QuEChERS净化—液相色谱串联质谱法测定血浆中8种抗凝血灭鼠剂

摘要:目的 建立QuEChERS净化，同位素稀释液相色谱串联质谱法（LC-MS/MS）测定血浆中大隆、溴敌隆、杀鼠醚、杀鼠灵、氯杀鼠灵、敌鼠、氯敌鼠、杀鼠酮抗凝血灭鼠剂的检测方法。方法 血浆样品经QuEChERS方法前处理，以杀鼠灵-D5作为同位素内标，采用BDS HYPERSIL C18(4.6 mm i.d×100 mm, 2.4 μm)分离，以甲醇-乙酸铵缓冲液梯度洗脱，流速为0.50 ml/min。采用电喷雾电离(ESI)负离子质谱在多反应监测(MRM)模式下检测。结果 8种抗凝血灭鼠剂在0.20 μg /L～25.0 μg /L(或者0.50 μg/L～25.0 μg/L)范围内有良好的线性关系(r＞0.999)，检测限为0.044～0.19 μg /L，平均加标回收率为70.4%～101.5%（n=7），相对标准偏差1.7%～8.9%（n=7）。结论 该方法快速、简单，灵敏度高，可应用于中毒病人血样中8种抗凝血灭鼠剂的同时检测。

Determination of 8 anticoagulant rodenticides in plasma by QuEChERS cleanup-liquid chromatography-tandem mass spectrometry

Abstract: Objective The study was conducted to develop a method for determination of 8 anticoagulant rodenticides including brodifacoum, bromadiolone, coumatetralyl, warfarin, coumachlor,diphenadione, chlorophacinone, and pindone, in plasma by QuEChERS cleanup coupled with isotope dilution and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods Plasma samples were pre-treated by QuEChERS method, and warfarin-D5 was adopted as the isotopic internal standard. The compounds were separated on a BDS HYPERSIL C18 column (4.6 mm i.d×100 mm, 2.4 μm) and eluted by methanol-ammonium acetate buffer gradient with a flow rate of 0.50 ml/min. The 8 anticoagulant rodenticides were detected by electrospray ionization (ESI)-negative ion mass spectrometry under the multiple-reaction monitoring (MRM) mode. Results A good linearity was observed in the range of 0.20 μg/L-25.0 μg/L (or 0.50 μg/L-25.0 μg/L)(r>0.999) of the anticoagulant rodenticides. The detection limit of the method was 0.044-0.19 μg/L, the average sample recovery rate ranged between 70.4% and 101.5% (n=7), and the relative standard deviation (RSD) was 1.7%-8.9% (n=7). Conclusion The method is fast, simple， highly sensitive, and can be adopted for simultaneous detection of the 8 anticoagulant rodenticides in plasma of intoxicated patients.