Identification of molecular species of oxidized triglyceride in plasma and its distribution in lipoproteins

Clinical Chemistry and Laboratory Medicine : CCLM
Rojeet ShresthaHitoshi Chiba

Abstract

The role of triglycerides carried in the triglyceride-rich lipoproteins (TRL) in the progression of atherosclerosis is uncertain. Identification of oxidized triglycerides and its possible association with atherosclerosis were largely ignored. Here we applied mass spectrometric approach to detect and identify triglyceride hydroperoxides (TGOOH) in human plasma and lipoproteins. EDTA plasma was collected from healthy human volunteers (n=9) after 14-16 h of fasting. Very low-density lipoprotein (VLDL) (d<1.006) and intermediate-density lipoprotein (IDL) (d=1.006-1.019) were isolated from the plasma (n=6) by sequential ultracentrifugation in KBr, followed by the isolation of LDL and high-density lipoprotein (HDL) using size-exclusion high-performance liquid chromatography (HPLC). Total lipids from the plasma and isolated lipoproteins were extracted, and analyzed for the detection and identification of TGOOH using liquid chromatography/LTQ ion trap Orbitrap mass spectrometry. All the processes, from specimen collection to the mass spectrometric analysis, were carried out at 4 °C in the presence of antioxidant to prevent oxidation of lipoproteins. We identified 11 molecular species of TGOOH in either plasma or VLDL and IDL, of which ...Continue Reading

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Methods Mentioned

BETA
size exclusion chromatography
electrophoresis

Software Mentioned

Qual Browser
LipoPhor

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