Rapid Simultaneous Determination of Five Amatoxins and Phallotoxins in Human Urine and Plasma by Ultra Performance Liquid Chromatography Coupled with Triple Quadrupole Mass Spectrometry
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-,βand γ-amanitin,phalloidin and phallacidin in human urine and plasma was first developed by ultra-performance 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 acetonitrile.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.