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Analysis of (O-acyl) alpha- and omega-hydroxy fatty acids in vernix caseosa by high-performance liquid chromatography-Orbitrap mass spectrometry

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Abstract

Fatty acid esters of long-chain hydroxy fatty acids or (O-acyl)-hydroxy fatty acids (OAHFAs) were identified for the first time in vernix caseosa and characterized using chromatography and mass spectrometry. OAHFAs were isolated from the total lipid extract by a two-step semipreparative TLC. The general structure of OAHFAs was established using high-resolution and tandem mass spectrometry of intact lipids and their transesterification and derivatization products. Two isomeric lipid classes were identified: O-acyl esters of ω-hydroxy fatty acids (ωOAHFA) and O-acyl esters of α-hydroxy fatty acids (αOAHFAs). To the best of our knowledge, αOAHFAs have never been detected in any biological sample before. Chromatographic separation and identification of OAHFAs species were achieved using non-aqueous reversed-phase HPLC coupled to electrospray ionization hybrid linear ion trap-Orbitrap mass spectrometry. The lipid species were detected as deprotonated molecules, and their structures were elucidated using data-dependent fragmentation in the negative ion mode. More than 400 OAHFAs were identified in this way. The most abundant ωOAHFAs species were 28:0/ω-18:2, 29:0/ω-18:2, 30:0/ω-18:2, 32:0/ω-18:2, and 30:0/ω-18:3, while αOAHFAs comprised saturated species 21:0/α-24:0, 22:0/α-24:0, 23:0/α-24:0, 24:0/α-24:0, and 26:0/α-24:0. OAHFAs were estimated to account for approximately 0.04% of vernix caseosa lipids.

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Abbreviations

CID:

Collision-induced dissociation

ECN:

Equivalent carbon number

FAHFA:

Fatty acid esters of hydroxy fatty acid

FAME:

Fatty acid methyl ester

HFA:

Hydroxy fatty acid

HFAME:

Hydroxy fatty acid methyl ester

OAHFA:

(O-Acyl)-hydroxy fatty acid

RDBE:

Ring and double bond equivalent

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Acknowledgments

This work was supported by the Grant Agency of Charles University in Prague (Project No. 1182216), the Charles University in Prague (Project SVV260440), and from European Regional Development Fund; OP RDE; Project: “Chemical biology for drugging undruggable targets (ChemBioDrug)” (No. CZ.02.1.01/0.0/0.0/16_019/0000729).

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Authors and Affiliations

  1. Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Femingovo nám. 2, 166 10, Prague 6, Czech Republic

    Aneta Vavrušová, Vladimír Vrkoslav & Josef Cvačka

  2. Department of Analytical Chemistry, Faculty of Science, Charles University in Prague, Hlavova 2030/8, 128 43, Prague 2, Czech Republic

    Aneta Vavrušová, Zuzana Bosáková & Josef Cvačka

  3. Department of Obstetrics and Gynaecology, General Faculty Hospital and 1st Faculty of Medicine, Charles University in Prague, Apolinářská 18, 128 00, Prague 2, Czech Republic

    Richard Plavka

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  1. Aneta Vavrušová
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Correspondence to Josef Cvačka.

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The study was approved by the Ethics Committee of the General University Hospital in Prague (910/09 S-IV).

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Published in the topical collection Current Progress in Lipidomics with guest editors Michal Holčapek, Gerhard Liebisch, and Kim Ekroos.

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Vavrušová, A., Vrkoslav, V., Plavka, R. et al. Analysis of (O-acyl) alpha- and omega-hydroxy fatty acids in vernix caseosa by high-performance liquid chromatography-Orbitrap mass spectrometry. Anal Bioanal Chem 412, 2291–2302 (2020). https://doi.org/10.1007/s00216-019-02348-2

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