Simultaneous analysis of plasma free fatty acids and their 3-hydroxy analogs in fatty acid beta-oxidation disorders.
We present a new derivatization procedure for the simultaneous gas chromatographic-mass spectrometric analysis of free fatty acids and 3-hydroxyfatty acids in plasma. Derivatization of target compounds involved trifluoroacetylation of hydroxyl groups and tert-butyldimethylsilylation of the carboxyl groups. This new derivatization procedure had the advantage of allowing the complete baseline separation of free fatty acids and 3-hydroxyfatty acids while the superior gas chromatographic and mass spectrometric properties of tert-butyldimethylsilyl derivatives remained unchanged, permitting a sensitive analysis of the target compounds. Thirty-nine plasma samples from control subjects and patients with known defects of mitochondrial fatty acid beta-oxidation were analyzed. A characteristic increase of long-chain 3-hydroxyfatty acids was observed for all of the long-chain 3-hydroxyacyl-CoA dehydrogenase-deficient and mitochondrial trifunctional protein-deficient plasma samples. For medium-chain acyl-CoA dehydrogenase deficiency and very-long-chain acyl-CoA dehydrogenase deficiency, decenoic and tetradecenoic acids, respectively, were the main abnormal fatty acids, whereas the multiple acyl-CoA dehydrogenase-deficient patients showed variable increases of these unusual intermediates. The results showed that this selective and sensitive method is a powerful tool in the diagnosis and monitoring of mitochondrial fatty acid beta-oxidation disorders.
Costa
C G
CG
Centro de Metabolismos e Genética, University of Lisboa, Portugal.
Dorland
L
L
Holwerda
U
U
de Almeida
I T
IT
Poll-The
B T
BT
Jakobs
C
C
Duran
M
M
D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000172:deficiency; D044942:Acyl-CoA Dehydrogenase, Long-Chain / Q000172:deficiency*; D002334:Carnitine O-Palmitoyltransferase / Q000172:deficiency; D002648:Child; D005230:Fatty Acids, Nonesterified / Q000097:blood*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006801:Humans; D006900:Hydroxylation; D017240:Mitochondrial Myopathies / Q000097:blood* / Q000201:enzymology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D005230:Fatty Acids, Nonesterified; D015094:3-Hydroxyacyl CoA Dehydrogenases; D044942:Acyl-CoA Dehydrogenase, Long-Chain; D002334:Carnitine O-Palmitoyltransferase