高速液体クロマトグラフィーによるL-シトルリン測定を指標としたマクロファージの一酸化窒素産生の評価 (<特集> バイオサイエンスと分析化学)

概要

論文の詳細を見る
Determination of L-citrulline (Cit) by high performance liquid chromatography (HPLC) using an on-column derivatization and a column-switching technique has been studied for estimation of nitric oxide (NO) production by macrophages. The reaction system consisted of a reversed-phase polymer-gel column (Asahipak ODP-50) and the eluant of alkaline borate buffer containing o-phthalaldehyde (OPA) and N, N-dimethyl-2- mercapto-ethylamine hydrochloride. The OPA-fluorophore formed was subsequently transferred to an analytical column (Asahipak ODP-50) using the column-switching system with the mobile phase of acetonitrile and alkaline borate buffer. Optimization of the HPLC conditions led to a simple and rapid analytical method without any purification procedure for the simultaneous determination of Cit and L-arginine (Arg), a precursor of Cit, in a macrophage culture supernatant. The detection limit was 5 μM for both Cit and Arg in the culture supernatant. The proposed method was successfully applied to the screening of foods for the estimation of nitric oxide production by macrophages.
社団法人日本分析化学会の論文
1995-10-05