Analysis of human plasma lipids by using comprehensive two‐dimensional gas chromatography with dual detection and with the support of high‐resolution time‐of‐flight mass spectrometry for structural elucidation

Salivo, Simona; Beccaria, Marco; Sullini, Giuseppe; Tranchida, Peter Q.; Dugo, Paola; Mondello, Luigi

doi:10.1002/jssc.201400844

Cited by 19 publications

(4 citation statements)

References 38 publications

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“…Other options exist for derivatization of human plasma such as acetyl chloride/methanol for the determination of FAMEs and the separation of positional and geometrical ( cis/trans ) isomers where special focus was paid to conjugated linoleic acid to avoid isomerization ; another derivatization such as pentaflurobenzyl bromide has been used for lipidomics analysis . However, the last approach, together with a report on the unsaponifiable lipid fraction analysis , focused mainly on GC–MS, GC x GC–quad MS–FID, or GC–HR‐TOF‐MS. Parallel, derivatives of tert ‐butyldimethylsilyl esters were characterized by GC–ESI‐MS analysis .…”

Section: Resultsmentioning

confidence: 99%

Quantification of fatty acids in erythrocytes and plasma by fast gas chromatography

Cruz‐Hernandez

Thakkar

Masserey-Elmelegy

et al. 2017

J of Separation Science

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As a result of the heterogeneous nature of lipid classes in complex biological matrices such as plasma and erythrocytes, it is imperative to have a robust and validated methodology for fatty acid quantification. The effective method presented here combines available methodology of fast gas chromatography and an improvement of the sample preparation methodology before injection into the gas chromatograph. This methodology ensures complete transesterification and quantification of total and individual fatty acids (and not only in relative amounts) by addition of internal standards. We considered sample preparation key, and we established the use of lysis buffer and ethanol for erythrocytes and plasma sample preparation, respectively. Fatty acid profile was determined by acid methylation and fast gas chromatography equipped with a flame ionization detector. The triacylglycerol 13:0, phosphatidylcholine 23:0, and methyl esters 21:0 were used as internal standards. Within the linearity of the calibration, the ratio of the peak area of each fatty acid over the peak area of the internal standard was constant (coefficient of variation ≤ 2.5). Satisfactory repeatability <15% and intermediate reproducibility < 15% were observed. Finally, this validated method was applied to a pre-clinical trial that investigated the impact of dietary fats on accretion of specific fatty acids in plasma and erythrocytes.

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Section: Resultsmentioning

confidence: 99%

Quantification of fatty acids in erythrocytes and plasma by fast gas chromatography

Cruz‐Hernandez

Thakkar

Masserey-Elmelegy

et al. 2017

J of Separation Science

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“…Up to now, LC–MS has become one of the most widely used methods for lipidomic analysis, while GC–MS is mainly accepted in fatty acid (FA) profiling and some specific analysis of less polar lipids . In addition, the appearance of 2D GC and 2D LC provides ultra‐high resolving power and peak capacity, which become more and more widely used to analyze lipids from complex samples. The third one is mass spectrometry imaging (MSI).…”

Section: Analytical Methods In Clinical Lipidomicsmentioning

confidence: 99%

Recent advances in lipidomics for disease research

Yang

Shan

et al. 2015

J of Separation Science

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Lipidomics is an important branch of metabolomics, which aims at the detailed analysis of lipid species and their multiple roles in the living system. In recent years, the development of various analytical methods for effective identification and characterization of lipids has greatly promoted the process of lipidomics. Meanwhile, as many diseases demonstrate a remarkable alteration in lipid profiles compared with that of healthy people, lipidomics has been extensively introduced to disease research. The comprehensive lipid profiling provides a chance to discover novel biomarkers for specific disease. In addition, it plays a crucial role in the study of lipid metabolism, which could illuminate the pathogenesis of diseases. In this review, after brief discussion of analytical methods for lipidomics in clinical research, we focus on the recent advances of lipidomics related to four types of diseases, including cancer, atherosclerosis, diabetes mellitus, and Alzheimer's disease.

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“…Lipids from 2 mL of plasma or 2 mL of an aqueous solution of 10% AlbuMAX I were extracted following a protocol based on the studies cited herein [ 40 , 89 , 90 ]. For this, samples were extracted with 18 mL of Chloroform/Methanol (2:1 by volume), mixed, and centrifuged at 1,000 x g for 2 minutes.…”

Section: Methodsmentioning

confidence: 99%

Beyond the MEP Pathway: A novel kinase required for prenol utilization by malaria parasites

Crispim,

Verdaguer,

Hernández

et al. 2024

PLoS Pathog

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A proposed treatment for malaria is a combination of fosmidomycin and clindamycin. Both compounds inhibit the methylerythritol 4-phosphate (MEP) pathway, the parasitic source of farnesyl and geranylgeranyl pyrophosphate (FPP and GGPP, respectively). Both FPP and GGPP are crucial for the biosynthesis of several essential metabolites such as ubiquinone and dolichol, as well as for protein prenylation. Dietary prenols, such as farnesol (FOH) and geranylgeraniol (GGOH), can rescue parasites from MEP inhibitors, suggesting the existence of a missing pathway for prenol salvage via phosphorylation. In this study, we identified a gene in the genome of P. falciparum, encoding a transmembrane prenol kinase (PolK) involved in the salvage of FOH and GGOH. The enzyme was expressed in Saccharomyces cerevisiae, and its FOH/GGOH kinase activities were experimentally validated. Furthermore, conditional knockout parasites (Δ-PolK) were created to investigate the biological importance of the FOH/GGOH salvage pathway. Δ-PolK parasites were viable but displayed increased susceptibility to fosmidomycin. Their sensitivity to MEP inhibitors could not be rescued by adding prenols. Additionally, Δ-PolK parasites lost their capability to utilize prenols for protein prenylation. Experiments using culture medium supplemented with whole/delipidated human plasma in transgenic parasites revealed that human plasma has components that can diminish the effectiveness of fosmidomycin. Mass spectrometry tests indicated that both bovine supplements used in culture and human plasma contain GGOH. These findings suggest that the FOH/GGOH salvage pathway might offer an alternate source of isoprenoids for malaria parasites when de novo biosynthesis is inhibited. This study also identifies a novel kind of enzyme related to isoprenoid metabolism.

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Analysis of human plasma lipids by using comprehensive two‐dimensional gas chromatography with dual detection and with the support of high‐resolution time‐of‐flight mass spectrometry for structural elucidation

Cited by 19 publications

References 38 publications

Quantification of fatty acids in erythrocytes and plasma by fast gas chromatography

Quantification of fatty acids in erythrocytes and plasma by fast gas chromatography

Recent advances in lipidomics for disease research

Beyond the MEP Pathway: A novel kinase required for prenol utilization by malaria parasites

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