Isolation and identification of the Amanita muscaria and Amanita pantherina toxins in human urine.
OBJECTIVE
Ibotenic acid, muscimol and muscarine were recognized as responsible for psychotropic effects of A. muscaria and A. pantherina. Demand for their specific and sensitive identification and quantitation in biological material lead to effort to develop reliable analytical method, but satisfactory solution is still lacking. Presented article describes liquid chromatography-mass spectrometry method suitable for isolation and identification of principal toxins of A. muscaria and A. pantherina in urine.


RESULTS
Dedicated liquid chromatography-mass spectrometry method is reported. Technique consists of an extraction of analytes on Strata X-CW and Discovery SCX SPE cartridges and separation is achieved using a Gemini C18 column (150 mm x 2.0 mm, 5 micron) and 8 mM heptafluorobutyric acid as mobile phase. Detection at m/z 159 for ibotenic acid, m/z 115 for muscimol and m/z 174 for muscarine was used. Retention times and LODs are 2.6 min and 50 ng.ml-1 for ibotenic acid, 4.6 min and 40 ng.ml-1 for muscimol and 14.2 min and 3 ng.ml-1 for muscarine.


CONCLUSIONS
A sensitive and specific liquid chromatography-mass spectrometry assay was developed for analysis of principal toxins of A. muscaria and A. pantherina in urine. Method was found to be sufficiently sensitive and specific for analysis of urine of intoxicated patients.
Merova
Barbora
B
Department of Medical Chemistry and Biochemistry, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic. baramer@centrum.cz
Ondra
Peter
P
Stankova
Marie
M
Valka
Ivo
I
D000545:Amanita / Q000737:chemistry*; D000546:Amanitins / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007051:Ibotenic Acid / Q000652:urine; D007202:Indicators and Reagents; D009116:Muscarine / Q000652:urine; D009118:Muscimol / Q000652:urine; D019384:Nucleic Acid Synthesis Inhibitors / Q000652:urine*; D052616:Solid Phase Extraction; D012997:Solvents; D021241:Spectrometry, Mass, Electrospray Ionization
D000546:Amanitins; D007202:Indicators and Reagents; D019384:Nucleic Acid Synthesis Inhibitors; D012997:Solvents; D007051:Ibotenic Acid; D009118:Muscimol; D009116:Muscarine