Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA.
J Anal Toxicol. 2012 Oct;36(8):541-7. doi: 10.1093/jat/bks063. Epub 2012 Jul 24.
When chronic pain patients are suspected of being non-compliant, their therapy can be withdrawn. Therefore, sensitive and specific confirmatory testing is important for identifying diversion and adherence. This work aimed to develop a novel liquid chromatography tandem mass spectrometry (LC-MS-MS) method to detect 14 opioids and six opioid glucuronide metabolites in urine with minimal sample preparation. Analytes included were morphine, oxymorphone, hydromorphone, oxycodone, hydrocodone, codeine, fentanyl, norfentanyl, 6-monoacetylmorphine, meperidine, normeperidine, propoxyphene, methadone, buprenorphine, morphine-3-glucuronide, morphine-6-glucuronide, oxymorphone glucuronide, hydromorphone glucuronide, codeine-6-glucuronide and norbuprenorphine glucuronide. Samples were processed by centrifugation and diluted in equal volume with a deuterated internal standard containing 14 opioids and four opioid glucuronides. The separation of all compounds was complete in nine minutes. The assay was linear between 10 and 1,000 ng/mL (fentanyl 0.25-25 ng/mL). Intra-assay imprecision (500 ng/mL, fentanyl 12.5 ng/mL) ranged from 1.0 to 8.4% coefficient of variation. Inter-assay precision ranged from 2.9 to 6.0%. Recovery was determined by spiking five patient specimens with opioid and opioid glucuronide standards at 100 ng/mL (fentanyl 2.5 ng/mL). Recoveries ranged from 82 to 107% (median 98.9%). The method correlated with our current quantitative LC-MS-MS assay for opioids, which employs different chromatography. Internal standards were not available for every analyte to critically evaluate for ion suppression. Instead, a novel approach was designed to achieve the most rigorous quality control possible, in which the recovery of each analyte was evaluated in each negative sample.
当怀疑慢性疼痛患者不遵医嘱时，可以停止其治疗。因此，进行敏感和特异的确认性检测对于识别药物滥用和用药依从性非常重要。本研究旨在开发一种新的液相色谱串联质谱（LC-MS-MS）方法，用于最小化样品制备检测尿液中的 14 种阿片类药物和 6 种阿片类葡萄糖醛酸代谢物。分析物包括吗啡、羟吗啡、氢吗啡酮、羟考酮、氢可酮、可待因、芬太尼、去甲芬太尼、6-单乙酰吗啡、哌替啶、去甲哌替啶、丙氧芬、美沙酮、丁丙诺啡、吗啡-3-葡萄糖醛酸、吗啡-6-葡萄糖醛酸、羟吗啡酮葡萄糖醛酸、氢吗啡酮葡萄糖醛酸、可待因-6-葡萄糖醛酸和去甲丁丙诺啡葡萄糖醛酸。样品经离心处理后，与含有 14 种阿片类药物和 4 种阿片类葡萄糖醛酸代谢物的氘代内标以等体积稀释。所有化合物在 9 分钟内完全分离。该测定法在 10 至 1000ng/mL（芬太尼 0.25-25ng/mL）范围内呈线性。批内精密度（500ng/mL，芬太尼 12.5ng/mL）为 1.0%-8.4%的变异系数。批间精密度为 2.9%-6.0%。通过向 5 份患者标本中加入阿片类药物和阿片类葡萄糖醛酸标准品（100ng/mL，芬太尼 2.5ng/mL）测定回收率。回收率为 82%-107%（中位数 98.9%）。该方法与我们目前使用不同色谱法的定量 LC-MS-MS 阿片类药物检测方法相关。由于不是每个分析物都有内标，因此无法严格评估离子抑制。相反，设计了一种新方法以尽可能严格地控制质量，在该方法中，在每个阴性样本中评估每个分析物的回收率。
