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在碱性介质中,发现血红蛋白对luminol-hydrazine弱化学发光体系有显著的增强作用。结合流动注射法提出了luminol-hydrazine化学发光(FI-CL)测定血红蛋白的新方法,优化了化学发光检测条件和FI-CL系统的运行参数,考察了实际样品中可能含有的共存物质对血红蛋白测定的影响。结果表明,血红蛋白的线性范围为5.0×10-9～6.0×10-5 g·mL-1,检出限是5.8×10-10 g·mL-1(9.0×10-12 mol·L-1),分别对5.0×10-7和3.0×10-6 g·mL-1的血红蛋白标准溶液平行八次测定所得RSDs分别为1.6%和1.5%。将提出的方法成功用于人血液和血清中血红蛋白含量的测定,加标回收率在83.0%～101.0%范围。结合化学发光光谱和紫外可见吸收光谱对血红蛋白催化luminol-hydrazine化学发光反应的机理进行了探讨。提出的方法具有灵敏度高、无需标记、操作简便、快速、易实现自动化操作等优点。
In alkaline media, hemoglobin was found to have a significant enhancement of the luminol-hydrazine weak chemiluminescence system. A new method for the determination of hemoglobin by luminol-hydrazine chemiluminescence (FI-CL) was proposed by flow injection method. The chemiluminescence detection conditions and the operating parameters of FI-CL system were optimized. The possible influence of coexisting substances on the determination of hemoglobin Impact. The results showed that the linear range of hemoglobin was 5.0 × 10-9 ~ 6.0 × 10-5 g · mL-1 with the detection limit of 5.8 × 10-10 g · mL-1 (9.0 × 10-12 mol·L-1) ). The RSDs of eight parallel determinations of 5.0 × 10-7 and 3.0 × 10-6 g · mL-1 hemoglobin standard solutions were 1.6% and 1.5%, respectively. The proposed method was successfully applied to the determination of hemoglobin in human blood and serum with the recovery of 83.0% ~ 101.0%. The mechanism of chemiluminescence reaction of luminol-hydrazine catalyzed by hemoglobin was studied by chemiluminescence and UV-Vis absorption spectra. The proposed method has the advantages of high sensitivity, no marking, easy operation, fast and easy to realize automatic operation.