LC-MS/MS分析有机磷农药中毒

Poison Analysis of Organophosphorus Pesticides by LC-MS/MS

关键词:  血浆  有机磷  高效液相色谱串联质谱法  检测

Key words:  plasma  organophosphorus pesticide  LC-MS/MS  determination

摘要:

目的 建立一种同时检测人血浆中9种有机磷农药的高效液相色谱串联质谱法(LC-MS/MS)。方法 色谱柱为Thermo C₁₈(4.6 mm×250 mm，5 μm)，柱温25 ℃，流动相为乙腈-水(含20 mmol·L^-1醋酸铵和0.1%甲酸)(85∶15)，流速0.3 mL·min^-1。血浆样品采用乙腈沉淀蛋白，质谱以正离子方式检测，采用多重反应监测模式(MRM)。结果 9种有机磷农药的线性范围为0.05~10 μg·mL^-1，定量下限为0.05 μg·mL^-1，检测限在0.1~5 ng·mL^-1之间，方法回收率在93.23%~107.5%之间，批内、批间精密度均<10%，稳定性良好。结论 该方法准确灵敏、简单快速，可用于筛查、测定患者体内的有机磷农药，有效辅助临床对中毒患者的诊断和疗效评价。

Abstract:

OBJECTIVE To establish a liquid chromatography/tandem mass spectrometry method(LC-MS/MS) to simultaneously determine nine organophosphorus pesticides in human plasma. METHODS The analytes were chromatographed on a Thermo C₁₈ (4.6 mm×250 mm, 5 μm) column at 25 ℃. The mobile phase was consisted of acetonitrile and water including 20 mmol·L^-1 ammonium acetate and 0.1% formic acid (85∶15) with a flow rate of 0.3 mL·min^-1. The plasma samples were precipitated by acetonitrile. LC-MS/MS detection was made with an electrospray positive ionization and multiple reaction monitoring. RESULTS The calibration curves of each organophosphorus pesticide showed a good linearity in a range of concentration between 0.05-10 μg·mL^-1. Lower limit of quantitation was 0.05 μg·mL^-1 and the limits of detection were ranged from 0.1 to 5 ng·mL^-1. The recoveries were between 93.23% and 107.5%. The intra- and inter- batch precisions were less than 10% and the stability was good. CONCLUSION The method is proved to be accurate, sensitive, simple and fast. It can be used to screen and determine the organophosphorus pesticides in patients, which can effectively support the diagnosis and efficacy evaluation of poisoned patients in clinical practice.