|
|
Study of selected mushroom toxins of Amanita muscaria.
Adámek, Michal ; Bosáková, Zuzana (advisor) ; Hložek, Tomáš (referee)
The presented bachelor's thesis was focused on the development of an HPLC-MS/MS method for the separation and eventual quantification of the main toxins of Amanita muscaria (fly agaric). After optimization of tandem mass detection conditions, MRM transitions were found for muscarine (174 → 57), ibotenic acid (159 → 113), muscimol (115 → 98) and cycloserine (103 → 75), which was used as an internal standard. Measurements were performed on Luna Omega 1.6 µm polar C18 column in RP-HPLC mode with an optimized mobile phase composition of 2:98 (v/v) acetonitrile/deionized water with the addition of 0.1% formic acid in both components and Ascentis Express RP- Amide in HILIC mode with an optimized mobile phase composed of 70:30 (v/v) methanol/deionized water with the addition of 0.5% formic acid in both components. Calibration curves with a range of 0.01 to 10 µg/ml for RP-HPLC and 0.01 to 25 µg/ml for HILIC were measured under optimized conditions. Urine and blood serum solutions of a patient with Amanita muscaria poisoning, including diluted versions of these solutions, were measured. Ibotenic acid and muscimol were found in the patient's urine, muscarine, ibotenic acid and muscimol in the blood serum. The RP-HPLC method gave narrower peaks with lower retentions and lower noise levels. The studied toxins...
|
guest ::
