Detection of N-nitroso-bile acids at 285 nm in reverse-phase HPLC |
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Authors: | Araki Yoshio Mukaisyo Ken-ichi Sugihara Hiroyuki Fujiyama Yoshihide Hattori Takanori |
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Affiliation: | Department of Pathology, Shiga University of Medical Science, Seta Tsukinowa, Shiga, Japan. yir0828@belle.shiga-med.ac.jp |
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Abstract: | 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. |
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Keywords: | 285 nm detection Carcinogenesis N‐Nitroso‐bile acid Reverse‐phase HPLC |
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