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Determination of stereochemical configuration of the glycerol moieties in glycoglycerolipids by chiral phase high-performance liquid chromatography

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Lipids

Abstract

This study reports a simple and sensitive method for determining the absolute configuration of the glycerol moieties in glycoglycerolipids. The method is based on chiral phase high-performance liquid chromatography (HPLC) separations of enantiomeric di- and monoacylglycerols released from glycosyldi- and monoacylglycerols, respectively, by periodate oxidation followed by hydrazinolysis. The released di- and monoacylglycerols were chromatographed as their 3,5-dinitrophenylurethane (3,5-DNPU), and bis(3,5-DNPU) derivatives, respectively. The derivatives were separated on two chiral phases of opposite configuration, (R)-and (S)-1-(1-naphthyl)ethylamine polymers for diacylglycerols and N-(R)-1-(1-naphthyl)ethylaminocarbonyl-(S)-valine and N-(S)-1-(1-naphyl)ethylamino-carbonyl-(R)-valine for monoacylglycerols. Clear enantiomer separations, which permit the assignment of the glycerol configuration, were achieved for sn-1,2(2,3)-dicyl- and sn-1(3)-monoacylglycerols generated from linseed oil triacylglycerols by partial Grignard degradation on all the chiral stationary phases employed. Using the method, we have determined the glycerol configuration in the glycosyl-diacylglycerols (monogalactosyl-, digalactosyl-, and sulfquinovo-syldiacylglycerols) and glycosylmonoacylglycerols (monogalactosyl-, digalactosyl-, and sulfoquinovosylmonoacylglycerols) isolated from spinach leaves and the coralline red alga Corallina pilulifera. The results clearly showed that the glycerol moieties in all the glycoglycerolipids examined have S-configuration sn-1,2-diacyl- and sn-1-monoacylglycerols). The new method demonstrates that chiral phase HPLC provides unambiguous information on the configuration of the glycerol backbone in natural glycosyldi- and monoacylglycerols, and that the two-step liberation of the free acylglycerols does not compromise glycerol chirality.

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Abbreviations

DGDG:

digalactosyldiacylglycerol

DGMG:

digalactosyl-monoacylglycerol

DNPU:

dinitrophenylurethane

ECN:

equivalent carbon number

ESI:

electrospray ionization

HPLC:

high-performance liquid chromatography

MGDG:

monogalactosyldiacylglycerol

MGMG:

monogalactosylmonoacylglycerol

MS:

mass spectrometry

NMR:

nuclear magnetic resonance

SQDG:

sulfoquinovosyldiacylglycerol

SQMG:

sulfoquinovosyl-monoacylglycerol

TLC:

thin-layer chromatography

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Authors and Affiliations

  1. Graduate School of Environmental Earth Science, Hokkaido University, 060-0810, Sapporo, Japan

    Minoru Suzuki

  2. Banting and Best Department of Medical Research, University of Toronto, M5G 1L6, Toronto, Ontario, Canada

    Arnis Kuksis

  3. Laboratory of Bioresources Chemistry Graduate School of Eisheries Sciences, Hokkaido University, 041-8611, Hakodate, Japan

    Yoshinori Takahashi & Yutaka Itabashi

Authors
  1. Yoshinori Takahashi
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  2. Yutaka Itabashi
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  3. Minoru Suzuki
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  4. Arnis Kuksis
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Correspondence to Yutaka Itabashi.

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Takahashi, Y., Itabashi, Y., Suzuki, M. et al. Determination of stereochemical configuration of the glycerol moieties in glycoglycerolipids by chiral phase high-performance liquid chromatography. Lipids 36, 741–748 (2001). https://doi.org/10.1007/s11745-001-0780-y

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  • DOI: https://doi.org/10.1007/s11745-001-0780-y

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