Enhancing biological analyses with three dimensional field asymmetric ion mobility, low field drift tube ion mobility and mass spectrometry (μFAIMS/IMS-MS) separations

Abstract

Multidimensional high throughput separations are ideal for analyzing distinct ion characteristics simultaneously in one analysis. We report on the first evaluation of a platform coupling a high speed field asymmetric ion mobility spectrometry microchip (μFAIMS) with drift tube ion mobility and mass spectrometry (IMS-MS). The μFAIMS/IMS-MS platform was used to analyze biological samples and simultaneously acquire multidimensional FAIMS compensation fields, IMS drift times, and accurate ion masses for the detected features. These separations thereby increased the overall measurement separation power, resulting in greater information content and more complete characterization of the complex samples. The separation conditions were optimized for sensitivity and resolving power by the selection of gas compositions and pressures in the FAIMS and IMS separation stages. The resulting performance provided three dimensional separations, benefitting both broad complex mixture studies and targeted analyses by improving isomeric separations and allowing detection of species obscured by interfering peaks.

Fucoxanthin, A Carotenoid Derived from Phaeodactylum tricornutum Exerts Antiproliferative and Antioxidant Activities In Vitro

“…The gradient used is shown in Table 1. The fucoxanthin was detected at 450 nm and analyzed in a mass spectrometer (LTQ XL, Thermo Scientific) using ESI in full scan mode from 200 to 1000 m/z at a temperature of 275 °C and −9.0 V. The m/z values (see supplementary material, S1) were compared to the analytical standard and to literature [30].…”

“…Anticoagulated blood was collected from healthy volunteers, approved by the local ethics committee (F-2015-064, Landesärztekammer Baden-Württemberg). Isolation of polymorphonuclear leukocytes (PML) and peripheral blood mononuclear cells (PBMCs) via dextran sedimentation and density gradient centrifugation was conducted as previously described by El Benna & Dang (2007) [30] and Neumann et al (2018) [31]. PMLs were resuspended in DPBS, PBMCs in RPMI medium with 10% fetal calf serum and 1% penicillin / streptomycin.…”

Coupling Front-End Separations, Ion Mobility Spectrometry, and Mass Spectrometry For Enhanced Multidimensional Biological and Environmental Analyses

“…However, this platform lacked measurement sensitivity due to the filtering nature of FAIMS and ion losses at the interface. Technological advances in the sensitivity of the ultra-FAIMS (μFAIMS) by the Owlstone firm (134, 135) prompted an additional evaluation of FAIMS-DTIMS-MS for 3D analyses of a bovine serum albumin tryptic digest and the separation of isomeric compounds (136). Although μFAIMS greatly improved the sensitivity of measurements, especially when utilizing a helium/nitrogen gas mixture as shown in Figure 6, its small size offered limited resolving power.…”

“…The red boxes in panels b and d illustrate the feature sensitivity differences between the two gas compositions. Reproduced with permission from Reference 136. Copyright 2015, Royal Society of Chemistry.…”

Greatly Increasing Trapped Ion Populations for Mobility Separations Using Traveling Waves in Structures for Lossless Ion Manipulations

“…Electrospray ionization (ESI) coupled with ion mobility-mass spectrometry (IM-MS) is an increasingly utilized and potentially powerful approach with broad applications in metabolomics, − glycomics, proteomics, etc. − Increases in the resolution, sensitivity, and dynamic range of IM-MS measurements are important for the analysis of complex samples. − The size of an ion population injected for IM separation is a key consideration, ultimately setting limits to the sensitivity and dynamic range and impacting essentially every aspect of platform performance, including ion utilization efficiency. For example, ions from electrospray ionization (ESI) are typically continuously generated, but conventional drift tube IM separations are discontinuous, requiring injection of a tightly bunched ion packet, and usually only a very small fraction of generated ions are utilized .…”