What do we measure with luminol-, lucigenin- and penicillin-amplified chemiluminescence? 1. Investigations with hydrogen peroxide and sodium hypochlorite

Evidence is provided that the amplifiers luminol and lucigenin react with d ifferent reactive oxygen species (ROS), depending on the ROS-generating sys tem used. H2O2 is used to produce calibration curves for luminol- and lucig enin-amplified chemiluminescence. With this chemiluminescence generator we characterized the specificity and sensitivity of luminol- and lucigenin-amp lified chemiluminescence and also studied penicillin G, a known enhancer of luminol-amplified chemiluminescence. The combination of luminol and lucige nin in reciprocally changing concentrations is effective in an additive man ner, but the weak amplifier penicillin increases luminol-amplified chemilum inescence distinctly more than in an additive manner in different combinati ons. Lucigenin-amplified chemiluminescence is increased by penicillin at ab out 1% of the optimum concentration of penicillin; increasing concentration s of penicillin are less and less effective. On the other hand, low lucigen in concentrations enhance penicillin-amplified chemiluminescence at optimum penicillin concentrations more than in an additive manner. Fe2+ does not a lter luminol-, lucigenin- or penicillin-amplified chemiluminescence. Co2+ i ncreases luminol-amplified chemiluminescence by a factor of 100. Lucigenin- and penicillin-amplified chemiluminescence are minimally enhanced by Co2+. Cu2+ enhances luminol-amplified chemiluminescence with increasing concentr ations by a factor of 1000. Lucigenin-amplified chemiluminescence increases also by the factor of 1000. but the concentration-reaction curve is not as steep. NaOCl enhances H2O2/Fe2+-driven luminol-amplified chemiluminescence in a concentration-dependent manner by a factor of 10(4) (in the highest c oncentration of 10 mmol/L) and lucigenin amplified chemiluminescence only b y a factor of about 25. Catalase (CAT) abolishes luminol-, lucigenin- and p enicillin-amplified chemiluminescence completely, whereas superoxide dismut ase (SOD) has no effect on luminol- or penicillin-amplified chemiluminescen ce, but enhances lucigenin-amplified chemiluminescence five-fold increasing ly with increasing SOD activity. (C) 1998 John Wiley & Sons, Ltd.