Wild mushroom poisoning: A case study of amatoxin-containing mushrooms and implications for public health.

Wild mushroom poisoning is a global public health concern, with mushrooms containing amatoxins being the main cause of fatalities. Mushrooms from the genus Amanita and Galerina contain amatoxins. Here we present a case of wild mushroom poisoning that affected three individuals, resulting in two fatalities. Within 10-15 hours after consumption, they experienced symptoms of gastroenteritis such as vomiting, abdominal pain, and diarrhea. One individual sought medical attention promptly and recovered, while the other two sought medical help nearly two or three days after the onset of symptoms, by which time their conditions had already worsened and led to their deaths. The mushrooms were identified belonging to genus Galerina, and laboratory test revealed variations in toxin levels among mushrooms collected from different parts of the decaying stump. The higher levels of α-amanitin, ß-amanitin, and γ-amanitin were detected near the base of the tree stump, but trace levels of α-amanitin were found near the top of the stump, while ß-amanitin and γ-amanitin were undetectable. This case emphasizes the importance of seeking immediate medical attention when experiencing delayed-onset gastrointestinal symptoms, as it may indicate more severe mushroom poisoning, particularly amatoxin poisoning. Timely and appropriate treatment is equally important. Additionally, consuming different units of the mushrooms in the same incident can lead to varying prognoses due to differences in toxin levels.

Acute hepatic and kidney injury after ingestion of Lepiota brunneoincarnata: Report of 2 cases.

Lepiota brunneoincarnata is a highly toxic mushroom species known to cause acute liver failure. However, there are limited reports investigating L. brunneoincarnata causing acute hepatic and renal damage. The present article reports 2 cases of L. brunneoincarnata poisoning in a mother and son from Chuxiong City, Yunnan Province, China. Both patients presented with gastrointestinal symptoms approximately 8-9 h after ingesting the suspect mushrooms and sought medical attention 27-28 h post-ingestion, both exhibiting acute hepatic and kidney injuries. Morphological and molecular biology studies confirmed the species of the mushrooms as L. brunneoincarnata. Liquid chromatography-tandem mass spectrometry analysis revealed mean fresh-weight concentrations of 123.5 µg/g α-amanitin and 45.7 µg/g ß-amanitin in the mushrooms. The patients underwent standard treatments, including multiple-dose activated charcoal, oral silibinin capsules, N-acetylcysteine, penicillin G, hemoperfusion, and plasma exchange. One patient recovered completely and was discharged after 16 days of hospitalization. The other patient exhibited gradual improvement in liver and renal function; however, renal function deteriorated 9 days after ingestion, and the patient declined renal replacement therapy and returned home 14 days post-ingestion. The patient was then re-hospitalized due to oliguria and edema in both lower extremities. Renal biopsy revealed acute tubular necrosis, inflammatory cell infiltration, minor glomerular capsular fibrosis, loss of microvilli in the renal tubular epithelial cells, and interstitial edema. The patient underwent 2 rounds of continuous renal replacement therapy, which eventually resulted in improvement, and was discharged 31 days after mushroom consumption. It is noteworthy that this patient had already progressed to chronic kidney insufficiency 11 months after intoxication.

Amatoxin poisoning caused by Galerina sulciceps, a species with no prior record of identification in Japan: a case report.

A 60-year-old man presented with acute gastroenteritis, hypovolemic shock, acute renal failure (BUN/Cr, 56.7/4.24 mg/dl), and aspiration pneumonia. The previous day, he ingested 30 caps of mushrooms of an unknown species. The patient was treated with a massive intravenous infusion, renal replacement therapy, and antimicrobial agents. Late-onset mild liver injury peaked on day 11 (AST/ALT, 62/67 IU/l). Acute renal failure improved once before worsening, with the worst symptoms on day 19 (BUN/Cr, 99/6.61 mg/dl). Thereafter, the patient showed gradual improvement, and renal replacement therapy was discontinued on day 23. His general condition improved fully and he was transferred to another hospital for rehabilitation on day 47. The mushrooms were later identified as Galerina sulciceps by the Basic Local Alignment Search Tool, and toxicologic analysis using liquid chromatography-tandem mass spectrometry revealed an average of 85 ppm α-amanitin and 330 ppm ß-amanitin in the tissue of the mushrooms brought in by the patient's family. Galerina sulciceps is distributed mainly in tropical and subtropical regions of Southeast Asia and had never been identified before in Japan. The heat of fermentation generated by the thick layer of wood chips on the ground or global warming may have contributed to its growth in Japan. Interestingly, our patient did not have liver dysfunction, which is one main and typical amatoxin poisoning symptom. Variation in clinical presentation may be attributed to the different ratios of α-amanitin to ß-amanitin in different mushroom species.

