A Sensitive Enzymatic Determination of Acylglycerol Hydroperoxides by Lipase and Glutathione Peroxidase with N-(9-Acrydinyl)maleimide fluorometry.

DOI DOI 被引用文献1件 オープンアクセス
  • KAMATA Tsuneo
    Division of Food and Nutrition, Jyumonji Gakuen Women's College
  • TANNO Hiroyuki
    Central Research Laboratory, Ajinomoto Co., Inc.
  • OHRUI Hiroshi
    Department of Food Chemistry, Faculty of Agriculture, Tohoku University
  • MEGURO Hiroshi
    Department of Food Chemistry, Faculty of Agriculture, Tohoku University

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説明

N-(9-Acrydinyl)maleimide (NAM) fluorometry was applied to the enzymatic determination of mono-, di- and triacylglycerol hydroperoxides (HPOs) by glutathione peroxidase (GPX) and lipase. Lipase hydrolysis made it possible to determine the acylglycerol HPOs. Two enzyme reactions were conducted simultaneously in one pot for 4hr at 25°C in 0.1 M phosphate buffer of pH 7.5 in a nitrogen atmosphere in the dark. The released glutathione-SH was measured by NAM fluorometry, the calibration curve giving a linear relationship in the range from 0.1 to 10nmol of HPO with a sample size of 0.2 to 20 mg. The sensitivity was about 250 times higher than that by conventional iodometry.

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