An analytical method for the detection of 40 benzodiazepines, (±)-zopiclone, zaleplon and zolpidem in blood and urine by solid-phase extraction liquid chromatography–tandem mass spectrometry was developed and validated. Twenty-nine of 43 analytes were quantified in 0.5 mL whole blood for investigating postmortem, drug-facilitated sexual assault (DFSA) and driving under the influence of drugs cases (DUID). The four different dynamic ranges of the seven-point, linear, 1/x weighted calibration curves with lower limits of quantification of 2, 5, 10 and 20 μg/L across the analytes encompassed the majority of our casework encountered in postmortem, DFSA and DUID samples. Reference materials were available for all analytes except α-hydroxyflualprazolam, a hydroxylated metabolite of flualprazolam. The fragmentation of α-hydroxyflualprazolam was predicted from the fragmentation pattern of α-hydroxyalprazolam, and the appropriate transitions were added to the method to enable monitoring for this analyte. Urine samples were hydrolyzed at 55°C for 30 min with a genetically modified β-glucuronidase enzyme, which resulted in >95% efficiency measured by oxazepam glucuronide. Extensive sample preparation included combining osmotic lysing and protein precipitation with methanol/acetonitrile mixture followed by freezing and centrifugation resulted in exceptionally high signal-to-noise ratios. Bias and between-and within-day imprecision for quality controls (QCs) were all within ±15%, except for clonazolam and etizolam that were within ±20%. All 29 of the 43 analytes tested for QC performance met quantitative reporting criteria within the dynamic ranges of the calibration curves, and 14 analytes, present only in the calibrator solution, were qualitatively reported. Twenty-five analytes met all quantitative reporting criteria including dilution integrity. The ability to analyze quantitative blood and qualitative urine samples in the same batch is one of the most useful elements of this procedure. This sensitive, specific and robust analytical method was routinely employed in the analysis of >300 samples in our laboratory over the last 6 months.

中文翻译：

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固相萃取检测血液和尿液中40种苯并二氮杂and和3种Z-药物的LC-MS-MS方法的开发和验证。

建立并验证了固相萃取液相色谱-串联质谱法检测血液和尿液中40种苯二氮卓类，（±）-佐匹克隆，扎来普隆和唑吡坦的分析方法。在0.5毫升全血中对29种43种分析物进行了定量分析，以研究死后，药物促成性侵犯（DFSA）和在药物病例的影响下驾驶（DUID）。七点线性1 / x的四个不同动态范围在分析物中定量下限分别为2、5、10和20μg/ L的加权校准曲线涵盖了我们在事后检验，DFSA和DUID样品中遇到的大多数案例。除α-羟基氟普拉唑仑（一种氟吡唑仑的羟基化代谢产物）外，所有其他分析物均可使用参考材料。根据α-羟基氟哌唑仑的断裂模式预测了α-羟基氟哌唑仑的断裂，并向该方法中添加了适当的跃迁以监测该分析物。用基因修饰的β-葡萄糖醛酸苷酶将尿液样品在55°C水解30分钟，经奥沙西m葡糖醛酸苷测定，效率> 95％。广泛的样品制备包括将渗透裂解和蛋白质沉淀与甲醇/乙腈混合物混合，然后进行冷冻和离心，从而获得极高的信噪比。质量控制（QC）的偏差和日间和日内不精确度均在±15％以内，但可乐唑仑和依替唑仑的误差均在±20％以内。经质量控制测试的43种分析物中的所有29种均符合校准曲线动态范围内的定量报告标准，并且定性报告了仅存在于校准溶液中的14种分析物。25种分析物符合所有定量报告标准，包括稀释完整性。分析同一批次中定量血液和定性尿液样品的能力是此过程中最有用的元素之一。这个敏感