April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
A New HPLC-ESI-MS/MS Method to Characterize and Quantify Phosphatidyl-Choline With VLC-PUFA: Application to Human Retina
O. Berdeaux; L. Martine; S. Cabaret; G. Thuret; S. Gregoire; P. Gain; C. P. Garcher; A. M. Bron; N. Acar; L. Bretillon
Author Affiliations & Notes
  • O. Berdeaux
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
    INRA, University of Burgundy, Lipid-Aroma Platform, Dijon, France
  • L. Martine
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
  • S. Cabaret
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
    INRA, University of Burgundy, Lipid-Aroma Platform, Dijon, France
  • G. Thuret
    University Hospital, Biology, Engineering and Imaging of Corneal Grafts, JE2521, University Jean Monnet, Saint Etienne, France
  • S. Gregoire
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
  • P. Gain
    University Hospital, Biology, Engineering and Imaging of Corneal Grafts, JE2521, University Jean Monnet, Saint Etienne, France
  • C. P. Garcher
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
    Ophthalmology, University Hospital, Dijon, France
  • A. M. Bron
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
    Ophthalmology, University Hospital, Dijon, France
  • N. Acar
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
  • L. Bretillon
    INRA, University of Burgundy, Eye & Nutrition Research Group, Dijon, France
  • Footnotes
    Commercial Relationships  O. Berdeaux, None; L. Martine, None; S. Cabaret, None; G. Thuret, None; S. Gregoire, None; P. Gain, None; C.P. Garcher, None; A.M. Bron, None; N. Acar, None; L. Bretillon, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 5954. doi:
  • Views
  • Share
  • Tools
    • Alerts
      User Alerts
      You are adding an alert for:
      A New HPLC-ESI-MS/MS Method to Characterize and Quantify Phosphatidyl-Choline With VLC-PUFA: Application to Human Retina
      You will receive an email whenever this article is corrected, updated, or cited in the literature. You can manage this and all other alerts in My Account
      The alert will be sent to:
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation
      Citation

      O. Berdeaux, L. Martine, S. Cabaret, G. Thuret, S. Gregoire, P. Gain, C. P. Garcher, A. M. Bron, N. Acar, L. Bretillon; A New HPLC-ESI-MS/MS Method to Characterize and Quantify Phosphatidyl-Choline With VLC-PUFA: Application to Human Retina. Invest. Ophthalmol. Vis. Sci. 2010;51(13):5954.

      Download citation file:

      © ARVO (1962-2015); The Authors (2016-present)

      ×
    • Get Permissions
  • Supplements
Abstract

Purpose: : Mutations in the ELOVL4 gene have been found in Stargardt-like macular dystrophy or STD3. Previous studies have shown that ELOVL4 is involved in the biosynthesis of very long chain polyunsaturated fatty acids (VLC-PUFA). The aim of this work was to develop a HPLC-ESI-MS/MS method of characterization and quantification of dipolyunsaturated phosphatidyl-choline (PC) molecular species containing VLC-PUFA and to apply it on retinas from human donors.

Methods: : Eyeballs were collected from calf as well as from nine human donors (body donation to Science). The neural retina was dissected from the RPE/choroid. Following lipid extraction, phosphorus content of total phospholipids was determined.Using a triple quadrupole MS instrument, PC molecular species were structurally characterized by collision-induced dissociation in the negative mode with a method based on normal-HPLC-ESI-MS/MS. PC molecular species were then quantified using precursor ion scanning of m/z 184amu in the positive mode.

Results: : The characterization of PC species was done on bovine retinas. Among them, 28 were dipolyunsaturated PC species containing one VLC-PUFA (C24 to C36) with three to six double bonds. VLC-PUFA were always in the sn-1 position whilst PUFA at the sn-2 position was exclusively docosahexaenoic acid (DHA, C22:6.n-3). Most of these VLC-PUFA-containing dipolyunsaturated PC were detected and quantified in human retinas. The main represented compounds were those having VLC-PUFA of 32 carbon atoms (C32:3, C32:4, C32:5 and C32:6) and 34 carbon atoms (C34:3, C34:4, C34:5 and C34:6). Dipolyunsaturated PC with 36:5 and 36:6 were detected in lower quantities.

Conclusions: : This new HPLC-ESI-MS/MS method is sensitive and specific enough to structurally characterize and quantify all molecular species of PC, including those esterified with VLC-PUFA. This technique is valuable for a precise characterization of PC containing VLC-PUFA in retina and may be useful for better understanding their implication in the pathogenesis of STD3.

Keywords: lipids • retina • clinical laboratory testing 
© 2010, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×
This site uses cookies. By continuing to use our website, you are agreeing to our privacy policy.Accept