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Separation and Identification of Odd Chain Triacylglycerols of the Protozoan Khawkinea quartana and the Mold Mortierella alpina Using LC–MS

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Lipids

Abstract

Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC–MS/APCI) with reversed- and chiral phases was used for separation of triacylglycerols (TAG) from protozoan and mold. This study describes the separation and identification of odd numbered chains of regioisomers and enantiomers of triacylglycerols from different natural sources, i.e., the protozoan Khawkinea quartana and the mold Mortierella alpina. Using the above-mentioned separation methods and the synthesis of appropriate standards of TAG, we identified regioisomers and enantiomers of both even and odd numbered TAG. The biosynthesis of odd numbered TAG was found to be strictly stereospecific and to depend on the production microorganism, one enantiomer predominating in the protozoan and the other in the mold. It was proved that even numbered TAG are synthesized in a higher optical purity, which can be explained by a higher affinity of acyltransferases to the respective substrate, i.e., to even chain PUFA.

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Abbreviations

ACN:

Acetonitrile

APCI:

Atmospheric pressure chemical ionization

CCF:

Culture Collection of Fungi, Faculty of Science, Charles University, Prague

DB:

Double bonds

DMAP:

4-Dimethylaminopyridine

EDCI:

1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide

ESI:

Electrospray ionization

FA:

Fatty acid(s)

FAME:

Fatty acid methyl ester(s)

iPrOH:

2-Propanol

L:

Linoleic acid

LC–MS/APCI:

Liquid chromatography-mass spectrometry/atmospheric pressure chemical ionization

Ln:

Linolenic acid

M:

Myristic acid

Ma:

Margaric acid

MEA:

Malt extract agar

Mo:

Margaroleic acid

MS2 :

Tandem mass spectrometry

NARP-LC:

Non aqueous reversed phase-liquid chromatography

NLS:

Neutral loss scan

O:

Oleic acid

ORD:

Optical rotatory dispersion

P:

Palmitic acid

PUFA:

Polyunsaturated fatty acid(s)

S:

Stearic acid

SAG:

Culture Collection of Algae at the University of Gottingen, Germany

TAG:

Triacylglycerol(s)

TIC:

Total ion current

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Acknowledgments

The research was supported by GACR projects P503/11/0215 and 14-00227S, and by Institutional Research Concept RVO61388971.

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Authors and Affiliations

  1. Institute of Microbiology, CAS, Vídeňská 1083, 142 20, Prague, Czech Republic

    Tomáš Řezanka, Alena Nováková & Karel Sigler

  2. Institute of Microbiology, CAS, Centre Algatech, Laboratory of Cell Cycles of Algae, Opatovický mlýn 237, 379 81, Třeboň, Czech Republic

    Milada Vítová

Authors
  1. Tomáš Řezanka
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  2. Milada Vítová
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  4. Karel Sigler
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Correspondence to Tomáš Řezanka.

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Řezanka, T., Vítová, M., Nováková, A. et al. Separation and Identification of Odd Chain Triacylglycerols of the Protozoan Khawkinea quartana and the Mold Mortierella alpina Using LC–MS. Lipids 50, 811–820 (2015). https://doi.org/10.1007/s11745-015-4042-8

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  • DOI: https://doi.org/10.1007/s11745-015-4042-8

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