[Determination of five amanita peptide toxins in poisonous mushrooms by ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high resolution mass spectrometry].

Food poisoning by toxic mushrooms occurs frequently worldwide. It is one of the most common food poisoning events and the main cause of death. Amanita peptide toxins are the most common lethal toxins in poisonous mushrooms. Presently, a novel method based on ultra performance liquid chromatography-quadrupole electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q/Orbitrap HRMS) was developed for the determination of five amanitapeptide toxins (α-amanitin, ß-amanitin, γ-amanitin, phalloidin, and phallacidin). Because the isotope summit of α-amanitin affects the detection of ß-amanitin, it cannot be distinguished by low resolution mass spectrometry. Therefore, experimental conditions including chromatography and mass spectrometry were explored in detail. The five peptide toxins were extracted from poisonous mushrooms with pure water and filtered through a 0.22 µm teflon microporous membrane. The procedure was rapid, simple, and environmentally friendly. Chromatographic separation was performed on a strong polarity HSS T3 column (100 mm×2.0 mm, 2.1 µm) with gradient elution using acetonitrile and 5 mmol/L ammonium acetate containing 0.1% (v/v) formic acid as mobile phases at a flow rate of 0.3 mL/min. The column temperature was set to 40 ℃. The analytes were ionized using a heating electrospray ionization source and collected in positive ion mode. Full scanning/data-dependent secondary mass spectrometry (Full mass-ddMS2) mode was used for qualitative analysis of the targets within 10 min. The target ion selective scan (Targeted-SIM) mode was used for quantification by external standard calibration. The measured and theoretical values of the exact mass and the MS2 fragment ions of the five compounds were within an error of 5×10-6. Method validation was performed according to the criteria recommended by the Chinese National Standard. All the compounds showed an excellent linear relationship in the range of 1.0-20.0 µg/L. The correlation coefficients (r) ranged from 0.9974 to 0.9989. The limit of detection was 0.006 mg/kg for all five compounds. Recoveries ranged from 81.8% to 102.4%. There was no matrix effect in the blank mushroom sample for the five compounds, and the relative standard deviations ranged from 3.2% to 8.3%. This method provides abundant compound characteristic mass information, such as retention time, exact mass, fragment ions, and other information. The data can be used to identify suspected compounds based on the extracted ion flow diagram and isotope distribution information. Comparison between the actual exact mass and the theoretical exact mass, combined with the fragment ions enables identification of the structures of unknown compounds and collision methods, which can be confirmed in the absence of standard materials. In this study, the isomer of γ-amanitin was identified as amaninamide. The novel method is simple, accurate, specific, and sensitive. The method permits the rapid qualitative and quantitative detection of compound in public health emergency settings and will provide reliable technical support for the rapid screening of such toxic compounds and the structural locking of unknown toxins in the future.

Ultrasensitive paper sensor for simultaneous detection of alpha-amanitin and beta-amanitin by the production of monoclonal antibodies.

Amanitin (AMA) is responsible for human fatalities after ingestion of poisonous mushrooms, thus, a rapid and accurate detection method is urgently needed. Here, gold nanoparticle-based immunosensor with monoclonal antibody against AMA was constructed for rapid detection of alpha- and beta-amanitin (α- and ß-AMA) in mushroom, serum and urine samples. Under optimized conditions, the visual limits of detection (vLOD) and calculated LOD for α-AMA and ß-AMA in mushroom were 10 ng/g, 20 ng/g, and 0.1 ng/g, 0.2 ng/g, respectively. Analysis of wild mushroom samples was also performed using a strip scan reader in the 10 min range. Furthermore, in mushrooms containing amatoxins results were confirmed and compared with those determined by liquid chromatography tandem mass spectrometry. Thus, this immunosensor provided a useful monitoring tool for rapid detection and screening of mushroom samples and in serum and urine from subjects who accidentally consumed AMA-containing mushrooms.

[Highly sensitive determination of three kinds of amanitins in urine and plasma by ultra performance liquid chromatography-triple quadrupole mass spectrometry coupled with immunoaffinity column clean-up].

