Laboratoire de sciences judiciaires et de médecine légale, Department of Toxicology, 1701 Parthenais Street, Montréal, Québec, H2K 3S7, Canada; Concordia University, Department of Chemistry & Biochemistry, 7141 Sherbrooke Street West, Montréal, Québec, H4B 1R6, Canada.
Laboratoire de sciences judiciaires et de médecine légale, Department of Toxicology, 1701 Parthenais Street, Montréal, Québec, H2K 3S7, Canada.
Forensic Sci Int. 2020 Dec;317:110506. doi: 10.1016/j.forsciint.2020.110506. Epub 2020 Sep 13.
A study of impaired driving rates in the province of Québec is currently planned following the legalization of recreational cannabis in Canada. Oral fluid (OF) samples are to be collected with a Quantisal® device and sent to the laboratory for analysis. In order to prepare for this project, a qualitative decision point analysis method monitoring for the presence of 97 drugs and metabolites in OF was developed and validated. This high throughput method uses incubation with a precipitation solvent (acetone:acetonitrile 30:70 v:v) to boost drug recovery from the collecting device and improve stability of benzodiazepines (e.g., α-hydroxyalprazolam, clonazepam, 7-aminoclonazepam, flunitrazepam, 7-aminoflunitrazepam, N-desmethylflunitrazepam, nitrazepam). The Quantisal® device has polyglycol in its stabilizing buffer, but timed use of the mass spectrometer waste valve proved sufficient to avoid the glycol interferences for nearly all analytes. Interferences from OF matrices and 140 potentially interfering compounds, carryover, ion ratios, stability, recovery, reproducibility, robustness, false positive rate, false negative rate, selectivity, sensitivity and reliability rates were tested in the validation process. Five of the targeted analytes (olanzapine, oxazepam, 7-aminoclonazepam, flunitrazepam and nitrazepam) did not meet the set validation criteria but will be monitored for identification purposes (no comparison to a cut-off level). Blind internal proficiency testing was performed, where six OF samples were tested and analytes were classified as "negative", "likely positive" or "positive" with success. The final validated OF qualitative decision point method covers 92 analytes, and the presence of 5 additional analytes is screened in this high throughput analysis.
安大略省拟在加拿大娱乐用大麻合法化后开展一项省内酒后驾车率研究。将使用 Quantisal®装置采集唾液（OF）样本并送至实验室进行分析。为了筹备该项目，开发并验证了一种定性检测点分析方法，以监测 OF 中 97 种药物和代谢物的存在情况。该高通量方法使用沉淀溶剂（丙酮：乙腈 30:70v:v）孵育，以提高从采集装置中提取药物的回收率并提高苯二氮䓬类药物（如 α-羟基阿普唑仑、氯硝西泮、7-氨基氯硝西泮、氟硝西泮、7-氨基氟硝西泮、去甲氟硝西泮、硝西泮）的稳定性。Quantisal®装置的稳定缓冲液中含有聚乙二醇，但经过定时使用质谱仪废液阀，可避免聚乙二醇对几乎所有分析物的干扰。在验证过程中测试了 Quantisal®装置的基质干扰和 140 种潜在干扰化合物、交叉污染、离子比、稳定性、回收率、重现性、稳健性、假阳性率、假阴性率、选择性、灵敏度和可靠性。有 5 种目标分析物（奥氮平、 地西泮、7-氨基氯硝西泮、氟硝西泮和硝西泮）未达到设定的验证标准，但将作为识别目的进行监测（不与截止值进行比较）。进行了盲法内部能力验证测试，对 6 个 OF 样本进行了测试，成功将分析物分类为“阴性”、“可能阳性”或“阳性”。最终验证的 OF 定性检测点方法涵盖 92 种分析物，同时该高通量分析还筛选了另外 5 种分析物的存在情况。
