The Effect of A High-Resolution Accurate Mass Spectrometer On Simultaneous Multiple Mushroom Toxin Detection

Yıl 2020, , 878 - 886, 15.10.2020

Yasemin Numanoğlu Çevik

https://doi.org/10.17714/gumusfenbil.680816

Cited By: 2

Öz

Amatoxins are deadly wild mushroom toxins that
cause severe poisoning in humans, from diarrhea to organ dysfunction. Mortality
can be as high as 80% if no specific treatment is applied. In this study,
separation and determination methods were developed for the simultaneous
determination of 7 toxins belonging to Amanita
phalloides. After extraction and purification of Amanita phalloides (death cap) wild mushrooms, toxins were detected
using HPLC-ESI MS and exact mass with time of flight MS (TOF-MS) in positive
ionization mode. In addition, it was observed that the toxins of α- and
β-amanitine could be differentiated from each other thanks to HR-MS detection
in case of their close retention (Rt) in LC. The presence of two toxins at the
same retention point in the chromatogram was detected by differentiating the molecular
ion mass (920.3514 DA) of the α-amanitine with the HR-MS (920.3696 DA). By
comparing both optimized methods with each other, faster and broad-spectrum
toxin identification was achieved compared to existing methods. In this study,
the rapid and accurate identification of seven toxins simultaneously with the
same method is of great importance. It will make important contributions to the
treatment process, especially by determining that poisoning is caused by wild
mushrooms.

Anahtar Kelimeler

Amanitin, Amatoxin, HPLC ESI/MS, LC-TOF MS, Mushroom Intoxication, Phalloidin

Destekleyen Kurum

TUBITAK BIDEB-2219 (International Post-Doctoral Research Fellowship Programme) and Ghent University ,Belgium

Proje Numarası

1059B191401045

Teşekkür

She is also deeply indebted to Frederic Lynen, who supervised this study, and to Ghent University, Belgium, for granting access to all available infrastructure. Many thanks also to Ertuğrul Kaya from Düzce University who provided the dried Amanita phalloides.

Yüksek Çözünürlüklü Kütle Spektrometrenin Eşzamanlı Çoklu Mantar Toksin Tayinleri Üzerindeki Etkisi

Öz

Amatoksinler, insanlarda
ishalden organ işlev bozukluğuna kadar ciddi zehirlenmelere neden olan ölümcül
yabani mantar toksinleridir. Spesifik bir tedavi uygulanmazsa mortalite %80
kadar yüksek olabilir. Bu çalışmada Amanita phalloides' e ait 7 tane toksinin
eş zamanda belirlenmesi için ayırma ve tayin yöntemleri geliştirildi. Amanita
phalloides (ölüm kapağı) yabani mantarının ekstraksiyonu ve saflaştırılmasının
ardından toksinler, HPLC-ESI MS ve LC-doğru kütle uçuş zamanlı MS (TOF-MS) pozitif
iyonizasyon modu kullanılarak tespit edildi. Ayrıca α- ve β-amanitin toksinlerinin
LC'de birbirlerine çok yakın alıkonmaları (Rt) durumunda HR-MS özellikteki
dedeksiyon sayesinde birbirlerinden ayırt edilmesinin mümkün olduğu görüldü. Kromatogramda
aynı alıkonma noktasında iki toksin varlığı HR-MS sayesinde α-amanitinin C¹³
izotop kütlesi (920.3696 DA) ile β-amanitinin moleküler iyon kütlesinin (920.3514 DA) farklandırılması
yapılarak tespit edildi. Optimize edilmiş her iki yöntem birbirleriyle
karşılaştırılarak mevcut yöntemlere göre daha hızlı ve geniş spektrumlu toksin
tanımlaması geçekleştirildi. Bu çalışmada aynı yöntem ile eş zamanda yedi adet
toksinin hızlı ve doğru tanımlanması büyük önem taşımaktadır. Özellikle zehirlenmenin
yabani mantardan kaynaklı olduğunun belirlenmesi ile tedavi sürecinde önemli
katkılar sağlayacaktır.

Anahtar Kelimeler

Amanitin, Amatoksin, HPLC ESI/MS, LC-TOF MS, Mantar Zehirlenmesi, Falloidin

Proje Numarası

1059B191401045

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