Assessment of the retention properties of poly(vinyl alcohol) stationary phase for lipid class profiling in liquid chromatography

Deschamps, Frantz S.; Chaminade, Pierre; Ferrier, D.; Baillet, A.

doi:10.1016/s0021-9673(01)01137-2

Cited by 38 publications

(30 citation statements)

References 24 publications

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“…Therefore, similar binary mobile phases were assessed with our mixture of lipids ( Table 1). As previously described [22], chloroform was indeed too strong for Cho retention. However, the separation between Cho and SA can be observed.…”

Section: Solvent Selection With Pva-sil Stationary Phasesupporting

confidence: 50%

“…This analysis required gradient elution, because of the large range of lipid class polarity in natural samples. For the solvent selection, we took advantage of a previous study with PVA-Sil column for lipid class analysis where a screening of solvents with binary gradients was performed [22]. This work studied lipid classes of various polarities from triglycerides to phospholipids.…”

Section: Solvent Selection With Pva-sil Stationary Phasementioning

confidence: 99%

“…Several other polar stationary phases were studied for SC class analysis, such as diol [11,19] or cyanopropyl [20,21]. A polymerized poly(vinyl alcohol) on silica gel (PVA-Sil) was recently used for lipid class analysis [20,22,23]. The PVA-Sil is chemically bonded on silica particles.…”

Section: Introductionmentioning

confidence: 99%

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Microanalytical systems for separations of stratum corneum ceramides

et al. 2006

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Original PaperMicroanalytical systems for separations of stratum corneum ceramidesThe small amount of lipids from human skin obtained with noninvasive sampling method led us to investigate microanalytical separation techniques. The lipid class analysis was performed with a micro polyvinyl alcohol-silica (PVA-Silm) column. The gradient elution was from heptane to acetone/butanol 90:10 v/v in 4%/min at 78 lL/ min. In addition an evaporative light scattering detector (ELSD) was modified for micro-LC. All solvents contained 0.1% of triethylamine and formic acid in stoichiometric amount, which increased the ELSD response. In these conditions, the cholesterol eluted before free fatty acid, and squalene and triglycerides close to the dead volume. The various ceramide classes eluted following the order of the increased number of hydroxyl groups. The LOD for ceramides was 2.2 ng. The advantages of this method are the use of a normal stationary phase more reliable due to its chemical stability, its surface homogeneity and its development in microchromatography without chlorinated solvents which offers small LOD and the whole profile of lipids present in stratum corneum (SC). A method using a narrow-bore PVA-Sil column was used to collect ceramide fraction. Then the molecular species were analysed with a porous graphitic carbon column in capillary LC using a gradient from CH 3 OH/CHCl 3 70:30 v/v to CHCl 3 at 2%/min with a flow rate at 5 lL/min. The LOD obtained for ceramide was 1 ng. Both methods were assessed with SC samples obtained by rinsing a 5.7 cm 2 area of the forearm with 25 mL of ethanol. IntroductionStratum corneum (SC) consists of corneocytes organized in a matrix of highly ordered multilamellar lipid sheets [1,2]. For the structural and physical properties of this lipid arrangement, SC is considered to play an essential role in the skin barrier function. Ceramides constitute 50% of this lipid barrier. Free fatty acid and cholesterol (Cho) constitute the remaining lipid classes composing this lipid barrier. The ceramide classes are highly heterogeneous in structure due to the sphingoid base and fatty acid moieties constituting these amide molecules. They differ by a different number of hydroxyl groups and carbon double bonds, and the chain length [3]. This structural heterogeneity is an important element to study the lipid barrier property. The nature of the structural difference between molecular species included in each lipid class is apolar. Therefore, apolar stationary phases are required for their discrimination such as octadecyl bonded silica [4][5][6] or porous graphitic carbon (PGC) [7]. This latter column provides original selectivity between molecular species. However, to apply such analytical methods to skin samples, the lipids should be previously extracted and the various lipid classes separated.Several noninvasive methods for extraction of lipids from SC exist, e.g. by stripping [8][9][10], scraping [11], skin rinsing [3,12] or with an extraction chamber [11,13]. Whatever the approach, the three...

