H2O2 is the most common reactive oxygen molecules in organisms. It is mainly produced by the
catalyzation of SOD and XOD and degraded by the catalyzation of CAT and POD. H2O2, which is not only
one of the important reactive oxygen, but also the hub of mutual conversion of reactive oxygen. On the one
hand, H2O2 can directly or indirectly oxidize intracellular nucleic acids, proteins and other biological
macromolecules, and damage cell membranes, thus accelerating the aging and disintegration of cells. On
the other hand, H2O2 is also a key regulatory factor in many oxidative emergency reactions.
H2O2 and titanium sulfate generate yellow titanium peroxide complex with the characteristic absorption at
415nm.
Reagents and Equipment Required but Not Provided.
Spectrophotometer, table centrifuge, pipettor, 1 mL glass cuvette, acetone, concentrated hydrochloric acid
(37% HCl), mortar and ice.
Sample preparation
1. H2O2 extraction
A. Bacteria, cells or tissue samples
The bacteria or cells were collected into a centrifuge tube, and discard the supernatant after centrifugation.
Add 1mL reagent I to per 5 million bacteria or cells, and use the ultrasonic to crush the bacteria or cells
(20% power, 3 seconds ultrasound, 10 seconds interval, and 30 times repetition). Centrifuge at 8000 g for
10 min with 4℃ and place the supernatant on the ice for test.
B. Tissue samples
Weigh and take about 0.1g of tissue and added with 1mL reagent I for ice bath homogenate. Centrifuge at
8000 g for 10 min with 4℃, take all that (note absorbs clean) supernatant fluid, place it on the ice for test.
