The actinobacterial test compound under study is slightly colored yellow. This seems to disturb the result of various assay performed at various wavelengths. At a wavelength of 405 where p-nitroaniline should be detected the compound color hinders it. And in hydrogen peroxide radical scavenging assay there is an increase in absorbance when a decrease in expected.
Since especially with radicals the spectrum of your compound may also change, so pretty much the only good way is to choose assay where the assay chromophore spectrum does not significantly overlap with your test compound spectrum (i.e. you want to avoid a compound that absorbs/fluoresces at the same wavelengths as the reporter dye in your assay).
If you take the reaction to completion, the interference due to the test compound can be minimized. Alternatively, you can subtract out the contribution of the test compound.
How about measuring the progress of the reaction by measuring the decrease in absorbance of the test compound?
If you do calibration and determination at at least 2 wavelength you will have 2 equations, where you can extract effect of yellow colouring. Alternatively, you may use derivative spectroscopy with sharp maxima.
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