SPE of 5-lipoxygenase metabolites and the effect of head-column field-amplified sample stacking in MEKC

Abromeit, Hans; Wu, Fang–Jing; Scriba, Gerhard K. E.

doi:10.1002/jssc.201300678

Cited by 4 publications

(5 citation statements)

References 27 publications

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“…Therefore, the authors compared the results of the C18 SPE of LTs and HETEs with those obtained from HPLC separations using a SAX (Hypersep Verify‐AX), a polymer‐based weak anionic exchanger (Strata X‐AW) and a polymer‐based mixed‐mode SAX (Bond Elut Plexa PAX). The authors demonstrated that higher recovery of the analytes could be obtained using anion‐exchange columns . The same research team wished to optimize the MEKC method to make it more suitable for use with more polar arachidonic acid metabolites and therefore set out to develop a MEEKC method.…”

Section: Free Fatty Acidsmentioning

confidence: 99%

A digest of capillary electrophoretic methods applied to lipid analyzes

Poinsot

Meang

et al. 2018

Electrophoresis

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Lipids are naturally occurring organic compounds that can be classified into a number of types based on their solubility in nonpolar organic solvents, and are generally insoluble in water. The great structural variety of these various types of lipids has led them to be components of many different biological substances such as oils, waxes, cellular membranes, tissues and biological fluids. The use of capillary electrophoresis (CE) for the study of lipids during the past 30 years has been relatively rare when compared to its use for other classes of biomolecules, primarily due to their insolubility in water. However, a number of interesting studies have been conducted, and as part of this review, we will present the different approaches that were used, which mainly consist of micellar kinetic chromatography and non-aqueous CE. The main advantages of the use of these techniques compared to GC is the very simple sample preparation that is required and, compared to LC, the very robust and quick separations that can be obtained. In this review, we present the various methods that have been reported in the literature that have been used for the study of fatty acids, phospholipids, glycerides, eicosanoids and sterols, with the inclusion of various tables presenting descriptions of the CE methods used as well as the methods of detection, including UV absorbance, fluorescence, mass spectrometry, and conductivity. This review aims to demonstrate that CE can be easily used for the analysis of lipids.

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Section: Free Fatty Acidsmentioning

confidence: 99%

A digest of capillary electrophoretic methods applied to lipid analyzes

Poinsot

Meang

et al. 2018

Electrophoresis

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“…Indeed, the results of our preliminary experiments using an anion-exchange SPE process revealed that the extraction efficiencies of prostaglandins and thromboxanes were <40 % (data not shown). To overcome these issues, we examined the lipid extraction conditions using mixed-mode SPE, which has been used in lipid biology to analyze fatty acid metabolites [27,28]. Although this approach is very attractive, its application has been limited because it can only be applied to a small number of lipids.…”

Section: Construction Of Mixed-mode Spementioning

confidence: 99%

“…With this in mind, reversed-phase/anion-exchange SPE (mixedmode SPE) was evaluated in the current study for the extraction of polar lipids from biological samples. Although mixed-mode SPE has been reported as a novel extraction approach for fatty acid metabolites [27,28], this technique has only been applied to a limited number of lipids and a detailed method has not yet been established.…”

Section: Introductionmentioning

confidence: 99%

Improvements in the High-Performance Liquid Chromatography and Extraction Conditions for the Analysis of Oxidized Fatty Acids Using a Mixed-Mode Spin Column

Fujihara¹

2015

Mod Chem Appl

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Lipidomics by liquid chromatography/mass spectrometry has been used for a better understanding of the roles of oxidized fatty acids in the development of various diseases. However, further work is required to improve the sample preparation process and the peak tailing of cysteinyl-leukotrienes. In this study, we evaluated various mobile phases and extraction conditions. The addition of phosphoric acid to the mobile phase improved the peak tailing of cysteinyl-leukotrienes. The extraction conditions were also optimized by spin-column possessing an anion-exchange and reversed-phase properties. The extraction efficiency of the modified extraction system was examined using 62 lipids, and 13 deuterated lipids were investigated to evaluate matrix effects and recovery from mouse lung homogenate samples. Extraction efficiencies of ≥70% were obtained for almost all of the lipids. Good results with standard deviations of <15% were obtained for the matrix effects and recovery. Finally, the efficiency of our extraction method was compared with those of several conventional methods, and those of leukotriene C4 was improved significantly using our method. Moreover, the proportion of variance between our method and the conventional methods was >0.99 for all the lipids tested. This newly developed method therefore represents a powerful tool to analyze lipids.

