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A low temperature flotation method to rapidly isolate lipoproteins from plasma

H Tong1, H R Knapp, M VanRollins

  • 1Department of Internal Medicine, College of Medicine, The University of Iowa, Iowa City 52242, USA.

Journal of Lipid Research
|August 26, 1998
PubMed
Summary

A new rapid flotation method isolates plasma lipoproteins in 18 hours, minimizing oxidative modification. This technique accurately quantifies cholesterol and triglyceride levels, offering a faster alternative for lipoprotein analysis.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Lipid Metabolism

Background:

  • Plasma lipoproteins are crucial for lipid transport but prone to oxidative modification.
  • Existing isolation methods can be time-consuming and may affect sample integrity.

Purpose of the Study:

  • To develop and validate a rapid, low-temperature method for isolating plasma lipoproteins.
  • To minimize oxidative damage during lipoprotein isolation.

Main Methods:

  • A sequential flotation method using a benchtop ultracentrifuge at low temperatures (4°C).
  • Characterization of protein and lipid content using electrophoresis, protein assays, gas chromatography-mass spectrometry, and enzymatic assays.
  • Validation against the established Havel et al. flotation method.

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Main Results:

  • The rapid method isolated identical lipoproteins and apolipoproteins compared to the Havel et al. method.
  • Summed recoveries of protein, cholesterol, and triglyceride were equivalent between methods.
  • Slightly higher triglyceride and cholesterol levels (9-12%) were observed in the rapid method's high-density lipoprotein (HDL) fraction, representing a minor portion of total plasma lipids.

Conclusions:

  • The 18-hour, low-temperature flotation method quantitatively isolates lipoproteins from human plasma.
  • This rapid technique offers a viable alternative for lipoprotein analysis, preserving sample integrity.
  • The method is suitable for accurate assessment of lipoprotein composition.