Luminol peroxidase-catalyzed

For example, peroxidase catalyzes the reaction of luminol derivatives with hydrogen peroxide and results in an increase of the CL reaction velocity and CL intensity. Therefore, intense CL can be obtained from the analyte labeled with luminol derivatives after HPLC separation, followed by reaction with peroxidase. 

As a hybrid between the isoluminol-based assays described in Section 3.1.4 and the horseradish peroxidase-catalyzed, luminol-based assays described here, Gun-dermann et al have described a high quantum-yield naphthalene cyclic hydrazide (7-dimethylaminonaphthalene-l,2-dicarboxylic acid hydrazide) and its use in assays catalyzed by horseradish peroxidase (G20). 

Fig. 17. Representative compounds that have been shown to enhance peroxidase-catalyzed luminescence from a mixture of luminol and peroxide.

Enhancement of Peroxidase-Catalyzed Luminol Chemiluminescence Effected by Benzothiazole Derivatives"... 

Peroxidase-catalyzed luminol oxidation, enhanced by phenols, naphthols, or luciferin analogs, has some attractive features for clinical assays. The emission is prolonged ( glow type) and constant, allowing for ease of measurement i.e., reactions can be carried out in batches away from the luminometer and read later as end-point determinations when the emission intensity is constant. The emission is also intense, so that detection is facilitated and can even be accomplished by silicon diode detectors or photographic film. 

M34. Misra, H. P., and Squatrito, P. M., The role of superoxide anion in peroxidase-catalyzed chemiluminescence of luminol. Arch. Biochem. Biophys. 215, 59-65 (1982). 

Naselund, B., Amer, P., Bemstrom, K., Bolinder, J., Hallander, L., and Lundin, A., Assays of free fatty acids and glucose by horseradish peroxidase catalyzed luminol reaction. In Bioluminescence and Chemiluminescence Current Status (P. E. Stanley and L. J. Kricka, eds.), pp. 385-388. Wiley, Chichester, 1991. 

NSselund, B. M. A., Berastrom, K., Lundin, A., and Amer, P., Free fatty acid determination by peroxidase catalyzed luminol chemiluminescence. J. Biotumin. Chemitumin. 3, 115-124 (1989). 

Detecting the presence of small, even invisible, amounts of blood is routine. Physical characteristics of dried stains give minimal information, however, as dried blood can take on many hues. Many of the chemical tests for the presence of blood rely on the catalytic peroxidase activity of heme (56,57). Minute quantities of blood catalyze oxidation reactions between colorless materials, eg, phenolphthalein, luco malachite green, luminol, etc, to colored or luminescent ones. The oxidant is typically hydrogen peroxide or sodium perborate (see Automated instrumentation,hematology). 

The emission yield from the horseradish peroxidase (HRP)-catalyzed luminol oxidations can be kicreased as much as a thousandfold upon addition of substituted phenols, eg, -iodophenol, -phenylphenol, or 6-hydroxybenzothiazole (119). Enhanced chemiluminescence, as this phenomenon is termed, has been the basis for several very sensitive immunometric assays that surpass the sensitivity of radioassay (120) techniques and has also been developed for detection of nucleic acid probes ia dot-slot. Southern, and Northern blot formats (121). 

The luminol reaction has been used for the determination of oxidizing agents such as hydrogen peroxide, for enzymes such as peroxidase and xanthine oxidase, and for metal ions such as copper or cobalt that catalyze this CL reaction [24],... 

Horseradish peroxidase HRP-catalyzed Luminol- H202 Borate buffer (pH 8.5) Microchip-based CE coupled with CL detection 7-35 nM (for HRP) 107... 

Recently, two major enzyme-catalyzed chemiluminescent reactions have become popular. These use either luminol as a substrate of peroxidase or 3-(2 -spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD) as a substrate of alkaline phosphatase (ALP). 

The development of optical immunosensors for the detection of antibody of HCY was performed by electrogeneration of the photoactive polypyrrole on ITO-modified optical fibers followed by internal light irradiation in the presence of the virus. The resulting layer of HCY can act as an antigen for the detection of the anti-HCV antibody. After the immunoreaction, the subsequent binding of a marker, a secondary peroxidase-labeled antibody, allowed to catalyze a chemiluminescence reaction in the presence of luminol and H202 [59]. 

Chemiluminescence of oxidized luminol has been the basis of several lumino-metric methods of estimation of TAC (Table 1). The mostcommon is to measure the induction time of the reaction. Often the chemiluminescence is first induced by an oxidant and then attenuated by addition of a sample, and the time to recover the initial fluorescence is measured. The enhanced chemiluminescent assay introduced a decade ago is based on the oxidation of luminol by perborate or by hydrogen peroxide in a reaction catalyzed by horseradish peroxidase. Enhancement (and stabilization) of luminescence is achieved by addition of p-iodophenol. The original procedure used a commercial reagent kit (ECL Anti-oxidant Detection Pack... 

CL detection is based on the optical emission of an excited species formed in a chemical reaction. For instance, an excited species is formed when luminol (or 3-aminophthalhydrazide) reacted with H202 as catalyzed by various substances, such as metal ions or the peroxidase enzyme. CL detection is achieved as in fluorescent detection, except that no excitation light is needed. 

Horseradish peroxidase (HRP)-catalyzed chemiluminescence detection of the intracellular ROS.3 The HRP-luminol solution was prepared by adding an adequate quantity of horseradish peroxidase (HRP) in luminol (0.1 mg/mL). The cuvette containing 2 mL of cell suspension (106 cells/mL) was placed in a homemade single-photon counter, and then, the photon emission from the cell suspension was recorded before and after addition of the HRP-luminol solution. The stationary photon emission reached after addition of HRP-luminol solution represents the ROS level in cell suspension. 

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