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Detection of N-nitroso-bile acids at 285 nm in reverse-phase HPLC
- Araki, Yoshio, Mukaisyo, Ken-ichi, Sugihara, Hiroyuki, Fujiyama, Yoshihide, Hattori, Takanori
- Journal of separation science 2008 v.31 no.15 pp. 2827-2830
- absorbance, pH, reversed-phase high performance liquid chromatography
- 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) (t₁/₂ = 0.96 h), but remained fairly stable under acidic (pH 2) (t₁/₂ = 12.8 h) and neutral (pH 7) (t₁/₂ = 7.8 h) conditions. This study is the first report, which simply and specifically analyzes N-nitroso-bile acids using an RP-HPLC system.