高效液相色谱-离子阱质谱法测定生物体液中的痕量敌鼠和氯敌鼠

建立了中毒应急检测样品生物体液内痕量敌鼠和氯敌鼠的高效液相色谱-离子阱质谱定性定量检测方法。全血样品由甲醇/乙腈(50/50, v/v)沉淀蛋白后再经Oasis HLB固相萃取小柱净化富集,尿液样品直接经同类小柱净化富集。采用Extend C18柱分离,以乙酸铵-乙酸(0.02 mol/L, pH 5.5)缓冲溶液和甲醇(15/85, v/v)溶液为流动相,使用电喷雾电离负离子多反应监测模式检测敌鼠和氯敌鼠。对全血样品,敌鼠和氯敌鼠分别在1.0～200.0 μg/L和0.5～100.0 μg/L范围内呈良好的线性关系,回收率分别在90.1%～92.2%和87.6%～93.4%范围内,日内相对标准偏差(RSD)分别小于6.8%和7.1%,日间RSD分别小于9.9%和10.9%,定量限分别为1.0 μg/L和0.5 μg/L。对尿液样品,敌鼠和氯敌鼠分别在0.2～ 40.0 μg/L和0.1～20.0 μg/L范围内呈良好的线性关系,回收率分别在90.1%～94.5%和90.0%～ 98.0%范围内,日内RSD分别小于6.1%和7.3%,日间RSD分别小于8.9%和11.2%,定量限分别为0.2 μg/L和0.1 μg/L。本方法简便、灵敏,能满足敌鼠和氯敌鼠中毒病人的临床快速诊断要求。

Simultaneous determination of trace diphacinone and chlorophacinone in biological samples by high performance liquid chromatography coupled with ion trap mass spectrometry

A rapid qualitative and quantitative method for the simultaneous determination of trace diphacinone and chlorophacinone in biological samples has been established.The method mainly serves for the emergent poisoning detection.The whole blood was treated with methanol-acetonitrile (50/50,v/v) and the urine was cleaned-up by Waters Oasis HLB SPE cartridges.The samples were separated on an Extend C18 column (150 mm×4.6 mm,5 μm) by using the mobile phase consisted of ammonium acetate-acetic acid (0.02 mol/L,pH 5.5)-methanol (15/85,v/v).The determination was performed by high performance liquid chromatography coupled with ion trap mass spectrometry (HPLC-IT-MS) using a negative electrospray ionization interface in the multiple reaction monitoring (MRM) mode.The transitions of m/z 339→167 for diphacinone and m/z 373→201 for chlorophacinone were selected for the quantificantions.For the whole blood samples,the calibration curves were linear within the ranges of 1.0-200.0 μg/L and 0.5-100.0 μg/L; the limits of quantification were 1.0 μg/L and 0.5 μg/L; the spike recoveries were 90.1% -92.2% and 87.6% -93.4% ,the intra-day relative standard deviations (RSDs) were less than 6.8% and 7.4% ,and the inter-day RSDs were less than 9.9% and 10.9% for diphacinone and chlorophacinone,respectively.For the urine samples,the calibration curves were linear within the ranges of 0.2-40.0 μg/L and 0.1-20.0 μg/L; the limits of quantification were 0.2 μg/L and 0.1 μg/L; the spike recoveries were 90.1% -94.5% and 90.0% -98.0% ,the intra-day RSDs were less than 6.1% and 7.3% ,and the inter-day RSDs were less than 8.9% and 11.2% for diphacinone and chlorophacinone,respectively.This method is simple and sensitive for the satisfactory determination of trace diphacinone and chlorophacinone residues in poisoned patients for the clinical diagnosis.