Department of Forensic Medicine, Huazhong University of Science and Technology, #13 Hangkong Road, Wuhan, 430030, China.
State Key Laboratory of Environment Health (Incubation), Key Laboratory of Environment and Health, Ministry of Education, Key Laboratory of Environment and Health (Wuhan), Ministry of Environmental Protection, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, #13 Hangkong Road, Wuhan, 430030, China.
Talanta. 2024 Dec 1;280:126767. doi: 10.1016/j.talanta.2024.126767. Epub 2024 Aug 24.
In this work, electromembrane extraction (EME) was used for the first time to separate aconitine (AC), mesaconitine (Mes-AC) and hypaconitine (Hyp-AC) from biological samples and Chinese herbal medicines. Efficient EME of polar and high molecular weight aconitine alkaloids from different sample matrices was achieved with the solvent of 1-ethyl-2-nitrobenzene (ENB). Under the optimal EME conditions, EME provided recoveries for all targets in the range of 72%-74 %, 85%-103 % and 92%-94 % for whole blood, urine and aqueous samples. The proposed EME systems combined with LC-MS/MS and HPLC-UV were evaluated using different sample matrices, and the methods displayed satisfactory analytical characteristic including negligible matrix effect. The LOD and LOQ of AC, Mes-AC, and Hyp-AC by EME-LC-MS/MS were in the range of 0.002-0.068 ng/mL and 0.005-0.228 ng/mL respectively. The LOD and LOQ of AC, Mes-AC, and Hyp-AC by EME-HPLC-UV were in the range of 0.06-0.26 μg/mL and 0.20-0.86 μg/mL, respectively. The coefficient of determination, R-value was ≥0.9926 for all cases, and the accuracy in the linear ranges was in the range of 91%-111 %. Finally, the method was successfully applied for the qualitative and quantitative analysis of AC and Mes-AC in the whole blood and herbal medicine dreg samples from an actual forensic case, and poisoning by aconitum alkaloids was identified as the cause of death. Therefore, we believe that EME could be a powerful tool to identify poisoning, and EME has great potential for efficient separation of polar and high molecular weight substances. These are of great importance in the fields of but not limited to forensic science, Traditional Chinese Medicine and clinics.
在这项工作中，首次使用电萃取（EME）从生物样品和中草药中分离出乌头碱（AC）、中乌头碱（Mes-AC）和次乌头碱（Hyp-AC）。用 1-乙基-2-硝基苯（ENB）作为溶剂，从不同的样品基质中高效萃取极性和高分子量的乌头生物碱。在最佳的 EME 条件下，EME 为全血、尿液和水样品中的所有目标物提供了 72%-74%、85%-103%和 92%-94%的回收率。结合 LC-MS/MS 和 HPLC-UV 对所提出的 EME 系统进行了评价，使用不同的样品基质，该方法具有令人满意的分析特性，包括可忽略的基质效应。EME-LC-MS/MS 测定 AC、Mes-AC 和 Hyp-AC 的检出限和定量限分别为 0.002-0.068ng/mL 和 0.005-0.228ng/mL。EME-HPLC-UV 测定 AC、Mes-AC 和 Hyp-AC 的检出限和定量限分别为 0.06-0.26μg/mL 和 0.20-0.86μg/mL。所有情况下的相关系数，R 值均≥0.9926，线性范围内的准确度在 91%-111%之间。最后，该方法成功应用于实际法医案例中全血和草药残渣样品中 AC 和 Mes-AC 的定性和定量分析，并鉴定出乌头碱类中毒是死亡的原因。因此，我们相信 EME 可以成为识别中毒的有力工具，并且 EME 具有高效分离极性和高分子量物质的巨大潜力。这在法医学、中药和临床等领域具有重要意义。
