Chemiluminescence luminol amplified

Voie, O. A., Wiik, P. and Fonnum, F., Ortho-substituted PCB activate respiratory burst measured as luminol-amplified chemiluminescence, Toxicol. Appl. Pharmacol., 150,369, 1998. 

Some authors used a completely different approaeh HA or the degradation products of HA were not directly detected, but the competition between HA and another (exogenously added) compound for the HO radicals was used as a measure of reactivity. In this context, the luminol-amplified chemiluminescence was often used [251] Luminol (5-amino-2,3-dihydro-phthalazine-l,4-dione) reacts with HO radicals under the emission of light. When luminol is present in excess over the generated radicals, the intensity of light... 

LUMINOL AMPLIFIED CHEMILUMINESCENCE IN THE MEASUREMENT OF PHAGOCYTE FUNCTION A METHOD... 

In a rat Uver microsomal system, both ascorbic acid and glutathione (GSH) as scavengers effectively diminish lucigenin ampUhed chemiluminescence in a concentration dependent manner to almost zero (Klinger et al. 1996). The spin trap substance N-t-butyl-a-phenylnitrone (C,H,5NO) diminished lucigenin ampUhed chemiluminescence more effectively than luminol amplified chemiluminescence. 

The different sensitivity of lucigenin and luminol to reactive oxygen species is reflected by some opposite results with certain compounds demonstrated by a low correlation coefficient for lucigenin- versus luminol-amplified chemiluminescence (Muller-Peddinghaus and Wurl 1987). 

Bryostatins stimulated phagocytosis of human granulocytes in vitro up to 233121 % in a concentration range from 1 mM to 10 nM (Eisemann et al. 1995). This high phagocytic potential could also be confirmed for bryostatin 1 in the in vivo carbon clearance model (cone, of 100 p-g/kg NMRI mouse injected intravenously, 350119%). In the luminol-amplified chemiluminescence test human granulocytes were stimulated by bryostatins 1, 2 and 5 between 1 pg and 10 ng to produce O2 radicals. The maximal O2 burst was measured after 8 min at a concentration of IpM. At 10 pM revealed a reduced burst capacity obviously due to the cytotoxic effect of the bryostatins (3719 %) at this concentration as estimated by trypan blue staining. 

Rost, M., Karge, E., and Klinger, W. (1998). What do we measure with luminol-, lucigenin- and penicillin-amplified chemiluminescence 1. Investigations with hydrogen peroxide and sodium hypochlorite. J. Biolumin. Chemilumin. 13 355-363. 

Another well-known CL amplifier, which is also frequently used for superoxide detection in biological systems, is luminol (5-amino-2,3-dihydro- 1,4-phthalazinedione). It has been proposed that luminol semiquinone reacts with superoxide to form the peroxide intermediate, whose decomposition is accompanied by chemiluminescence [62]. 

Oosthuizen and Greyling [93] recently investigated the possibility of using chemiluminescent methods for hydroxyl radical detection. These authors concluded that the lifetime of hydroxyl radicals (10 9 s) is too short to produce a meaningful level of CL. However, in the presence of carbonate the significant levels of luminol- and MCLA-amplified CL were observed supposedly due to Reaction (18), in which the formed much more stable radical C03- is capable of interacting with luminol or MCLA. 

Since the emission spectra of enhanced and unenhanced luminol oxidations are similar, it seems clear that emission is from the excited-state aminophthalate derived from luminol itself and not from the enhancer. The enhancement is also specific for peroxidase i.e., it does not occur under conditions in which the heme would dissociate from the enzyme. Furthermore, luminol chemiluminescence triggered by heme-containing compounds, such as hemoglobin or cytochrome c, is actually reduced by enhancers such as p-iodophenol (T7). This latter phenomenon explains the potential usefulness of enhanced chemiluminescence, i.e., an amplified signal combined with a reduced background and reduced interference from the reagents as well as from endogenous heme compounds. 

Our experience in measuring ohemiliminescent events by peripheral blood monocytes stimulated with phagocytizable particles has shown that consistent and precise data can be acquired if technical aspects are given special attention. The failure to acclimate monocytes to the media and to the dark, and the presence of phenol red in the media will result in suboptimal results. The suspension of cells in gelatin and the addition of luminol to amplify the response are unnecessary. Glass vials hare lower background chemiluminescence and provide more consistent data than that obtained with plastic vials. 

Ascorbic acid is the major water-soluble antioxidant present in cells and plasma. It will quench reactive oxygen species as 02 (Nishikimi 1975), HO (Bielski et aL 1975), and O2 (Bodannes and Chan 1979). On the other hand, it reduces Fe to Fe and thus will stimulate Fenton catalysis of H2O2 —> HO. Hydroperoxide-dependent lipid peroxidation in rat liver microsomes was enhanced by ascorbic acid (Laudicina and Marnett 1990). Ascorbic acid protected cardiac microsomes against lipid peroxidation and oxidative damage (Mukhopadhyay et al. 1993). It diminished both luminol- and lucigenin-amplified H2O2 derived chemiluminescence in concentrations > 10" (Klinger et al. 1996). 

The production of superoxide fit)m PMA stimulated HL-60 cells was determined by lucigenin-amplified chemiluminescence (CL) and luminol dependent CL as described 18). CL was assessed by luminometers (models 1251, LKB, Bromma, Sweden) and recorded as mV 19). HL-60 cells (2.3x10 ) in HBSS with lucigenin 200 pM or luminol 20 pM were preincubated with various compounds for 3 minutes, readings were started by 100 ng/mL PMA and CL was measured and recorded for 120 min. The total amount of superoxide produced during the assay period was determined by integrating the area under the curve (in mV.min). 

Kitagawa, R. R. Raddi, M. S. G. Khalil, N. M. Vilegas, W. Marcos da Fonseca, L. Effect of the isocoumarin paepalantine on the luminol and lucigenin amplified chemiluminescence of rat neutrophils. Biol. Pharm. Bull. 2003, 26, 905-908. 

---