Analyzing DNA barcoding and identifying toxins caused by neurotoxic mushroom poisoning using liquid chromatography tandem mass spectrometry
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Abstract
Background: Neurotoxic mushroom poisoning often exhibits rapid symptom onset, typically attributed to compounds such as Ibotenic acid, which affect the central nervous system. This study addresses a new case of mushroom-related food poisoning in southern Thailand.
Objective: The objectives are to determine the presence of ibotenic acid in cases of mushroom-related food poisoning utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and to identify toxic Amanita species implicated in these cases.
Materials and methods: Remnant mushroom samples obtained from three clinically reported cases were used. Nucleotide similarity was compared against the rRNA/ ITS databases using NCBI BLAST search. Phylogenetic analyses were conducted using maximum likelihood (ML) and FastTree approaches. LC-MS/MS was employed to separate of Ibotenic acid, determine its molecular weight and perform precursor ion fragmentation.
Results: Analysis of the rRNA/ITS databases revealed a high nucleotide similarity between suspected mushroom samples and Amanita digitosa. Detailed phylogenetic analysis confirmed that mushroom samples from the three poisoning cases clustered with A. digitosa. LC-MS/MS analysis showed the presence of ibotenic acid, with precursor ion (m/z 159) and product ion (m/z 113.1) as the major toxic substances. Clinically, patients poisoned by ibotenic acid-containing mushrooms exhibited a short latent period with symptoms of nausea, vomiting, vertigo, delirium, confusion, and fatigue.
Conclusion: The genus Amanita comprises both edible and inedible species that produce several lethal toxins. The report of ibotenic acid in A. digitosa is a novel finding, valuable for food safety monitoring and healthcare decision-maker. This is especially notable due to the accuracy and rapidity of the analytical process.
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