Department of Chemistry and Biochemistry, The University of Texas at Arlington, 700 Planetarium Pl., Arlington, TX, 76019, USA.
Scottsdale Research Institute, 12815 N. Cave Creek Rd., Phoenix, AZ, 85022, USA.
Anal Chim Acta. 2024 Feb 1;1288:342161. doi: 10.1016/j.aca.2023.342161. Epub 2023 Dec 18.
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.
建立了一种使用液相色谱-串联质谱（LC-MS/MS）测定迷幻蘑菇品种裸盖菇素和赛洛西宾临床效价的方法。获得并分析了五株干燥完整的蘑菇：蓝魔、爬行者、B-Plus、德克萨斯黄和泰国裸盖菇。开发了一种提取方案；包括评估样品研磨技术、提取溶剂和回收率/稳定性。使用内部标准校准每个分析物的氘代同量异位物，在熔融核颗粒相上开发了反相色谱法来测定这两种分析物。分离时间不到 5 分钟。通过比较校准样品在纯溶液和几种蘑菇基质中的信号响应，研究了基质效应；未观察到明显的基质效应。裸盖菇素的检测限为 1.5ng/mL（1.5pg 柱上；300ng/g 蘑菇），赛洛西宾的检测限为 0.15ng/mL（0.15pg 柱上；30ng/g 蘑菇），使用 Shimadzu LCMS-8050 三重四极杆质谱仪。方法准确性和精密度的评估表明，所有浓度水平的相对误差和 RSD 均<6%。从每个菌株中单独分析三个完整的、完整的蘑菇，以获得菌株之间和同一菌株的蘑菇之间的平均含量差异。从最有效到最无效，研究发现，爬行者、蓝魔、B+、德克萨斯黄和泰国裸盖菇素菌株的总裸盖菇素和赛洛西宾的平均浓度分别为 1.36%、1.221%、1.134%、1.103%和 0.879%（w/w）。这些蘑菇的一部分也在另一个独立的实验室进行了测试，两个实验室的结果具有可比性。第二个实验室的结果表明，在提取前将多个蘑菇混合均匀可以提高精密度。
