Abstract：
A method for clinical potency determination of psilocybin and psilocin in hallucinogenic mushroom species Psilocybe cubensis was developed using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Five strains of dried, intact mushrooms were obtained and analyzed: Blue Meanie, Creeper, B-Plus, Texas Yellow, and Thai Cubensis. An extraction protocol was developed; this included an evaluation of sample milling technique, extraction solvents, and recovery/stability. Reversed phase chromatography on fused-core particle phases was developed for the determination of the two analytes using internal standard calibration with deuterated isotopologues of each analyte. The separation takes less than 5 min. Matrix effects were investigated by comparing signal response of calibration samples in neat solution and several mushroom matrices; no significant matrix effects were observed. The limit of detection for psilocybin was 1.5 ng/mL (1.5 pg on-column; 300 ng/g mushroom) and for psilocin was 0.15 ng/mL (0.15 pg on-column; 30 ng/g mushroom) using a Shimadzu LCMS-8050 triple quadrupole mass spectrometer. Assessment of the accuracy and precision of the method indicated percent error and RSD were <6 % at all concentration levels. Three whole, intact mushrooms from each strain were analyzed individually to obtain average content differences both between strains and between mushrooms of the same strain. From most to least potent, the study found that the average total psilocybin and psilocin concentrations for the Creeper, Blue Meanie, B+, Texas Yellow, and Thai Cubensis strains were 1.36, 1.221, 1.134, 1.103, and 0.879 % (w/w), respectively. A subset of these mushrooms was also tested in a separate non-affiliated laboratory, and the results were comparable between the two laboratories. Results from the secondary laboratory showed improved precision when multiple mushrooms were homogenized together, prior to extraction.
摘要：
建立了一种使用液相色谱-串联质谱（LC-MS/MS）测定致幻蘑菇中psilocybin和psilocin的临床效价的方法。五株干的，获得并分析了完整的蘑菇：蓝色Meanie，爬山虎,B-Plus,德州黄,和泰国古巴。开发了提取方案；这包括对样品研磨技术的评估，萃取溶剂，和恢复/稳定性。开发了在融合核心颗粒相上的反相色谱，用于使用每种分析物的氘代同位素进行内标校准来测定两种分析物。分离需要少于5分钟。通过比较纯溶液和几种蘑菇基质中校准样品的信号响应来研究基体效应；没有观察到明显的基体效应。使用ShimadzuLCMS-8050三重四极杆质谱仪，psilocybin的检测限为1.5ng/mL（柱上1.5pg；300ng/g蘑菇），而psilocin的检测限为0.15ng/mL（柱上0.15pg；30ng/g蘑菇）。该方法的准确度和精密度的评估表明,在所有浓度水平下,误差百分比和RSD均<6%。三个整体，分别分析每个菌株的完整蘑菇，以获得菌株之间和同一菌株的蘑菇之间的平均含量差异。从最强大到最不强大，研究发现，爬山虎的平均总psilocybin和psilocin浓度，蓝色Meanie,B+,德州黄,和泰国Cubensis菌株分别为1.36、1.221、1.134、1.103和0.879%(w/w)，分别。这些蘑菇的一部分也在一个独立的非附属实验室进行了测试，两个实验室的结果具有可比性。二级实验室的结果显示，当多个蘑菇一起均质时，精度得到了提高，在提取之前。