Cases of toxic mushroom poisoning occur frequently in China every year. In particular, mushrooms containing amanitins can cause acute liver damage, with high mortality rates. The symptoms of acute liver damage are experienced 9-72 h after consumption of the mushrooms. At this time, the concentration of amanitins in blood and urine is too low to be detected even by the highly sensitive ultra performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS), thus rendering clinical diagnosis and treatment difficult. To this end, a method was developed for the determination of α-amanitin, ß-amanitin and γ-amanitin in urine and plasma by UPLC-MS/MS. Urine and plasma samples were extracted and cleaned up by using an immunoaffinity column. A sample of 2.00 mL urine or 1.00 mL of plasma was diluted with 8.00 mL of phosphate buffer solution (PBS) and then loaded onto the immunoaffinity column at a flow rate of 0.5-1.0 mL/min. After washing the column with 10 mL of PBS and 13 mL of water successively, the bound amanitins were eluted with 3.00 mL of methanol-acetone (1∶1, v/v). The eluent was dried under nitrogen at 55 ℃. The residue was dissolved in 100 µL of 10% (v/v) methanol aqueous solution. The amanitins in urine were concentrated 20 times, while those in plasma were concentrated 10 times. Chromatographic separation was performed on a Kinetex Biphenyl column (100 mm × 2.1 mm, 1.7 µm) with gradient elution using methanol and 0.005% (v/v) formic acid aqueous solution as mobile phases. The three amanitins were detected by negative electrospray ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode and quantified by the solvent standard curve external standard method. Method validation was performed as recommended by the European Drug Administration (EMEA). Four levels of quality control (QC) samples were prepared, which covered the calibration curve range, viz., the limit of quantification (LOQ), within three times the LOQ (low QC), medium QC, and at 85% of the upper calibration curve range (high QC), and used to test the accuracy, precision, matrix effect, extraction recovery, and stability. The calibration curves for the three amanitins showed good linear relationships in the range of 0.1-200 ng/mL, and the correlation coefficients (r) were greater than 0.999. The matrix effects and extraction efficiencies of the three amanitins in urine and plasma were 92%-108% and 90%-103%, respectively, and the coefficients of variation were less than 13%. The accuracies of the three amanitins in urine were within -9.4%-8.0%. The repeatability and intermediate accuracies were 3.0%-14% and 3.5%-18%, respectively. When the sampling volume was 2.00 mL, the limits of detection of the three amanitins in urine were 0.002 ng/mL. The accuracies of the three amanitins in plasma were within -13%-8.0%. The repeatability and intermediate accuracies were 3.9%-9.7% and 5.5%-12%, respectively. When the sampling volume was 1.00 mL, the limits of detection of the three amanitins in plasma were 0.004 ng/mL. The developed method is simple, sensitive, and accurate. During toxic mushroom poisoning detection, 0.0067 ng/mL of α-amanitin and 0.0059 ng/mL of ß-amanitin were detected in the urine of poisoned patients 138 h after ingesting poisonous mushrooms. This method has successfully solved the problem of detecting ultra-trace levels of amanitins in the urine and plasma of poisoned patients. It has important practical significance for the early diagnosis, early treatment, and mortality reduction of suspected poisoning patients. This method can also provide reliable technical support for future research on the toxicological effects and in vivo metabolism of these toxins.

Toxin components and toxicological importance of Galerina marginata from Turkey.

Amatoxins, most of which are hepatotoxic, can cause fatal intoxication. While mushrooms in the amatoxin-containing Galerina genus are rare, they can poison humans and animals worldwide. Few studies have profiled the toxicity of Galerina marginata. In addition, many studies indicate that macrofungi can have different characteristics in different regions. In this study, the quantities of toxins present in G. marginata from different provinces in Turkey were analysed using reversed-phase high-performance liquid chromatography with ultraviolet detection (RP-HPLC-UV) and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). G. marginata samples were collected from three different regions of Turkey. The taxonomic categorization of mushrooms was based on their micro- and macroscopic characteristics. The presence of toxins α-amanitin (AA), ß-amanitin (BA), γ-amanitin (GA), phalloidin (PHD) and phallacidin (PHC) quantities were measured using RP-HPLC-UV and then were confirmed using LC-ESI-MS/MS. BA levels were higher than AA levels in G. marginata mushrooms collected from all three regions. Moreover, the levels of GA were below the detection limit and no phallotoxins were detected. This is the first study to identify and test the toxicity of G. marginata collected from three different regions of Turkey using RP-HPLC-UV. This is also the first study to confirm the UV absorption of amatoxins in G. marginata using LC-ESI-MS/MS, which is a far more sensitive process. More studies evaluating the toxicity of G. marginata in other geographic regions of the world are needed.

