Supramolecular solvents with restricted-access properties (SUPRAS-RAMs) are proposed as a new approach for integrating extraction and sample cleanup in the quantification of benzodiazepines (BDZs) in urine by liquid chromatography–tandem mass spectrometry (LC–MS-MS). The SUPRAS-RAM was synthesized in situ in the urine by the addition of 1-hexanol (154 µL) and tetrahydrofuran (THF) (600 µL). BDZ extraction was driven by both hydrogen bonds and dispersion interactions. Removal of proteins and polar macromolecules was performed by the action of the SUPRAS through chemical and physical mechanisms. Phospholipids were removed by precipitation during SUPRAS extract evaporation. A multivariate method was used for the optimization of the extraction process by applying Box–Behnken response surface design. The proposed method was validated according to the guiding principles of the European Commission Decision (2002/657/EC). Method detection and quantification limits for the target BDZs were in the intervals 0.21–0.85 and 0.67–2.79 ng/mL, respectively. The repeatability and reproducibility (expressed as relative standard deviations) were in the range 2–6% and 3–8%, respectively. The method enabled the simultaneous extraction of BDZs (recoveries in the range 84–105%) and the removal of matrix effects. The method was applied to the analysis of 13 urine samples using external calibration. Five out of 13 samples tested positive in alprazolam and lorazepam at concentrations in the range 5.4–74 ng/mL. The method allows simple and quick sample treatment with minimal solvent consumption while delivering accurate and precise data.

中文翻译：

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基于限制使用超分子溶剂和 LC-MS-MS 的无干扰测定尿液中苯二氮卓类药物的方法

具有限制访问特性的超分子溶剂 (SUPRAS-RAM) 被提出作为一种新方法，用于通过液相色谱-串联质谱 (LC-MS-MS) 对尿液中的苯二氮卓 (BDZ) 进行定量分析，将萃取和样品净化相结合。通过添加 1-己醇 (154 µL) 和四氢呋喃 (THF) (600 µL) 在尿液中原位合成 SUPRAS-RAM。BDZ 提取由氢键和分散相互作用驱动。蛋白质和极性大分子的去除是通过 SUPRAS 的作用通过化学和物理机制进行的。在 SUPRAS 提取物蒸发过程中通过沉淀去除磷脂。通过应用 Box-Behnken 响应面设计，使用多元方法优化提取过程。提议的方法根据欧盟委员会决定 (2002/657/EC) 的指导原则进行了验证。目标 BDZ 的方法检测和定量限分别为 0.21–0.85 和 0.67–2.79 ng/mL。重复性和再现性（表示为相对标准偏差）分别在 2-6% 和 3-8% 范围内。该方法能够同时提取 BDZ（回收率在 84-105% 范围内）和去除基质效应。将该方法应用于使用外标法对 13 份尿样进行分析。13 个样本中有 5 个在 5.4–74 ng/mL 范围内的阿普唑仑和劳拉西泮检测呈阳性。该方法允许以最少的溶剂消耗进行简单快速的样品处理，同时提供准确和精确的数据。