Determination of a guanosine-malonaldehyde adduct in urine by high-performance liquid chromatography with a thiobarbituric acid reaction detector.
A new method for the determination of a guanosine-malonaldehyde adduct, beta-D-ribofuranosylpyrimido[1,2-a]purin-10(3H)-one (GMA), in rat and human urine is described. The method involves rapid pretreatment using, in sequence, polyamide, ion-exchange and reversed-phase cartridges; determination is by means of high-performance liquid chromatography with a thiobarbituric acid reactor in series with a fluorescence detector. This device can quantitatively determine the adduct at the sub-picomole level. This rapid, selective and sensitive method is suitable for the determination of guanine-malonaldehyde adducts in biological samples, such as human and rat urine. A semi-preparative method for the extraction and purification of these adducts from rat urine and for their identification by mass spectrometry and high-performance liquid chromatography with ultraviolet detection is also reported.
Seto
H
H
Tokyo Metropolitan Research Laboratory of Public Health, Japan.
Ohkubo
T
T
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006151:Guanosine / Q000031:analogs & derivatives* / Q000652:urine; D006801:Humans; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D011741:Pyrimidine Nucleosides / Q000652:urine; D051381:Rats; D011916:Rats, Inbred F344; D013056:Spectrophotometry, Ultraviolet; D013858:Thiobarbiturates / Q000737:chemistry*
D007202:Indicators and Reagents; D011741:Pyrimidine Nucleosides; D013858:Thiobarbiturates; D006151:Guanosine; C073391:3-ribofuranosyl-pyrimido(1,2-a)purin-10(3H)-one; C029684:thiobarbituric acid