Reverse-phase/phenylboronic-acid-type magnetic microspheres to eliminate the matrix effects in amatoxin and phallotoxin determination via ultrahigh-performance liquid chromatography-tandem mass spectrometry.

In this study, we present the preparation of a new reverse-phase/phenylboronic-acid (RP/PBA)-type mixed-mode magnetic solid-phase extraction (MSPE) adsorbent for use in the cleanup of amatoxin- and phallotoxin-containing samples intended for ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. Further, the RP/PBA magnetic microspheres have phenyl and phenylboronic acid groups on their surfaces that selectively adsorb amatoxins and phallotoxins through hydrophobic, π-π, and boronate affinity, significantly reducing matrix effects in UPLC-MS/MS analysis. After systematic optimization, all the standard calibration curves expressed satisfactory linearity (r > 0.9930), limits of detection (0.3 µg/kg), and recovery (97.6%-114.2%). Compared with other reported methods, this method also has the advantages of simple, fast, and efficient operation using relatively small amounts of the MSPE adsorbent. Furthermore, the method was successfully applied in a poisoning incident caused by Lepiota brunneoincarnata Chodat & C. Martín ingestion.

Lateral flow immunoassay (LFIA) for the detection of lethal amatoxins from mushrooms.

The mushroom poison that causes the most deaths is the class of toxins known as amatoxins. Current methods to sensitively and selectively detect these toxins are limited by the need for expensive equipment, or they lack accuracy due to cross-reactivity with other chemicals found in mushrooms. In this work, we report the development of a competition-based lateral flow immunoassay (LFIA) for the rapid, portable, selective, and sensitive detection of amatoxins. Our assay clearly indicates the presence of 10 ng/mL of α-AMA or γ-AMA and the method including extraction and detection can be completed in approximately 10 minutes. The test can be easily read by eye and has a presumed shelf-life of at least 1 year. From testing 110 wild mushrooms, the LFIA identified 6 out of 6 species that were known to contain amatoxins. Other poisonous mushrooms known not to contain amatoxins tested negative by LFIA. This LFIA can be used to quickly identify amatoxin-containing mushrooms.

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

Amatoxins (AMAs) are lethal toxins found in a variety of mushroom species. Detection methods are needed to determine the occurrence of AMAs in mushroom species suspected in mushroom poisonings. In this manuscript, we report the generation of novel monoclonal antibodies (mAbs, AMA9G3 and AMA9C12) and the development of a competitive, enzyme-linked immunosorbent assay (cELISA) that is sensitive at 1 ng mL-1 and shows selectivity for α-amanitin (α-AMA) and γ-amanitin (γ-AMA), and less for ß-amanitin (ß-AMA). In order to decrease the overall time needed for analysis, the extraction procedure for mushrooms was also simplified. A rapid (1 min) extraction procedure of AMAs using solvents as simple as water alone was successfully demonstrated using Amanita mushrooms. Together, the extraction method and the mAb-based ELISA represent a simple and rapid method that readily detects AMAs extracted from mushroom samples.

Simultaneous identification and characterization of amanita toxins using liquid chromatography-photodiode array detection-ion trap and time-of-flight mass spectrometry and its applications.

Rapid and accurate identification of multiple toxins for clinical diagnosis and treatment of mushroom poisoning cases is still a challenge, especially with the lack of authentic references. In this study, we developed an effective method for simultaneous identification of amanita peptide toxins by liquid chromatography coupled with photodiode array detection and ion trap time-of-flight mass spectrometry. The accuracy and selectivity of the methodology were validated through similar multiple fragmentation patterns and characteristic ions of standard α- and ß-amanitin. The developed method could successfully separate and identify major toxic constituents in Amanita mushrooms. Two amatoxins and three phallotoxins were confirmed in a single run through their fragmentation patterns and characteristic ions, which can be used as diagnostic fragment ions to identify mushroom toxins in complex samples. Furthermore, the performance of the developed method was verified by using real biological samples, including plasma and urine samples collected from rats after intraperitoneal administration of toxins. Thus, the development methodology could be crucial for the accurate detection of mushroom toxins without standard references.

A New Conjugation Method Used for the Development of an Immunoassay for the Detection of Amanitin, a Deadly Mushroom Toxin.

