Nontargeted Quantitation of Lipid Classes Using Hydrophilic Interaction Liquid Chromatography–Electrospray Ionization Mass Spectrometry with Single Internal Standard and Response Factor Approach

Cífková, Eva; Holčapek, Michal; Lísa, Miroslav; Ovčačíková, Magdaléna; Lyčka, Antonı́n; Lynen, Frédéric; Sandra, Patrick

doi:10.1021/ac3024476

Cited by 123 publications

(98 citation statements)

References 46 publications

(86 reference statements)

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“…Different chromatographic systems have been described for analysis of multiple lipid classes on the same platform. Normal-phase LC or hydrophilic interaction LC (HILIC) have been described to separate different lipid classes (16)(17)(18)(19). However, run times are often more than 30 min, which is not favorable for high-throughput analysis, and additionally, separation of one lipid class into different molecular species is not possible.…”

Section: Uplc Analysismentioning

confidence: 99%

Enhanced lipid isomer separation in human plasma using reversed-phase UPLC with ion-mobility/high-resolution MS detection

Damen

Isaac

Langridge

et al. 2014

Journal of Lipid Research

113

105

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This article is available online at http://www.jlr.orgLipids are the building blocks of all cell membranes and are thus essential for various biological functions varying from membrane traffi cking to signal transduction. Disorders in lipid metabolism play a key role in various diseases including cardiovascular disease, cancer, diabetes mellitus, and infl ammation ( 1, 2 ). Lipids have been classifi ed into eight categories (fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids, prenol lipids, saccharolipids, and polyketides) by the International Committee for the Classifi cation and Nomenclature of Lipids in conjunction with the LIPID MAPS consortium, based on their chemically distinct functional backbones and hydrophobic nature ( 3, 4 ).As the study of lipids attracts more and more attention, lipidomics, as a branch of metabolomics, is a quickly emerging fi eld. It has been defi ned as "the full characterization of lipid molecular species and of their biological roles with respect to expression of proteins involved in lipid metabolism and function, including gene regulation" ( 5 ).In order to achieve the goal of full characterization, different MS-based approaches have been described. Some groups describe shotgun lipidomics using direct infusion into the mass spectrometer ( 6-11 ). Direct infusion MS has the advantage of short analysis times and consuming very small amounts of sample. However, structural elucidation of lipid isomers is not possible. In order to elucidate structures Abstract An ultraperformance LC (UPLC) method for the separation of different lipid molecular species and lipid isomers using a stationary phase incorporating charged surface hybrid (CSH) technology is described. The resulting enhanced separation possibilities of the method are demonstrated using standards and human plasma extracts. Lipids were extracted from human plasma samples with the Bligh and Dyer method. Separation of lipids was achieved on a 100 × 2.1 mm inner diameter CSH C 18 column using gradient elution with aqueous-acetonitrile-isopropanol mobile phases containing 10 mM ammonium formate/0.1% formic acid buffers at a fl ow rate of 0.4 ml/min. A UPLC run time of 20 min was routinely used, and a shorter method with a 10 min run time is also described. The method shows extremely stable retention times when human plasma extracts and a variety of biofl uids or tissues are analyzed [intra-assay relative standard deviation (RSD) <0.385% and <0.451% for 20 and 10 min gradients, respectively (n = 5); interassay RSD <0.673% and <0.763% for 20 and 10 min gradients, respectively (n = 30)]. The UPLC system was coupled to a hybrid quadrupole orthogonal acceleration time-of-fl ight mass spectrometer, equipped with a traveling wave ion-mobility cell. Besides demonstrating the separation for different lipids using the chromatographic method, we demonstrate the use of the ion-mobility MS platform for the structural elucidation of lipids. The method can now be used to elucidate structures of a wide variety of lipid...

