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.
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, SP, Brazil.
R E
RE
Ummus
J
J
Onuki
D
D
Dörnemann
M H
MH
Medeiros
P
P
Di Mascio
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