We evaluated the performance of three different single-phase extraction methods to be used before untargeted lipidomics analysis by Liquid Chromatography High-Resolution Mass Spectrometry. Lipids were extracted from a pool of healthy human donors’ plasma in triplicates and run in both positive and negative ESI. The most satisfactory results were attained using methanol/chloroform (2:1, v/v) mixture. Eventually, we evaluated whether a filtration of the samples could be beneficial to yield cleaner and more mass-friendly extracts. Instead of using syringes, we set up a method we called tip-tip filtration, which requires the usage of a filtrating pipette tip. This way of purification led to superior results than the solvent extraction method alone. This additional procedure not only increased reproducibility but also allowed the same lipid coverage. In addition, it permitted to spare time and money, as tip-tip filtration is not particularly expensive nor time-consuming and hopefully it may be useful to increase analytical column lifetime.

Tip-tip filtration ameliorates single-phase extraction methods for plasma large-scale lipidomics analysis / C. Morano, G. Roda, R. Paroni, M. Dei Cas. - In: JOURNAL OF CHROMATOGRAPHY. B. - ISSN 1570-0232. - 1189:(2022 Jan 15), pp. 123099.1-123099.6. [10.1016/j.jchromb.2022.123099]

Tip-tip filtration ameliorates single-phase extraction methods for plasma large-scale lipidomics analysis

C. Morano
Primo
;
G. Roda
Secondo
;
R. Paroni
Penultimo
;
M. Dei Cas
2022

Abstract

We evaluated the performance of three different single-phase extraction methods to be used before untargeted lipidomics analysis by Liquid Chromatography High-Resolution Mass Spectrometry. Lipids were extracted from a pool of healthy human donors’ plasma in triplicates and run in both positive and negative ESI. The most satisfactory results were attained using methanol/chloroform (2:1, v/v) mixture. Eventually, we evaluated whether a filtration of the samples could be beneficial to yield cleaner and more mass-friendly extracts. Instead of using syringes, we set up a method we called tip-tip filtration, which requires the usage of a filtrating pipette tip. This way of purification led to superior results than the solvent extraction method alone. This additional procedure not only increased reproducibility but also allowed the same lipid coverage. In addition, it permitted to spare time and money, as tip-tip filtration is not particularly expensive nor time-consuming and hopefully it may be useful to increase analytical column lifetime.
lipid extraction; mass spectrometry; sample pre-treatment; untargeted lipidomics
Settore BIO/10 - Biochimica
Settore BIO/12 - Biochimica Clinica e Biologia Molecolare Clinica
Settore CHIM/08 - Chimica Farmaceutica
15-gen-2022
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/895448
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