FIA-Chemiluminescence Determination of Protein

The chemiluminescence (CL) reaction of the system of luminol-K3Fe(CN)6-protein,taking protein as a variable,and designing a FIA manifold for this CL reaction were studied. It was found that the CL reaction between luminol and K3Fe(CN)6 was catalyzed by the presence of certain protein and the intensity of CL was enhanced. Linear relationship was found to be kept between the increase of CL intensity and the concentration of protein. Based on this phenomenon,a thorough study on the factors affecting the CL reaction for the determination of protein,including the flow-rate of various solutions,the concentration of the CL reagents,pH of solution and the use of sensitizer etc,were made and a new method with optimized conditions was proposed. Linearity for OVA was found in the range of 1.8×10-10-2.2×10-7mol·L-1 (r=0.996 4) and for BSA in the range of 1.5×10-10-3.8×10-7mol·L-1 (r=0.998 8),with their respective detection limit (S/N=3) of 1.8×10-10mol·L-1 and 1.5×10-10mol·L-1. Test for precision was made at the concentration level of 7.6×10-8mol·L-1 of standard BSA,value of RSD (n=6) obtained were 0.88%. Results of total protein in human serum samples found were in consistency with the results obtained by the biuret method.