Rapid profiling and quantification of phospholipid molecular species in human plasma based on chemical derivatization coupled with electrospray ionization tandem mass spectrometry

Wei, Fang; Wang, Xiang; Ma, Huifang; Lv, Xiongwen; Dong, Xuyan; Hong, Chen

doi:10.1016/j.aca.2018.04.012

Cited by 37 publications

(29 citation statements)

References 24 publications

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“…For enrichment of particular lipid classes, or for separation of a lipid extract into its constituent lipid classes, solid-phase extraction (SPE) [42] or thin-layer chromatography [43] can be employed. Furthermore, chemical derivatization can be used for enhanced ionization and improved detection of low abundant lipid classes [44,45]. Derivatization and SPE are time-consuming procedures, which are typically only necessary if the analysis is targeted toward extremely low abundant lipid classes, while for most applications, a single-phase precipitation or a two-phase extraction method is sufficient.…”

Section: Analytical Considerationsmentioning

confidence: 99%

Analytical Considerations of Stable Isotope Labelling in Lipidomics

Triebl

Wenk

2018

Biomolecules

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Over the last two decades, lipids have come to be understood as far more than merely components of cellular membranes and forms of energy storage, and are now also being implicated to play important roles in a variety of diseases, with lipid biomarker research one of the most widespread applications of lipidomic techniques both in research and in clinical settings. Stable isotope labelling has become a staple technique in the analysis of small molecule metabolism and dynamics, as it is the only experimental setup by which biosynthesis, remodelling and degradation of biomolecules can be directly measured. Using state-of-the-art analytical technologies such as chromatography-coupled high resolution tandem mass spectrometry, the stable isotope label can be precisely localized and quantified within the biomolecules. The application of stable isotope labelling to lipidomics is however complicated by the diversity of lipids and the complexity of the necessary data analysis. This article discusses key experimental aspects of stable isotope labelling in the field of mass spectrometry-based lipidomics, summarizes current applications and provides an outlook on future developments and potential.

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Section: Analytical Considerationsmentioning

confidence: 99%

Analytical Considerations of Stable Isotope Labelling in Lipidomics

Triebl

Wenk

2018

Biomolecules

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“…Under acidic conditions, aminophospholipids with phosphate moieties behave as strong Lewis bases (electron donors), which can interact with Zr atoms coated on the surface of silica in the HybridSPE-PL column; under basic conditions, the bound aminophospholipids can be eluted with a basic solution. The SPE process is based on the steps in our previously published article (15). The HybridSPE-PL column was first activated with 3 ml of methanol, and then the lipid extract reconstituted in 300 l of methanol with 4% (v/v) formic acid was loaded onto the SPE column.…”

Section: Lipid Extraction and Purificationmentioning

confidence: 99%

“…The MRM mode is widely used in targeted metabolomics in LC-MS analysis, which has good repeatability, sensitivity, and broad dynamic range. For comparison, the d0-acetonelabeled aminophospholipid standards were also analyzed by direct infusing-NLS-MS, which is commonly used for lipid analysis (12,15). Table 2 shows the LODs, LOQs, and…”

Section: Enhancement Of Detection Sensitivity Upon Acetone Labelingmentioning

confidence: 99%

“…Recently, we developed an absolute quantification method for determination of PL molecular species in biological samples. Two or more PL standards were selected for each PL class and derivatized with TMSCHN 2 , which were then used as the internal standards for construction of a calibration curve to normalize the nonuniformity response resulting from the distinct chemical constitution of individual PL species in the biological samples (15). It is a promising methodology for profiling and accurate quantification of complex lipid molecules in biological samples.…”

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confidence: 99%

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Profiling and quantification of aminophospholipids based on chemical derivatization coupled with HPLC-MS

Wei

et al. 2019

Journal of Lipid Research

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and metabolic roles (1). PLs consist of a glycerol backbone esterified with two fatty acids at the sn-1 and sn-2 positions. The backbone's sn-3 position is occupied by a phosphate group attached to a polar head of different nature. Aminophospholipids [phosphatidylethanolamine (PE) and phosphatidylserine (PS)] belong to the category of PLs. PE and PS account for approximately 20% and 3-15% of total PLs in eukaryotic cells, respectively. They play a critical role in regulating various biological processes, including intraand inter-cellular signaling, phagocytic recognition, apoptotic cell clearance, cell division, angiogenesis, and vascular remodeling under normal physiological conditions in eukaryotic cells (2). Furthermore, aminophospholipids are associated with various human diseases, including cancer, diabetes, obesity, and neurodegeneration (3, 4). Hence, profiling of the individual aminophospholipid molecular species in cellular components is critical to link physiological or pathophysiological changes to the underlying biochemistry and thus target therapeutic interventions. However, precise profiling of aminophospholipids is not easy because of the sheer number and the high complexities of their structures. Direct infusion MS equipped with an ESI source holds much promise for the characterization of PLs (5). Through effective intra-source separation of predetermined groups of lipid classes according to their intrinsic electrical propensities, lipid extracts can be analyzed directly without preseparation. However, the disadvantages of the direct infusion MS method are ion suppression for lipid species with low proton affinities, negative impact of unavoidable matrix effects on the analysis of minor content of lipid species in biological samples, and serious contamination to Abstract In this study, a novel strategy based on acetone stable-isotope derivatization coupled with HPLC-MS for profiling and accurate quantification of aminophospholipids (phosphatidylethanolamine and phosphatidylserine) in biological samples was developed. Acetone derivatization leads to alkylation of the primary amino groups of aminophospholipids with an isopropyl moiety; the use of deuterium-labeled acetone (d6-acetone) introduced a 6 Da mass shift that was ideally suited for profiling and quantification analysis with high selectivity and accuracy. After derivatization, significantly increased column efficiency for chromatographic separation and detection sensitivity for MS analysis of aminophospholipids was observed. Furthermore, an accuracy quantification method was developed. Aminophospholipids in biological samples were derivatized with d0-acetone; while more than two aminophospholipid standards were selected for each class of aminophospholipid and derivatized with d6-acetone, which were then used as the internal standards to typically construct a calibration curve for each class to normalize the nonuniformity response caused by the differential fragmentation kinetics resulting from the distinct chemical constitution of individual...

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“…PE 16:0_18:1 in the pooled human plasma sample was 2.11±0.26 μM, which was consistent with the reported concentration ranges (1.20 to 5.95 μM). [24]…”

mentioning

confidence: 99%

A Polymer Coating Transfer Enrichment Method for Direct Mass Spectrometry Analysis of Lipids in Biofluid Samples

Zhang

Chiang

et al. 2019

Angew Chem Int Ed

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Ap orous polymer coating transfer enrichment method is developed for the direct mass spectrometry (MS) analysis of lipids.T he enrichment is fast (ca. 1min) and enables the profiling and quantitation of lipids in small-volume biofluid samples.C oupled with ap hotochemical Paternò-Büchi reaction, this method enables the fast determination of lipid structure at the C = Clocation level and point-of-care lipid biomarker analysis.Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

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Rapid profiling and quantification of phospholipid molecular species in human plasma based on chemical derivatization coupled with electrospray ionization tandem mass spectrometry

Cited by 37 publications

References 24 publications

Analytical Considerations of Stable Isotope Labelling in Lipidomics

Analytical Considerations of Stable Isotope Labelling in Lipidomics

Profiling and quantification of aminophospholipids based on chemical derivatization coupled with HPLC-MS

A Polymer Coating Transfer Enrichment Method for Direct Mass Spectrometry Analysis of Lipids in Biofluid Samples

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