Analytical and Bioanalytical Chemistry

, Volume 402, Issue 7, pp 2407–2415 | Cite as

Particle size measurement of lipoprotein fractions using diffusion-ordered NMR spectroscopy

  • Roger Mallol
  • Miguel A. Rodríguez
  • Mercedes Heras
  • Maria Vinaixa
  • Núria Plana
  • Lluís Masana
  • Gareth A. Morris
  • Xavier Correig
Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

The sizes of certain types of lipoprotein particles have been associated with an increased risk of cardiovascular disease. However, there is currently no gold standard technique for the determination of this parameter. Here, we propose an analytical procedure to measure lipoprotein particles sizes using diffusion-ordered nuclear magnetic resonance spectroscopy (DOSY). The method was tested on six lipoprotein fractions, VLDL, IDL, LDL1, LDL2, HDL2, and HDL3, which were obtained by sequential ultracentrifugation from four patients. We performed a pulsed-field gradient experiment on each fraction to obtain a mean diffusion coefficient, and then determined the apparent hydrodynamic radius using the Stokes–Einstein equation. To validate the hydrodynamic radii obtained, the particle size distribution of these lipoprotein fractions was also measured using transmission electron microscopy (TEM). The standard errors of duplicate measurements of diffusion coefficient ranged from 0.5% to 1.3%, confirming the repeatability of the technique. The coefficient of determination between the hydrodynamic radii and the TEM-derived mean particle size was r 2 = 0.96, and the agreement between the two techniques was 85%. Thus, DOSY experiments have proved to be accurate and reliable for estimating lipoprotein particle sizes.

Figure

 

Keywords

Lipoprotein NMR DOSY TEM 

Abbreviations

CVD

Cardiovascular disease

DOSY

Diffusion-ordered NMR spectroscopy

DSTE

Double-stimulated echo

GGE

Gradient gel electrophoresis

HDL

High-density lipoprotein

IDL

Intermediate density lipoprotein

LDL

Low-density lipoprotein

LED

Longitudinal eddy current delay

LS

Light scattering

NMR

Nuclear magnetic resonance

PFG

Pulsed-field gradient

RMSPE

Root mean squared percentage error

sdLDL

Small, dense LDL

SE

Standard error

SNR

Signal-to-noise ratio

TEM

Transmission electron microscopy

TSP

3-Trimethylsilyl[2,2,3,3-d4]propionate

VLDL

Very low-density lipoprotein

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Notes

Acknowledgments

We acknowledge CIBER de Diabetes y Enfermedades Metabólicas Asociadas (ISCIII, Ministerio de Ciencia e Innovación), for partially funding this work, as well as the FIS (project PI 081409). This work was partly supported by the Engineering and Physical Sciences Research Council (Grant Numbers EP/H024336/1 and EP/I007989/1). We also acknowledge Dr. M. Moncusí and Dr. R. Marimon for their assistance with the TEM analysis of the lipoprotein fractions as well as Dr. S. Pujol for her assistance with viscosity measurements.

Supplementary material

216_2011_5705_MOESM1_ESM.pdf (878 kb)
ESM 1 (PDF 878 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Roger Mallol
    • 1
    • 2
    • 3
  • Miguel A. Rodríguez
    • 2
    • 3
  • Mercedes Heras
    • 3
    • 4
  • Maria Vinaixa
    • 1
    • 2
    • 3
  • Núria Plana
    • 3
    • 4
  • Lluís Masana
    • 3
    • 4
  • Gareth A. Morris
    • 5
  • Xavier Correig
    • 1
    • 2
    • 3
  1. 1.Department of Electronic EngineeringUniversitat Rovira i VirgiliTarragonaSpain
  2. 2.Metabolomics Platform, IISPVUniversitat Rovira i VirgiliTarragonaSpain
  3. 3.CIBERDEMBarcelonaSpain
  4. 4.Lipids and Atherosclerosis Research Unit, Sant Joan University Hospital, IISPVUniversitat Rovira i VirgiliReusSpain
  5. 5.School of ChemistryUniversity of ManchesterManchesterUK

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