, Volume 52, Issue 1, pp 11–26 | Cite as

Identification of Oxidized Phosphatidylinositols Present in OxLDL and Human Atherosclerotic Plaque

  • Devin Hasanally
  • Andrea Edel
  • Rakesh Chaudhary
  • Amir Ravandi
Original Article


Oxidized low-density lipoprotein (OxLDL) plays an important role in initiation and progression of atherosclerosis. Proatherogenic effects of OxLDL have been attributed to bioactive phospholipids generated during LDL oxidation. It is unknown what effect oxidation has on the phosphatidylinositol (PtdIns) molecules in LDL, even though PtdIns is 6% of the total LDL phospholipid pool. We sought to identify and quantitate oxidized phosphatidylinositol (OxPtdIns) species in OxLDL and human atherosclerotic plaque. Bovine liver PtdIns was subjected to non-enzymatic and lipoxygenase-catalyzed oxidation. Reversed-phase liquid chromatography with negative ESI–MS identified and confirmed compounds by fragmentation pattern analysis from which an OxPtdIns library was generated. Twenty-three OxPtdIns molecules were identified in copper-oxidized human LDL at 0, 6, 12, 24, 30, and 48 h, and in human atherosclerotic plaque. In OxLDL, OxPtdIns species containing aldehydes and carboxylates comprised 17.3 ± 0.1 and 0.9 ± 0.2%, respectively, of total OxPtdIns in OxLDL at 48 h. Hydroperoxides and isoprostanes at 24 h (68.5 ± 0.2 and 22.8 ± 0.2%) were significantly greater than 12 h (P < 0.01) without additional changes thereafter. Hydroxides decreased with increased oxidation achieving a minimum at 24 h (5.2 ± 0.3%). Human atherosclerotic plaques contained OxPtdIns species including aldehydes, carboxylates, hydroxides, hydroperoxides and isoprostanes, comprising 18.6 ± 4.7, 1.5 ± 0.7, 16.5 ± 7.4, 33.3 ± 1.1 and 30.2 ± 3.3% of total OxPtdIns compounds. This is the first identification of OxPtdIns molecules in human OxLDL and atherosclerotic plaque. With these novel molecules identified we can now investigate their potential role in atherosclerosis.


Oxidized phospholipids Lipoproteins OxLDL Mass spectrometry Phosphatidylinositol Atheroma Atherosclerosis 



2:1 Chloroform:methanol


Butylated hydroxytoluene


Ethylenediaminetetraacetic acid


Embolic protection device








Levuglandin E2




Lipoprotein-associated phospholipase A2


Multiple reaction monitoring




Oxidized low density lipoprotein


Oxidized phosphocholine


Oxidized phosphatidylinositol


Platelet-activating factor




Percutaneous coronary intervention










Phosphatidylinositol 4-phosphate


Phosphatidylinositol 4,5-bisphosphate


Phosphatidylinositol 3,4,5-trisphosphate

PtdIns 3K

Phosphoinositide 3-kinases


Protein kinase B








Reactive oxygen species












Saphenous vein graft


Total ion chromatogram


Tumor necrosis factor alpha



This research was supported by operating grants from the Heart and Stroke foundation of Canada and Research Manitoba. DH was a recipient of a Research Manitoba graduate scholarship.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Supplementary material

11745_2016_4217_MOESM1_ESM.xlsx (66 kb)
Supplementary material 1 (XLSX 65 kb)


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Copyright information

© AOCS 2016

Authors and Affiliations

  • Devin Hasanally
    • 1
  • Andrea Edel
    • 1
  • Rakesh Chaudhary
    • 1
  • Amir Ravandi
    • 1
    • 2
  1. 1.Institute of Cardiovascular Sciences, St. Boniface Hospital, Albrechtsen Research CentreUniversity of ManitobaWinnipegCanada
  2. 2.Section of Cardiology, Department of Internal Medicine, Bergen Cardiac Care Centre, St. Boniface General HospitalUniversity of ManitobaWinnipegCanada

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