12种剧毒鹅膏菌的肽类毒素成分鉴定及其相对含量差异比较研究

采用高效液相色谱-高分辨质谱联用技术对12种剧毒鹅膏菌的肽类毒素成分及其相对含量进行了比较研究.样品经含0.5%甲酸-50%甲醇溶液提取, 采用Agilent 300Extend-C18色谱柱, 含0.05% TFA的20 mmol/L NH4Ac溶液-甲醇为流动相, 梯度洗脱分离.12种鹅膏菌的高效液相色谱-质谱联用分析结果表明, 各鹅膏菌所含毒素种类及相对含量差异较大, 呈现显著的色谱指纹特征信息, 12种鹅膏菌中共检测出19种化合物, 13种属于已知鹅膏肽类毒素, 5种为未知鹅膏肽类毒素, 1种为未知小分子化合物组分.本研究为准确识别鹅膏菌的种类、鹅膏肽类毒素的鉴定、预防和鉴别鹅膏毒肽中毒, 提供了科学依据.

Identification and Content Difference of Toxic Peptidesin 12 Kinds of Lethal Amanita Species

The types and relative content difference of the toxic peptides in twelve species of Amanita were evaluated by the high performance liquid chromatography (HPLC) and ultra performnace liquid chromatography-electrospray-quadrupole-time-of-flight-mass spectrometry (HPLC-ESI-QTOF-MS).The efficient toxic compounds in mushroom were extracted using 50% methanol in water containing 0.5% formic acid.Then, separation of various components in mushroom was carried out using Agilent 300 Extend-C18 column with 0.05% (V/V) trifluoroacetic acid in 20 mmol/L ammonium acetate-methanol as mobile phase under a gradient program.The results indicated the significant difference in types and relative content of toxic peptides among all investigated mushrooms.In addition, we generated a chromatographic fingerprint based on the toxic peptide profiles.Finally 19 compositions were detected in the Amanita species, including 13 Amanita toxic peptides, 5 unknown toxic peptides and 1 unknown compound.These results play an important role in the discrimination of different Amanita species, and provide evidence in prevention and diagnosis of Amanitin poisoned events.