关键词: 超高效液相色谱-四级杆飞行时间串联质谱法, 鹅膏肽类毒素, 野生菌, 食物中毒

Abstract: Objective Establish an analytical method for the qualitative and quantitative detection of Amanita peptide toxins in poisonous mushrooms using ultra performance liquid chromatography mass spectrometer detector quadrupole time of flight（UPLC-Q-TOF）. Method The samples were homogenized with methanol and extracted by ultrasonic centrifugation. The liquid to be measured was separated by ACQUITY UPLC?? HSS T₃ 1.8 μm column（2.1×50 mm）,and gradient elution with 0.1% ammonia solution-methanol（0 min,2%B;0-0.5 min,2%B;0.5-6.0 min,95%B;6.0-7.0 min,95%B;7.0-8.0 min,2%B）,with flow rate of 0.3 mL/min,column temperature of 40 ℃,and sample intake of 2.0 μL. UPLC-Q-TOF was used to isolate and identify the toxin components of poisonous mushrooms. Result No amanita toxins were detected in samples 1,2,3,5,6 and 7 of the toxic mushroom samples. In sample 4,α-amanitin、β-amanitin、γ-amanitin、phallacidin、phalloidin and amanullin were detected. Further establish the rapid quantification method of Amanita toxins,5 kinds of Amanita toxins showed a good linear relationship within 25-1 000 μg/L,the correlation coefficient was 0.991 2-0.998 9,the average standard recovery rate was 89.5%-110.5%,the relative standard deviation（RSD） was 3.80%-7.80%. The content of α-amanitin、β-amanitin、γ-amanitin、phalloidin and phallacidin was （19.701±0.175）,（0.098±0.014）,（1.548±0.001）,（0.331±0.002） and （0.108±0.060）mg/kg respectively,and the relative content of amanullin and phallacidin was 0.754 mg/kg. Conclusion This method is simple,fast,and reliable for the determination of various amanita peptide toxins in poisonous mushrooms. It provides an effective scientific basis for the rapid clinical diagnosis and timely treatment of patients with sudden poisoning caused by eating poisonous mushrooms.

Key words: Ultra performance liquid chromatography/mass spectrometer detector quadrupole time of flight, Amanita toxins, Wild mushroom, Food poisoning