One of the deadliest mushrooms is the death cap mushroom, Amanita phalloides. The most toxic constituent is α-amanitin, a bicyclic octapeptide, which damages the liver and kidneys. To develop a new tool for detecting this toxin, polyclonal antibodies were generated and characterized. Both α- and β-amanitin were coupled to carrier proteins through four different linking chemistries, one of which has never before been described. These conjugates were evaluated for their effectiveness in generating antibodies specific for the free toxin, as well as their utility in formatting heterogeneous assays with high sensitivity. Ultimately, these efforts yielded a newly described conjugation procedure utilizing periodate oxidation followed by reductive amination that successfully resulted in generating sensitive immunoassays (limit of detection (LOD), ~1.0 µg/L). The assays were characterized for their selectivity and were found to equally detect α-, β-, and γ-amanitin, and not cross-react with other toxins tested. Toxin detection in mushrooms was possible using a simple sample preparation method. This enzyme-linked immunosorbent assay (ELISA) is a simple and fast test, and readily detects amatoxins extracted from A. phalloides.

[Determination of amanitins and phallotoxins in wild mushrooms by online liquid chromatography-diode array detector-tandem mass spectrometry].

A fast and wide linear range method was established for the determination of mushroom toxins amanitins (α-amanitin,ß-amanitin and γ-amanitin) and phallotoxins (phallacidin and phalloidin) in wild mushrooms by online liquid chromatography-diode array detector-tandem mass spectrometry (LC-DAD-MS/MS). The mushroom toxins were extracted with methanol, and diluted with water. The extracts were separated on an XBridgeTM BEH C18 column (150 mm×3.0 mm, 2.5 µm) under pH 10.7, measured by DAD and then analyzed by MS/MS. Basic mobile phase conditions were applied to improve the ionization efficiency of hydrogen ion adducts. The baseline separation of the analytes was obtained within 15 min. The limits of detection (LODs) of the sample matrix were 0.005-0.02 mg/kg. The toxins were quantified by the results measured by MS/MS when the toxin contents less than 2 mg/kg, and quantified by the results obtained from DAD when the contents more than 2 mg/kg. The linear range was 0.05-500 mg/kg for the whole method in one injection. The method was successfully applied to the analyses of amanitins and phallotoxins in Lepiota brunneoincarnata and white Amanita.

Production of a broad-specificity monoclonal antibody and application as a receptor to detection amatoxins in mushroom.

In this study, we report the production of a monoclonal broad-specificity monoclonal antibody (mAb) specific for amatoxins and development of an indirect competitive immunoassay for detection of amatoxins in mushroom samples. In the assay, the complete antigen (α-amanitin-OVA) was used as coating antigen, and amatoxins as competitor competes with coating antigen to bind with mAb. Using this approach, The half-maximum inhibition concentrations (IC50) of α-amanitin, ß-amanitin and γ-amanitin, and limits of detection (LODs, IC15) were 66.3, 97.4, 163.1 ng/mL and 0.91, 0.98, 0.89 ng/mL, respectively. The LODs for α-amanitin, ß-amanitin and γ-amanitin in mushroom samples were 4.55, 4.9, and 4.45 ng/mL. The spiked results were also confirmed by HPLC, which showed a good correlation (R2 = 0.996) between the two methods. The results indicated that the developed assay was reliable and suitable for the detection of amatoxins in mushroom samples.

A Case Study: What Doses of Amanita phalloides and Amatoxins Are Lethal to Humans?

There are few data estimating the human lethal dose of amatoxins or of the toxin level present in ingested raw poisonous mushrooms. Here, we present a patient who intentionally ingested several wild collected mushrooms to assess whether they were poisonous. Nearly 1 day after ingestion, during which the patient had nausea and vomiting, he presented at the emergency department. His transaminase levels started to increase starting from hour 48 and peaking at hour 72 (alanine aminotransferase 2496 IU/L; aspartate aminotransferase 1777 IU/L). A toxin analysis was carried out on the mushrooms that the patient said he had ingested. With reversed-phase high-performance liquid chromatography analysis, an uptake of approximately 21.3 mg amatoxin from nearly 50 g mushroom was calculated; it consisted of 11.9 mg alpha amanitin, 8.4 mg beta amanitin, and 1 mg gamma amanitin. In the urine sample taken on day 4, 2.7 ng/mL alpha amanitin and 1.25 ng/mL beta amanitin were found, and there was no gamma amanitin. Our findings suggest that the patient ingested approximately 0.32 mg/kg amatoxin, and fortunately recovered after serious hepatotoxicity developed.

