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ルミノール化学発光を用いた,抗酸化物質の次亜塩素酸イオン消去能測定のためのシーケンシャルインジェクション分析(sequential injection analysis, SIA)法を開発した.同消去能は,抗酸化物質の次亜塩素酸イオン消去により減少するルミノール化学発光の減少率により算出した.50 mMホウ酸塩緩衝液(pH 9.5)をキャリヤーに用い,試薬濃度,流量,試薬体積等について最適化を行った.本SIA法によりL-アスコルビン酸,α-トコフェロール及びトロロッ クスの次亜塩素酸イオン消去能測定に応用した.構築したSIA法は,3検体当たり4分以内で測定可能であり,2 nM L-アスコルビン酸の消去能測定における相対標準偏差は,日内で2.5%,日間で3.8%と良好であった(n=3).本法は迅速,高感度かつ高精度であ り,測定時間及び試薬消費量も従来のバッチ法に比べ大幅に減少した.本SIA法は,次亜塩素酸イオン消去能を有する抗酸化物質の迅速なスクリーニングに有 用である.A sequential injection analysis (SIA) with chemiluminescence detection for measuring the antioxidative activity against hypochlorite ion was developed. The antioxidative activity was expressed as the percentage inhibition of luminol chemiluminescence due to the scavenging of hypochlorite ion by an antioxidant. Using a 50 mM borate buffer (pH 9.5) as a carrier, the SIA system was optimized with respect to the reagent concentrations, volume and flow rates. The SIA system could measure 3 samples within 4 min. The relative standard deviations of the antioxidative activity for 2 nM ascorbic acid were 2.5% (within-day, n=3) and 3.7% (between-day, n=3). Several antioxidants, such as ascorbic acid, α-tocopherol and Trolox, were success-fully measured by the SIA system, and the relationships between their antioxidative activity and the logarithm of concentrations of antioxidants were linear (r=0.983-0.999). The proposed SIA system was rapid, sensitive and reproducible with minimum consumption of the reagents, and was thus useful for the rapid screening of antioxidants possessing antioxidative activity against hypochlorite ion

Measurement of antioxidative activity against hypochlorite ion by sequential injection analysis with luminol chemiluminescence detection