I need to measure the H2O2 content in cotton anthers. However, due to the high pigment content, I cannot obtain stable and repeatable results. I have tested methods based on Fe2+, I-, and titanium. I found that Fe2+ is the most sensitive ion. To remove pigments, I followed the method proposed in this article (A simple colorimetric method for determination of hydrogen peroxide in plant tissues, doi: 10.1007/s10725-006-9000-2). This paper suggested that activated charcoal can be used to remove pigments and antioxidants. However, I found that hydrogen peroxide was degraded and I could not determine any hydrogen peroxide at all (see figure). We used different solvents to extract hydrogen peroxide. The solvent I used was acetone, which is common in the literature. Maybe I need to try 5% TCA. Can you offer me any advice or experiences?
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