Determination of amatoxin concentration in heat-treated samples of Amanita phalloides by high-performance liquid chromatography: A forensic approach

Sharma, Spriha; Aydın, Meryem; Bansal, Gulshan; Kaya, Ertuğrul; Singh, Rajinder

doi:10.1016/j.jflm.2020.102111

Cited by 5 publications

(6 citation statements)

References 27 publications

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“…Freshness of the samples analyzed also influences the results of HLPC, but little information exists about the degradation of amatoxins and phallotoxins during storage. Stijve & Seeger [99] observed a decrease in amanitins (α-and β-) and phalloidin contents over time in specimens collected in the same location in different years and suggest that phallotoxins might be less stable during storage than amanitins, whose stability is known to depend on treatment and storage of the samples [100][101][102]. Results obtained in the present work support this hypothesis, since at least αand β-amanitins seem less abundant in older specimens (i.e., A. vidua from 1987 vs. A. verna from 2001).…”

Section: Discussionmentioning

confidence: 99%

Amanita Section Phalloideae Species in the Mediterranean Basin: Destroying Angels Reviewed

Alvarado¹,

Gasch-Illescas

Morel

et al. 2022

Biology

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In Europe, amatoxin-containing mushrooms are responsible for most of the deadly poisonings caused by macrofungi. The present work presents a multidisciplinary revision of the European species of Amanita sect. Phalloideae based on morphology, phylogeny, epidemiology, and biochemistry of amatoxins and phallotoxins. Five distinct species of this section have been identified in Europe to date: A. phalloides, A. virosa, A. verna, the recently introduced North American species A. amerivirosa, and A. vidua sp. nov., which is a new name proposed for the KOH-negative Mediterranean species previously described as A. verna or A. decipiens by various authors. Epitypes or neotypes are selected for species lacking suitable reference collections, namely A. verna and A. virosa. Three additional taxa, Amanita decipiens, A. porrinensis, and A. virosa var. levipes are here considered later heterotypic synonyms of A. verna, A. phalloides, and A. amerivirosa, respectively.

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Section: Discussionmentioning

confidence: 99%

Amanita Section Phalloideae Species in the Mediterranean Basin: Destroying Angels Reviewed

Alvarado¹,

Gasch-Illescas

Morel

et al. 2022

Biology

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“…These results are consistent with the view that α-amanitin is thermally stable. In contrast, the results reported by Sharma et al [31] and Kaya et al [32] did not measure toxin activity. In both cases, they used HPLC to quantify α-amanitin after heat treatment in mushroom materials and purified toxin in sealed vials, respectively.…”

Section: Discussionmentioning

confidence: 74%

“…In this study, it was demonstrated that the biological activity of α-amanitin remains after a heat treatment. This question was raised after a recent publication by Sharma et al [31] reported that α-amanitin is thermally degraded in mushrooms subjected to common cooking methods, i.e., boiling, baking, grilling, and frying. They found the greatest reduction in boiled mushrooms but noted that this was likely to be partly due to the toxin's high water solubility.…”

Section: Discussionmentioning

confidence: 99%

“…These commonly used detection methods can only detect the presence of amatoxins and not their activity. It has recently been reported by Sharma et al [31], using high-performance liquid chromatography (HPLC), that α-amanitin is thermally degraded in mushrooms that have been subjected to common cooking methods, i.e., boiling, baking, grilling, and frying. Kaya et al [32] reported that the remaining level of α-amanitin was only 5% after 6 h in boiling water.…”

Section: Introductionmentioning

confidence: 99%

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A Sensitive, Cell-Based Assay for Measuring Low-Level Biological Activity of α-Amanitin

Rasooly,

Do,

et al. 2023

IJMS

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α-Amanitin is one of the primary toxins produced by the poisonous mushroom genus, Amanita. Because it is odorless and tasteless, it is an important cause of death from the consumption of misidentified mushrooms. To study the thermal stability of α-amanitin, novel cell-based assays were developed to measure the toxin’s activity, based on the inhibition of RNA polymerase II by α-amanitin. First, an MTT–formazan cell viability assay was used to measure the biological activity of α-amanitin through the inhibition of cellular activity. This method can detect 10 μg/mL of α-amanitin in a time-dependent manner. Second, a more sensitive quantitative PCR approach was developed to examine its inhibition of viral replication. The new RT-qPCR assay enabled the detection of 100 ng/mL. At this level, α-amanitin still significantly reduced adenovirus transcription. Third, a simpler GFP expression-based assay was developed with an equal sensitivity to the RT-qPCR assay. With this assay, aqueous α-amanitin heated at 90 °C for 16 h or treated in the microwave for 3 min retained its biological activity when tested in HEK293 cells, but a slight reduction was observed when tested in Vero cells. Beyond detecting the activity of α-amanitin, the new method has a potential application for detecting the activity of other toxins that are RNA polymerase inhibitors.

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“…In recent investigations by Sharma and coworkers [28], death cap samples were treated with various heat and technical conditions (frying, grilling, boiling, baking). The duration of these treatments was 30 min, but the temperatures used were not published.…”

Section: Compoundsmentioning

confidence: 99%

Amanitins: The Most Poisonous Molecules of the Fungal World

Vetter

2023

Molecules

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Among the toxic metabolites of the fungal world, those that, due to their strong biological effect, can seriously (even fatally) damage the life processes of humans (and certain groups of animals) stand out. Amatoxin-containing mushrooms and the poisonings caused by them stand out from the higher fungi, the mushrooms. There are already historical data and records about such poisonings, but scientific research on the responsible molecules began in the middle of the last century. The goals of this review work are as follows: presentation of the cosmopolitan mushroom species that produce amanitins (which are known from certain genera of four mushroom families), an overview of the chemical structure and specific properties of amanitins, a summary of the analytical methods applicable to them, a presentation of the “medical history” of poisonings, and a summary of the therapeutic methods used so far. The main responsible molecules (the amanitins) are bicyclic octapeptides, whose structure is characterized by an outer loop and an inner loop (bridge). It follows from the unusual properties of amanitins, especially their extreme stability (against heat, the acidic pH of the medium, and their resistance to human, and animal, digestive enzymes), that they are absorbed almost without hindrance and quickly transported to our vital organs. Adding to the problems is that accidental consumption causes no noticeable symptoms for a few hours (or even 24–36 h) after consumption, but the toxins already damage the metabolism of the target organs and the synthesis of nucleic acid and proteins. The biochemical catastrophe of the cells causes irreversible structural changes, which lead to necrotic damage (in the liver and kidneys) and death. The scientific topicality of the review is due to the recent publication of new data on the probable antidote molecule (ICR: indocyanine green) against amanitins. Further research can provide a new foundation for the therapeutic treatment of poisonings, and the toxicological situation, which currently still poses a deadly threat, could even be tamed into a controllable problem. We also draw attention to the review conclusions, as well as the mycological and social tasks related to amanitin poisonings (prevention of poisonings).

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Determination of amatoxin concentration in heat-treated samples of Amanita phalloides by high-performance liquid chromatography: A forensic approach

Cited by 5 publications

References 27 publications

Amanita Section Phalloideae Species in the Mediterranean Basin: Destroying Angels Reviewed

Amanita Section Phalloideae Species in the Mediterranean Basin: Destroying Angels Reviewed

A Sensitive, Cell-Based Assay for Measuring Low-Level Biological Activity of α-Amanitin

Amanitins: The Most Poisonous Molecules of the Fungal World

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