Protein-binding capacity of microquantities of tannins

The physiological effect of tannins is studied in terms of their protein-binding or precipitation capacity. A number of assays based on binding of hemoglobin or bovine serum albumin (BSA) and subsequent determination of unbound protein in supernatant or tannin in a protein-tannin complex are available but with various limitations. These methods are unable to estimate protein-binding capacity, if the quantity of tannin available is low. In the method reported here, tannins or other phenolics were applied on chromatography paper and reacted with BSA and unbound BSA was washed off. The protein in the tannin-protein complex was measured spectrophotometrically after staining with Ponceau S. It required microquantities of sample. Using this method the protein-binding capacity of total leaf extract and hydrolyzable and condensed tannins of Quercus incana, Q. semecarpofolia, and Q. dilatata was determined. The protein binding capacities of ellagic acid and quercetin (microgram BSA/mg) were 297.3 and 78.0, respectively.