Detection of N-nitroso-bile acids at 285 nm in reverse-phase HPLC.
In the present study, we developed a novel, simple, and specific detection method using an RP-HPLC at UV 285 nm for the separation and quantification of N-nitroso-bile acids. First, we found that N-nitroso-bile acids have a specific spectrophotometric absorbance at 285 nm. Using this 285 nm detection system, we could especially measure N-nitroso-bile acids, even in co-existence of non-N-nitroso-bile acids. Next, we observed the decomposition of N-nitroso-glychocholate under alkaline, acidic, and neutral conditions. N-nitroso-glychocholate rapidly decomposed under alkaline conditions (pH 9) (t(1/2) = 0.96 h), but remained fairly stable under acidic (pH 2) (t(1/2) = 12.8 h) and neutral (pH 7) (t(1/2) = 7.8 h) conditions. This study is the first report, which simply and specifically analyzes N-nitroso-bile acids using an RP-HPLC system.
Araki
Yoshio
Y
Department of Pathology, Shiga University of Medical Science, Seta Tsukinowa, Shiga, Japan. yir0828@belle.shiga-med.ac.jp
Mukaisyo
Ken-ichi
K
Sugihara
Hiroyuki
H
Fujiyama
Yoshihide
Y
Hattori
Takanori
T
D001647:Bile Acids and Salts / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D006863:Hydrogen-Ion Concentration; D008968:Molecular Conformation; D009603:Nitroso Compounds / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation / Q000379:methods; D013237:Stereoisomerism; D013997:Time Factors
D001647:Bile Acids and Salts; D009603:Nitroso Compounds