Rapid Solution-Phase Hydrogen/Deuterium Exchange for Metabolite Compound Identification

Majuta, Sandra N.; Li, Chong; Jayasundara, Kinkini; Karanji, Ahmad Kiani; Attanayake, Kushani; Ranganathan, Nandhini; Valentine, Stephen J.

doi:10.1007/s13361-019-02163-0

Cited by 17 publications

(44 citation statements)

References 93 publications

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“…The performance of in - droplet HDX using cVSSI benefits from groundbreaking experiments suggesting that reaction rates for different reactions could be increased by orders of magnitude when occurring in microdroplets as opposed to a bulk solution environment. − The most influential experiments to this work are those that utilized theta capillaries to monitor processes such as complex formation, HDX, protein unfolding, and ion supercharging using mass spectrometry. − In one study, the HDX rates of different amino hydrogens were monitored for a small molecule using mass spectrometry . This study, in particular, prompted recent experiments using cVSSI in a manner similar to nano-electrospray ionization (nESI) with theta capillaries to obtain unique isotopic distributions for different low-molecular-weight compounds . A problem with using the theta capillary approach to obtain exchangeable hydrogen information for metabolomics experiments is that HDX cannot be instantaneously turned on and off, making it difficult to effectively sample compounds eluting from LC columns.…”

Section: Introductionmentioning

confidence: 99%

“…Finally, although the primary goal of LC-HDX-MS in metabolomics analyses is to determine the number of heteroatom sites on different molecules, it may also be possible to obtain additional distinguishing information. One example discussed in previous cVSSI work is the ability to generate unique isotopic distributions for compounds based on differences in exchange site reactivities . In this scenario, it can be envisioned that such isotopic distributions could be matched to those in databases similar to searches performed with GC–MS data.…”

Section: Introductionmentioning

confidence: 99%

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Characterizing Multidevice Capillary Vibrating Sharp-Edge Spray Ionization for In-Droplet Hydrogen/Deuterium Exchange to Enhance Compound Identification

et al. 2021

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Multidevice capillary vibrating sharp-edge spray ionization (cVSSI) source parameters have been examined to determine their effects on conducting in-droplet hydrogen/deuterium exchange (HDX) experiments. Control experiments using select compounds indicate that the observed differences in mass spectral isotopic distributions obtained upon initiation of HDX result primarily from solution-phase reactions as opposed to gas-phase exchange. Preliminary studies have determined that robust HDX can only be achieved with the application of same-polarity voltage to both the analyte and the deuterium oxide reagent (D 2 O) cVSSI devices. Additionally, a similar HDX reactivity dependence on the voltage applied to the D 2 O device for various analytes is observed. Analyte and reagent flow experiments show that, for the multidevice cVSSI setup employed, there is a nonlinear dependence on the D 2 O reagent flow rate; increasing the D 2 O reagent flow by 100% results in only an ∼10−20% increase in deuterium incorporation for this setup. Instantaneous (subsecond) response times have been demonstrated in the initiation or termination of HDX, which is achieved by turning on or off the reagent cVSSI device piezoelectric transducer. The ability to distinguish isomeric species by in-droplet HDX is presented. Finally, a demonstration of a three-component cVSSI device setup to perform multiple (successive or in combination) in-droplet chemistries to enhance compound ionization and identification is presented and a hypothetical metabolomics workflow consisting of successive multidevice activation is briefly discussed.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterizing Multidevice Capillary Vibrating Sharp-Edge Spray Ionization for In-Droplet Hydrogen/Deuterium Exchange to Enhance Compound Identification

et al. 2021

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“…Presumably, this produces a more expansive area from which solvent streaming may occur. Notably, the employment of the sharp edges in VSSI and cVSSI is what allows for such high efficiency nebulization with very little power requirements (see below). − …”

