Reporting of lipidomics data should be standardized

Liebisch, Gerhard; Ekroos, Kim; Hermansson, Martin; Ejsing, Christer S.

doi:10.1016/j.bbalip.2017.02.013

Cited by 83 publications

(73 citation statements)

References 47 publications

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“…Validation of the novel method demonstrated its suitability for large-scale studies despite the higher variations observed for some samples. These variations are related to inhomogeneity of samples and lipolytic activity that requires further investigations considering pre-analytical issues as an essential part of lipidomic workflows and their standardization [29,30,41]. Therefore, we recommend performing measurements in triplicate, when high accuracy is needed.…”

Section: Resultsmentioning

confidence: 99%

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Quantification of diacylglycerol and triacylglycerol species in human fecal samples by flow injection Fourier transform mass spectrometry

et al. 2020

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The intestinal microbiome plays an important role in human health and disease and fecal materials reflect the microbial activity. Thus, analysis of fecal metabolites provides insight in metabolic interactions between gut microbiota and host organism. In this work, we applied flow injection analysis coupled to Fourier transform mass spectrometry (FIA-FTMS) to identify and quantify lipid species in human fecal samples. Fecal homogenates were subjected to lipid extraction and analyzed by FIA-FTMS. The analysis of different subjects revealed a vast heterogeneity of lipid species abundance. The majority of samples displayed prominent signals of triacylglycerol (TG) and diacylglycerol (DG) species that could be verified by MS2 spectra. Therefore, we focused on the quantification of TG and DG. Method validation included limit of quantification, linearity, evaluation of matrix effects, recovery, and reproducibility. The validation experiments demonstrated the suitability of the method, with exception for approximately 10% of samples, where we observed coefficients of variation higher than 15%. Impaired reproducibility was related to sample inhomogeneity and could not be improved by additional sample preparation steps. Additionally, these experiments demonstrated that compared with aqueous samples, samples containing isopropanol showed higher amounts of DG, presumably due to lysis of bacteria and increased TG lipolysis. These effects were sample-specific and substantiate the high heterogeneity of fecal materials as well as the need for further evaluation of pre-analytic conditions. In summary, FIA-FTMS offers a fast and accurate tool to quantify DG and TG species and is suitable to provide insight into the fecal lipidome and its role in health and disease.

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Section: Resultsmentioning

confidence: 99%

“…In an important next step within method development [29,30], we evaluated the performance of the FIA-FTMS method. Due to sample heterogeneity, intra-and inter-day precisions were evaluated in five different samples (Tables 2 and 3).…”

Section: Reproducibilitymentioning

confidence: 99%

Quantification of diacylglycerol and triacylglycerol species in human fecal samples by flow injection Fourier transform mass spectrometry

et al. 2020

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“…These and other intriguing results have spurred interest in adopting lipidomics capabilities across research communities. A major challenge in the field of lipidomics, however, is the large disparity in methodologies and technologies, resulting in discrepancies in published data and broader issues of irreproducibility 8 . Common problems include improper annotation of lipid species (despite the publication of an accurate shorthand annotation for lipid species in 2013 9 ), misidentifications and overreporting, likely caused by incorrect mapping of mass spectral features to potential lipid molecules due to software errors combined with the a lack of manual data inspection or curation.…”

Section: Current State Of Lipidomicsmentioning

confidence: 99%

Lipidomics needs more standardization

et al. 2019

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“…ESI is commonly used in MS but is particularly sensitive to ion suppression and matrix effects. Therefore, coextraction of interferents, such as nonlipid biomolecules, leads to increased complexity of MS spectra, decreased sensitivity, and reduced lipid identification (2,6,7). The importance of sample preparation prior to lipidomic analysis is critical, as it can negatively influence the analytical measurements.…”

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confidence: 99%

“…To overcome this difficulty, alternative extraction methods have been developed using solvents such as methyl-tert-butyl ether (7) and butanol/methanol (15), in which the density of the organic layer is less then water, making it the upper phase (UP). These methods have been validated in a variety of sample matrices and with some lipid classes but may be limited with untested conditions for lipid species (6,16). If new matrices are used, methods must be revalidated regarding lipid class diversity, total lipid extraction, and reproducibility, and the results must be compared with the extraction efficiency of well-established methods such as the mBD and Folch methods.…”

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Three-phase liquid extraction: a simple and fast method for lipidomic workflows

Vale

Martin

Mitsche

et al. 2019

Journal of Lipid Research

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This article is available online at http://www.jlr.org hydrophobic lipid species. Ceramides and diacylglycerols (DAGs) have polarities that fall in between the previous examples. This range of hydrophobicity adds complexity to the lipid analysis. For this reason, an effective method that decreases bias while isolating a representative sample of lipids from biological matrices is critical for a lipidomic analysis.Lipidomic profiling by MS has become a widely used tool in lipid biochemistry. Recent advances in MS have led to better sensitivity, improved mass accuracy, and novel methods for broad lipid profiling. ESI is commonly used in MS but is particularly sensitive to ion suppression and matrix effects. Therefore, coextraction of interferents, such as nonlipid biomolecules, leads to increased complexity of MS spectra, decreased sensitivity, and reduced lipid identification (2, 6, 7). The importance of sample preparation prior to lipidomic analysis is critical, as it can negatively influence the analytical measurements. Ensuring the quality of lipid extracts is an important step because improper sample preparation will yield questionable or misleading results. Liquid-liquid extractions (LLEs) and solid-phase extractions (SPEs) are the most common sample preparation techniques used in lipidomics and are implemented depending on analytical strategy; LLE is favored for global lipidomic analysis (8-10), while SPE is predominately used for targeted methods when the fractionation and/or isolation of a specific lipid class or classes is necessary. However, SPE can be variable, time-consuming, and challenging to automate. The efficiency of LLE depends on the partitioning of lipids into an organic phase. The most commonly Abstract An unbiased sample preparation free of interferents (i.e., competing analytes, detergents, plastics) is critical to any lipid MS workflow. Here we present a novel three-phase lipid extraction (3PLE) technique using a single-step liquid-liquid extraction (LLE) that allows both extraction and fractionation of lipids by polarity. 3PLE is composed of one aqueous and two organic phases. The upper organic phase is enriched in neutral lipids (triacylglycerols and cholesteryl esters), while the middle organic phase contains the major glycerophospholipids. Thin-layer chromatography, radioactive labeling, and MS were used to confirm lipid partitioning. 3PLE efficiency was demonstrated for bovine liver, human pooled plasma, mouse liver, mouse brain, and mouse white adipose tissue. Compared with the gold-standard Bligh/Dyer LLE, 3PLE showed significant advantages. For direct-infusion workflows, there was a decrease in ion suppression with a corresponding increased number of lipid species identified. For LC/MS workflows, increased signal intensities were observed for lower-abundance lipid species such as phosphatidic acid and phosphatidylserine. 3PLE also proved to be a valuable tool for fatty acid profiling by GC/MS, allowing for the separate identification of neutral and polar fatty acids.-Vale, G., S. A. ...

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Reporting of lipidomics data should be standardized

Cited by 83 publications

References 47 publications

Quantification of diacylglycerol and triacylglycerol species in human fecal samples by flow injection Fourier transform mass spectrometry

Quantification of diacylglycerol and triacylglycerol species in human fecal samples by flow injection Fourier transform mass spectrometry

Lipidomics needs more standardization

Three-phase liquid extraction: a simple and fast method for lipidomic workflows

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