Rapid Simultaneous Determination of Five Amatoxins and Phallotoxins in Human Urine and Plasma by Ultra Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry / 分析化学

ABSTRACT

Specific detection of amatoxins and phallotoxins in body fluids is necessary for an early diagnosis of an intoxication with mushrooms.In this study, a rapid method for the simultaneous determination of α-, (β-and γ-amanitin, phalloidin and phallacidin in human urine and plasma was first developed by ultra-perform ance liquid chromatography-tandem mass spectrometry.Urine sample was directly injected into the separation system and plasma sample was initially prepared by precipitation of proteins with 1% acetic acid in acetoni trile.The toxin was analyzed on an ACQUITY UPLC HSS T3 column using a gradient program with a cycle time of 9 min, and detected by positive electrospray ionization tandem mass spectrometry in the MRM mode, and quantified by matrix-match standard solution.The detection limits (S/N = 3) of the toxins were within 0.2-1 μg/L and 0.1-0.5 μg/L for urine and plasma, respectively.The standard curves were linear in the range of 2-100 μg/L for urine and 1-100 μg/L for plasma.The average recoveries were 92.0%-108.0% and 85.0%-100.0% for the toxins spiked in urine and plasma, with RSDs of 1.0%-22.0% and 2.0%-22.0% (n = 6), respectively.The method was simple, selective and sensitive to detect the amatoxins and phallotoxins in urine and plasma for both clinical and forensic purposes.