Center for Innovative Technology, Department of Chemistry, Institute of Chemical Biology, Institute for Integrative Biosystems Research and Education, Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, Tennessee 37235, United States.
Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee 37235, United States.
Anal Chem. 2020 Nov 3;92(21):14648-14656. doi: 10.1021/acs.analchem.0c03172. Epub 2020 Oct 13.
Routine small-molecule analysis is challenging owing to the need for high selectivity and/or low limits of quantification. This work reports a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify 14 antiepileptic drugs (AEDs) in human serum. For the optimized LC-MS/MS method described herein, we applied the guidelines outlined in the Clinical and Laboratory Standards Institute (CLSI) LC-MS C62-A document and the U.S. Food and Drug Administration (FDA) Bioanalytical Method Validation Guidance for Industry to evaluate the quality of the assay. In these studies, AED linearity, analyte recovery, matrix effects, precision, and accuracy were assessed. Using liquid chromatography-drift tube ion mobility-mass spectrometry (LC-DTIM-MS), a qualitative method was also used to increase confidence in AED identification using accurate mass and collision cross section (CCS) measurements. The LC-DTIM-MS method was also used to assess the ability of drift tube CCS measurements to aid in the separation and identification of AED structural isomers and other AEDs. These data show that another dimension of information, namely CCS measurements, provides an orthogonal dimension of structural information needed for AED analysis. Multiplexed AED measurements using LC-MS/MS and LC-DTIM-MS have the potential to enable better optimization of dosing owing to the high precision capabilities available in these types of analytical studies. Taken together, these data also show the ability to increase confidence in small-molecule identification and quantification using these analytical technologies.
由于需要高选择性和/或低定量下限，常规小分子分析具有挑战性。本工作报道了一种用于定量人血清中 14 种抗癫痫药物（AED）的液相色谱-串联质谱（LC-MS/MS）方法。对于本文中描述的优化 LC-MS/MS 方法，我们应用了临床和实验室标准协会（CLSI）LC-MS C62-A 文件和美国食品和药物管理局（FDA）生物分析方法验证行业指南中概述的指南，以评估该测定方法的质量。在这些研究中，评估了 AED 的线性、分析物回收率、基质效应、精密度和准确度。使用液相色谱-漂移管离子迁移质谱（LC-DTIM-MS），还使用定性方法来提高使用准确质量和碰撞截面（CCS）测量值鉴定 AED 的可信度。LC-DTIM-MS 方法还用于评估漂移管 CCS 测量值在辅助 AED 结构异构体和其他 AED 的分离和鉴定方面的能力。这些数据表明，另一个维度的信息，即 CCS 测量值，为 AED 分析提供了所需的结构信息的正交维度。使用 LC-MS/MS 和 LC-DTIM-MS 进行的多重 AED 测量具有由于这些类型的分析研究中具有高精度能力而更好地优化剂量的潜力。综上所述，这些数据还表明，使用这些分析技术可以提高小分子鉴定和定量的可信度。
