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首页> 外文期刊>Journal of Analytical Toxicology >Complete Automation of Solid-Phase Extraction with Subsequent Liquid Chromatography-Tandem Mass Spectrometry for the Quantification of Benzoylecgonine, m-Hydroxybenzoylecgonine, p-Hydroxybenzoylecgonine, and Norbenzoylecgonine in Urine-Application to a High-Throughput Urine Analysis Laboratory
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Complete Automation of Solid-Phase Extraction with Subsequent Liquid Chromatography-Tandem Mass Spectrometry for the Quantification of Benzoylecgonine, m-Hydroxybenzoylecgonine, p-Hydroxybenzoylecgonine, and Norbenzoylecgonine in Urine-Application to a High-Throughput Urine Analysis Laboratory

机译：固相萃取与随后的液相色谱-串联质谱法完全自动化，用于定量分析尿液中的苯甲酰芽子碱，间羟基苯甲酰芽子碱，对羟基苯甲酰芽子碱和正苯甲酰芽子碱-在高通量尿液分析实验室中的应用

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摘要

A fully automated system utilizing a liquid handler and an online solid-phase extraction (SPE) device coupled with liquid chromatography-tandem mass spectrometry (LC-MS-MS) was designed to process, detect, and quantify benzoylecgonine (BZE), meta-hydroxybenzoylecgonine (m-OH BZE), para-hydroxybenzoylecgonine (p-OH BZE), and norbenzoylecgonine (nor-BZE) metabolites in human urine. The method was linear for BZE, m-OH BZE, and p-OH BZE from 1.2 to 10,000 ng/mL with limits of detection (LOD) and quantification (LOQ) of 1.2 ng/mL. Nor-BZE was linear from 5 to 10,000 ng/mL with an LOD and LOQ of 1.2 and 5 ng/mL, respectively. The intrarun precision measured as the coefficient of variation of 10 replicates of a 100 ng/mL control was less than 2.6%, and the interrun precision for 5 replicates of the same control across 8 batches was less than 4.8% for all analytes. No assay interference was noted from controls containing cocaine, cocaethylene, and ecgonine methyl ester. Excellent data concordance (R2 > 0.994) was found for direct comparison of the automated SPE-LC-MS-MS procedure and an existing gas chromatography-MS procedure using 94 human urine samples previously determined to be positive for BZE. The automated specimen handling and SPE procedure, when compared to the traditional extraction schema, eliminates the human factors of specimen handling, processing, extraction, and derivatization, thereby reducing labor costs and rework resulting from batch handling issues, and may reduce the number of fume hoods required in the laboratory.
机译：设计了一个利用液体处理机和在线固相萃取（SPE）装置以及液相色谱-串联质谱（LC-MS-MS）的全自动系统，以处理，检测和定量苯甲酰芽子碱（BZE），间苯二酚人体尿液中的羟基苯甲酰芽子碱（m-OH BZE），对羟基苯甲酰芽子碱（p-OH BZE）和正苯甲酰芽子碱（nor-BZE）代谢产物。该方法对于BZE，m-OH BZE和p-OH BZE为1.2至10,000 ng / mL呈线性，检测限（LOD）和定量限（LOQ）为1.2 ng / mL。 Nor-BZE在5至10,000 ng / mL范围内呈线性，LOD和LOQ分别为1.2和5 ng / mL。对于所有分析物，以100 ng / mL对照的10个重复样品的变异系数测量的批内精密度小于2.6％，同一对照在8个批次中重复5个重复的样品间精密度小于4.8％。含有可卡因，可卡乙烯和芽子碱甲酯的对照组未发现分析干扰。对于自动SPE-LC-MS-MS程序和使用先前确定为BZE阳性的94个人尿样品的现有气相色谱-MS程序的直接比较，发现极好的数据一致性（R2> 0.994）。与传统的提取方案相比，自动样品处理和SPE程序消除了样品处理，处理，提取和衍生化的人为因素，从而减少了因批次处理问题而导致的人工成本和返工，并可以减少烟尘数量实验室所需的通风柜。

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