Spectrophotometric determination of total protein in serum using a novel near-infrared cyanine dye, 5,5'-dicarboxy-1,1'-disulfobutyl-3,3,3',3'-tetramethylindotricarbocyanine.

The application of near-infrared (NIR) dyes (lambda (em) > 750 nm) to the analysis of biological samples shows much promise, because the long emission wavelengths of such dyes allow interferences from biomolecule matrices to be minimized. In this paper, a novel NIR dye, 5,5'-dicarboxy-1,1'-disulfobutyl-3,3,3',3'-tetramethylindotricarbocyanine (DCDSTCY) has been developed for the spectrophotometric determination of total protein in serum. Under acidic conditions, the binding of DCDSTCY to proteins caused a new peak at 878 nm, the height of which was proportional to the concentration of protein. The linear range of the method was found to be 0.04-0.5 microg mL(-1) for bovine serum albumin (BSA) and human serum albumin (HSA), and detection limits of 5 ng mL(-1) were obtained for these substances. The maximum binding number of BSA with DCDSTCY was measured to be 133. The method proposed here has been applied to the quantitation of total protein in serum, and recoveries of 96.6-104% were achieved. Figure Near-infrared probe for protein determination.