The Journal of Lipid Research, Vol. 40, 260-266, February 1999
Copyright © 1999 by Lipid Research, Inc.
Analysis of pristanic acid ß-oxidation intermediates in plasma from healthy controls and patients affected with peroxisomal disorders by stable isotope dilution gas chromatography mass spectrometry
N. M. Verhoevena,
D. S. M. Schora,
E. A. Struysa,
E. E. W. Jansena,
H. J. ten Brinka,
R. J. A. Wandersb, and
C. Jakobsa
a Department of Clinical Chemistry, Metabolic Unit, Free University Hospital, Amsterdam, The Netherlands
b Departments of Clinical Biochemistry and Pediatrics, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
Correspondence to:
C. Jakobs
In this paper we report the development of highly sensitive, selective, and accurate stable isotope dilution gas chromatography negative chemical ionization mass spectrometry (GC-NCI-MS) methods for quantification of peroxisomal ß-oxidation intermediates of pristanic acid in human plasma: 2,3-pristenic acid, 3-hydroxypristanic acid, and 3-ketopristanic acid. The carboxylic groups of the intermediates were converted into pentafluorobenzyl esters, whereas hydroxyl groups were acetylated and ketogroups were methoximized. Hereafter, the samples were subjected to clean-up by high performance liquid chromatography. Analyses were performed by selected monitoring of the carboxylate anions of the derivatives. Control values of all three metabolites were established (2,3-pristenic acid: 2–48 nM, 3-hydroxypristanic acid: 0.02–0.81 nM, 3-ketopristanic acid: 0.07–1.45 nM). A correlation between the concentrations of pristanic acid and its intermediates in plasma was found. The diagnostic value of the methods is illustrated by measurements of the intermediates in plasma from patients with peroxisomal disorders. It is shown that in generalized peroxisomal disorders, the absolute concentrations of 2,3-pristenic acid, 3-hydroxypristanic acid, and 3-ketopristanic acid were comparable to those in the controls, whereas relative to the pristanic acid concentrations these intermediates were significantly decreased. In bifunctional protein deficiency, elevated levels of 2,3-pristenic acid and 3-hydroxypristanic acid were found. 3-Ketopristanic acid, although within the normal range, was relatively low when compared to the high pristanic acid levels in these patients.—Verhoeven, N. M., D. S. M. Schor, E. A. Struys, E. E. W. Jansen, H. J. ten Brink, R. J. A. Wanders, and C. Jakobs. Analysis of pristanic acid ß-oxidation intermediates in plasma from healthy controls and patients affected with peroxisomal disorders by stable isotope dilution gas chromatography mass spectrometry. J. Lipid Res. 1999. 40: 260–266.
Supplementary key words:
2,3-pristenic acid, 3-hydroxypristanic acid, 3-ketopristanic acid, Zellweger syndrome, bifunctional protein, diagnosis, stable isotopes
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