超高效液相色谱质谱法快速测定血浆中6种蘑菇毒素

目的 建立超高效液相色谱质谱法快速测定血浆中异绒白乳菇醛、α-鹅膏毒肽、β-鹅膏毒肽、γ-鹅膏毒肽、二羟鬼笔毒肽和羧基二羟鬼笔毒肽的含量。方法 取血浆样本，以1∶3的比例加入乙腈，旋涡混匀1min后，以10 000转/min、4℃离心3min。吸取上清液，以0.03%氨水和乙腈进行超高效液相分离，在电喷雾正离子模式下进行多反应监测检测。结果 一次进样4min即可实现血浆中异绒白乳菇醛、α-鹅膏毒肽、β-鹅膏毒肽、γ-鹅膏毒肽、二羟鬼笔毒肽和羧基二羟鬼笔毒肽的分离检测。血浆中6种蘑菇毒素的线性范围为40~500μg/L，检出限分别为10.0μg/L、10.0μg/L、10.0μg/L、6.0μg/L、6.0μg/L和6.0μg/L（S/N=3），相对标准偏差为0.60%~1.90%（n=11），回收率为90.8%~127.0%。结论 本方法可实现血浆中包括异绒白乳菇醛在内的6种蘑菇毒素的快速同时测定，操作简单，结果准确，为中毒患者的蘑菇毒素快速确证提供技术平台。

Rapid determination of six mushroom toxins in plasma by ultra performance liquid chromatography-mass spectrometry

Objective To establish a method for rapid determination of isovelleral, α-amanitin, β-amanitin, γ-amanitin, dihydroxy-phallotoxin and carboxy-dihydroxy-phallotoxin in plasma by ultra performance liquid chromatography-mass spectrometry. Methods Take plasma samples and add acetonitrile solution in a 1 : 3 ratio. The sample was vortex mixed for 1 min, then centrifugated at 10 000r/min and 4℃ for 3min. The supernatant was injected, while 0.03% ammonia water and acetonitrile were used as mobile phases for ultra high performance liquid phase separation. Multi reaction monitoring was carried out in the positive ion mode of electric spray. Results The separation and detection of isovelleral, α-amanitin, β-amanitin, γ-amanitin, dihydroxy-phallotoxin and carboxyl-dihydroxy-phallotoxin in plasma could be achieved within 4 min by one injection. The linear range of six mushroom toxins in plasma was 40-500μg/L, and the detection limits were 10.0μg/L 10.0μg/L, 10.0μg/L, 6.0μg/L, 6.0μg/L and 6.0μg/L (S/N=3), respectively. The relative standard deviations were 0.60%-1.90% (n=11), and the recovery rates were 90.8%-127.0%. Conclusion This method realizes the rapid and simultaneous determination of 6 kinds of mushroom toxins in plasma, including isovelleral. The operation is simple and the result is accurate, which provides a technical platform for the rapid confirmation of mushroom toxins in poisoned patients.