[Determination of protein concentration in human saliva]

Seven colorimetric methods (Biuret, Lowry, a modified Lowry technique using bicinchoninic acid, Coomassie brilliant blue G (CBBG) dye-binding in phosphoric acid, perchloric acid or hydrochloric acid, and bromophenol blue dye-binding) were evaluated for determination of protein concentration in human whole saliva. Using bovine serum albumin (BSA), mucin, and thyroglobulin as protein standards to calculate salivary protein concentration gave different results among the assays used and even with the different standards of the same method. Among the seven methods used, the CBBG dye-binding assay in hydrochloric acid appeared to provide the most accurate estimation of protein concentration in human saliva. Using BSA as a standard, the mean values of human salivary proteins from 39 healthy individuals ranged from 0.72 to 2.45 mg/ml. The optimum conditions for the CBBG dye-binding assay in hydrochloric acid are: (1) the absorbance at 595 nm was measured between 15 to 30 minutes after addition of reagent. (2) the optimum concentration of dye in hydrochloric acid was 0.06-0.12% (w/v) for the assay. (3) the dye reagent was stable within one month.