Clinical Chemistry
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Clinical Chemistry 44: 1459-1465, 1998;
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(Clinical Chemistry. 1998;44:1459-1465.)
© 1998 American Association for Clinical Chemistry, Inc.

Lipids and Lipoproteins

Fluorometric determination of total retinyl esters in triglyceride-rich lipoproteins

Matthias Orth1,2,a, Monika Hanisch2, Gerd Kröning1, Mustafa Porsch-Özcürümez1, Heinrich Wieland2, and Claus Luley1,2

1 Institut für Klinische Chemie, Klinikum der Otto-von-Guericke-Universität, 39120 Magdeburg, Germany.

2 Abteilung Klinische Chemie, Albert-Ludwigs Universität, 79106 Freiburg, Germany.
a Address correspondence to this author at: Institut für Klinische Chemie, Klinikum der Otto-von-Guericke-Universität, Haus 39, Leipziger Str. 44, D-39120 Magdeburg, Germany. Fax 49-391-671-3902; e-mail matthias.orth{at}medizin.uni-magdeburg.de.

A time-consuming sample preparation and measuring procedure is required for the quantitation of retinyl palmitate by HPLC. We developed a fluorometric method for the determination of total retinyl esters in chylomicrons, chylomicron remnants, and VLDL. This method is precise, sensitive, rapid, simple, and particularly useful for large-scale studies of postprandial lipid metabolism. Because the turbidity of postprandial lipemic samples interferes with the fluorescence measurement, all samples were incubated for 10 min with a clearing buffer containing esterase and detergents. This buffer eliminates the turbidity and hydrolyzes all retinyl esters to retinol. The fluorescence signal (excitation wavelength, 330 nm; emission wavelength, 490 nm) was linear from 0.1 mg/L up to 4 mg/L retinyl palmitate, and the CVs were 3.6% within-run and 5.1% within-series. A first application studied postprandial lipoproteins, which were first separated by ultracentrifugation and then subjected to size exclusion chromatography. Fluorescence analysis revealed that the chylomicron density fraction contains large amounts of chylomicron remnants.




The following articles in journals at HighWire Press have cited this article:


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M. Porsch-Ozcurumez, S. Westphal, and C. Luley
Measurement of Low Apolipoprotein Concentrations by Optimized Immunoturbidimetric Applications
Clin. Chem., March 1, 2001; 47(3): 594 - 597.
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S. Westphal, M. Orth, A. Ambrosch, K. Osmundsen, and C. Luley
Postprandial chylomicrons and VLDLs in severe hypertriacylglycerolemia are lowered more effectively than are chylomicron remnants after treatment with n-3 fatty acids
Am. J. Clinical Nutrition, April 1, 2000; 71(4): 914 - 920.
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M. Orth, W. Weng, H. Funke, A. Steinmetz, G. Assmann, M. Nauck, J. Dierkes, A. Ambrosch, K. H. Weisgraber, R. W. Mahley, et al.
Effects of a Frequent Apolipoprotein E Isoform, ApoE4Freiburg (Leu28->Pro), on Lipoproteins and the Prevalence of Coronary Artery Disease in Whites
Arterioscler Thromb Vasc Biol, May 1, 1999; 19(5): 1306 - 1315.
[Abstract] [Full Text] [PDF]


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