Liquid Chromatography Techniques in Lipidomics Research

Lange, M.; Ni, Zhixu; Criscuolo, Angela; Fedorova, Maria

doi:10.1007/s10337-018-3656-4

Cited by 46 publications

(48 citation statements)

References 132 publications

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“…It becomes increasingly clear that a high number of lipid standards is mandatory for accurate -omics type of quantification by RP-LC based methods. 3,15,32,34,35 In this work, a FI/LC-HRMS workflow integrated isotopically labeled yeast (LILY) as internal standards (see Figure 1) with the aim of expanding the number of lipids amenable to absolute accurate quantification as defined by the FDA guideline. 30 Several proof-of-concept studies showed the potential of LILY.…”

Section: Resultsmentioning

confidence: 99%

“…24 It is the method of choice when aiming at in-depth characterization of lipidomes. Reversed phase chromatography provides efficient matrix separation 32 together with excellent chromatographic selectivity and retentivity for lipids, resulting in high sensitivity and dynamic range when combined to MS detection. While the method is unrivalled in terms of lipid separation and thus identification (in combination with high resolution MS (HRMS)), proper standardization approaches imply the use of multiple standards per lipid class.…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, quantification accuracy is compromised when only few standards per lipid class are implemented. 3,15,32,34,35 In this work we propose a RP-LC-HRMS workflow enlarging the number of standards in a cost saving manner, thereby improving the quantification capability of the most powerful lipidomics technique. More specifically, the presented standardization strategy is based on a lipid library provided by 13 C fully labelled biomass, coined as lipidome isotope labeling of yeast (LILY).…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, quantification accuracy is compromised when only few standards per lipid class are implemented. 3,15,32,34,35…”

Section: Introductionmentioning

confidence: 99%

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A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with¹³C- internal standards

Rampler

Abiead

Hildebrand

et al. 2020

Preprint

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We propose a fully automated novel workflow for lipidomics based on flow injection- followed by liquid chromatography high resolution mass spectrometry (FI/LC-HRMS). The workflow combined in-depth characterization of the lipidome achieved via reversed phase LC-HRMS with absolute quantification as obtained by a high number of lipid species-specific- and/or retention time (RT) matched/class-specific calibrants. The lipidome of 13C labelled yeast (LILY) provided a cost efficient, large panel of internal standards covering triacylglycerols (TG), steryl esters (SE), free fatty acids (FA), diacylglycerols (DG), sterols (ST), ceramides (Cer), hexosyl ceramides (HexCer), phosphatidylglycerols (PG), phosphatidylethanolamines (PE), phosphatidic acids (PA), cardiolipins (CL), phosphatidylinositols (PI), phosphatidylserines (PS), phosphatidylcholines (PC), lysophosphatidylcholines (LPC) and lysophosphatidylethanolamines (LPE). In order to exploit the full potential of isotopically enriched biomass, LILY was absolutely quantified on demand via reversed isotope dilution analysis using FI-HRMS. Subsequent LC-HRMS analysis integrated different calibration strategies including lipid species-specific standards for >90 lipids. Extensive measures on quality control allowed to rank the calibration strategies and to automatically selected the calibration strategy of highest metrological order for the respective lipid species. Overall, the workflow enabled a streamlined analysis pipeline (identification and quantification in separate analytical runs) and provided validation tools together with absolute concentration values for > 350 lipids in human plasma on a species level with an analytical run-time of less than 25 min per sample.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…Consequently, quantification accuracy is compromised when only few standards per lipid class are implemented. 3,15,32,34,35…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with¹³C- internal standards

Rampler

Abiead

Hildebrand

et al. 2020

Preprint

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show abstract

“…In addition to these benefits, the retention time ( ) of an analyte provides complementary information to the mass-to-charge (m/z), as it derives from a broader set of physiochemical properties of the analyte. This complementary information can be especially beneficial in metabolomics where many of the analytes are isobaric 4,5 .…”

Section: Introductionmentioning

confidence: 99%

Generalized Calibration Across Liquid Chromatography Setups for Generic Prediction of Small-Molecule Retention Times

2020

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Motivation: Accurate prediction of liquid chromatographic retention times from small molecule structures is useful for reducing experimental measurements and for improved identification in targeted and untargeted MS. However, different experimental setups (e.g. differences in columns, gradients, solvents, or stationary phase) have given rise to a multitude of prediction models that only predict accurate retention times for a specific experimental setup. In practice this typically results in the fitting of a new predictive model for each specific type of setup, which is not only inefficient but also requires substantial prior data to be accumulated on each such setup. Results: Here we introduce the concept of generalized calibration, which is capable of the straightforward mapping of retention time models between different experimental setups. This concept builds on the database-controlled calibration approach implemented in PredRet, and fits calibration curves on predicted retention times instead of only on observed retention times. We show that this approach results in significantly higher accuracy of elution peak prediction than is achieved by setup-specific models. .

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Liquid Chromatography – and Supercritical Fluid Chromatography – Mass Spectrometry

Holčapek¹,

Peterka²,

Chocholoušková³

et al. 2023

Mass Spectrometry for Lipidomics

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Liquid Chromatography Techniques in Lipidomics Research

Cited by 46 publications

References 132 publications

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with¹³C- internal standards

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with¹³C- internal standards

Generalized Calibration Across Liquid Chromatography Setups for Generic Prediction of Small-Molecule Retention Times

Liquid Chromatography – and Supercritical Fluid Chromatography – Mass Spectrometry

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Liquid Chromatography Techniques in Lipidomics Research

Cited by 46 publications

References 132 publications

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with13C- internal standards

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with13C- internal standards

Generalized Calibration Across Liquid Chromatography Setups for Generic Prediction of Small-Molecule Retention Times

Liquid Chromatography – and Supercritical Fluid Chromatography – Mass Spectrometry

Contact Info

Product

Resources

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A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with¹³C- internal standards

A combined flow injection/reversed phase chromatography – high resolution mass spectrometry workflow for accurate absolute lipid quantification with¹³C- internal standards