Supercritical fluid chromatography coupled to mass spectrometry for lipidomics

Chollet, Catherine; Boutet-Mercey, Stéphanie; Laboureur, Laurent; Rincón, Carlos; Méjean, Marie; Jouhet, Juliette; Fenaille, François; Colsch, Benoît; Touboul, David

doi:10.1002/jms.4445

Cited by 29 publications

(22 citation statements)

References 43 publications

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“…The optimized chromatographic gradient starting with pure non-polar CO 2 , which is then followed by the addition of up to 51% methanol/water (99:1, v/v) containing 30 mM of ammonium acetate, is key to separate both polar and non-polar lipids. Further applications of analytical SFC in lipidomics are comprehensively summarized elsewhere [15][16][17][18]. However, the lipid class-specific SFC separation introduced by Lísa et al remains unrivaled in both separation speed and coverage up to date.…”

Section: Introductionmentioning

confidence: 99%

Preparative supercritical fluid chromatography for lipid class fractionation—a novel strategy in high-resolution mass spectrometry based lipidomics

et al. 2020

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In this work, a lipidomics workflow based on offline semi-preparative lipid class-specific fractionation by supercritical fluid chromatography (SFC) followed by high-resolution mass spectrometry was introduced. The powerful SFC approach offered separation of a wide polarity range for lipids, enabled enrichment (up to 3 orders of magnitude) of lipids, selective fractionation of 14 lipid classes/ subclasses, and increased dynamic range enabling in-depth characterization. A significantly increased coverage of low abundant lipids improving lipid identification by numbers and degree (species and molecular level) was obtained in Pichia pastoris when comparing high-resolution mass spectrometry based lipidomics with and without prior fractionation. Proof-of-principle experiments using a standard reference material (SRM 1950, NIST) for human plasma showed that the proposed strategy enabled quantitative lipidomics. Indeed, for 70 lipids, the consensus values available for this sample could be met. Thus, the novel workflow is ideally suited for lipid class-specific purification/isolation from milligram amounts of sample while not compromising on omics type of analysis (identification and quantification). Finally, compared with established fractionation/pre-concentration approaches, semi-preparative SFC is superior in terms of versatility, as it involved only volatile modifiers and salt additives facilitating any follow-up use such as qualitative or quantitate analysis or further purification down to the single lipid species level.

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Section: Introductionmentioning

confidence: 99%

Preparative supercritical fluid chromatography for lipid class fractionation—a novel strategy in high-resolution mass spectrometry based lipidomics

et al. 2020

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“…In addition, a supercritical fluid chromatography (SFC) has recently received a lot of attention for lipid analysis, thanks to the development of robust commercial solutions and high separation efficiency and short analysis time compared to commonly used LC‐MS. [ 93,94 ] In general, the specified separation techniques are powerful in reducing the matrix effects to minimize the ion‐suppression and therefore maximize the MS signal intensity. They will also maximize the measurement specificity by providing the complementary data, such as retention time, for metabolite identification.…”

Section: “One Size Doesn't Fit All”—resolving Metabolite Chemical DIVmentioning

confidence: 99%

“…It was straight off applied in clinical practice during surgical interventions (i.e., iKnife) for supervised cancerous tissue removal. [88,89] Although MS imaging offers this lateral resolution on local distribution of metabolites within tissue (vs. the analysis of a homogenized tissue extracts as a whole) and its spatial resolution was improved remarkably to allow for imaging at single-cell and even subcellular level, [90][91][92] several challenges, mainly regarding the low ionization yields of still broad-range of metabolites, identification and quantification issues remain to be addressed prior to its wider application in biomedical and clinical research.…”

Section: Surface Ionization Techniques: Mass Spectrometry Imagingmentioning

confidence: 99%

Metabolomics meets lipidomics: Assessing the small molecule component of metabolism

2020

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Metabolomics, including lipidomics, is emerging as a quantitative biology approach for the assessment of energy flow through metabolism and information flow through metabolic signaling; thus, providing novel insights into metabolism and its regulation, in health, healthy ageing and disease. In this forward-looking review we provide an overview on the origins of metabolomics, on its role in this postgenomic era of biochemistry and its application to investigate metabolite role and (bio)activity, from model systems to human population studies. We present the challenges inherent to this analytical science, and approaches and modes of analysis that are used to resolve, characterize and measure the infinite chemical diversity contained in the metabolome (including lipidome) of complex biological matrices. In the current outbreak of metabolic diseases such as cardiometabolic disorders, cancer and neurodegenerative diseases, metabolomics appears to be ideally situated for the investigation of disease pathophysiology from a metabolite perspective.

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“…Taking into account all these technical limitations, for untargeted lipidomic profiling, in our facility, we chose to work with a supercritical fluid chromatography system using CO 2 under pressure as an eluent. This technique is especially suitable for lipids and allows for a shorter separation than liquid chromatography [ 11 , 12 ]. Recently, different studies [ 8 , 13 ] presented the potential of Supercritical Fluid Chromatography (SFC) for profiling lipids in a very short time, especially for clinical data.…”

Section: Introductionmentioning

confidence: 99%

Untargeted Lipidomic Profiling of Dry Blood Spots Using SFC-HRMS

et al. 2021

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Lipids are essential cellular constituents that have many critical roles in physiological functions. They are notably involved in energy storage and cell signaling as second messengers, and they are major constituents of cell membranes, including lipid rafts. As a consequence, they are implicated in a large number of heterogeneous diseases, such as cancer, diabetes, neurological disorders, and inherited metabolic diseases. Due to the high structural diversity and complexity of lipid species, the presence of isomeric and isobaric lipid species, and their occurrence at a large concentration scale, a complete lipidomic profiling of biological matrices remains challenging, especially in clinical contexts. Using supercritical fluid chromatography coupled with high-resolution mass spectrometry, we have developed and validated an untargeted lipidomic approach to the profiling of plasma and blood. Moreover, we have tested the technique using the Dry Blood Spot (DBS) method and found that it allows for the easy collection of blood for analysis. To develop the method, we performed the optimization of the separation and detection of lipid species on pure standards, reference human plasma (SRM1950), whole blood, and DBS. These analyses allowed an in-house lipid data bank to be built. Using the MS-Dial software, we developed an automatic process for the relative quantification of around 500 lipids species belonging to the 6 main classes of lipids (including phospholipids, sphingolipids, free fatty acids, sterols, and fatty acyl-carnitines). Then, we compared the method using the published data for SRM 1950 and a mouse blood sample, along with another sample of the same blood collected using the DBS method. In this study, we provided a method for blood lipidomic profiling that can be used for the easy sampling of dry blood spots.

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Supercritical fluid chromatography coupled to mass spectrometry for lipidomics

Cited by 29 publications

References 43 publications

Preparative supercritical fluid chromatography for lipid class fractionation—a novel strategy in high-resolution mass spectrometry based lipidomics

Preparative supercritical fluid chromatography for lipid class fractionation—a novel strategy in high-resolution mass spectrometry based lipidomics

Metabolomics meets lipidomics: Assessing the small molecule component of metabolism

Untargeted Lipidomic Profiling of Dry Blood Spots Using SFC-HRMS

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