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Comparison of sample preparation methods for the quantitative analysis of eicosanoids and other oxylipins in plasma by means of LC-MS/MS

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Abstract

Oxylipins are potent lipid mediators. For the evaluation of their biological roles, several LC-MS based methods have been developed. While these methods are similar, the described sample preparation procedures for the extraction of oxylipins differ considerably. In order to deduce the most appropriate method for the analysis of non-esterified oxylipins in human plasma, we evaluated the performance of seven established sample preparation procedures. Six commonly used solid phase extraction (SPE) and one liquid-liquid extraction (LLE) protocol were compared based on the recovery of 13 added internal standards, extraction efficacy of oxylipins from plasma and reduction of ion-suppressing matrix. Dramatic differences in the performance in all three parameters were found. LLE with ethyl acetate was overall not a sufficient sample preparation strategy. The protocols using Oasis- and StrataX-material insufficiently removed interfering matrix compounds. Extraction efficacy of oxylipins on anion-exchanging BondElut cartridges was low, while removal of matrix was nearly perfect. None of the protocols led to a high extraction efficacy of analytes while removing all interfering matrix components. However, SPE on a C18-material with removal of matrix by water and n-hexane prior elution with methyl formate showed the best performance for the analysis of a broad spectrum of oxylipins in plasma.

TOC art: Scheme of oxylipin extraction from plasma and ion suppression analysis of PGE2

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Abbreviations

AA:

Arachidonic acid

ACN:

Acetonitrile

CID:

Collision-induced dissociation

COX:

Cyclooxygenase

DHA:

Docosahexaenoic acid

DiHDPE:

Dihydroxy docosapentaenoic acid

DiHETE:

Dihydroxy eicosatetraenoic acid

DiHETrE:

Dihydroxy eicosatrienoic acid

DiHODE:

Dihydroxy octadecadienoic acid

DiHOME:

Dihydroxy octadecenoic acid

EA:

Ethyl acetate

EDTA:

Ethylenediaminetetraacetic acid

EPA:

Eicosapentaenoic acid

EpDPE:

Epoxy docosapentaenoic acid

EpETE:

Epoxy eicosatetraenoic acid

EpETrE:

Epoxy eicosatrienoic acid

EpODE:

Epoxy octadecadienoic acid

EpOME:

Epoxy octadecenoic acid

ESI:

Electrospray ionization

HAc:

Acetic acid

HDHA:

Hydroxy docosahexaenoic acid

HEPE:

Hydroxy eicosapentaenoic acid

HETE:

Hydroxy eicosatetraenoic acid

HODE:

Hydroxy octadecadienoic acid

HOTrE:

Hydroxy octadecatrienoic acid

IS:

Internal standard

LC-MS:

Liquid chromatography mass spectrometry

LLE:

Liquid-liquid extraction

LOD:

Limit of detection

LOX:

Lipoxygenase

LTB:

Leukotriene

LXA:

Lipoxin

MeOH:

Methanol

oxo-ETE:

Oxo eicosatetraenoic acid

PG:

Prostaglandin

PUFA:

Polyunsaturated fatty acid

RP:

Reversed phase

RSD:

Relative standard deviation

SD:

Standard deviation

SPE:

Solid phase extraction

SRM:

Selected reaction monitoring

TriHOME:

Trihydroxy-ocadecenoic acid

Tx:

Thromboxane

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Acknowledgments

This study was supported the Fonds der Chemischen Industrie, a Marie Curie Career Integration Grant (CIG 293536) of the European Union and a Grant of the German Research Foundation (SCHE 1801).

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Authors and Affiliations

  1. Institute for Food Toxicology and Analytical Chemistry, University of Veterinary Medicine Hannover, Bischofsholer Damm 15, 30173, Hannover, Germany

    Annika I. Ostermann, Ina Willenberg & Nils Helge Schebb

  2. Institute of Food Chemistry, University of Wuppertal, Gaußstr. 20, 42119, Wuppertal, Germany

    Nils Helge Schebb

Authors
  1. Annika I. Ostermann
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  2. Ina Willenberg
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  3. Nils Helge Schebb
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Correspondence to Nils Helge Schebb.

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Ostermann, A.I., Willenberg, I. & Schebb, N.H. Comparison of sample preparation methods for the quantitative analysis of eicosanoids and other oxylipins in plasma by means of LC-MS/MS. Anal Bioanal Chem 407, 1403–1414 (2015). https://doi.org/10.1007/s00216-014-8377-4

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  • DOI: https://doi.org/10.1007/s00216-014-8377-4

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