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

鉴于世界范围内对止痛药、阿片类药物和其他类别处方药上瘾的流行，迫切需要新技术来更准确、更可靠地筛查滥用药物(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、生产力丧失，以及因药物过量而死亡的人数惊人地增加，如高度有效的合成阿片类药物芬太尼。该项目将引入一个高通量平台，用于使用多段注射-毛细管电泳质谱(MSI-CE-MS)对尿样中DOA进行多路筛选，该平台适合在经认可的临床或法医毒理学实验室内实施。目前用于DOA初步筛选的方法依赖于免疫分析，但由于选择性较差，容易出现假阳性和假阴性，而由于无法获得或长期延迟获得仅针对单一药物的有效抗体试剂，这些方法不适用于新兴药物。虽然LC-MS/MS(LC-MS/MS)用于筛查阳性尿样的确证检测，但由于其样品吞吐量较低和化学分析的针对性，不适合进行初步筛查。我们将通过优化和严格验证基于MSI-CE-MS的专利筛查技术来克服这些限制，该筛查技术依赖于在一次运行(&lt；3分钟/样本)中连续注射多达10个尿样，其中多路电泳分离与使用飞行时间质谱仪进行全扫描数据采集的高分辨率、准确的MS相结合。这种方法将为具有高数据保真度的初步筛查提供更具选择性、更精确和成本更低的格式，这也将使几乎不受限制的高于可接受截止值的DOA小组能够明确地识别和检测。基于MSI-CE-MS的非靶向药物筛查还将能够在终端用户高风险消费的掺假或掺入的街头药物中发现新的致命阿片类似物。这项创新技术将为更有效的治疗药物监测计划和改善患者护理所需的传统免疫分析引入一个优越的筛查平台，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.