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Section: Uplc Analysismentioning

confidence: 99%

Enhanced lipid isomer separation in human plasma using reversed-phase UPLC with ion-mobility/high-resolution MS detection

Damen

Isaac

Langridge

et al. 2014

Journal of Lipid Research

113

105

View full text Add to dashboard Cite

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“…In one analysis, such a wealth of information is not given by conventional univariate tests such as T-test or F-test. Investigating the reasons for the underlying causes of such breed-specific differences is beyond the scope of this study as it is discussed elsewhere [13,14]. What we have attempted to do here is to provide the scientific community with possible statistical tools to improve their analysis and interpretation of datasets similar to ours.…”

Section: Resultsmentioning

confidence: 99%

“…While techniques to measure and characterize fatty acids are well established [3,4,13,14], they produce large amounts of accurate data, which have to be interpreted. Most methods use pair-wise comparisons; comparing one fatty-acid at a time [2].…”

Section: Introductionmentioning

confidence: 99%

Are Breed-specific Differences in Beef Fatty-acid Profiles a "Needle in the Haystack"?

Ranganathan¹,

Hulová²,

Schreiner³

et al. 2014

AJFST

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A healthy human nutrition stems from macronutrients that provide energy and also from micronutrients that are indispensable for basic physiological processes. Essential fatty-acids have been proved and commonly agreed in the recent years as important elements of a healthy diet. This gives impetus to look for sources of essential fatty-acids in the diet. Beef is not only considered a complete protein, but also a source of essential fatty-acids such as linoleic and linolenic acids. The levels of these fatty-acids are however variable depending on the diet and breed of cattle. In this work, we analyzed intra-muscular fatty acid profiles of two breeds (Aberdeen Angus and Blonde d'Aquitaine) as a part of long-term complex and comprehensive comparison of breed differences. Using a novel statistical algorithm we found interesting breed-specific fatty-acid profiles. The advantages of this approach over conventional approaches are discussed as well as the specific differences in fatty acid profiles between the breeds.

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“…3. Alternative chromatographic techniques for lipid separations 25 are hydrophilic interaction LC, 26 supercritical fluid chromatography, 27,28 mixed mode columns and chiral stationary phases, 28 silver ion high performance LC, and gas chromatography. Gas chromatography MS (GC-MS) is mainly used for volatile analytes.…”

Section: Separation Of Lipid Speciesmentioning

confidence: 99%

“…The absolute concentration can be calculated by the ratio of the lipid species to the IS multiplied by the response factor and the known concentration of the IS. 26 However, 1 IS may not be sufficient for nonpolar lipid classes, such as TG and polar lipids, in concentrations above the critical micellar concentration. …”

Section: Quantitation Of Lipid Speciesmentioning

confidence: 99%

Lipidomics

Hinterwirth

Stegemann

Mayr

2014

Circ Cardiovasc Genet

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Abstract-Lipidomics is the comprehensive analysis of molecular lipid species, including their quantitation and metabolic pathways. The huge diversity of native lipids and their modifications make lipidomic analyses challenging. The method of choice for sensitive detection and quantitation of molecular lipid species is mass spectrometry, either by direct infusion (shotgun lipidomics) or coupled with liquid chromatography. Although shotgun lipidomics allows for high-throughput analysis, low-abundant lipid species are not detected. Previous separation of lipid species by liquid chromatography increases ionization efficiency and is better suited for quantifying low abundant and isomeric lipid species. In this review, we will discuss the potential of lipidomics for cardiovascular research. To date, cardiovascular research predominantly focuses on the role of lipid classes rather than molecular entities. An in-depth knowledge about the molecular lipid species that contribute to the pathophysiology of cardiovascular diseases may provide better biomarkers and novel therapeutic targets for cardiovascular disease. (Circ Cardiovasc Genet. 2014;7:941-954.)

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Nontargeted Quantitation of Lipid Classes Using Hydrophilic Interaction Liquid Chromatography–Electrospray Ionization Mass Spectrometry with Single Internal Standard and Response Factor Approach

Cited by 123 publications

References 46 publications

Enhanced lipid isomer separation in human plasma using reversed-phase UPLC with ion-mobility/high-resolution MS detection

Enhanced lipid isomer separation in human plasma using reversed-phase UPLC with ion-mobility/high-resolution MS detection

Are Breed-specific Differences in Beef Fatty-acid Profiles a "Needle in the Haystack"?

Lipidomics

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