Simultaneous determination of mushroom toxins α-amanitin, β-amanitin and muscarine in human urine by solid-phase extraction and ultra-high-performance liquid chromatography coupled with ultra-high-resolution TOF mass spectrometry

Tomková, Jana; Ondra, Peter; Válka, Ivo

doi:10.1016/j.forsciint.2015.04.007

Cited by 27 publications

(11 citation statements)

References 18 publications

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“…Although several methods have been described to determine amatoxins and/or phallotoxins in biological fluids, including radioimmunoassay (RIA) [ 11 , 12 , 13 ], enzyme-linked immunosorbent assay (ELISA) [ 14 , 15 ], capillary zone electrophoresis (CZE) [ 16 , 17 ], and liquid chromatography (LC) combined with ultraviolet (UV) [ 7 , 18 , 19 ] or electrochemical detection (ECD) [ 20 , 21 , 22 ], various problems, such as false positives, unstable reproducibility, poor confirmatory ability and laborious procedures, have limited their application in practice [ 19 , 23 ]. Methods combining LC with mass spectrometry (MS), such as triple quadrupole tandem MS (MS/MS) [ 24 , 25 , 26 , 27 , 28 ], time-of-flight (TOF) MS [ 29 ] and matrix-assisted laser desorption ionization time-of-flight MS (MALDI-TOF MS) [ 30 ], have been reported for the detection of amatoxins and phallotoxins. LC-MS/MS is a powerful technique allowing high sensitivity, reproducibility and specificity, and is less expensive than high-resolution MS.…”

Section: Introductionmentioning

confidence: 99%

A Simple and High-Throughput Analysis of Amatoxins and Phallotoxins in Human Plasma, Serum and Urine Using UPLC-MS/MS Combined with PRiME HLB μElution Platform

Zhang

Zhao

et al. 2016

Toxins

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Amatoxins and phallotoxins are toxic cyclopeptides found in the genus Amanita and are among the predominant causes of fatal food poisoning in China. In the treatment of Amanita mushroom poisoning, an early and definite diagnosis is necessary for a successful outcome, which has prompted the development of protocols for the fast and confirmatory determination of amatoxins and phallotoxins in human biological fluids. For this purpose, a simple, rapid and sensitive multiresidue UPLC-MS/MS method for the simultaneous determination of α-amanitin, β-amanitin, γ-amanitin, phalloidin (PHD) and phallacidin (PCD) in human plasma, serum and urine was developed and validated. The diluted plasma, serum and urine samples were directly purified with a novel PRiME technique on a 96-well μElution plate platform, which allowed high-throughput sample processing and low reagent consumption. After purification, a UPLC-MS/MS analysis was performed using positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. This method fulfilled the requirements of a validation test, with good results for the limit of detection (LOD), lower limit of quantification (LLOQ), accuracy, intra- and inter-assay precision, recovery and matrix effects. All of the analytes were confirmed and quantified in authentic plasma, serum and urine samples obtained from cases of poisoning using this method. Using the PRiME μElution technique for quantification reduces labor and time costs and represents a suitable method for routine toxicological and clinical emergency analysis.

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Section: Introductionmentioning

confidence: 99%

A Simple and High-Throughput Analysis of Amatoxins and Phallotoxins in Human Plasma, Serum and Urine Using UPLC-MS/MS Combined with PRiME HLB μElution Platform

Zhang

Zhao

et al. 2016

Toxins

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“…Normalized collision energy (NCE) of 28% provided the highest abundance of this fragment ion. The best sensitivity in terms of LOIs was achieved when the isolation window was widened to m/z 4.5 for inclusion of α- and β-amanitin and their most common 13 C-isotopes [ 43 ]. In order to discriminate α- from β-amanitin, baseline separation in LC was mandatory, which was achieved ( Figure 2 ) by using an Accucore PhenylHexyl column and a gradient elution ( Table 1 ).…”

Section: Resultsmentioning

confidence: 99%

Analysis of α- and β-amanitin in Human Plasma at Subnanogram per Milliliter Levels by Reversed Phase Ultra-High Performance Liquid Chromatography Coupled to Orbitrap Mass Spectrometry

