜱和TCPyr的线性范围分别为0.2616-13.08μg/mL和0.137 4-6.870μg/mL,相关系数良好;最低检测限分别为105.0ng/mL和62.5 ng/mL;日内及日间精密度RSD均小于10.0%,平均加标回收率分别为67.0%和85.0%。结论此方法灵敏准确,简便实用,可用于尿样中毒死蜱与TCPyr的含量分析。
Purpose To develop a simple and sensitive HPLC method for simultaneous determination of the insecticide chlorpyrifos (O, O-diethyl-O-[-3, 5, 6-trichloro-2-pyridinyl]phosphorothioate), and its specific metabolite TCPyr (3, 5, 6-trichloro-2-pyridinol) in human urine samples by HPLC. Methods The method was based on using solvent extraction and HPLC with reversed-phase ODS-3 column. The mobile phase was gradient mixture of acetonitrile-methanol-water containing 0.05 % (V/ V) trifluoroacetic acid (TFA). The compounds were identified by diode array detector (DAD) at 290 nm for chlorpyrifos and at 298 nm for TCP, respectively. Chlorpyrifos and TCPyr were extracted by a mixed solvent of ethyl acetate-n-hexane (3: 1, V/V). After purification and centrifugation, the organic layer was evaporated with a stream of nitrogen at 45 ℃, and the residue was dissolved in mixed solvent acetonitrile-methanol of (1: 1, V/V) and injected into the HPLC system Results Precise separation of the chlorpyrifos and TCPyr was achieved within15 minutes. The relationship between peak areas and concentration showed a good linearity over a range from 0. 261 6 to 13. 080 μg /mL for chlorpyrifos, and from 0. 137 4 to 6.87 0 μg/mL for TCPyr, respectively. The limits of detection were 105.0 ng/ mL for chlorpyrifos and 62.5 ng/mL for TCPyr, respectively. Percentage recoveries of this procedure for fortified samples in different matrix were between 65.2% - 68.9% for chlorpyrifos, and between 71.5% - 96.8% for TCPyr. The RSD of the intraday and the inter-day variation were both lower than 10.0%. Conclusions These results demonstrated that the proposed method is simple, reliable and sensitive for simultaneous determination of chlorpyrifos and its metabolite TCPyr in human urine samples with satisfactory results.
出处
《复旦学报(医学版)》
CAS
CSCD
北大核心
2007年第3期423-426,共4页
Fudan University Journal of Medical Sciences
基金
国家自然科学基金(30571554)
上海市曙光计划资助项目