Amanitinas / Amanitines

La intoxicación por consumo de hongos es un fenómeno estacional que se produce con relativa frecuencia en áreas geográficas donde es habitual su consumo, en especial de especies silvestres. Dependiendo del tipo de hongo ingerido pueden aparecer distintos cuadros clínicos (gastrointestinal, nefrotóxico, alucinatorio, etc.). El cuadro más grave es el hepatotóxico, asociado a una alta mortalidad, y causado por hongos que contienen amatoxinas (síndrome ciclopeptídico). Presentamos una revisión actualizada de las características de las amatoxinas, su cinética y mecanismo de acción, los métodos utilizados para su determinación analítica, así como las diferentes opciones para el tratamiento de la intoxicación (AU)

Mushroom poisoning is a seasonal phenomenon that occurs relatively frequently in geographical areas where its consumption is common. Depending on the type of fungus ingested different clinical symptoms (gastrointestinal, nephrotoxic, hallucinatory, etc.) can occur. Hepatotoxic syndrome caused by fungi containing amatoxins is the most serious condition, associated to high mortality. We present an updated review of amatoxins characteristics, kinetics, mechanism of action, methods used for analytical determination, as well as the different options for the treatment of poisoning (AU)

Amatoxin-containing mushroom (Lepiota brunneoincarnata) familial poisoning.

Serious to fatal toxicity may occur with amanitin-containing mushrooms ingestions. A Lepiota brunneoincarnata familial poisoning with hepatic toxicity is reported. In such poisonings, acute gastroenteritis may be firstly misdiagnosed leading to delay in preventing liver dysfunction by silibinin or penicillin G. Mushroom picking finally requires experience and caution.

Amatoxin and phallotoxin concentration in Amanita phalloides spores and tissues.

Most of the fatal cases of mushroom poisoning are caused by Amanita phalloides. The amount of toxin in mushroom varies according to climate and environmental conditions. The aim of this study is to measure α-, ß-, and γ-amanitin with phalloidin and phallacidin toxin concentrations. Six pieces of A. phalloides mushrooms were gathered from a wooded area of Düzce, Turkey, on November 23, 2011. The mushrooms were broken into pieces as spores, mycelium, pileus, gills, stipe, and volva. α-, ß-, and γ-Amanitin with phalloidin and phallacidin were analyzed using reversed-phase high-performance liquid chromatography. As a mobile phase, 50 mM ammonium acetate + acetonitrile (90 + 10, v/v) was used with a flow rate of 1 mL/min. C18 reverse phase column (150 × 4.6 mm; 5 µm particle) was used. The least amount of γ-amanitin toxins was found at the mycelium. The other toxins found to be in the least amount turned out to be the ones at the spores. The maximum amounts of amatoxins and phallotoxin were found at gills and pileus, respectively. In this study, the amount of toxin in the spores of A. phalloides was published for the first time, and this study is pioneering to deal with the amount of toxin in mushrooms grown in Turkey.

Profiling of amatoxins and phallotoxins in the genus Lepiota by liquid chromatography combined with UV absorbance and mass spectrometry.

Species in the mushroom genus Lepiota can cause fatal mushroom poisonings due to their content of amatoxins such as α-amanitin. Previous studies of the toxin composition of poisonous Lepiota species relied on analytical methods of low sensitivity or resolution. Using liquid chromatography coupled to UV absorbance and mass spectrometry, we analyzed the spectrum of peptide toxins present in six Italian species of Lepiota, including multiple samples of three of them collected in different locations. Field taxonomic identifications were confirmed by sequencing of the internal transcribed spacer (ITS) regions. For comparison, we also analyzed specimens of Amanita phalloides from Italy and California, a specimen of A. virosa from Italy, and a laboratory-grown sample of Galerina marginata. α-Amanitin, ß-amanitin, amanin, and amaninamide were detected in all samples of L. brunneoincarnata, and α-amanitin and γ-amanitin were detected in all samples of L. josserandii. Phallotoxins were not detected in either species. No amatoxins or phallotoxins were detected in L. clypeolaria, L. cristata, L. echinacea, or L. magnispora. The Italian and California isolates of A. phalloides had similar profiles of amatoxins and phallotoxins, although the California isolate contained more ß-amanitin relative to α-amanitin. Amaninamide was detected only in A. virosa.