|Xu, Ruping - DPT FD SCI, UC DAVIS, CA|
|Walzem, Rosemary - VET SCHL, UC DAVIS, CA|
|German, Bruce - DPT FD SCI, UC DAVIS, CA|
Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: October 20, 2003
Publication Date: May 1, 2005
Citation: Yokoyama, W.H., Xu, R., Walzem, R., German, B. 2005. Size-Exclusion Chromatography and Ultracentrifugation of Lipoproteins. Book Chapter. Chapter 17 - HPLC of Acyl Lipids. p. 521-534. Interpretive Summary: The measurement of serum cholesterol in lipoprotein fractions (LDL, HDL, and VLDL) is often necessary for medical and research applications. New size exclusion chromatography was compared to traditional ultracentrifugation methods for measuring these fractions in both hamster and human plasma. Although the two methods use different physical characteristics, size and density, respectively, the results are similar. Chromatographic methods require about 1% of the sample (10 ul vs 1000 ul) and only a fraction of the time (45 min vs 3 days) required for centrifugation methods.
Technical Abstract: The limited volume of plasma available from small animal favors micro methods of analysis. Hamster and human plasma lipoproteins were fractionated using size exclusion chromatography followed by an inline post column enzymatic reactor and results compared with the sequential density ultracentrifugation method. Correlation coefficients between the two methods for hamster and human HDL, LDL and VLDL ranged from 0.90 to 0.95. Enzymatic post column detection of cholesterol increases the sensitivity of the method over UV detection of apolipoprotein. This method is especially suitable for small animals where only microliters of plasma may be available.