Determination of amatoxins and phallotoxins in Amanita phalloides mushrooms from northeastern Portugal by HPLC-DAD-MS

Abstract

Amanita phalloides is a toxic mushroom responsible for the majority of deaths occurring after mushrooms ingestion, mainly due to amatoxins. In the present study the contents and distribution of the major amatoxins and phallotoxins in different tissues of A. phalloides from two different sites of Portugal were analyzed by liquid chromatography (LC) coupled to diode array (DAD) and mass spectrometry (MS) detection. The main toxins were separated by LC and its chemical structures confirmed by MS. a-Amanitin contents in caps, stipe and volva tissues were quantified by RP-HPLC. The results show that caps have the highest content of amatoxins, whereas the volva was richest in phallotoxins. Moreover variability in the toxins composition from different geographic sites was also observed. This study provides for the first time the content of toxins in A. phalloides from Portugal.

A Rapid Extraction Method Combined with a Monoclonal Antibody-Based Immunoassay for the Detection of Amatoxins

“…Maceration of dried mushroom samples increases exposure of the researcher to the toxin-containing dust. In our previous work, and that of many others, the mushroom tissue was ground to a powder [11][12][13][14]19,20,25]. In this study, however, we did not grind the samples and still achieved sufficient toxin extraction, suitable for cELISA detection.…”

“…Furthermore, the detection of AMAs in the 27,000-fold dilution samples of these AMA-containing sample extracts provided evidence that we were obtaining a high recovery of AMA. Other investigators performed a second extraction following a 1 h extraction (using an acetonitrile solution) and did not recover any detectable amounts of residual toxin in the second extract [14]. While most of the AMAs appear to be easily extracted, further work is needed to determine the recovery coefficient for the rapid extraction procedure.…”

“…A single dried mushroom typically contains around 1-2 mg g −1 of α-AMA [8,10,11]. The most common method for the detection of AMAs extracted from mushrooms is liquid chromatography (LC), coupled with UV detection or mass spectrometry (MS) [8,[12][13][14]. Although these methods are sensitive and provide a high resolution of individual analytes, they are time-consuming and require expensive, laboratory-based instrumentation and highly trained personnel to interpret the results.…”

“…Regardless of the method used to detect the toxin, extraction of the AMA is required before identification. Over the years, the extraction procedure has been streamlined from 24 h [8,10,19] to one hour [12,14,16,20]. Most of these methods have utilized an extraction solution consisting of methanol, acid, and water.…”

Mushrooms Intoxications

“…Death cap (Amanita phalloides) poisoning is a serious medical condition, causing organ failure with potentially fatal outcome [3]. It is a toxic mushroom responsible for the majority of deaths occurring after mushrooms ingestion [4,5]. Amanita phalloides is the most dangerous, poisonous mushroom species.…”