A High Throughput Platform for Multiplexed Screening of Drugs of Abuse by Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry

鉴于止痛类阿片和其他类别处方药成瘾在世界范围内的流行，迫切需要新技术来更准确、更可靠地筛选滥用药物（DoA）。加拿大处方药滥用问题的社会经济影响对公共卫生保健和刑事司法系统造成了毁灭性的影响，包括精神疾病和药物滥用、生产力损失，以及因药物过量（如强效合成阿片类药物芬太尼）导致的死亡人数惊人地增加。该项目将引入一个高通量平台，使用多段注射-毛细管电泳-质谱（MSI-CE-MS）对尿液样本中的DoA进行多路筛选，适用于经认可的临床或法医毒理学实验室。目前对DoA进行初步筛选的方法依赖于免疫测定，由于选择性差，容易出现假阳性和假阴性，由于无法获得或长时间延迟获得仅针对单一药物的可行抗体试剂，这种方法不适用于新兴药物类别。虽然液相色谱-串联质谱（LC-MS/MS）用于筛查阳性尿液样本的确认性测试，但由于其样品通量较低和化学分析的针对性，它不适合用于初步筛选。我们将通过优化和严格验证基于MSI-CE-MS的专利筛选技术来克服这些限制，该技术依赖于在单次运行中连续注射多达10个尿液样本（< 3分钟/样本），其中多路电泳分离与高分辨率相结合，精确的MS使用飞行时间质量分析仪进行全扫描数据采集。这种方法将为具有高数据保真度的初级筛选提供更有选择性、更精确和成本更低的格式，这也将使在可接受的截止范围以上的几乎无限的DoA面板的明确识别和检测成为可能。基于MSI-CE-MS的非靶向药物筛选也将有助于在最终用户冒着极大风险消费的掺假或掺假街头毒品中发现新的致命阿片类类似物。这项创新技术将为传统的免疫分析提供一个更好的筛选平台，以实现更有效的治疗药物监测计划和改善患者护理，同时防止药物转移和意外药物过量的悲惨案例。

New technologies are urgently required for more accurate yet reliable screening of drugs of abuse (DoA) given the worldwide epidemic of addiction to pain relief opioids and other classes of prescription medications. The socioeconomic impacts to the prescription drug abuse problem in Canada has been devastating to public health care and the criminal justice system, including mental illness and substance abuse, lost productivity, and an alarming increase in deaths due to drug overdose, such as the highly potent synthetic opioid, fentanyl. This project will introduce a high throughput platform for multiplexed screening of DoA in urine samples using multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS) that is suitable for implementation within an accredited clinical or forensic toxicology laboratory. Current approaches for primary screening of DoA rely on immunoassays that are prone to false positives and false negatives due to poor selectivity, which are not applicable to emerging classes of drugs due to the unavailability or long delays to access of viable antibody reagents that target only a single drug. Although liquid chromatography-tandem mass spectrometry (LC-MS/MS) is used for confirmatory testing of screen-positive urine samples, it is not suitable for primary screening due its lower sample throughput and targeted nature of chemical analysis. We will overcome these limitations by optimizing and rigorously validating a patented screening technology based on MSI-CE-MS that relies on serial injections of up to ten urine samples within a single run (< 3 min/sample), where multiplexed electrophoretic separations are coupled to high resolution, accurate MS with full-scan data acquisition using a time-of-flight mass analyzer. This approach will offer a more selective, precise and less costly format for primary screening with high data fidelity, which will also enable unambiguous identification and detection of virtually an unlimited panel of DoA above accepted cut-off limits. Non-targeted drug screening based on MSI-CE-MS will also enable the discovery of new classes of lethal opioid analogs in adulterated or laced street drugs consumed at great risk by end-users. This innovative technology will introduce a superior screening platform to conventional immunoassays as required for more effective therapeutic drug monitoring programs and improved patient care while preventing drug diversion and tragic cases of accidental drug overdose.