Dual cationic–anionic profiling of metabolites in a single identified cell in a live <i>Xenopus laevis</i> embryo by microprobe CE-ESI-MS

Portero, Erika P.; Nemes, Péter

doi:10.1039/c8an01999a

Cited by 47 publications

(34 citation statements)

References 45 publications

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“…Recently, Portero et al. developed a method that allows sequential cationic and anionic analysis of metabolites in a single cell off a live vertebrate embryo of the South African frog Xenopus laevis . The low‐flow SL CE–MS interface was custom‐built and supplemented with a nitrogen gas filled chamber to minimize electrical discharges and produce a stable ESI spray in both positive and negative ion mode.…”

Section: Methodological Developmentsmentioning

confidence: 99%

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CE–MS for anionic metabolic profiling: An overview of methodological developments

2019

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The efficient profiling of highly polar and charged metabolites in biological samples remains a huge analytical challenge in metabolomics. Over the last decade, new analytical techniques have been developed for the selective and sensitive analysis of polar ionogenic compounds in various matrices. Still, the analysis of such compounds, notably for acidic ionogenic metabolites, remains a challenging endeavor, even more when the available sample size becomes an issue for the total analytical workflow. In this paper, we give an overview of the possibilities of capillary electrophoresis‐mass spectrometry (CE–MS) for anionic metabolic profiling by focusing on main methodological developments. Attention is paid to the development of improved separation conditions and new interfacing designs in CE–MS for anionic metabolic profiling. A complete overview of all CE–MS‐based methods developed for this purpose is provided in table format (Table 1) which includes information on sample type, separation conditions, mass analyzer and limits of detection (LODs). Selected applications are discussed to show the utility of CE–MS for anionic metabolic profiling, especially for small‐volume biological samples. On the basis of the examination of the reported literature in this specific field, we conclude that there is still room for the design of a highly sensitive and reliable CE–MS method for anionic metabolic profiling. A rigorous validation and the availability of standard operating procedures would be highly favorable in order to make CE–MS an alternative, viable analytical technique for metabolomics.

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Section: Methodological Developmentsmentioning

confidence: 99%

“…Representative mass spectra of a V1 cell extract revealing simplified spectral complexity during ESI−. Reproduced from with permission.…”

Section: Methodological Developmentsmentioning

confidence: 99%

CE–MS for anionic metabolic profiling: An overview of methodological developments

2019

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“…Additionally, 100s of volatiles from single lung cancer cells with various p53 genetic status and normal lung cells were captured using an innovative n‐Tube Extraction (ITEX)‐GC‐MS extraction‐analysis method; where18 VOCs showed significant changes in their concentration levels in tumor cells versus control in this volatilomics approach . Detailed reviews are available on single cell metabolomics that use MS‐based technologies and methods and live single cell metabolomics using microprobe CE‐ESI‐MS . New MS technologies include structures for lossless ion manipulations (SLIM) and hydrogen flame desorption ionization mass spectrometer (HFDI‐MS), among others .…”

Section: Spatial Resolution—a Solution To Heterogeneity and Increasedmentioning

confidence: 99%

Challenges and Opportunities in Cancer Metabolomics

Kumar

Misra

2019

Proteomics

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Challenges in metabolomics for a given spectrum of disease are more or less comparable, ranging from the accurate measurement of metabolite abundance, compound annotation, identification of unknown constituents, and interpretation of untargeted and analysis of high throughput targeted metabolomics data leading to the identification of biomarkers. However, metabolomics approaches in cancer studies specifically suffer from several additional challenges and require robust ways to sample the cells and tissues in order to tackle the constantly evolving cancer landscape. These constraints include, but are not limited to, discriminating the signals from given cell types and those that are cancer specific, discerning signals that are systemic and confounded, cell culture‐based challenges associated with cell line identities and media standardizations, the need to look beyond Warburg effects, citrate cycle, lactate metabolism, and identifying and developing technologies to precisely and effectively sample and profile the heterogeneous tumor environment. This review article discusses some of the current and pertinent hurdles in cancer metabolomics studies. In addition, it addresses some of the most recent and exciting developments in metabolomics that may address some of these issues. The aim of this article is to update the oncometabolomics research community about the challenges and potential solutions to these issues.

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“…For example, Portero et al. complemented a home‐made low‐flow sheath‐liquid interface design for CZE–MS with a nitrogen gas filled chamber to minimize electrical discharges and to obtain a stable ESI spray in the negative ion mode . In order to avoid corona discharge, Dodbiba et al.…”

Section: Introductionmentioning

confidence: 99%

“…This effect can be attenuated by the addition of organic solvents, nitrogen gas, oxygen or other reagents acting as electron scavengers. For example, Portero et al complemented a home-made low-flow sheath-liquid interface design for CZE-MS with a nitrogen gas filled chamber to minimize electrical discharges and to obtain a stable ESI spray in the negative ion mode [15]. In order to avoid corona discharge, Dodbiba et al analyzed nucleotides in the positive ion mode by employing different cationic ion-pairing reagents that associate with nucleotides resulting in overall positively charged complexes [16].…”

Section: Introductionmentioning

confidence: 99%

Profiling nucleotides in low numbers of mammalian cells by sheathless CE–MS in positive ion mode: Circumventing corona discharge

et al. 2020

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Negative ion mode nano‐ESI–MS is often considered for the analysis of acidic compounds, including nucleotides. However, under high aqueous separation conditions, corona discharge is frequently observed at emitter tips, which may result in low ion abundances and reduced nano‐ESI needle emitter lifetimes. In this work, we introduce a sheathless CE‐MS method for the highly efficient and sensitive analysis of nucleotides employing ESI in positive ion mode, thereby fully circumventing corona discharge. By using a background electrolyte of 16 mM ammonium acetate (pH 9.7) a mixture of 12 nucleotides, composed of mono‐, di‐, and tri‐phosphates, could be efficiently analyzed with plate numbers per meter above 220 000 and with LODs in the range from 0.06 to 1.3 nM, corresponding to 0.4 to 8.6 attomole, when using an injection volume of about 6.5 nL only. The utility of the method was demonstrated for the profiling of nucleotides in low numbers of mammalian cells using HepG2 cells as a model system. Endogenous nucleotides could be efficiently analyzed in extracts from 50 000 down to 500 HepG2 cells only. Moreover, apart from nucleotides, also some nicotinamide‐adenine dinucleotides and amino acids could be analyzed under these conditions, thereby clearly illustrating the utility of this approach for metabolic profiling of low amounts of biological material.

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Dual cationic–anionic profiling of metabolites in a single identified cell in a live Xenopus laevis embryo by microprobe CE-ESI-MS

Abstract: In situ capillary microsampling with capillary electrophoresis (CE) electrospray ionization (ESI) mass spectrometry (MS) enabled the characterization of cationic and anionic metabolites in single cells in complex tissues and organisms, such as the live frog (X. laevis) embryo.

Cited by 47 publications

References 45 publications

CE–MS for anionic metabolic profiling: An overview of methodological developments

CE–MS for anionic metabolic profiling: An overview of methodological developments

Challenges and Opportunities in Cancer Metabolomics

Profiling nucleotides in low numbers of mammalian cells by sheathless CE–MS in positive ion mode: Circumventing corona discharge

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