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一种使用液相色谱串联质谱结合快速固相萃取技术,对人血清中 12 种植物毒素进行高灵敏度定量分析的方法。

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.

机构信息

Nagoya City Public Health Research Institute, 2266-132 Shimoshidami, Moriyama-ku, Nagoya, 463-0003, Japan; Department of Legal Medicine & Bioethics, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan.

Gifu Prefectural Research Institute for Health and Environmental Sciences, 1-1 Naka-Fudogaoka, Kagamigahara City, Gifu, 504-0838, Japan.

出版信息

J Pharm Biomed Anal. 2021 Jan 5;192:113676. doi: 10.1016/j.jpba.2020.113676. Epub 2020 Oct 9.

DOI:10.1016/j.jpba.2020.113676
PMID:33099112
Abstract

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 (r) 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 serum via dilution 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.

摘要

我们开发了一种使用液相色谱串联质谱(LC/MS/MS)对人血清中的 12 种植物毒素进行高灵敏度定量的方法。在本文中,我们选择了石蒜碱、加兰他敏、原藜芦碱 A、原藜芦碱 B、藜芦碱、藜芦定、白屈菜碱、澳洲茄碱、刻叶紫堇碱、α-茄碱、α-卡茄碱和茄啶作为目标分析物。ADME 柱用于 LC 分离,整体 SPE 柱(MonoSpin® C18)用于分析物提取。SPE 净化和 LC/MS/MS 分析的总时间在 30 分钟内完成。方法验证结果如下:每个校准曲线的线性(r)均大于 0.99;日内和日间准确度分别为 92.7%-116%和 91.6%-106%;日内和日间精密度分别低于 14%和 11%。此外,检测限和定量下限分别为 0.0071-0.15 和 0.022-0.46 ng/mL,表明该方法具有高灵敏度。最后,为了确认其可行性,我们的方法应用于两个模型样品:(1)市售人血清和(2)用人血清稀释小鼠血清得到的假性中毒血清。我们能够在血清中定量 0.84±0.02 ng/mL 的α-卡茄碱(案例 1)和 0.15±0.032 ng/mL 的原藜芦碱 A(案例 2),证明了我们方法的实用性。这是首次同时对人血清中的 12 种植物毒素进行定量分析。我们的方法可以调查涉及有毒植物的意外中毒,从而能够及时决定患者的治疗方案。

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