Department of Analytical Chemistry, Institute of Fine Chemistry and Nanochemistry, Universidad de Córdoba, Marie Curie-Annex Building, Campus de Rabanales, Córdoba 14071, Spain.
J Anal Toxicol. 2022 Mar 21;46(3):285-294. doi: 10.1093/jat/bkab023.
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.
超分子溶剂具有受限访问属性 (SUPRAS-RAMs)，被提议作为一种新方法，用于整合提取和样品净化，以通过液相色谱-串联质谱 (LC-MS-MS) 定量尿液中的苯二氮䓬类药物 (BDZs)。SUPRAS-RAM 通过在尿液中加入 1-己醇 (154 µL) 和四氢呋喃 (THF) (600 µL) 原位合成。BDZ 的提取是由氢键和分散相互作用共同驱动的。通过 SUPRAS 的化学和物理作用去除蛋白质和极性大分子。通过 SUPRAS 萃取蒸发过程中的沉淀去除磷脂。通过应用 Box-Behnken 响应面设计，采用多元方法优化萃取过程。根据欧洲委员会决定 (2002/657/EC) 的指导原则对该方法进行了验证。目标 BDZs 的方法检测限和定量限分别为 0.21-0.85 和 0.67-2.79 ng/mL。重复性和重现性（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 BDZs（回收率在 84-105%范围内）并去除基质效应。该方法应用于 13 个尿液样本的外部校准分析。在浓度为 5.4-74 ng/mL 的范围内，13 个测试样本中有 5 个检测到阿普唑仑和劳拉西泮呈阳性。该方法允许在最小溶剂消耗的情况下进行简单快速的样品处理，同时提供准确和精确的数据。
