Division of Clinical Pharmacology and Toxicology, Department of Biomedicine, University Hospital Basel and University of Basel, Basel, Switzerland; Division of Clinical Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.
Division of Clinical Pharmacology and Toxicology, Department of Biomedicine, University Hospital Basel and University of Basel, Basel, Switzerland; Division of Clinical Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Feb 1;1164:122486. doi: 10.1016/j.jchromb.2020.122486. Epub 2020 Dec 7.
Psilocin is the active metabolite of psilocybin, a serotonergic psychedelic substance. It is used recreationally and investigated in substance-assisted psychotherapy. The pharmacokinetic properties of psilocin are only partially characterized. Therefore, we developed and validated a rapid LC-MS/MS method to quantify psilocin and its metabolite 4-hydroxyindole-3-acetic acid (4-HIAA) in human plasma. Plasma samples were processed by protein precipitation using methanol. The injected sample was mixed with water in front of a C analytical column to increase retention of the analytes. Psilocin and 4-HIAA were detected by multiple reaction monitoring (MRM) in positive and negative electrospray ionisation mode, respectively. An inter-assay accuracy of 100-109% and precision of ≤8.7% was recorded over three validation runs. The recovery was near to complete (≥94.7%) and importantly, consistent over different concentration levels and plasma batches (CV%: ≤4.1%). The plasma matrix caused negligible ion suppression and endogenous interferences could be separated from the analytes. Psilocin and 4-HIAA plasma samples could be thawed and re-frozen for three cycles, kept at room temperature for 8 h or 1 month at -20 °C without showing degradation (≤10%). The linear range (R ≥ 0.998) of the method covered plasma concentrations observed in humans following a common therapeutic oral dose of 25 mg psilocybin and was therefore able to assess the pharmacokinetics of psilocin and 4-HIAA. The LC-MS/MS method was convenient and reliable for measuring psilocin and 4-HIAA in plasma and will facilitate the clinical development of psilocybin.
裸盖菇素是裸盖菇碱的活性代谢物，一种血清素能致幻物质。它被娱乐性使用，并在物质辅助心理疗法中进行研究。裸盖菇素的药代动力学特性仅部分得到描述。因此，我们开发并验证了一种快速 LC-MS/MS 方法，用于定量人血浆中的裸盖菇素及其代谢物 4-羟基吲哚-3-乙酸（4-HIAA）。使用甲醇通过蛋白沉淀处理血浆样品。在 C 分析柱前将注入的样品与水混合，以增加分析物的保留。裸盖菇素和 4-HIAA 通过正、负离子电喷雾电离模式的多重反应监测（MRM）分别进行检测。在三个验证运行中，记录了 100-109%的批内准确度和≤8.7%的精密度。回收率接近完全（≥94.7%），重要的是，在不同浓度水平和血浆批次之间保持一致（CV%：≤4.1%）。血浆基质几乎没有引起离子抑制，并且可以将内源性干扰与分析物分离。裸盖菇素和 4-HIAA 血浆样品可以解冻并重新冷冻三个循环，可以在室温下 8 小时或在-20°C 下保存 1 个月而不会降解（≤10%）。该方法的线性范围（R≥0.998）涵盖了人类在接受 25mg 裸盖菇碱常见治疗口服剂量后血浆中观察到的浓度，因此能够评估裸盖菇素和 4-HIAA 的药代动力学。LC-MS/MS 方法方便可靠，可用于测量血浆中的裸盖菇素和 4-HIAA，将有助于裸盖菇素的临床开发。
