Complementing reversed-phase selectivity with porous graphitized carbon to increase the metabolome coverage in an on-line two-dimensional LC-MS setup for metabolomics

Ortmayr, Karin; Koellensperger, Gunda

doi:10.1039/c5an00206k

Cited by 30 publications

(27 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Because the combination of different chromatography methods could increase metabolome coverage, MDLC approaches, which have been commonly adopted in proteomics, are also of interest in metabolomics . Ortmayr et al developed an on‐line two‐dimensional LC‐MS method, which combined RP and PGC, for metabolomics . When tested with a mixture of 82 metabolite standards, the number of retained metabolites could be doubled compared to RPLC alone; the use of PGC extends the metabolite coverage of those analytically challenging compounds like sugar phosphates.…”

Section: Metabolome Coverage Expansionmentioning

confidence: 99%

“…125 Ortmayr et al developed an on-line two-dimensional LC-MS method, which combined RP and PGC, for metabolomics. 126 When tested with a mixture of 82 metabolite standards, the number of retained metabolites could be doubled compared to RPLC alone; the use of PGC extends the metabolite coverage of those analytically challenging compounds like sugar phosphates. Good retention-time stability (average 0.4%) was achieved with cell-culture extracts to show that the method might be suitable for untargeted metabolomics.…”

Section: Expand Metabolome Detection Via Different Types Of Liquid mentioning

confidence: 99%

See 1 more Smart Citation

Challenges and emergent solutions for LC‐MS/MS based untargeted metabolomics in diseases

Liang

Lee

2018

Mass Spectrometry Reviews

247

169

View full text Add to dashboard Cite

In the past decade, advances in liquid chromatography-mass spectrometry (LC-MS) have revolutionized untargeted metabolomics analyses. By mining metabolomes more deeply, researchers are now primed to uncover key metabolites and their associations with diseases. The employment of untargeted metabolomics has led to new biomarker discoveries and a better mechanistic understanding of diseases with applications in precision medicine. However, many major pertinent challenges remain. First, compound identification has been poor, and left an overwhelming number of unidentified peaks. Second, partial, incomplete metabolomes persist due to factors such as limitations in mass spectrometry data acquisition speeds, wide-range of metabolites concentrations, and cellular/tissue/temporal-specific expression changes that confound our understanding of metabolite perturbations. Third, to contextualize metabolites in pathways and biology is difficult because many metabolites partake in multiple pathways, have yet to be described species specificity, or possess unannotated or more-complex functions that are not easily characterized through metabolomics analyses. From a translational perspective, information related to novel metabolite biomarkers, metabolic pathways, and drug targets might be sparser than they should be. Thankfully, significant progress has been made and novel solutions are emerging, achieved through sustained academic and industrial community efforts in terms of hardware, computational, and experimental approaches. Given the rapidly growing utility of metabolomics, this review will offer new perspectives, increase awareness of the major challenges in LC-MS metabolomics that will significantly benefit the metabolomics community and also the broader the biomedical community metabolomics aspire to serve.

show abstract

Section: Metabolome Coverage Expansionmentioning

confidence: 99%

Section: Expand Metabolome Detection Via Different Types Of Liquid mentioning

confidence: 99%

Challenges and emergent solutions for LC‐MS/MS based untargeted metabolomics in diseases

Liang

Lee

2018

Mass Spectrometry Reviews

247

169

View full text Add to dashboard Cite

show abstract

“…A Vanquish UHPLC (Thermo Fisher Scientific) equipped with an additional pump was used for HILIC and RP coupling. A previously published RP-PGC metabolite setup 55 was adapted and further developed for lipid separation. The void volume including non-polar lipids was transferred from the firstdimension column (HILIC) to the second dimension (RP) using a separate two positional 6-port valve controlled by the mass spectrometer.…”

Section: Setup For Hilic-rp Liquid Chromatographymentioning

confidence: 99%

“…By exploiting hydrophobic and hydrophilic lipid interactions, (1) the use of a short HILIC column and direct transfer of the non-polar lipids without further dilution, (2) short gradient programs and (3) a relatively simple instrumental setup is enabled. The instrumental setup applied was adapted from a previously published RP-PGC metabolite method 55 and further developed for lipid separation. In position A (Fig.…”

Section: On-line Coupled Hilic-rp Liquid Chromatographymentioning

confidence: 99%

Simultaneous non-polar and polar lipid analysis by on-line combination of HILIC, RP and high resolution MS

Rampler

Schoeny

Mitic

et al. 2018

Analyst

Self Cite

View full text Add to dashboard Cite

Given the chemical diversity of lipids and their biological relevance, suitable methods for lipid profiling and quantification are demanded to reduce sample complexity and analysis times. In this work, we present a novel on-line chromatographic method coupling hydrophilic interaction liquid chromatography (HILIC) dedicated to class-specific separation of polar lipid to reversed-phase chromatography (RP) for non-polar lipid analysis. More specifically, the void volume of the HILIC separation-consisting of non-polar lipids- is transferred to the orthogonal RP column enabling the on-line combination of HILIC with RP without any dilution in the second dimension. In this setup the orthogonal HILIC and RP separations were performed in parallel and the effluents of both columns were combined prior to high-resolution MS detection, offering the full separation space in one analytical run. Rapid separation for both polar and non-polar lipids within only 15 min (including reequilibration time) was enabled using sub-2 μm particles and UHPLC. The method proved to be robust with excellent retention time stability (RSDs < 1%) and LODs in the fmol to pmol (absolute on column) range even in the presence of complex biological matrix such as human plasma. The presented high-resolution LC-MS/MS method leads to class-specific separation of polar lipids and separation of non-polar lipids which is lost in conventional HILIC separations. HILIC-RP-MS is a promising tool for targeted and untargeted lipidomics workflows as three interesting features are combined namely (1) the decreased run time of state of the art shotgun MS methods, (2) the elevated linear dynamic range inherent to chromatographic separation and (3) increased level of identification by separation of polar and non-polar lipid classes.

show abstract

“…Sample preparation is therefore the most time-consuming and challenging step in environmental analysis. The primary goal of sample preparation is to isolate one or several target analytes from the other components of the sample mixture [2,3]. Several methods have been presented for sample preparation [4,5].…”

Section: Introductionmentioning

confidence: 99%

Biocoacervation extraction combined with dispersive solid phase extraction using a reversed-phase core–shell magnetic molecularly imprinted sorbent for 2,4-dichlorophenoxyacetic acid prior to its determination by HPLC

Haeri

Abbasi

2016

J IRAN CHEM SOC

View full text Add to dashboard Cite

Complementing reversed-phase selectivity with porous graphitized carbon to increase the metabolome coverage in an on-line two-dimensional LC-MS setup for metabolomics

Abstract: A novel on-line combination of reversed phase and porous graphitized carbon liquid chromatography increases the versatility in non-targeted metabolomics.

Cited by 30 publications

References 46 publications

Challenges and emergent solutions for LC‐MS/MS based untargeted metabolomics in diseases

Challenges and emergent solutions for LC‐MS/MS based untargeted metabolomics in diseases

Simultaneous non-polar and polar lipid analysis by on-line combination of HILIC, RP and high resolution MS

Biocoacervation extraction combined with dispersive solid phase extraction using a reversed-phase core–shell magnetic molecularly imprinted sorbent for 2,4-dichlorophenoxyacetic acid prior to its determination by HPLC

Contact Info

Product

Resources

About