Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device.
Using Dicationic Ion-Pairing Compounds To Enhance the Single Cell Mass Spectrometry Analysis Using the Single-Probe: A Microscale Sampling and Ionization Device.
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DOI：
10.1021/acs.analchem.6b01284
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发表时间：
2016-07-05
影响因子：
7.4
通讯作者：
Yang Z
中科院分区：
文献类型：
--
作者：
Pan N;Rao W;Standke SJ;Yang Z
我们开发了一种独特的质谱 (MS) 方法，可使用正电离模式确定活单细胞中带负电荷的物质。该方法通过微型多功能装置（单探针）利用双离子对化合物，在环境条件下对活单细胞进行反应性 MS 分析。在本研究中，将两种二甲基试剂，1,5-戊二基-双(1-丁基吡咯烷鎓)二氟化物(C5(bpyr)2F2)和1, 3-丙二基-双(三丙基鏻)二氟化物(C3(triprp)2F2)添加到溶剂中并引入单细胞中以提取细胞内容物用于实时MS分析。带负电（1− 带电）的细胞代谢物与双离子化合物（带 2+ 带电）形成稳定的离子对（带 1+ 带电），在正电离模式下进行检测，灵敏度大大提高。在正离子模式下的三个独立的 SCMS 实验中，我们暂时将 192 种和 70 种带负电的常见代谢物分别指定为与 (C5(bpyr)2F2) 和 (C3(triprp)2F2) 的加合物。 C5(bpyr)2F2 的暂定代谢物总数为 285 个，C3(triprp)2F2 的暂定代谢物总数为 143 个。此外，复合物形成中双离子化合物的选择性允许区分低丰度的重叠离子峰。对选定的加合物离子进行单细胞水平的串联 (MS/MS) 分析，以进行分子鉴定。在单探针 MS 技术中使用二价化合物为在单细胞水平上检测多种代谢物提供了一种有效的方法。
A unique mass spectrometry (MS) method has been developed to determine the negatively charged species in live single cells using the positive ionization mode. The method utilizes dicationic ion-pairing compounds through the miniaturized multifunctional device, the single-probe, for reactive MS analysis of live single cells under ambient conditions. In this study, two dicationic reagents, 1,5-pentanediyl-bis(1-butylpyrrolidinium) difluoride (C5(bpyr)2F2) and 1, 3-propanediyl-bis-(tripropylphosphonium) difluoride (C3(triprp)2F2), were added in the solvent and introduced into single cells to extract cellular contents for real-time MS analysis. The negatively charged (1− charged) cell metabolites, which form stable ion-pairs (1+ charged) with dicationic compounds (2+ charged), were detected in positive ionization mode with a greatly improved sensitivity. We have tentatively assigned 192 and 70 negatively charged common metabolites as adducts with (C5(bpyr)2F2) and (C3(triprp)2F2), respectively, in three separate SCMS experiments in the positive ion mode. The total number of tentatively assigned metabolites is 285 for C5(bpyr)2F2 and 143 for C3(triprp)2F2. In addition, the selectivity of dicationic compounds in the complex formation allows for the discrimination of overlapped ion peaks with low abundances. Tandem (MS/MS) analyses at the single cell level were conducted for selected adduct ions for molecular identification. The utilization of the dicationic compounds in the single-probe MS technique provides an effective approach to the detection of a broad range of metabolites at the single cell level.