A highly sensitive quantification method for 12 plant toxins in human serum using liquid chromatography tandem mass spectrometry with a quick solid-phase extraction technique
A highly sensitive quantification method for 12 plant toxins in human serum using liquid chromatography tandem mass spectrometry with a quick solid-phase extraction technique
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采用液相色谱串联质谱法和快速固相萃取技术对人血清中 12 种植物毒素进行高灵敏度定量方法
DOI：
10.1016/j.jpba.2020.113676
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发表时间：
2021
影响因子：
3.4
通讯作者：
Kei Zaitsu
中科院分区：
文献类型：
--
作者：
Masaru Taniguchi;Tomiaki Minatani;Hitoshi Miyazaki;Hitoshi Tsuchihashi;Kei Zaitsu
建立了人血清中12种植物毒素的LC/MS/MS高灵敏定量方法。在本论文中，我们选择石蒜碱、加兰他明、原藜芦碱A、原藜芦碱B、藜芦碱、藜芦碱、芦荟碱、环丙胺、西瓦定、α-茄宁、α-查可宁和茄尼定作为目标分析物。ADME色谱柱用于LC分离，整体固相萃取柱(MonoSpin®C18)用于提取分析物。固相萃取净化和LC/MS/MS分析的总时间在30min内完成。方法验证结果为：各标准曲线的线性关系(R2)均在0.99以上，日内和日间精密度分别为92.7%~116%和91.6%~106%，日间和日间精密度分别小于14%和11%。该方法的检测下限为0.0071~0.15 ng/m L，定量下限为0.022~0.46 ng/m L，具有较高的灵敏度。最后，为了验证该方法的可行性，我们将该方法应用于两个模型样本：(1)市售人血清和(2)用人血清稀释小鼠血清。病例1血清中α-Chaconine含量为0.84±0.02 ng/m L，假性中毒血清中原藜芦碱A含量为0.15±0.032 ng/m L，证明了本方法的实用性。这是首次对人体血清中的12种植物毒素进行同时定量。我们的方法可以调查涉及有毒植物的意外中毒事件，使患者能够迅速做出治疗决定。
We developed a highly sensitive quantification method using liquid chromatography tandem mass spectrometry (LC/MS/MS) for 12 plant toxins in human serum. In this paper, we selected lycorine, galanthamine, protoveratrine A, protoveratrine B, veratramine, veratridine, jervine, cyclopamine, cevadine, α-solanine, α-chaconine, and solanidine as targeted analytes. The ADME column was utilized for LC separation and a Monolithic SPE column (MonoSpin® C18) for analyte extraction. The total time for SPE clean-up and LC/MS/MS analysis was completed within 30 min. The method validation results were as follows: the linearity (r2) of each calibration curve was over 0.99; the inter- and intra-day accuracies were 92.7 %–116 % and 91.6 %–106 %, respectively; and the inter- and intra-day precisions were below 14 % and 11 %, respectively. Also, the lower limits of detection and quantification were 0.0071–0.15 and 0.022–0.46 ng/mL, respectively, indicating the method’s high sensitivity. Finally, to confirm its feasibility, our method was applied to two model samples: (1) commercially available human serum and (2) pseudo poisoning serumviadilution of mouse serum with human serum. We were able to quantify α-chaconine at 0.84 ± 0.02 ng/mL in the serum (Case 1) and protoveratrine A at 0.15 ± 0.032 ng/mL in the pseudo poisoning serum (Case 2), demonstrating our method’s practicality. This is the first time that the 12 plant toxins in human serum were simultaneously quantitated. Our method can investigate accidental poisonings involving toxic plants, enabling prompt decisions on patient treatment.