Integration of stable isotope labeling derivatization and magnetic dispersive solid phase extraction for measurement of neurosteroids by in vivo microdialysis and UHPLC-MS/MS
Integration of stable isotope labeling derivatization and magnetic dispersive solid phase extraction for measurement of neurosteroids by in vivo microdialysis and UHPLC-MS/MS
复制标题
整合稳定同位素标记衍生化和磁分散固相萃取，通过体内微透析和 UHPLC-MS/MS 测量神经类固醇
DOI：
10.1016/j.talanta.2019.02.011
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发表时间：
2019
期刊：
影响因子：
6.1
通讯作者：
Liu Huwei
中科院分区：
文献类型：
--
作者：
Xu Yanqiu;Sun Luping;Wang Xin;Zhu Shuyun;You Jinmao;Zhao Xian En;Bai Yu;Liu Huwei
在这项工作中，提出了一种稳定同位素标记衍生化（SILD）结合磁分散固相萃取（MDSPE）的新策略，通过体内微透析同时监测与帕金森病（PD）相关的神经甾体变化。该方法基于多反应监测（MRM）模式的超高效液相色谱-串联质谱（UHPLC-MS/MS）检测。本研究设计并合成了一对新的稳定同位素标记试剂sd0-/d3-3- n -甲基-2 ' -羧基罗丹明6G (d0-/d3-MCR6G)，分别用于大鼠血液微透析物和神经类固醇标准物的衍生化。采用d3-MCR6G标记的神经甾体标准物作为内标进行后续预处理和UHPLC-MS/MS定量，以减少质谱分析中复杂基质和离子抑制效应带来的偏差。在优化的衍生化和提取条件下，8种神经甾体的线性关系良好，相关系数r2值为 > 0.98。检测限（lod）和定量限（loq）分别为0.06 ~ 0.12 pg/mL和0.30 ~ 0.40 pg/mL。结果表明，该方法灵敏度高，选择性好，准确度高，操作方便，效率高。应用于同时动态测定正常和PD大鼠血液微透析液中多种神经甾体的含量。该方法有望在PD及相关神经系统疾病的监测和药物治疗中发挥潜在的作用。
In this work, a novel strategy of stable isotope labeling derivatization (SILD) combined with magnetic dispersive solid-phase extraction (MDSPE), has been proposed for simultaneous monitoring of neurosteroids changes linked to Parkinson's disease (PD) by in vivo microdialysis. The developed method was based on ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) detection using multiple-reaction monitoring (MRM) mode. In this study, a new pair of stable isotope labeling reagentsd0-/d3-3-N-methyl-2′-carboxyl Rhodamine 6G (d0-/d3-MCR6G), were designed and synthesized for derivatizing neurosteroids in rat blood microdialysates and neurosteroid standards, respectively.d3-MCR6G labeled neurosteroids standards were used as internal standard for the following pretreatment and UHPLC-MS/MS quantification to minimize the deviations caused by complex matrix and ion suppression effects in mass spectrometry analysis. Under the optimized derivatization and extraction conditions, good linearities of eight neurosteroids were obtained with correlation coefficients R2values > 0.98. The limits of detection (LODs) and quantitation (LOQs) ranged from 0.06 to 0.12 pg/mL and 0.30–0.40 pg/mL, respectively. Taken together, the established method exhibited high sensitivity and selectivity, excellent accuracy, convenience and high efficiency. It was applied for the simultaneous and dynamic measurement of multiple neurosteroids in normal and PD rat blood microdialysates. This method would be expected to be potentially useful for the monitoring and drug treatment of PD and related neurological disorders in the future.