Modern Methods of Sample Preparation for the Analysis of Oxylipins in Biological Samples

Liakh, Ivan; Pakiet, Alicja; Śledziński, Tomasz; Mika, Adriana

doi:10.3390/molecules24081639

Cited by 44 publications

(36 citation statements)

References 170 publications

(232 reference statements)

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“…The presence of most oxylipins in low concentrations, as well as their enormous heterogeneity and the emergence of many structurally similar oxylipins, makes their qualitative and quantitative determination difficult due to the low sensitivity of traditional methods. In the previous review [12], we described the current methods of sample preparation from various biological materials preceding the analysis of oxylipins. This paper included a description of the stages of sample collection and storage and a summary of the used pre-extraction additives and standards.…”

Section: Introductionmentioning

confidence: 99%

Methods of the Analysis of Oxylipins in Biological Samples

et al. 2020

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Oxylipins are derivatives of polyunsaturated fatty acids and due to their important and diverse functions in the body, they have become a popular subject of studies. The main challenge for researchers is their low stability and often very low concentration in samples. Therefore, in recent years there have been developments in the extraction and analysis methods of oxylipins. New approaches in extraction methods were described in our previous review. In turn, the old analysis methods have been replaced by new approaches based on mass spectrometry (MS) coupled with liquid chromatography (LC) and gas chromatography (GC), and the best of these methods allow hundreds of oxylipins to be quantitatively identified. This review presents comparative and comprehensive information on the progress of various methods used by various authors to achieve the best results in the analysis of oxylipins in biological samples.

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Section: Introductionmentioning

confidence: 99%

Methods of the Analysis of Oxylipins in Biological Samples

et al. 2020

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“…and human tissues as well as solid food samples requires usually quick freezing in liquid nitrogen and grinding before the addition of organic solvents [65,66]. Samples derived from cell cultures must be first thoroughly washed from serum-borne lipids.…”

Section: Sample Collection and Storagementioning

confidence: 99%

“…Samples derived from cell cultures must be first thoroughly washed from serum-borne lipids. Depending on the further procedure used, the cells can be homogenised [48] or more commonly the lipids are extracted from the cell pellet directly by the addition of cold solvents [65,67]. In turn, liquid samples, such as serum or plasma as well as foodstuffs, e.g.…”

Section: Sample Collection and Storagementioning

confidence: 99%

“…This approach involves protein removing from the samples by the addition of a watermiscible solvent such as acetonitrile, methanol, ethanol or 2-propanol, which in contrast to LLE technique leads to the formation of a monophasic system. If the concentration of analytes in the sample is high enough for the detection method used, the sample can be directly injected after protein precipitation (PPT) [65]. Sarafia et al compared 4-PPT methods using different watermiscible solvents (methanol, acetonitrile, 2-propanol and mixture of 2-propanol/acetonitrile (1:2 v/v)) with LLE methods employing four different solvent systems (methanol combined with chloroform, dichloromethane or MTBE as well as isopropanol with hexane).…”

Section: Isolation Of Pl Fractionsmentioning

confidence: 99%

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Qualitative analysis of phospholipids and their oxidised derivatives – used techniques and examples of their applications related to lipidomic research and food analysis

Parchem

Sasson

Ferreri

et al. 2019

Free Radical Research

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Phospholipids (PLs) are important biomolecules that not only constitute structural building blocks and scaffolds of cell and organelle membranes but also play a vital role in cell biochemistry and physiology. Moreover, dietary exogenous PLs are characterised by high nutritional value and other beneficial health effects, which are confirmed by numerous epidemiological studies. For this reason, PLs are of high interest in lipidomics that targets both the analysis of membrane lipid distribution as well as correlates composition of lipids with their effects on functioning of cells, tissues and organs. Lipidomic assessments follow-up the changes occurring in living organisms, such as free radical attack and oxidative modifications of the polyunsaturated fatty acids (PUFAs) build in PL structures. Oxidised PLs (oxPLs) can be generated exogenously and supplied to organisms with processed food or formed endogenously as a result of oxidative stress. Cellular and tissue oxPLs can be a biomarker predictive of the development of numerous diseases such as atherosclerosis or neuroinflammation. Therefore, suitable high-throughput analytical techniques, which enable comprehensive analysis of PL molecules in terms of the structure of hydrophilic group, fatty acid (FA) composition and oxidative modifications of FAs, have been currently developed. This review addresses all aspects of PL analysis, including lipid isolation, chromatographic separation of PL classes and species, as well as their detection. The bioinformatic tools that enable handling of a large amount of data generated during lipidomic analysis are also discussed. In addition, imaging techniques such as confocal microscopy and mass spectrometry imaging for analysis of cellular lipid maps, including membrane PLs, are presented.

