Screening method for the analysis of blood and urine for the presence of naturally occurring toxic compounds from mushrooms (fungi) using the LC-MS method

A qualitative method using solid phase extraction and liquid chromatography with quadrupole time-of-flight mass spectrometry (LC/QTOF) was developed to detect five fungal toxins—α-amanitin, β-amanitin, γ-amanitin, muscarine, and psilocin—in blood and urine. The method uses HLB columns and acetonitrile-based extraction, with detection limits ranging from 0.3 to 2.1 ng/mL depending on the compound and matrix. It enables efficient identification of these toxins in forensic toxicology, addressing challenges in analyzing biological material from autopsies or living humans for mushroom poisoning.

Study at a glance

Abstract

Analysis of biological material collected during autopsies and even from living humans for the presence of amanitins and other fungal toxins remains a challenge in forensic toxicology. A qualitative method for the detection of α-amanitin, β-amanitin, γ-amanitin, muscarine, and psilocin in blood and urine has been developed. To achieve this goal, solid phase extraction HLB 3cc 60mg columns were used. Blood and urine samples were purified with water and aqueous methanol solution, and then extracted with acetonitrile. An LC/QTOF system equipped with a C18 column was applied to identify the analytes. Acetonitrile and water with formic acid were utilized as mobile phases. The developed method was validated. The detection limits for α-amanitin, β-amanitin, γ-amanitin, muscarine and psilocin are, respectively, 1.4 ng/ml, 0.3 ng/ml, 1.2 ng/ml, 1.8 ng/ml, and 0.3 ng/ml in blood, and 1.5 ng/ml, 2.1 ng/ml, 1.5 ng/ml, 1.6 ng/ml, and 1.1 ng/ml in urine. The developed method allows for efficient, qualitative identification of all the above-mentioned compounds in a toxicological laboratory.