Analysis of postmortem samples with the presence of morphine can sometimes be challenging to interpret. Tolerance complicates interpretation of intoxications and causes of death due to overlap in therapeutic and fatal concentrations. Determination of metabolites and metabolic ratios can potentially differentiate between abstinence, continuous administration, and perhaps time of administration. The purpose of this study was to (a) develop and validate a method for quantitation of morphine-3β-D-glucuronide, morphine-6β-D-glucuronide, normorphine, codeine-6β-D-glucuronide, norcodeine, codeine, 6-acetylmorphine, and ethylmorphine in urine using liquid chromatography-tandem mass spectrometry; (b) apply the method to opiate related deaths; (c) compare metabolic ratios in urine in different causes of death (CoD) and after different drug intakes and (d) compare heroin intoxications in rapid and delayed deaths. Validation parameters such as precision, bias, matrix effects, stability, process efficiency, and dilution integrity were assessed and deemed acceptable. Lower limits of quantitation ranged from 0.01-0.2 μg/mL for all analytes. Autopsy cases (n=135) with paired blood and urine samples were analyzed. Cases were divided into three groups based on CoD; opiate intoxication, intoxication with other drugs than opiates, and other CoD. The cases were classified by intake: codeine (n=42), heroin (n=36), morphine (n=49), and ethylmorphine (n=3). Five cases were classified as mixed intakes and excluded. Heroin intoxications (n=35) were divided into rapid (n=15) or delayed (n=20) deaths. Parent drug groups were compared using metabolic ratio morphine-3β-D-glucuronide/morphine and significant differences were observed between codeine vs morphine (p=0.005) and codeine vs heroin (p≤0.0001). Urine and blood concentrations, and metabolic ratios in rapid and delayed heroin intoxications were compared and determined a significant difference for morphine (p=0.001), codeine (p=0.009), 6-acetylmorphine (p=0.02) in urine, and morphine (p=0.02) in blood, but there was no significant difference (p=0.9) between metabolic ratios. Morphine-3β-D-glucuronide results suggested a period of abstinence prior to death in 25% of the heroin intoxications.

中文翻译：

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使用LC-MS-MS在死后样品中与海洛因有关的化合物和代谢率

在存在吗啡的情况下对尸体样本进行分析有时可能难以解释。宽容使中毒和由于治疗浓度和致命浓度重叠造成的死亡原因的解释复杂化。代谢物和代谢率的确定可能会在禁欲，持续给药和给药时间之间进行区分。这项研究的目的是（a）开发和验证定量吗啡3β-D-葡糖醛酸，吗啡-6β-D-葡糖醛酸，正吗啡，可待因-6β-D-葡糖醛酸，正典，可待因6-的定量方法液相色谱-串联质谱法测定尿液中的乙酰吗啡和乙基吗啡；（b）将这种方法用于鸦片相关的死亡；（c）比较不同死亡原因（CoD）和不同药物摄入后尿液中的代谢率，以及（d）比较快速死亡和迟发性死亡中的海洛因中毒。评估参数，如精度，偏差，基质效应，稳定性，过程效率和稀释完整性，被认为是可以接受的。所有分析物的定量下限为0.01-0.2μg/ mL。尸检病例（n = 135）具有成对的血液和尿液样本。根据CoD将病例分为三组。鸦片中毒，除鸦片以外的其他药物和其他CoD的中毒。病例按摄入量分类：可待因（n = 42），海洛因（n = 36），吗啡（n = 49）和乙基吗啡（n = 3）。五例被分类为混合摄入，被排除在外。海洛因中毒（n = 35）分为快速死亡（n = 15）或延迟死亡（n = 20）。使用吗啡-3β-D-葡萄糖醛酸/吗啡的代谢率比较了母体药物组，在可待因与吗啡（p = 0.005）和可待因与海洛因（p≤0.0001）之间观察到显着差异。比较快速和延迟海洛因中毒时的尿液和血液浓度以及代谢率，确定尿液中吗啡（p = 0.001），可待因（p = 0.009），6-乙酰吗啡（p = 0.02）和吗啡（ p = 0.02），但代谢率之间无显着差异（p = 0.9）。吗啡3β-D-葡糖醛酸苷的结果表明，在死亡前有25％的海洛因中毒戒断。比较快速和延迟海洛因中毒时的尿液和血液浓度以及代谢率，确定尿液中吗啡（p = 0.001），可待因（p = 0.009），6-乙酰吗啡（p = 0.02）和吗啡（ p = 0.02），但代谢率之间无显着差异（p = 0.9）。吗啡3β-D-葡糖醛酸苷的结果表明，在死亡前有25％的海洛因中毒戒断。比较快速和延迟海洛因中毒时的尿液和血液浓度以及代谢率，确定尿液中吗啡（p = 0.001），可待因（p = 0.009），6-乙酰吗啡（p = 0.02）和吗啡（ p = 0.02），但代谢率之间无显着差异（p = 0.9）。吗啡3β-D-葡糖醛酸苷的结果表明，在死亡前有25％的海洛因中毒戒断。