Thomann Jan, Kraus Selina, Erne Livio, Vogt Severin B, Liechti Matthias E, Luethi Dino
Division of Clinical Pharmacology and Toxicology, Department of Biomedicine, University Hospital Basel, Basel, Switzerland.
Division of Clinical Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.
Biomed Chromatogr. 2025 Sep;39(9):e70171. doi: 10.1002/bmc.70171.
Ketamine, a well-established dissociative anesthetic, has recently gained significant attention for its rapid-acting antidepressant effects, particularly in treatment-resistant depression. In this study, we developed and validated a state-of-the-art liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the bioanalysis of ketamine and its metabolites, norketamine, dehydronorketamine (DHNK), and (2R,6R)-hydroxynorketamine (HNK), in human plasma. The method features a small sample volume, a streamlined protein precipitation protocol, and a rapid sample runtime. The mobile phase gradient is composed of an aqueous ammonium hydrogen carbonate solution and pure acetonitrile. Using positive electrospray ionization, linear quantification ranges of 1-1,000 ng/mL were established for ketamine and norketamine, while ranges of 0.25-100 ng/mL for DHNK and 2.5-1,000 ng/mL for (2R,6R)-HNK were achieved. The method demonstrated high accuracy, precision, selectivity, and sensitivity, along with consistent matrix effects, efficient extraction recovery, and analyte stability. Finally, the method was successfully applied to assess the pharmacokinetics of six clinical trial participants. Overall, this LC-MS/MS method offers a robust and efficient approach for the achiral quantification of ketamine and its metabolites in human plasma. Its minimal sample preparation and reduced analytical runtime make it particularly well-suited for clinical studies, drug monitoring, and forensic investigations.
氯胺酮是一种成熟的解离麻醉剂，最近因其快速起效的抗抑郁作用而备受关注，尤其是在难治性抑郁症的治疗中。在本研究中，我们开发并验证了一种先进的液相色谱-串联质谱（LC-MS/MS）方法，用于生物分析人血浆中的氯胺酮及其代谢物去甲氯胺酮、脱氢去甲氯胺酮（DHNK）和（2R,6R）-羟基去甲氯胺酮（HNK）。该方法具有进样量小、简化的蛋白沉淀方案和快速的样品运行时间等特点。流动相梯度由碳酸铵水溶液和纯乙腈组成。采用正电喷雾电离，建立了氯胺酮和去甲氯胺酮1-1000 ng/mL的线性定量范围，而DHNK为0.25-100 ng/mL，（2R,6R）-HNK为2.5-1000 ng/mL。该方法具有高准确度、精密度、选择性和灵敏度，同时具有一致的基质效应、高效的提取回收率和分析物稳定性。最后，该方法成功应用于评估6名临床试验参与者的药代动力学。总体而言，这种LC-MS/MS方法为非手性定量人血浆中的氯胺酮及其代谢物提供了一种强大而有效的方法。其最少的样品制备和缩短的分析运行时间使其特别适合临床研究、药物监测和法医调查。