Bambauer

Wagmann

Weber

et al. 2020

Toxins

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Amatoxins are known to be one of the main causes of serious to fatal mushroom intoxication. Thorough treatment, analytical confirmation, or exclusion of amatoxin intake is crucial in the case of any suspected mushroom poisoning. Urine is often the preferred matrix due to its higher concentrations compared to other body fluids. If urine is not available, analysis of human blood plasma is a valuable alternative for assessing the severity of intoxications. The aim of this study was to develop and validate a liquid chromatography (LC)-high resolution tandem mass spectrometry (HRMS/MS) method for confirmation and quantitation of α- and β-amanitin in human plasma at subnanogram per milliliter levels. Plasma samples of humans after suspected intake of amatoxin-containing mushrooms should be analyzed and amounts of toxins compared with already published data as well as with matched urine samples. Sample preparation consisted of protein precipitation, aqueous liquid-liquid extraction, and solid-phase extraction. Full chromatographical separation of analytes was achieved using reversed-phase chromatography. Orbitrap-based MS allowed for sufficiently sensitive identification and quantification. Validation was successfully carried out, including analytical selectivity, carry-over, matrix effects, accuracy, precision, and dilution integrity. Limits of identification were 20 pg/mL and calibration ranged from 20 pg/mL to 2000 pg/mL. The method was applied to analyze nine human plasma samples that were submitted along with urine samples tested positive for amatoxins. α-Amanitin could be identified in each plasma sample at a range from 37–2890 pg/mL, and β-amanitin was found in seven plasma samples ranging from <20–7520 pg/mL. A LC-HRMS/MS method for the quantitation of amatoxins in human blood plasma at subnanogram per milliliter levels was developed, validated, and used for the analysis of plasma samples. The method provides a valuable alternative to urine analysis, allowing thorough patient treatment but also further study the toxicokinetics of amatoxins.

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“…İnsanlar için muskarinin öldürücü dozu kesin olarak bilinmemekle beraber 40 mg-495 mg arasında (150 g taze Inocybe türü mantara eşdeğer) olduğu tahmin edilmektedir (13). Muskarin, yüksek performanslı sıvı kromatografisi ve kütle spektrometresi gibi analitik teknikler ile idrarda saptanabilse de kullanımları yaygın değildir (15,16). Bu olguda hastanın yakınlarından temin edilen mantar örnekleri mikolojik inceleme amacıyla otolize uğramaması için kurutularak Düzce Üniversitesi Tıp Fakültesi Farmakoloji Laboratuvarı'na gönderildi.…”

Section: Discussionunclassified

Early Stage Muscarinic Findings and Syndromic Classification in Wild Mushroom Poisoning

Yılmaz¹,

Kazbek²,

Kaya³

et al. 2018

tybd

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Mushroom poisoning is a frequently encountered health problem around the globe. Depending on its type and toxin, it results in a wide spectrum of issues that range from asymptomatic findings to fatal organ failure. Since the toxin of the mushroom leading to poisoning is often unidentified, syndromic classification may benefit clinical management according to the findings of target organ system toxicity. A 64-year-old female patient with comorbidities such as diabetes mellitus, coronary artery disease, hypertension and hyperlipidemia was admitted to the intensive care unit. Approximately four hours after consuming wild mushrooms the patient's diarrhea, muscle cramps, cold sweating, blurry vision, coughing and shortness of breath complaints have been begun. Her respiratory system examination revealed tachypnea, generalized bilateral rhonchus, bronchorrhea and wheezing. Patient was normotensive and had bradycardia (heart rate 35 beats/minute). She had anxiety and the Glasgow Coma score was 15. Her pupillary bilateral isochoric, miotic and light reflexes were diminished. In her arterial blood gas pH level 7.26, partial carbon dioxide pressure: 34 mmHg, potassium: 5.3 mmol/L, sodium: 130 mmol/L, lactate: 5.3 mmol/L, bicarbonate: 16 mEq/L and base excess:-10,3 mmol/L was found. Target organ system toxicity findings and timing of occurrence indicated early-onset cholinergic syndrome. Treatments that consisted of intravenous atropine, fluid, electrolytes and sodium bicarbonate alleviated the toxicity in a short period of time. Late mycologic examination results that followed the treatment revealed that the patient was poisoned by Inocybe lacera, a mushroom type that contains muscarine. With this case presentation, we summarized the syndromic classification that we use in the clinical management in the light of current information in the literature.

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Simultaneous determination of mushroom toxins α-amanitin, β-amanitin and muscarine in human urine by solid-phase extraction and ultra-high-performance liquid chromatography coupled with ultra-high-resolution TOF mass spectrometry

Cited by 27 publications

References 18 publications

A Simple and High-Throughput Analysis of Amatoxins and Phallotoxins in Human Plasma, Serum and Urine Using UPLC-MS/MS Combined with PRiME HLB μElution Platform

A Simple and High-Throughput Analysis of Amatoxins and Phallotoxins in Human Plasma, Serum and Urine Using UPLC-MS/MS Combined with PRiME HLB μElution Platform

Analysis of α- and β-amanitin in Human Plasma at Subnanogram per Milliliter Levels by Reversed Phase Ultra-High Performance Liquid Chromatography Coupled to Orbitrap Mass Spectrometry

Early Stage Muscarinic Findings and Syndromic Classification in Wild Mushroom Poisoning

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