Repeatability and reproducibility of lipoprotein particle profile measurements in plasma samples by ultracentrifugation

Sandra Monsonis-Centelles 1 , 2 , Huub C.J. Hoefsloot 1 , Søren B. Engelsen 2 , Age K. Smilde 1 , 2  and Mads V. Lindhttp://orcid.org/https://orcid.org/0000-0002-4999-1218 3
  • 1 Biosystems Data Analysis, Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands
  • 2 Department of Food Science, Chemometrics and Analytical Technology, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
  • 3 Department of Nutrition, Exercise and Sports, University of Copenhagen, Rolighedsvej 26, DK-1958 Frederiksberg C, Denmark
Sandra Monsonis-Centelles
  • Biosystems Data Analysis, Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands
  • Department of Food Science, Chemometrics and Analytical Technology, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
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, Huub C.J. Hoefsloot
  • Biosystems Data Analysis, Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands
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, Søren B. Engelsen
  • Department of Food Science, Chemometrics and Analytical Technology, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
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, Age K. Smilde
  • Biosystems Data Analysis, Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands
  • Department of Food Science, Chemometrics and Analytical Technology, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark
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and Mads V. LindORCID iD: https://orcid.org/0000-0002-4999-1218

Abstract

Background

Characterization of lipoprotein particle profiles (LPPs) (including main classes and subclasses) by means of ultracentrifugation (UC) is highly requested given its clinical potential. However, rapid methods are required to replace the very labor-intensive UC method and one solution is to calibrate rapid nuclear magnetic resonance (NMR)-based prediction models, but the reliability of the UC-response method required for the NMR calibration has been largely overlooked.

Methods

This study provides a comprehensive repeatability and reproducibility study of various UC-based lipid measurements (cholesterol, triglycerides [TGs], free cholesterol, phospholipids, apolipoprotein [apo]A1 and apoB) in different main classes and subclasses of 25 duplicated fresh plasma samples and of 42 quality control (QC) frozen pooled plasma samples of healthy individuals.

Results

Cholesterol, apoA1 and apoB measurements were very repeatable in all classes (intraclass correlation coefficient [ICC]: 92.93%–99.54%). Free cholesterol and phospholipid concentrations in main classes and subclasses and TG concentrations in high-density lipoproteins (HDL), HDL subclasses and low-density lipoproteins (LDL) subclasses, showed worse repeatability (ICC: 19.21%–99.08%) attributable to low concentrations, variability introduced during UC and assay limitations. On frozen QC samples, the reproducibility of cholesterol, apoA1 and apoB concentrations was found to be better than for the free cholesterol, phospholipids and TGs concentrations.

Conclusions

This study shows that for LPPs measurements near or below the limit of detection (LOD) in some of the subclasses, as well as the use of frozen samples, results in worsened repeatability and reproducibility. Furthermore, we show that the analytical assay coupled to UC for free cholesterol and phospholipids have different repeatability and reproducibility. All of this needs to be taken into account when calibrating future NMR-based models.

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Clinical Chemistry and Laboratory Medicine ( CCLM) publishes articles on novel teaching and training methods applicable to laboratory medicine. CCLM welcomes contributions on the progress in fundamental and applied research and cutting-edge clinical laboratory medicine. It is one of the leading journals in the field, with an impact factor of over three. CCLM is the official journal of nine national clinical societies and associated with EFLM.

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