I am estimating the total phenolic content in a herbal formulation (Tablet or liquid Formulation), in this case what blank I should used (Placebo as such dissolved in water or placebo treated with Folin–Ciocalteu and all other reagents). What should I do in case if there is an absorbance in blank? Please help. I am taking pyrogallol as standard.
Tt is a relevant problem when your reagent and sample both are colored - your blank should be all components of the reaction mixture without the sample - using this blank also take a reading of your sample without reagents and check for absorbance - you may substract this from your sample + reagent reading - but generally sample reacts with reagents so ideally if there is a complete reaction of sample with reagent, the sample OD does not matter at all. Alternatively, you can use a 0min reading of sample + reagent. Folin-ciocalteu is widely used but only gives approximates - also be careful about experimental procedures - always keep all the reaction mixtures at same temp. and same duration for reliable results
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