采用流动注射化学发光法(FI-CL)研究了鲁米诺-铁氰化钾-蛋白质化学发光反应.试验发现某些蛋白质对鲁米诺-铁氰化钾发光体系有增强作用,对流速、发光试剂浓度、pH值以及增敏剂等条件进行了试验和优化并建立了测定蛋白质的FI-CL方法.测得卵清白蛋白(OVA)、牛血清蛋白(BSA)线性范围分别为1.8×10-10~2.2×10-7mol·L-1(r=0.996 4)和1.5×10-10~3.8×10-7mol·L-1(r=0.998 8),检出限(S/N=3)分别为1.8×10-10mol·L-1和1.5×10-10mol·L-1;对7.6×10-8mol·L-1牛血清蛋白标准的相对标准偏差为0.88%(n=6).用于人体血清样品总蛋白的测定,与双缩脲法的结果间无显著差异.

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.