Amanitin determination in bile samples by UHPLC-MS: LR-MS and HR-MS analytical performance.

Autor: Leite M; University of Coimbra, Faculty of Pharmacy, Health Science Campus, Azinhaga de Santa Comba, Coimbra 3000-548, Portugal; National Institute for Agricultural and Veterinary Research (INIAV), Rua dos Lágidos, Lugar da Madalena, Vila do Conde 4485-655, Portugal; REQUIMTE/LAQV, R. D. Manuel II, Apartado, Porto 55142, Portugal., Freitas A; National Institute for Agricultural and Veterinary Research (INIAV), Rua dos Lágidos, Lugar da Madalena, Vila do Conde 4485-655, Portugal; REQUIMTE/LAQV, R. D. Manuel II, Apartado, Porto 55142, Portugal., Mitchell T; Dominican Hospital, Santa Cruz, CA, USA., Barbosa J; National Institute for Agricultural and Veterinary Research (INIAV), Rua dos Lágidos, Lugar da Madalena, Vila do Conde 4485-655, Portugal; REQUIMTE/LAQV, R. D. Manuel II, Apartado, Porto 55142, Portugal., Ramos F; University of Coimbra, Faculty of Pharmacy, Health Science Campus, Azinhaga de Santa Comba, Coimbra 3000-548, Portugal; REQUIMTE/LAQV, R. D. Manuel II, Apartado, Porto 55142, Portugal. Electronic address: framos@ff.uc.pt.
Jazyk: angličtina
Zdroj: Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 2024 Sep 01; Vol. 247, pp. 116253. Date of Electronic Publication: 2024 May 24.
DOI: 10.1016/j.jpba.2024.116253
Abstrakt: Consumption of misidentified foraged mushrooms containing bicyclic amanitin octapeptides is a worldwide public health and veterinary problem, being considered one of the deadliest accidental human and canine food ingestion due to acute liver failure (ALF). Reversal of advanced ALF and complete clinical recovery can be achieved following definitive removal of accumulated amatoxin laden bile from the gallbladder. An accurate means of quantifying amanitin content in aspirated bile is, therefore, urgently needed. Building on our prior work validating a method to detect and quantify amanitin in hepatic autopsy tissue, the development of an accurate method of measuring α- and β-amanitin in aspirated gallbladder bile was performed to evaluate the efficiency of this emergency procedure applied as a clinical treatment for intoxicated patients. A solid-phase extraction (SPE) procedure was optimized followed by detection based on ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS). Low resolution mass spectrometry (LRMS) was compared with high resolution (HRMS) by the validation of UHPLC-MS/MS (triple quadrupole MS) and UHPLC-ToF-MS (time-of-flight MS). Both methods were able to detect amatoxins in bile with limits of detection and quantification ranging from 2.71 to 3.46 µg.kg -1 , and 8.36-9.03 µg.kg -1 for α-amanitin and, 0.32-1.69 µg.kg -1 and 0.55-5.62 µg.kg -1 for β-amanitin, respectively. Validation was completed with the evaluation of linearity, specificity, robustness, recovery, and precision following the ICH guidelines and CIR 808/2021. The validated methods were finally applied to bile samples obtained 48-96 hours + post-ingestion from 4 amatoxin poisoning patients who underwent gallbladder drainage procedures in Vietnam, Canada, and California. Gallbladder bile from patients with amatoxin mushroom poisoning contained significant amanitin content, even when aspirated several days post-ingestion, thus confirming the important role of enterohepatic circulation in amatoxin hepatotoxicity. This work represents a high and unique analytical throughput in amanitin poisoning allowing to efficiently respond to this fatal health problem.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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