Quantitative acylcarnitine profiling in fibroblasts using [U-13C] palmitic acid: an improved tool for the diagnosis of fatty acid oxidation defects.
A method was developed for the investigation of mitochondrial fatty acid beta-oxidation in cultured fibroblasts. Monolayer cultures were incubated without foetal calf serum with commercially available [U-13C] palmitic acid and L-carnitine for 96 h. The acylcarnitines produced by the cells were extracted from the cell suspension and analysed either by quantitative stable isotope dilution gas chromatography chemical ionization mass spectrometry, or by fast atom bombardment mass spectrometry. Characteristic acylcarnitine profiles were obtained for all the different enzyme deficiencies investigated, with the exception of carnitine palmitoyltransferase II deficiency and carnitine/acylcarnitine carrier deficiency which showed similar patterns. Comparison between this method and the 3H-myristate and 3H-palmitate tritium release assays revealed that the method described here is superior, allowing unequivocal identification of patients.
Ventura
F V
FV
Department of Clinical Biochemistry, University Hospital Amsterdam, The Netherlands.
Costa
C G
CG
Struys
E A
EA
Ruiter
J
J
Allers
P
P
Ijlst
L
L
Tavares de Almeida
I
I
Duran
M
M
Jakobs
C
C
Wanders
R J
RJ
D002247:Carbon Isotopes; D002331:Carnitine / Q000031:analogs & derivatives* / Q000378:metabolism; D002478:Cells, Cultured; D005227:Fatty Acids / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D008052:Lipid Metabolism, Inborn Errors / Q000175:diagnosis*; D010084:Oxidation-Reduction; D019308:Palmitic Acid; D016339:Spectrometry, Mass, Fast Atom Bombardment
D002247:Carbon Isotopes; D005227:Fatty Acids; C116917:acylcarnitine; D019308:Palmitic Acid; D002331:Carnitine