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。

A sensitive method to detect hemoglobin (Hb) by flow injection (FI) coupled with chemiluminescence (CL) detection was described, which based on Hb strong enhancing effect on weak luminol-hydrazine CL system in alkaline medium. Parameters affecting the CL detection conditions and FI-CL system were optimized. The effects of possible coexisting substances in human blood and serum of detection Hb were evaluated. Under the optimum conditions, the net CL intensity versus Hb concentration was linear in the range of 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 relative standard deviations (RSDs) for 8 replicate determinations of 5.0×10-7 and 3.0×10-6 g·mL-1 Hb were 1.6% and 1.5%, respectively. In addition, the recoveries of Hb in human blood and serum were carried out and varied from 83.0% to 101.0%. The proposed method has been successfully applied for the determination of Hb in healthy human blood and serum. The possible mechanism of Hb enhancing the weak CL emission of luminol-hydrazine system in NaOH solution was discussed by fluorescence spectrophotometer and UV-Vis spectrophotometer.