Citation

Yang S, Wen D, Zheng F, Pu S, Chen Z, Chen M, Di B, Liu W, Shi Y. J. Anal. Toxicol. 2023; ePub(ePub): ePub.

Copyright

(Copyright © 2023, Preston Publications)

DOI

10.1093/jat/bkad081

PMID

37929913

Abstract

Amatoxins and phallotoxins are toxic cyclopeptides found in the genus Amanita and are among the predominant causes of foodborne sickness and poisoning-related fatalities in China. This study introduces and validates a simple, rapid, and cost-effective ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) method for the simultaneous determination and quantification of α-amanitin, β-amanitin, γ-amanitin, phallisacin (PSC), phallacidin (PCD), and phalloidin (POD) in human blood and urine. Quick therapeutic decision-making is supported by a 9 min chromatographic separation performed on a Waters Acquity UPLC HSS T3 column (100 mm×2.1 mm, 1.8 µm) using a gradient of HPLC-grade water and methanol:0.005% formic acid. The analyte LOQs were 1-3 ng/mL in blood and 0.5-2 ng/mL in urine. Calibrations curves, prepared by spiking drug-free blood and urine, demonstrated acceptable linearity with mean correlation coefficients (r) greater than 0.99 for all phallotoxins and amatoxins. Acceptable intraday and interday precision (RSD <15%) and accuracy (bias, -4.8% to 13.0% for blood and -9.0% to 14.7% for urine) were achieved. The validated method was successfully applied to analyze 9 blood samples and 2 urine samples testing positive for amatoxins and/or phallotoxins. Amatoxins and/or phallotoxins were identified in each whole blood sample at a range of 1.12-5.63 ng/mL and in two urine samples from 1.01-9.27 ng/mL. The method has the benefits of simple sample preparation (protein precipitation) and wide analyte coverage, making it suitable for emergency quantitative surveillance toxicological analysis in clinics and forensic poisoning practice.

Language: en

Keywords

Amatoxin; Mushroom poisoning; Phallotoxin; ultra-performance liquid chromatography–mass spectrometry