Ayahuasca tea is an entheogen hallucinogenic beverage used for shamanic and spiritual purposes, prepared by the decoction of different Amazonian plants containing N,N-dimethyltryptamine (DMT) and harmala alkaloids. Since the therapeutic potential of this tea has been broadly studied in recent years, mainly for the treatment of psychiatric disorders, the determination of the ayahuasca tea components in human and animal matrices is of utmost importance. In order to avoid the use of large amounts of toxic solvents, typically employed in traditional sample preparation methods, hollow fiber liquid-phase microextraction (HF-LPME) presents a greener and time-saving alternative. The present study aims to fully develop and apply an HF-LPME method for the determination of DMT, harmine (HRM), harmaline (HRL), and tetrahydroharmine (THH) in human urine samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Fractional factorial and Box–Behnken designs were used to identify and optimize significant method variables. Once optimized, validation has shown a limit of detection (LoD) of 1.0 ng/ml for DMT and 2.0 ng/ml for the harmala alkaloid. The limit of quantification (LoQ) was of 5.0 ng/ml for all analytes. The method has shown to be linear over a concentration range of 5–200 ng/ml (r² ≥ 0.99). Intra/inter-day precision and accuracy met the acceptance criteria at the three quality control (QC) levels studied (15.0, 90.0, and 170.0 ng/ml, n = 6, each). Matrix effect evaluation showed predominant ion enhancement and recovery values were above 80%. Dilution factors of 10- and 20-fold have shown acceptable values of accuracy. Selectivity studies showed no interferences. Analysis of eight authentic samples collected from four subjects proved method feasibility. A simple, time-saving and green alternative for the analysis of DMT and harmala alkaloids in human urine samples was developed, optimized using design of experiments, fully validated and applied to authentic samples.

Ayahuasca茶是一种用于致巫术和精神目的的致幻剂饮料，由含有N，N-二甲基色胺（DMT）和harmala生物碱的亚马逊不同植物水煎制而成。由于近年来对这种茶的治疗潜力进行了广泛的研究，主要用于治疗精神疾病，因此，确定人和动物基质中的hua花茶成分至关重要。为了避免使用通常在传统样品制备方法中通常使用的大量有毒溶剂，中空纤维液相微萃取（HF-LPME）提供了一种绿色环保且节省时间的替代方法。本研究旨在全面开发和应用HF-LPME方法测定DMT，甜菜碱（HRM），甜菜碱（HRL），液相色谱-串联质谱法（LC-MS / MS）测定人尿液中的四氢大麻酚（THH）。分数阶乘和Box–Behnken设计用于识别和优化重要的方法变量。优化后，验证显示DMT的检出限（LoD）为1.0 ng / ml，而harmala生物碱的检出限（LoD）为2.0 ng / ml。所有分析物的定量限（LoQ）为5.0 ng / ml。该方法在5–200 ng / ml的浓度范围内呈线性关系（所有分析物为0 ng / ml。该方法在5–200 ng / ml的浓度范围内呈线性关系（所有分析物为0 ng / ml。该方法在5–200 ng / ml的浓度范围内呈线性关系（[R² ≥0.99）。在研究的三个质量控制（QC）水平（15.0、90.0和170.0 ng / ml，ñ= 6，每个）。基质效应评估表明，主要的离子增强和回收率值均高于80％。稀释倍数为10倍和20倍，显示出可接受的精度值。选择性研究表明没有干扰。分析从四个受试者收集的八个真实样品证明了该方法的可行性。开发了一种简单，省时且绿色的分析方法，用于分析人类尿液样品中的DMT和harmala生物碱，并通过实验设计对其进行了优化，充分验证并应用于真实样品。