Section: Introductionmentioning

confidence: 99%

Combining Field-Enabled Capillary Vibrating Sharp-Edge Spray Ionization with Microflow Liquid Chromatography and Mass Spectrometry to Enhance ‘Omics Analyses

Majuta

DeBastiani

et al. 2021

J. Am. Soc. Mass Spectrom.

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Field-enabled capillary vibrating sharp-edge spray ionization (cVSSI) has been combined with high-flow liquid chromatography (LC) and mass spectrometry (MS) to establish current ionization capabilities for metabolomics and proteomics investigations. Comparisons are made between experiments employing cVSSI and a heated electrospray ionization probe representing the state-of-the-art in microflow LC-MS methods for 'omics studies. For metabolomics standards, cVSSI is shown to provide an ionization enhancement by factors of 4 ± 2 for both negative and positive ion mode analyses. For chymotryptic peptides, cVSSI is shown to provide an ionization enhancement by factors of 5 ± 2 and 2 ± 1 for negative and positive ion mode analyses, respectively. Slightly broader high-performance liquid chromatography peaks are observed in the cVSSI datasets, and several studies suggest that this results from a slightly decreased post-split flow rate. This may result from partial obstruction of the pulled-tip emitter over time. Such a challenge can be remedied with the use of LC pumps that operate in the 10 to 100 μL•min −1 flow regime. At this early stage, the proof-of-principle studies already show ion signal advantages over stateof-the-art electrospray ionization (ESI) for a wide variety of analytes in both positive and negative ion mode. Overall, this represents a ∼20−50-fold improvement over the first demonstration of LC-MS analyses by voltage-free cVSSI. Separate comparisons of the ion abundances of compounds eluting under identical solvent conditions reveal ionization efficiency differences between cVSSI and ESI and may suggest varied contributions to ionization from different physicochemical properties of the compounds. Future investigations of parameters that could further increase ionization gains in negative and positive ion mode analyses with the use of cVSSI are briefly presented.

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“…In recent years, the Valentine group has conducted a series of systematic HDX studies on small molecules. This work ranges from predicting the HDX pattern of functional groups to the proof-of-concept droplet HDX study for metabolomic studies [18,20,23]. Motivated by this past work, we were inspired to integrate liquid-phase HDX with AIMS to develop a simple, robust, and cost-effective method for distinguishing isobaric ions in untargeted metabolomic studies.…”

Section: Introductionmentioning

confidence: 99%

Distinguishing between Isobaric Ions Using Microdroplet Hydrogen–Deuterium Exchange Mass Spectrometry

Song

Mofidfar

et al. 2021

Metabolites

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Isobaric ions having the same mass-to-charge ratio cannot be separately identified by mass spectrometry (MS) alone, but this limitation can be overcome by using hydrogen–deuterium exchange (HDX) in microdroplets. Because isobaric ions may contain a varied number of exchangeable sites and different types of functional groups, each one produces a unique MS spectral pattern after droplet spray HDX without the need for MS/MS experiments or introduction of ion mobility measurements. As an example of the power of this approach, isobaric ions in urinary metabolic profiles are identified and used to distinguish between healthy individuals and those having bladder cancer.

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Rapid Solution-Phase Hydrogen/Deuterium Exchange for Metabolite Compound Identification

Cited by 17 publications

References 93 publications

Characterizing Multidevice Capillary Vibrating Sharp-Edge Spray Ionization for In-Droplet Hydrogen/Deuterium Exchange to Enhance Compound Identification

Characterizing Multidevice Capillary Vibrating Sharp-Edge Spray Ionization for In-Droplet Hydrogen/Deuterium Exchange to Enhance Compound Identification

Combining Field-Enabled Capillary Vibrating Sharp-Edge Spray Ionization with Microflow Liquid Chromatography and Mass Spectrometry to Enhance ‘Omics Analyses

Distinguishing between Isobaric Ions Using Microdroplet Hydrogen–Deuterium Exchange Mass Spectrometry

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