Phenylmethanesulfonyl fluoride pretreatment stabilizes plasma lipidome in lipidomic and metabolomic analysis

Wang, Xiaolin; Gu, Xiao Li; Song, Huahua; Song, Qing; Gao, Xiaoling; Yang, Lu; Chen, Hongzhuan

doi:10.1016/j.aca.2015.08.049

Cited by 11 publications

(10 citation statements)

References 30 publications

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“…The lipids of plasma (50 uL) and liver samples (200 mg) were extracted using liquid-liquid methyl tert-butyl ether (MTBE) extraction protocols developed by our group [ 15 ]. Briefly, endogenous free lipids including PE (17:0/17:0), PC (19:0/19:0), LPE 14:0, LPC 17:0 were selected as the internal standard for their corresponding lipid classes.…”

Section: Methodsmentioning

confidence: 99%

“…However, the alternation of individual PC/PE specie in both plasma and liver of APAP-injured mice and correlations of PCs/PEs between plasma and liver remains unknown. We already developed an LC-MS based lipidomic method for simultaneous detection of diverse lipids [ 15 ]. In the present study, using this high-throughput method, we compared the relative concentrations of phosphatidylcholine and phosphatidylethanolamine in the plasma and liver of the APAP-induced liver injured mice and saline-treated control mice at series of time-points, and then measured the expressions of genes involved in PC/PE metabolism in liver.…”

Section: Introductionmentioning

confidence: 99%

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Liquid chromatography mass spectrometry-based profiling of phosphatidylcholine and phosphatidylethanolamine in the plasma and liver of acetaminophen-induced liver injured mice

et al. 2017

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BackgroundAcetaminophen (APAP) overdose is one of the most common causes of acute liver failure in many countries. The aim of the study was to describe the profiling of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in the plasma and liver of Acetaminophen -induced liver injured mice.MethodsA time course study was carried out using C57BL/6 mice after intraperitoneal administration of 300 mg/kg Acetaminophen 1 h, 3 h, 6 h, 12 h and 24 h. A high-throughput liquid chromatography mass spectrometry (LC-MS) lipidomic method was utilized to detect phosphatidylcholine and phosphatidylethanolamine species in the plasma and liver. The expressions of phosphatidylcholine and phosphatidylethanolamine metabolism related genes in liver were detected by quantitative Reverse transcription polymerase chain reaction (qRT-PCR) and Western-blot.ResultsFollowing Acetaminophen treatment, the content of many PC and PE species in plasma increased from 1 h time point, peaked at 3 h or 6 h, and tended to return to baseline at 24 h time point. The relative contents of almost all PC species in liver decreased from 1 h, appeared to be lowest at 6 h, and then return to normality at 24 h, which might be partly explained by the suppression of phospholipases mRNA expressions and the induction of choline kinase (Chka) expression. Inconsistent with PC profile, the relative contents of many PE species in liver increased upon Acetaminophen treatment, which might be caused by the down-regulation of phosphatidylethanolamine N-methyltransferase (Pemt).ConclusionsAcetaminophen overdose induced dramatic change of many PC and PE species in plasma and liver, which might be caused by damaging hepatocytes and interfering the phospholipid metabolism in Acetaminophen -injured liver.Electronic supplementary materialThe online version of this article (doi:10.1186/s12944-017-0540-4) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Liquid chromatography mass spectrometry-based profiling of phosphatidylcholine and phosphatidylethanolamine in the plasma and liver of acetaminophen-induced liver injured mice

et al. 2017

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“…Lipid instability can be caused by a variety of factors such as chemical changes ( e.g ., oxidation) biochemical/microbial enzymatic actions (Jones et al, 2007; Wang et al, 2015), or nonoxidative heating ( e.g ., thermal decomposition) (Lorenz et al, 2011; Nawar, 1969). Chemical changes can trigger the release of hydrolyzed or oxidized lipid species.…”

Section: Forms Of Lipid Degradationmentioning

confidence: 99%

“…Additives can be applied to reduce both physical degradation ( e.g ., oxidation) and enzymatic degradation. As an example of reducing enzymatic degradation, Wang et al proposed the use of 5 mM phenylmethanesulfonyl fluoride (PMSF) as a sample pretreatment, prior to sample extraction, to increase the stability of glycerophospholipids, glycerolipids, and sphingolipids (Wang et al, 2015). The authors demonstrated higher levels of phosphatidylcholine and phosphatidylethanolamine compared to the respective lysoglycerophospholipids, thus suggesting the inhibition of phospholipase (PLA) activity.…”

Section: Strategies For Improving Lipid Stabilitymentioning

confidence: 99%

A Review of Efforts to Improve Lipid Stability during Sample Preparation and Standardization Efforts to Ensure Accuracy in the Reporting of Lipid Measurements

Ulmer

Koelmel

Jones

et al. 2020

Lipids

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Lipidomics is a rapidly growing field, fueled by developments in analytical instrumentation and bioinformatics. To date, most researchers and industries have employed their own lipidomics workflows without a consensus on best practices. Without a community‐wide consensus on best practices for the prevention of lipid degradation and transformations through sample collection and analysis, it is difficult to assess the quality of lipidomics data and hence trust results. Clinical studies often rely on samples being stored for weeks or months until they are analyzed, but inappropriate sampling techniques, storage temperatures, and analytical protocols can result in the degradation of complex lipids and the generation of oxidized or hydrolyzed metabolite artifacts. While best practices for lipid stability are sample dependent, it is generally recommended that strategies during sample preparation capable of quenching enzymatic activity and preventing oxidation should be considered. In addition, after sample preparation, lipid extracts should be stored in organic solvents with antioxidants at −20 °C or lower in an airtight container without exposure to light or oxygen. This will reduce or eliminate sublimation, and chemically and physically induced molecular transformations such as oxidation, enzymatic transformation, and photon/heat‐induced degradation. This review explores the available literature on lipid stability, with a particular focus on human health and/or clinical lipidomic applications. Specifically, this includes a description of known mechanisms of lipid degradation, strategies, and considerations for lipid storage, as well as current efforts for standardization and quality insurance of protocols.

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“…The integrity of samples is maintained by remaining in a frozen state until chemical extraction; however, it is often necessary to weigh samples prior to extraction where thawing is undoubtedly occurring. At this point, certain lipids can be susceptible to changes via enzymatic activity [8] and chemical degradation due to pH and oxidation [9], thus complicating the interpretation of the observed lipid profile. A study by Wang et al analyzed the lipidome in mouse and rat plasma and noted that several lipids significantly increased or decreased after 4 h on a benchtop [8].…”

mentioning

confidence: 99%

Examining Heat Treatment for Stabilization of the Lipidome

et al. 2018

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Phenylmethanesulfonyl fluoride pretreatment stabilizes plasma lipidome in lipidomic and metabolomic analysis

Cited by 11 publications

References 30 publications

Liquid chromatography mass spectrometry-based profiling of phosphatidylcholine and phosphatidylethanolamine in the plasma and liver of acetaminophen-induced liver injured mice

Liquid chromatography mass spectrometry-based profiling of phosphatidylcholine and phosphatidylethanolamine in the plasma and liver of acetaminophen-induced liver injured mice

A Review of Efforts to Improve Lipid Stability during Sample Preparation and Standardization Efforts to Ensure Accuracy in the Reporting of Lipid Measurements

Examining Heat Treatment for Stabilization of the Lipidome

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