Lipidomic Analysis of Red Blood Cell Membrane

Fatty-acid-based membrane lipidome analyses were performed by the Lipidomic Laboratory of Lipinutragen (Bologna, Italy). Blood samples (0.5 mL) collected in vacutainer tubes with ethylenediaminetetraacetic acid (EDTA) were treated according to the ISO17025 certified procedure (accredited Lab. #1836L) by robotic equipment and processed as described in previous studies [21 (link),22 (link),36 (link),46 (link)]. Briefly, the mature cell fraction was isolated based on the higher density of the aged cells with control of diameter controlled by cell counter (Scepter 2.0 with Scepter Software Pro, EMD Millipore, Darmstadt, Germany) [47 (link)]. After phospholipid extraction, derivatization to fatty acid methyl esters (FAME) was performed, transforming membrane glycerophospholipids (mainly phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidyl inositol, and plasmalogens) to examine up to 80% of the RBC membrane lipidome [48 (link)]. Fatty acids analysis was performed by gas chromatography (GC), and percentages are given as % relative quantitative (% rel. quant.), as previously described [21 (link),22 (link),36 (link),46 (link)], comparing with the benchmark of the interval values of each fatty acid and index [15 (link)].

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Del Duca E., Sansone A., Sgrulletti M., Di Nolfo F., Chini L., Ferreri C, & Moschese V. (2022). Fatty-Acid-Based Membrane Lipidome Profile of Peanut Allergy Patients: An Exploratory Study of a Lifelong Health Condition. International Journal of Molecular Sciences, 24(1), 120.

Publication 2022

Scepter 2.0 Scepter Software Pro

Blood Cell Esters Ethylenediaminetetraacetic acid Fatty acid Gas chromatography Glycerophospholipids Lipidome Membrane Phosphatidyl inositol Phosphatidylcholine Phosphatidylethanolamine Phosphatidylserine Phospholipid Plasmalogens

Corresponding Organization : Institute of Organic Synthesis and Photoreactivity

Other organizations : Policlinico Tor Vergata, University of Rome Tor Vergata

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Protocol cited in 2 other protocols

Variable analysis

independent variables

Fatty-acid-based membrane lipidome analyses

dependent variables

Percentages of fatty acids (as % relative quantitative)

control variables

Blood samples collected in vacutainer tubes with ethylenediaminetetraacetic acid (EDTA)
ISO17025 certified procedure (accredited Lab. #1836L) by robotic equipment
Diameter of mature cell fraction controlled by cell counter (Scepter 2.0 with Scepter Software Pro, EMD Millipore, Darmstadt, Germany)
Comparison with benchmark interval values of each fatty acid and index

controls

Benchmark interval values of each fatty acid and index as a positive control

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