Development of ultra trace sensing method of vital substances based on new chemical amplification

本研究的目的是建立一种以新型化学扩增为指示反应的高性能超痕量传感方法，该方法具有典型的l型或s型反应曲线。特别是对分析需求高的重要物质。两年内的成果如下：催化剂与指示剂同时分解的l型曲线的应用。血红蛋白的测定：本方法可测定血红蛋白浓度范围为12 ~ 258 μg/1，检出限（3σ）为8.13 μg/1。变异系数为2.0%（测定值），表明采用l型曲线的传感方法提高了分析精度。与已知的许多分光光度法测定血红蛋白的方法相比，该方法具有最佳的灵敏度。基于自催化反应的s型曲线的应用。(1)采用钴吡啶偶氮染料配合物与过氧化物单硫酸根自催化反应，建立了过氧化物的简单视觉感应方法。用秒表和肉眼就可以测定重要物质。此外，该方法还应用于过氧化物酶等标记的酶免疫分析。(2)亚硫酸盐与过氧化氢的自催化反应采用了新的传感方法。由于指示剂反应中的pH值信号是用溴thumol蓝（BTB）等pH指示剂和石蕊试纸可视化的，因此该方法可以用秒表法和肉眼法测定人血清中的肿瘤坏死因子。总之，本研究按照原计划获得了满意的结果。在不久的将来，这种有用的方法将广泛应用于实际样本。

The aim of this study is that creates the high-performance ultra trace sensing method using new chemical amplification as a indicator reaction, which has a characteristic L-shape or S-shape reaction curve. Especially, the target is vital substances which have high analytical needs.The produced fruits during the term of two years are as follows.1. The utilization of L-shape curve based on simultaneous decomposition of catalyst and indicator.Determination of hemoglobin : This established method is able to determine the hemoglobin of concentration range of 12-258 μg/1 and the detection limits (3σ) were 8.13 μg/1. Moreover, the coefficient of variation was 2.0% (determinations), and this result showed that the sensing method using L-shape curve promotes the improvement of analytical precision. The proposed method is that has the best sensitivity compared with already-known many spectrophotometric determinations of hemoglobin.2. The utilization of S-shape curve based on autocatalytic reaction.(1) The simple visual sensing method of peroxidaze was developed using autocatalytic reaction with cobalt-pyridylazo dye complex and peroxomonosulfate. The determination of vital substances is possible by using stop-watch and naked eye. Moreover, this method was applied to the enzyme immunoassay using the label such as peroxidaze.(2) The autocatalytic reaction with sulfite and hydrogen peroxide was used new sensing method. Because of pH signal in indicator reaction are visualized by the use of pH indicator such as bromothumol blue(BTB) and litmus paper, this method has enable determination of tumor necrosis factor in human serum using stop-watch and naked eye.In conclusion, the satisfactory results were obtained in this study according to the original plan. In the near future, this useful method will be applied to wide range of practical sample.