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Simultaneous determination of the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their corresponding hydroperoxides obtained by lipoxygenase treatment

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Lipids

Abstract

A method for the simultaneous determination of the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their corresponding hydroperoxides is described. Hydroperoxides were formed by incubation of phospholipids with lipoxygenase at pH 9.2. Silicic acid column chromatography (silica Sep-Pak column) was used to separate the phospholipids into phosphatidylcholine and phosphatidylethanolamine. A single C−18 reverse-phase column was employed to separate the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their hydroperoxides by high-performance liquid chromatography. The mobile phase consisted of 5% 10 mM ammonium acetate at pH 5 and 95% methanol. The molecular species of phosphatidylcholine and phosphatidylethanolamine were detected at 205 nm; the eluate was mixed with a chemiluminescence reagent (isoluminol and microperoxidase) and monitored by fluorometry. Under the experimental conditions used, three individual molecular species of both soybean phosphatidylethanolamine and phosphatidylcholine (18∶3/18∶2, 18∶2/18∶2 and 16∶0/18∶2), together with their corresponding hydroperoxides, were identified and quantitated.

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Abbreviations

CL:

chemiluminescence

GLC:

gas-liquid chromatography

15-HPETE:

15-hydroperoxyeicosatetraenoic acid

HPLC:

high-performance liquid chromatography

MTBE:

methyl tertiarybutyl ether

PC:

phosphatidylcholine

PCOOH:

phosphatidylcholine hydroperoxides

16∶0/18∶2 PC:

1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine

18∶0/20∶4 PC:

1-stearoyl-2-arachidonoyl-sn-glycero-3-phosphocholine

18∶2/18∶2 PC:

1,2-dilinoleoyl-sn-glycero-3-phosphocholine

18∶3/18∶2 PC:

1-linolenoyl-2-linoleoyl-sn-glycero-3-phosphocholine

PE:

phosphatidylethanolamine

PEOOH:

phosphatidylethanolamine hydroperoxides

RP:

reverse-phase

UV:

ultraviolet

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Authors and Affiliations

  1. Unité de Recherche d'Hépatologie Pédiatrique (INSERM U347), Hôpital Bicêtre, 80 rue du Général Leclerc, 94276, Le Kremlin Bicêtre, France

    Martine Couturier & Frédérique Lemonnier

  2. Laboratoire de Biochimie, Hôpital R. Debré, 75019, Paris, France

    Patrice Therond & Jean-François Demelier

Authors
  1. Patrice Therond
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  2. Martine Couturier
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  3. Jean-François Demelier
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  4. Frédérique Lemonnier
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Therond, P., Couturier, M., Demelier, JF. et al. Simultaneous determination of the main molecular species of soybean phosphatidylcholine or phosphatidylethanolamine and their corresponding hydroperoxides obtained by lipoxygenase treatment. Lipids 28, 245–249 (1993). https://doi.org/10.1007/BF02536647

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  • DOI: https://doi.org/10.1007/BF02536647

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