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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
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.
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.
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.