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Section: Solvent Selection With Pva-sil Stationary Phasesupporting

confidence: 50%

Section: Solvent Selection With Pva-sil Stationary Phasementioning

confidence: 99%

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Microanalytical systems for separations of stratum corneum ceramides

et al. 2006

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“…Due to the complexity of biological lipid extracts, numerous separation techniques have been used such as thin layer chromatography (TLC) [6], gas chromatography (GC) [7], normal-phase liquid chromatography (NPLC) [8], or reverse-phase liquid chromatography (RPLC) [9]. In order to detect and elucidate the structure of minor lipid species, mass spectrometry has been largely used, and liquid chromatography/mass spectrometry has become the gold standard for lipidomic analysis thanks to its sensitivity and specificity.…”

Section: Introductionmentioning

confidence: 99%

Atmospheric Pressure Photoionization as a Powerful Tool for Large-Scale Lipidomic Studies

Gaudin

Imbert

Libong

et al. 2012

J. Am. Soc. Mass Spectrom.

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Lipidomic studies often use liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) for separation, identification, and quantification. However, due to the wide structural diversity of lipids, the most apolar part of the lipidome is often detected with low sensitivity in ESI. Atmospheric pressure (APPI) can be an alternative ionization source since normal-phase solvents are known to enhance photoionization of these classes. In this paper, we intend to show the efficiency of APPI to identify different lipid classes, with a special interest on sphingolipids. In-source APPI fragmentation appears to be an added value for the structural analysis of lipids. It provides a detailed characterization of both the polar head and the non polar moiety of most lipid classes, and it makes possible the detection of all lipids in both polarities, which is not always possible with ESI.

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“…On this conventional normal phase (NP) column they achieved a comprehensive separation of up to 16 lipid classes in one single run. Further development was achieved with a polyvinyl alcohol-bonded phase (PVA), that was used for the separation of sphingolipids, glycoglycerolipids, phospholipids and the more polar lipid constituents of marine particulate matter [9,10]. Other authors presented methods for the separation of neutral and polar lipid classes using diol-NP columns [11,12].…”

Section: Introductionmentioning

confidence: 99%

Improved separation and quantification of neutral and polar lipid classes by HPLC–ELSD using a monolithic silica phase: Application to exceptional marine lipids

Graeve

Janssen

2009

Journal of Chromatography B

122

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a b s t r a c tAn improved HPLC method is presented, which allows separation and quantification of a broad range of lipid classes of marine zooplankton with special regard to neutral lipids. Marine zooplankton species often produce high amounts of exceptional lipids, especially at high latitudes, in order to cope with the harsh environmental conditions and strong seasonality in food supply. Major neutral lipid classes are wax esters, triacylglycerols, diacylglycerol ethers, free fatty alcohols and sterols. Neutral and polar lipids were separated and identified on a monolithic silica column (Chromolith ® Performance-Si) using high performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). The method resolves a broad spectrum of lipids, varying in polarity from squalene to lysophosphatidylcholine in a single run. The total run time was 35 min including column re-equilibration. The calibration was made at levels of 0.1-60 g lipid/injection, but a 10-15-fold greater amount can be injected if single lipid classes need to be separated, e.g. for further determination of individual fatty acids. The method was applied to representative Arctic zooplankton species (copepods, pteropods, euphausiids and ctenophores) that are known to biosynthesize in particular neutral lipids like diacylglycerol ethers and free fatty alcohols.

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Assessment of the retention properties of poly(vinyl alcohol) stationary phase for lipid class profiling in liquid chromatography

Cited by 38 publications

References 24 publications

Microanalytical systems for separations of stratum corneum ceramides

Microanalytical systems for separations of stratum corneum ceramides

Atmospheric Pressure Photoionization as a Powerful Tool for Large-Scale Lipidomic Studies

Improved separation and quantification of neutral and polar lipid classes by HPLC–ELSD using a monolithic silica phase: Application to exceptional marine lipids

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