Cannabis is still the most widely used illegal plant in the world. However, cannabis use is prohibited in many countries. After cannabis use, Δ9-tetrahydrocannabinol is metabolized in the liver to 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) and most undergo glucuronidation. THC-COOH and THC-COOH glucuronide are excreted in the urine. The total concentration of THC-COOH in the urine sample is measured to determine cannabis use. The total concentration is determined after enzymatic or alkaline hydrolysis. In this study, comparing enzymatic hydrolysis efficiency is presented comprehensively together with the method developed for the determination of total THC-COOH in the urine. Also, the method was validated according to the European Medicines Agency Guidelines on bioanalytical method validation. The method has rapid hydrolysis time (20 min), rapid analysis time (5 min) and simple sample preparation. The lower limit of quantitation of the developed method was 1 ng/mL for THC-COOH. The calibration curve of THC-COOH was between 1 and 2,000 ng/mL with a correlation coefficient >0.99. Also, the method was applied to real patient’s urine. We think that the results will provide a new perspective on enzymatic hydrolysis optimization studies.

中文翻译：

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UPLC-MS-MS 测定大麻使用检测中的酶水解效率

大麻仍然是世界上使用最广泛的非法植物。然而，许多国家禁止使用大麻。使用大麻后，Δ9-四氢大麻酚在肝脏中代谢为 11-nor-9-羧基-Δ9-四氢大麻酚 (THC-COOH)，并且大多数会发生葡萄糖醛酸化。THC-COOH 和 THC-COOH 葡糖苷酸通过尿液排出。测量尿液样本中 THC-COOH 的总浓度以确定大麻的使用情况。总浓度在酶解或碱解后测定。在这项研究中，比较酶水解效率与开发用于测定尿液中总 THC-COOH 的方法一起全面介绍。此外，该方法根据欧洲药品管理局生物分析方法验证指南进行了验证。该方法具有快速水解时间（20 min），快速分析时间（5 分钟）和简单的样品制备。对于 THC-COOH，所开发方法的定量下限为 1 ng/mL。THC-COOH 的校准曲线在 1 和 2,000 ng/mL 之间，相关系数＞0.99。此外，该方法应用于真实患者的尿液。我们认为该结果将为酶水解优化研究提供新的视角。