Sensitive Determination of Proteins with Naphthol Green B by Resonance Light Scattering Technique

A new quantitative determination method for trace proteins using naphthol green B (NGB) by resonance light scattering (RLS) spectroscopy has been developed. The method is based on the interaction of protein and NGB at pH 3.00, which causes a substantial enhancement of the resonance scattering signal of NGB in the wavelength range 300-550 nm with the maximum RLS at 392.0 nm. Under optimum conditions, the linear range is 0.010-28.2 μg/ml for bovine serum albumin (BSA) and 0.010-31.3 μg/ml for human serum albumin (HSA). The detection limits (S/N=3) are 8.2 ng/ml for BSA and 7.9 ng/ml for HSA, respectively. There is little or no interference from amino acids, most of the metal ions, or other coexisting substances. The easy-to-use method, with high sensitivity and good reproducibility, was satisfactorily applied to the determination of total protein in human serum samples. The determination results for human serum samples are identical to those provided by clinical physicians.