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“…Ideally, plasma oxylipin measurements provide an accurate representation of in vivo intravascular levels. However, the labile nature of oxylipins and related compounds presents unique challenges for accurate identification, quantitation, and interpretation of concentrations in human plasma and other tissues [12][13][14][15]. There is potential to generate artifact during each step of collection, transport, pre-processing delay, processing, storage, and/or analysis due to: (1) degradation, inactivation, or metabolic conversion, and (2) enzymatic and/or nonenzymatic biosynthesis from precursor fatty acids present in the specimen.…”

Section: Introductionmentioning

confidence: 99%

Temperature and time-dependent effects of delayed blood processing on oxylipin concentrations in human plasma

Ramsden

Yuan

Horowitz

et al. 2019

Prostaglandins, Leukotrienes and Essential Fatty Acids

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Background: Oxidized derivatives of polyunsaturated fatty acids, collectively known as oxylipins, are labile bioactive mediators with diverse roles in human physiology and pathology. Oxylipins are increasingly being measured in plasma collected in clinical studies to investigate biological mechanisms and as pharmacodynamic biomarkers for nutrient-based and drug-based interventions. Whole blood is generally stored either on ice or at room temperature prior to processing. However, the potential impacts of delays in processing, and of temperature prior to processing, on oxylipin concentrations are incompletely understood. Objective: To evaluate the effects of delayed processing of blood samples in a timeframe that is typical of a clinical laboratory setting, using typical storage temperatures, on concentrations of representative unesterified oxylipins measured by liquid chromatography-tandem mass spectrometry. Design: Whole blood (drawn on three separate occasions from a single person) was collected into 5 mL purpletop potassium-EDTA tubes and stored for 0, 10, 20, 30, 60 or 120 min at room temperature or on wet ice, followed by centrifugation at 4°C for 10 min with plasma collection. Each sample was run in duplicate, therefore there were six tubes and up to six data points at each time point for each oxylipin at each condition (ice/room temperature). Representative oxylipins derived from arachidonic acid, docosahexaenoic acid, and linoleic acid were quantified by liquid chromatography tandem mass spectrometry. Longitudinal models were used to estimate differences between temperature groups 2 h after blood draw. Results: We found that most oxylipins measured in human plasma in traditional potassium-EDTA tubes are reasonably stable when stored on ice for up to 2 h prior to processing, with little evidence of auto-oxidation in either condition. By contrast, in whole blood stored at room temperature, substantial time-dependent increases in the 12-lipoxygenase-derived (12-HETE, 14-HDHA) and platelet-derived (thromboxane B2) oxylipins were observed. Conclusion: These findings suggest that certain plasma oxylipins can be measured with reasonable accuracy despite delayed processing for up to 2 h when blood is stored on ice prior to centrifugation. 12-Lipoxygenaseand platelet-derived oxylipins may be particularly sensitive to post-collection artifact with delayed processing at room temperature. Future studies are needed to determine impacts of duration and temperature of centrifugation on oxylipin concentrations.

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Modern Methods of Sample Preparation for the Analysis of Oxylipins in Biological Samples

Cited by 44 publications

References 170 publications

Methods of the Analysis of Oxylipins in Biological Samples

Methods of the Analysis of Oxylipins in Biological Samples

Qualitative analysis of phospholipids and their oxidised derivatives – used techniques and examples of their applications related to lipidomic research and food analysis

Temperature and time-dependent effects of delayed blood processing on oxylipin concentrations in human plasma

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