Estimation of hydrogen peroxide in leaf using spectrophotometric analysis
Answers
Crush 0.1 g of tissue (leaf, root or shoot) (previously collected under ice cold conditions) in 0.1 % tri-chloro-acetic acid (TCA) and homogenize it at 4 °C. After centrifugation at 10 000 g for 15 min, keep the supernatant in dark for 1 h after mixing with phosphate buffer (10 mM, pH 7.0) and potassium iodide (1 M) (Add in the ratio 0.5 ml: 0.5 ml: 1 ml). Record the absorbance of the resulting solution at 390 nm.
All the steps were performed at 4 °C except absorbance measurement.
This is the simplest method for detection of H2O2. We have been doing it for last 5-6 years from different plant parts.
Also see the following publications of our lab (attached as pdfs):
(1) J Agron Crop Sci (2014) 200; 273–289
(2) IJBB 50 (2) (2013) 139-149
I hope this help
Answer:
A rapid and sensitive spectrophotometric method was described for the determination of hydrogen peroxide in aqueous solution. This method was based on the reduction of copper(II) ions by hydrogen peroxide in the presence of excess of 2,9-dimethyl-1,10-phenanthroline (DMP) to form the copper(I)-DMP complex.