Three-phase extraction for enhancement of lipidomic analysis

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F25%3A00639576" target="_blank" >RIV/61388963:_____/25:00639576 - isvavai.cz</a>

Výsledek na webu

<a href="https://hdl.handle.net/11104/0370001" target="_blank" >https://hdl.handle.net/11104/0370001</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Three-phase extraction for enhancement of lipidomic analysis

Popis výsledku v původním jazyce

Due to the large diversity of lipids in different biological materials, extraction is a critical step in the sample preparation process. Currently used two-phase systems extract all lipids in one phase, which often leads to a loss of the number of annotated lipid species. We optimized and evaluated a three-phase extraction system. The optimized three-phase extraction system was compared with the commonly used two-phase methyl-tert-butyl ether (MTBE) extraction. Untargeted lipidomics was performed using LC-MS with heated electrospray ionization. A total of 72 different solvent mixtures were tested to create three-phase systems. Suitable three-phase systems were formed from 21 mixtures. The ability of 12 of these mixtures to separate lipid standards was investigated in detail using a commercially available mixture of lipid standards. Lipidomic analysis showed how selected three-phase systems separate lipid classes into individual phases of the extract ion system. Finally, two solvent systems were selected for the lipidomic analysis of blood plasma. The extraction systems IO:MTBE:ACN:H2O (2:2:2:1, v/v) and IO:EtOAc:ACN:H2O (4:3:4:4, v/v) resulted in the lowest lipid loss to the aqueous phase. Moreover, they enabled the identification of a higher number of lipid species compared to the traditionally used two-phase MTBE extraction.

Název v anglickém jazyce

Three-phase extraction for enhancement of lipidomic analysis

Popis výsledku anglicky

Due to the large diversity of lipids in different biological materials, extraction is a critical step in the sample preparation process. Currently used two-phase systems extract all lipids in one phase, which often leads to a loss of the number of annotated lipid species. We optimized and evaluated a three-phase extraction system. The optimized three-phase extraction system was compared with the commonly used two-phase methyl-tert-butyl ether (MTBE) extraction. Untargeted lipidomics was performed using LC-MS with heated electrospray ionization. A total of 72 different solvent mixtures were tested to create three-phase systems. Suitable three-phase systems were formed from 21 mixtures. The ability of 12 of these mixtures to separate lipid standards was investigated in detail using a commercially available mixture of lipid standards. Lipidomic analysis showed how selected three-phase systems separate lipid classes into individual phases of the extract ion system. Finally, two solvent systems were selected for the lipidomic analysis of blood plasma. The extraction systems IO:MTBE:ACN:H2O (2:2:2:1, v/v) and IO:EtOAc:ACN:H2O (4:3:4:4, v/v) resulted in the lowest lipid loss to the aqueous phase. Moreover, they enabled the identification of a higher number of lipid species compared to the traditionally used two-phase MTBE extraction.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10406 - Analytical chemistry

Projekt

<a href="/cs/project/LX22NPO5104" target="_blank" >LX22NPO5104: Národní institut pro výzkum metabolických a kardiovaskulárních onemocnění</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů