Zhongguo shipin weisheng zazhi (Mar 2022)
Fast determination of 9 mushroom toxins in wild fungus by liquid chromatography-tandem mass spectrometry
Abstract
ObjectiveTo establish an analytical method for 9 mushroom toxins in wild fungus by liquid chromatography-tandem mass spectrometry （LC-MS/MS） including 5 acute liver failure toxins （α-amanitin， β-amanitin， γ-amanitin， phallacidin and phalloidin） and 4 psycho-neurological disorder toxins （ibotenic acid， musimol， muscarine and psilocybin）.MethodsThe dried sample was extracted with 50%（V/V）methanol /water mixture containing 0.5%（V/V）formic acid， protein was precipitated with acetonitrile and diluted with water. The toxins were separated with XBridgeTM BEH C18 column （150 mm×3.0 mm， 2.5 μm）， gradient eluted with the mobile phase of 0.005%（V/V） formic acid aqueous solution–methanol， and measured by MS/MS with multiple reaction monitoring （MRM） mode.ResultsThe conditions of sample extraction， dilution ratio and chromatographic separation were optimized. 9 mushroom toxins in different wild fungus could be qualitatively and quantitatively measured based on the pure solvent standard calibration. The limits of detection （LODs） and quantification （LOQs） in sample matrix were 6-15 mg/kg and 20-50 mg/kg， respectively. The linear ranges were 0.004-2 mg/L with correlation coefficients （r） of between 0.9973-0.9995. The average recoveries at three spiking levels （50 mg/kg， 500 mg/kg and 5000 mg/kg） were 78.6%-109.7% with relative standard deviations （RSDs） of 2.7%-9.0%. The toxin compositions and contents varied greatly in the wild mushrooms sampled in some poisoning cases.ConclusionThe method is simple， rapid and accurate. It has been applied to the analysis of mushroom toxins in common poisonous wild mushrooms and the disposal of poisoning emergencies. It can provide rapid and accurate technical support for the etiology identification of poisoning cases and the timely rescue of patients.
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