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首页> 外文期刊>Journal of Analytical Toxicology >Development and Validation of a Novel LC-MS/MS Opioid Confirmation Assay: Evaluation of beta-glucuronidase Enzymes and Sample Cleanup Methods
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Development and Validation of a Novel LC-MS/MS Opioid Confirmation Assay: Evaluation of beta-glucuronidase Enzymes and Sample Cleanup Methods

机译：新型LC-MS / MS阿片类药物确证试验的开发和验证：β-葡萄糖醛酸苷酶的评估和样品净化方法

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With the rise in the use and misuse of prescription opioids, there is an increasing need for the confirmed identification of opioid analgesics in toxicology laboratories. The goals of this study were to (i) systematically evaluate the hydrolysis efficiency of four beta-glucuronidase enzymes under optimized condition; (ii) evaluate compound recovery, matrix effects and precision of three protein precipitation plates and (iii) develop and validate a qualitative liquid-chromatography mass spectrometry (LC-MS/MS) assay to identify 13 opioids in urine. A recombinant beta-glucuronidase exhibited the best overall hydrolysis efficiency for seven opioid glucuronide conjugates compared with beta-glucuronidase from red abalone, Escherichia coli and Patella vulgata. One of the protein precipitation plates tested exhibited overall better recovery of the opioids and lower ion suppression compared with the other two plates. An ESI positive mode LC-MS/MS assay for qualitative opioid analysis was developed and validated. Linearity, LOD, precision, matrix effect, recovery, carryover and interference of the method were evaluated. Sixty-two patient samples were analyzed by both a legacy GC-MS opioid method and the LC-MS/MS method, and 22 samples were analyzed by the LC-MS/MS and an LC-MS/MS reference method. The results of the comparisons showed good concordance. Overall, we described an efficient sample preparation procedure for a sensitive qualitative opioid confirmation assay in urine.
机译：随着处方阿片类药物的使用和滥用的增加，越来越需要在毒理学实验室中确认阿片类镇痛药的鉴定。这项研究的目的是（i）在最佳条件下系统地评估四种β-葡萄糖醛酸苷酶的水解效率; （ii）评估三个蛋白质沉淀板的化合物回收率，基质效应和精密度;（iii）开发和验证定性液相色谱质谱法（LC-MS / MS），以鉴定尿液中的13种阿片类药物。与来自红鲍鱼，大肠杆菌和寻常cher虫的β-葡糖醛酸苷酶相比，重组β-葡糖醛酸苷酶对七个类鸦片葡糖醛酸苷缀合物表现出最佳的总水解效率。与其他两个板相比，测试的蛋白质沉淀板之一显示出阿片样物质的总体更好的回收率和较低的离子抑制。开发并验证了用于定性阿片样物质分析的ESI阳性模式LC-MS / MS测定法。评价了该方法的线性，LOD，精密度，基质效应，回收率，残留和干扰。通过传统的GC-MS阿片类药物方法和LC-MS / MS方法分析了62个患者样品，并通过LC-MS / MS和LC-MS / MS参考方法分析了22个样品。比较结果显示出良好的一致性。总的来说，我们描述了一种有效的样品制备程序，用于尿液中敏感的定性阿片样物质确证测定。

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