Hydrogen Peroxide → Luminol

Participants

Reaction Steps

The pyridopyridazine compound (L-012) as obtained in Example 3 and luminol were dissolved at the concentration of 0.2 mM in 0.2 M Tris buffer (pH 8.6) containing 0.1 mM EDTA. In test tubes were put 990 μl respectively of the solutions, and to each of the solutions was added 10 μl of a dimethylsulfoxide (DMSO) solution of 40 mM p-iodophenol as a chemiluminescent enhancer, followed by mixing. In other test tubes were put 50 μl of each of the mixtures, whereto 100 μl of a 0.2 M Tris buffer (pH 8.6) containing horseradish peroxidase at the concentrations of 10 pg/ml, 100 pg/ml 1 ng/ml or 10 ng/ml. By further adding 50 μl of Tris buffer (pH 8.6) containing of 0.6 mM H2O2 to each of the tubes, chemiluminescent reaction was initiated. The measurement was carried out for the luminous amounts for 10 seconds between 60 seconds later and 70 seconds later after addition of H2O2. The results are tabulated in Table 3. From Table 3, it was shown that the pyridopyridazine compound of Example 3 (L-012) provides about 4 to 20 times as great chemiluminescent amounts as luminol. The sodium salt of L-012 showed the same chemiluminescence activity as L-012.

Provenance & Evidence