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“…Various online sample preconcentration techniques without modification of the instrument provide convenient approaches for sensitivity improvement in CE. − Field-amplified sample stacking (FASS) is simple yet effective, only requiring samples with low conductivity. − Sample may be injected electrokinetically (field-amplified sample injection (FASI)) or hydrodynamically (FASS and large-volume sample stacking (LVSS)). , FASI (also called head-column FASS (HC FASS)) is not limited by sample injection volume and thus often provides greater sensitivity enhancements than FASS and LVSS. It has generated over 1000-fold sensitivity enhancements for drug analysis of clinical samples. − It has been implemented in other applications utilizing CZE, − MEKC, , CE-LIF, CE-mass spectrometry, − CE-flow injection, , DNA, oligosaccharides analysis, and microfluidic electrophoresis. − …”

mentioning

confidence: 99%

“…It has generated over 1000-fold sensitivity enhancements for drug analysis of clinical samples. 33−37 It has been implemented in other applications utilizing CZE, 38−41 MEKC, 42,43 CE-LIF, 44 CE-mass spectrometry, 45−48 CE-flow injection, 49,50 DNA, 51 oligosaccharides analysis, 52 and microfluidic electrophoresis. 53 Various sample stacking approaches have been explored in various separation modes of CE for protein analysis.…”

mentioning

confidence: 99%

Sample Stacking Provides Three Orders of Magnitude Sensitivity Enhancement in SDS Capillary Gel Electrophoresis of Adeno-Associated Virus Capsid Proteins

Zhang

Meagher

2017

Anal. Chem.

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Size-based protein analysis utilizing only 25 ng of total proteins has been realized by sodium dodecyl sulfate capillary gel electrophoresis (SDS CGE) with head-column field-amplified sample stacking as an online sample preconcentration technique. This method has been used as a replacement of SDS-PAGE for purity analysis of adeno-associated virus (AAV) therapeutic products of different serotypes and transgenes. A limit of detection of 0.2 ng/mL (3.3 pM) capsid proteins was achieved with convenient UV absorbance detection at 214 nm, equivalent to 20 pg of protein (330 attomole) loaded in the autosampler vial. For purity analysis, only 25 ng of total AAV capsid proteins (4.3 femtomole virus particles) were loaded to the autosampler vial. The sensitivity is comparable to silver-stained SDS-PAGE. The RSD of purity measurement was 0.0-0.8%, comparable to conventional SDS CGE utilizing 0.1-0.5 mg proteins. The new method provided 3 orders of magnitude sensitivity enhancement as compared to conventional SDS CGE. It shares all the advantages of conventional SDS CGE (labor-saving, easy automation, and convenient quantitation) and also the high sensitivity of silver stained SDS-PAGE. The sample stacking SDS CGE technique can be adopted for size-based analysis of other types of proteins. It is especially useful when protein quantity or concentration is not sufficient for regular SDS CGE or SDS-PAGE assay.

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SPE of 5-lipoxygenase metabolites and the effect of head-column field-amplified sample stacking in MEKC

Cited by 4 publications

References 27 publications

A digest of capillary electrophoretic methods applied to lipid analyzes

A digest of capillary electrophoretic methods applied to lipid analyzes

Improvements in the High-Performance Liquid Chromatography and Extraction Conditions for the Analysis of Oxidized Fatty Acids Using a Mixed-Mode Spin Column

Sample Stacking Provides Three Orders of Magnitude Sensitivity Enhancement in SDS Capillary Gel Electrophoresis of Adeno-Associated Virus Capsid Proteins

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