Department of Forensic Science, School of Basic Medical Sciences, Fudan University, Shanghai, P.R. China.
Rapid Commun Mass Spectrom. 2013 Feb 28;27(4):513-20. doi: 10.1002/rcm.6477.
Bromadiolone and brodifacoum, two common anticoagulant rodenticides, are involved in the majority of anticoagulant rodenticide poisoning cases in humans in China. Hair analysis can provide long-term information on drug exposure. A method using liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) was developed and validated for the measurement of bromadiolone and brodifacoum in human hair.
A 1 mL aliquot of a phosphate buffer solution (pH 6.8) was added to 20 mg of pulverized hair followed by ultrasonication and liquid-liquid extraction. Liquid chromatography was performed using a C(18) column with a mobile phase gradient of ammonium acetate (10 mM) and methanol. A tandem mass spectrometer in multiple reaction monitoring mode with a negative electrospray ionization source was employed for detection. Warfarin-d(5) was used as an internal standard for both analytes.
The limits of detection (LODs) for bromadiolone and brodifacoum were 0.010 and 0.025 ng/mg, respectively. The calibration curves for both analytes were linear from 0.025 to 1 ng/mg. The accuracy ranged from 90.3 to 109.3%, and the intra-day and inter-day imprecisions were less than 15%.
The established method was found effective when applied to the analyses of bromadiolone or brodifacoum in five cases, indicating that segmental hair analysis could be useful for clinical and forensic purposes by identifying the time of ingestion.
在中国，绝大多数人类抗凝血灭鼠剂中毒案例涉及溴敌隆和大隆两种常见的抗凝血灭鼠剂。毛发分析可以提供药物暴露的长期信息。本研究建立并验证了一种使用液相色谱-串联质谱（LC/MS/MS）测定人发中溴敌隆和大隆的方法。
取 1mL 磷酸盐缓冲液（pH6.8）加入 20mg 粉碎的毛发，然后进行超声提取和液-液萃取。采用 C（18）柱，以乙酸铵（10mM）和甲醇为流动相梯度洗脱，进行液相色谱分离。串联质谱采用负离子电喷雾源多反应监测模式进行检测，同时使用华法林-d（5）作为内标。
溴敌隆和大隆的检测限（LOD）分别为 0.010 和 0.025ng/mg。两种分析物的校准曲线在 0.025 至 1ng/mg 范围内均呈线性。准确度在 90.3%至 109.3%之间，日内和日间精密度均小于 15%。
该方法应用于 5 例案例中溴敌隆或大隆的分析，结果有效，表明分段毛发分析通过确定摄入时间，可用于临床和法医学目的。
