Ammonium bicarbonate buffers combined with hybrid surface technology columns improve the peak shape of strongly tailing lipids.

Autor: Nilsson JM; Department of Medicinal Chemistry, Uppsala Biomedical Centre, Uppsala University, Box 574, 75123 Uppsala, Sweden., Balgoma D; Department of Medicinal Chemistry, Uppsala Biomedical Centre, Uppsala University, Box 574, 75123 Uppsala, Sweden; Instituto de Biomedicina y Genética Molecular (IBGM), CSIC-Universidad de Valladolid, C/ Sanz y Forés 3, 47003, Valladolid, Spain., Pettersson C; Department of Medicinal Chemistry, Uppsala Biomedical Centre, Uppsala University, Box 574, 75123 Uppsala, Sweden., Lennernäs H; Department of Pharmaceutical Biosciences, Uppsala Biomedical Centre, Uppsala University, Box 591, 75123 Uppsala, Sweden., Heindryckx F; Department of Medical Cell Biology, Uppsala Biomedical Centre, Uppsala University, Box 571, 75123 Uppsala, Sweden., Hedeland M; Department of Medicinal Chemistry, Uppsala Biomedical Centre, Uppsala University, Box 574, 75123 Uppsala, Sweden. Electronic address: mikael.hedeland@ilk.uu.se.
Jazyk: angličtina
Zdroj: Analytica chimica acta [Anal Chim Acta] 2024 Aug 08; Vol. 1316, pp. 342811. Date of Electronic Publication: 2024 Jun 08.
DOI: 10.1016/j.aca.2024.342811
Abstrakt: Background: Lipids such as phosphatidic acids (PAs) and cardiolipins (CLs) present strongly tailing peaks in reversed phase liquid chromatography, which entails low detectability. They are usually analyzed by hydrophilic interaction liquid chromatography (HILIC), which hampers high-throughput lipidomics. Thus, there is a great need for improved analytical methods in order to obtain a broader coverage of the lipidome in a single chromatographic method. We investigated the effect of ammonium bicarbonate (ABC) on peak asymmetry and detectability, in comparison with ammonium formate (AFO) on both a conventional BEH C18 column and an HST-CSH C18 column.
Results: The combination of 2.5 mM ABC buffer pH 8 with an HST-CSH C18 column produced significantly improved results, reducing the asymmetry factor at 10 % peak height of PA 16:0/18:1 from 8.4 to 1.6. Furthermore, on average, there was up to a 54-fold enhancement in the peak height of its [M - H] - ion compared to AFO and the BEH C18 column. We confirmed this beneficial effect on other strongly tailing lipids, with accessible phosphate moieties e.g., cardiolipins, phosphatidylinositol phosphate, phosphatidylinositol bisphosphate, phosphorylated ceramide and phosphorylated sphingosine. Furthermore, we found an increased detectability of phospho- and sphingolipids up to 28 times in negative mode when using an HST-CSH C18 column. The method was successfully applied to mouse liver samples, where previously undetected endogenous phospholipids could be analyzed with improved chromatographic separation.
Significance: In conclusion, the use of 2.5 mM ABC substantially improved the peak shape of PAs and enhanced the detectability of the lipidome in negative mode on an RPLC-ESI-Q-TOF-MS system on both BEH C18 and HST-CSH C18 columns. This method provides a wider coverage of the lipidome with one single injection for future lipidomic applications in negative mode.
Competing Interests: Declaration of competing interest There is no conflict of interest from the authors of this article.
(Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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