A procedure based on gas chromatography–mass spectrometry was developed for the analysis of benzodiazepines (nordiazepam, oxazepam, lormetazepam, lorazepam, clonazepam, bromazepam and alprazolam) in postmortem human ribs. Powdered bone samples, including marrow remains inside, with the internal standard diazepam-d₅ were subjected to enzymatic hydrolysis with 100 μL of β-glucoronidase and were incubated in sodium hydroxide for 1 h in a 70°C oven. Samples underwent liquid phase extraction and ethyl acetate was used as eluent. Chromatography was performed on a fused silica capillary column and the selected-ion-monitoring mode was used for analytes determination. The method was validated in the range 0.1–0.5 ng/mg (depending on the benzodiazepine) to 100 ng/mg with average values of recovery, matrix effect and process efficiency ranged from 83.2 to 94.3%, from 97.3 to 102.1% and from 80.5 to 91.2%, respectively. The intra- and inter-day accuracy was <15%. The procedure was tested in rib specimens obtained during routine autopsies from 20 cases where these benzodiazepines were found in blood. Benzodiazepines were detected in the combined bone and marrow samples in 60% of cases. Lorazepam was detected in bone in the range of 0.3–0.7 ng/mg, nordiazepam at 1.3–4.2 ng/mg and oxazepam at 1.1–1.2 ng/mg. To our knowledge, this protocol for the simultaneous analysis of these benzodiazepines is the first performed and validated using human ribs.

中文翻译：

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气相色谱-质谱法对人体骨骼中的苯二氮卓类进行事后分析。

开发了一种基于气相色谱-质谱的方法来分析死后人肋骨中的苯二氮卓类药物（去甲西ze，奥沙西m，氯甲西epa，劳拉西m，氯硝西,、溴马西m和阿普唑仑）。骨粉样品（包括骨髓）保留在内部，内标地西epa-d ₅用100μLβ-葡萄糖苷酸酶进行酶水解，并在70°C的烤箱中于氢氧化钠中孵育1小时。样品进行液相萃取，乙酸乙酯用作洗脱液。在熔融石英毛细管柱上进行色谱分析，并将选定的离子监测模式用于分析物测定。验证该方法的范围为0.1–0.5 ng / mg（取决于苯二氮卓类）至100 ng / mg，回收率，基质效应和工艺效率的平均值分别为83.2至94.3％，97.3至102.1％和80.5。分别达到91.2％。日内和日间准确度<15％。该程序在例行尸检期间从20例血液中发现了这些苯二氮卓类药物的肋骨标本中进行了测试。60％的病例在合并的骨髓和骨髓样本中检出了苯二氮卓类药物。在骨骼中检出劳拉西m的范围为0.3–0.7 ng / mg，去甲西p的范围为1.3–4.2 ng / mg，奥沙西m的范围为1.1–1.2 ng / mg。据我们所知，这种同时分析这些苯并二氮杂pine的方案是首次使用人肋骨进行验证。