高效液相色谱-串联质谱法测定野生蘑菇中毒样品中5种鹅膏肽类毒素

Determination of 5 kinds of peptide toxins in wild mushroom poisoning samples by high performance liquid chromatography-tandem mass spectrometry

中文关键词:  高效液相色谱-串联质谱法  食物中毒；鹅膏肽类毒素；鹅膏毒肽  鬼笔毒肽

英文关键词:high performance liquid chromatography-tandem mass spectrometry  food poisoning  peptide toxins  amatoxins  phallotoxins

中文摘要:

      目的 建立高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry, HPLC-MS/MS)快速测定野生蘑菇中毒样品中5种鹅膏肽类毒素的分析方法。方法 剩余蘑菇和患者尿液经甲醇提取, 250 mg N-丙基乙二胺(N-propyl ethylenediamine, PSA)吸附剂净化, 采用Waters CORTECS C18+液相色谱柱分离待测物, 以甲醇-2 mmol/L乙酸铵(含0.1%甲酸)为流动相, 进行梯度洗脱。采用HPLC-MS/MS检测。结果 5种鹅膏肽类毒素在10.0~500.0 μg/L质量浓度范围内线性关系良好(r>0.997), 蘑菇样品中5种毒素检出限(limits of detection, LODs)为2.0~3.0 μg/kg, 定量限(limits of quantification, LOQs)为5.0~10.0 μg/kg, 尿液样品中5种毒素LODs为0.8~1.2 μg/L, LOQs为2.0~4.0 μg/L。方法回收率范围为60.1%~94.9%, 相对标准偏差为2.0%~17.7% (n=6)。结论 该方法准确、快速, 适用于食用野生蘑菇中毒事件剩余蘑菇样品和患者尿液样品中鹅膏肽类毒素的检测。

英文摘要:

      Objective To establish an analytical method for the rapid determination of 5 kinds of peptide toxins in samples from wild mushroom poisoning incidents by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods The remaining mushrooms and patient urine samples were extracted by methanol, purified by 250 mg N-propyl ethylenediamine (PSA), and separated by Waters CORTECS C18+ liquid chromatography column. The mobile phase consisted of methanol and 2 mmol/L ammonium acetate (containing 0.1% formic acid) under gradient elution. They were determined by HPLC-MS/MS. Results The 5 kinds of peptide toxins had a good linear relationship (r>0.997) at the concentration 10.0–500.0 μg/L. The limits of detection (LODs) of 5 kinds of peptide toxins in the mushroom samples were 2.0–3.0 μg/kg, the limits of quantification (LOQs) were 5.0–10.0 μg/kg. In the urine samples, the LODs of 5 kinds of peptide toxins were 0.8–1.2 μg/L, the LOQs were 2.0–4.0 μg/L. The recoveries of method were 60.1%–94.9%, and the relative standard deviations were 2.0%–17.7% (n=6). Conclusion This method is accurate and rapid, and suitable for determination of peptide toxins in residual mushroom and patient urine samples from wild mushroom poisoning incidents.