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The quantification of reactive oxygen species and their scavengers by photon emission Yumiko Yoshiki*; Yoshinobu Akiyama; Kiyoshi Abe; Kazuyoshi Okubo Tohoku University, 1-1 tsutsumidori Amamiyamachi Aobaku, Sendai, Miyagi, 981-8555, Japan      *(yoshiki@bios.tohoku.ac.jp) The reactive oxygen species/hydrogen donor/mediator system (XYZ system) produced photon emission. The photon intensity of the XYZ system has a high concentration dependency to the reactive oxygen species (X), hydrogen donor (Y) and mediator (Z), which is indicated by [P] = k[X][Y][Z] (k = photon constant). Based on this XYZ system, we have developed a screening system for biomaterial, whereby a chemiluminescence analyzer is connected to HPLC system (Photon-HPLC system), in order to quantify the X, Y, Z species. After the HPLC-fractionation, the elution is mixed with two out of three of the following solvents (X solvent; 196 mM H2O2, Y solvent; 10 mM gallic acid dissolved in 10 % MeOH, Z solvent; sat. KHCO3 dissolved in 360 mM MeCHO) to produce the photon emission. Using this system, the detection limits were found to be 78 nmol for H2O2 (X), 50 pmol for gallic acid (Y). The results showed detection limits of lipoxygenase (Z) to be influenced by X solvent, such as, 116*102 U/ml used 196 mM H2O2, and 12*102 U/ml used 68 mM tert-BuOOH. In addition, we were able to quantify X species from cigarette smoke, Y species from Chinese tea and Z species from soybean seedlings. [Poster: yoshiki.yumiko.37372]

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