Polarographic Behavior of Co(II)-BSA or -HSA Complex in the Presence of a Guanidine Modifier

An extremely sensitive adsorptive wave of BSA at about -1.73 V (vs SCE) has been obtained in the solution containing 8 × 10^-7 mol/L CoCl2, 0.2 mol/L guanidine hydrochloride, and 0.2 mol/L NaOH by using single-sweep polarography. The interaction of BSA with guanidine in an alkaline solution results in its Co(II) complex with a positive excess charge. Thus, the complex is strongly adsorbed by the DME as a result of static electrical attraction. The adsorption efficiently accumulates the electrochemical active complex of Co(II)-BSA onto the DME. In the following potential scan, the complex produces a sensitive adsorptive reduction peak, which can be used to determine low-level BSA. In the absence of guanidine, the complex of Co(II)-BSA with a negative excess charge is repulsed by the DME. The reduction current is very small. The peak current depends on both BSA and Co(II) ion concentrations. HSA is similar to BSA in polarographic behavior. At the optimal conditions, the peak height is linearly proportional to the BSA or HSA concentration in the range of 0.005-20 mg/L (correlation coefficient 0.999). The detection limit for BSA or HSA is 0.002 mg/L. Lysozyme, common amino acids, and metal ions have no interference with the protein determination. The new method could be useful in protein studies.