Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, China.
Key Laboratory of Pharmaceutical Intermediates and Analysis of Natural Medicine, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu 273165, Shandong, China.
Talanta. 2019 Jul 1;199:97-106. doi: 10.1016/j.talanta.2019.02.011. Epub 2019 Feb 10.
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 reagents d-/d-3-N-methyl-2'-carboxyl Rhodamine 6G (d-/d-MCR6G), were designed and synthesized for derivatizing neurosteroids in rat blood microdialysates and neurosteroid standards, respectively. d-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 R values > 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.
在这项工作中，提出了一种新的稳定同位素标记衍生化（SILD）策略，结合磁分散固相萃取（MDSPE），通过体内微透析监测与帕金森病（PD）相关的神经甾体变化。所开发的方法基于超高效液相色谱串联质谱（UHPLC-MS/MS）检测，采用多反应监测（MRM）模式。在这项研究中，设计并合成了一对新的稳定同位素标记试剂 d-/d-3-N-甲基-2'-羧基罗丹明 6G（d-/d-MCR6G），分别用于衍生大鼠血液微透析液和神经甾体标准品中的神经甾体。将 d-MCR6G 标记的神经甾体标准品用作内标，用于以下预处理和 UHPLC-MS/MS 定量分析，以最大限度地减少复杂基质和质谱分析中离子抑制效应对偏差的影响。在优化的衍生化和提取条件下，八种神经甾体均获得了良好的线性关系，相关系数 R 值均大于 0.98。检出限（LOD）和定量限（LOQ）范围分别为 0.06-0.12 pg/mL 和 0.30-0.40 pg/mL。总之，所建立的方法表现出高灵敏度和选择性、优异的准确性、方便性和高效率。它被应用于正常和 PD 大鼠血液微透析液中多种神经甾体的同时和动态测量。该方法有望在未来用于 PD 及相关神经障碍的监测和药物治疗。
