Measurement of 4,5-dioxovaleric acid by high-performance liquid chromatography and fluorescence detection.
In this work we describe a sensitive method for the detection of 4,5-dioxovaleric acid (DOVA). 4,5-Dioxovaleric acid is derivatized with 2,3-diaminonaphthalene to form 3-(benzoquinoxalinyl-2)propionic acid (BZQ), a product with favorable UV absorbance and fluorescence properties. The high-performance liquid chromatographic method with UV absorbance and fluorescence detection is simple and its detection limit is approximately 100 fmol. This method was used to detect 4,5-dioxovaleric acid formation during metal-catalyzed 5-aminolevulinic acid (ALA) oxidation. Iron and ferritin were active in the formation of 4,5-dioxovaleric acid in the presence of 5-aminolevulinic acid. In addition, HPLC-MS-MS assay was used to characterize BZQ. The determination of 4,5-dioxovaleric acid is of great interest for the study of the mechanism of the metal-catalyzed damage of biomolecules by 5-aminolevulinic acid. This reaction may play a role in carcinogenesis after lead intoxication. The high frequency of liver cancer in acute intermittent porphyria patients may also be due to this reaction.
Ummus
R E
RE
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, SP, Brazil.
Onuki
J
J
Dörnemann
D
D
Medeiros
M H
MH
Di Mascio
P
P
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006580:Genetic Carrier Screening; D006801:Humans; D013058:Mass Spectrometry; D011164:Porphyrias / Q000175:diagnosis; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014631:Valerates / Q000032:analysis*
D014631:Valerates; C007509:4,5-dioxovaleric acid