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

本研究的目的是利用新型的化学放大反应作为指示反应，建立具有L式或S式反应曲线特征的高性能超痕量传感方法。特别是以分析需求较高的重要物质为研究对象，两年来取得的成果如下。基于催化剂和指示剂同时分解的L曲线的利用。血红蛋白的测定：建立的方法能够测定浓度范围为12-258μg/1的血红蛋白，检出限(3σ)为8.13μg/1，变异系数为2.0%(测定)，这表明利用L曲线的传感方法促进了分析精度的提高。与已知的多种血红蛋白分光光度测定方法相比，该方法具有最好的灵敏度。基于自催化反应的S曲线的利用。(1)建立了钴-吡啶偶氮染料络合物与硫代过氧磷酸盐的自催化反应，建立了过氧化氢的简便目视传感方法。用秒表和肉眼测定生命物质是可能的。并将该方法应用于过氧化氢等标记物的酶免疫分析。(2)采用亚硫酸盐与过氧化氢的自催化反应作为传感新方法。由于指示剂反应中的pH信号是用溴苏莫蓝(BTB)和石蕊试纸等pH指示剂可视化的，因此该方法可以用秒表和肉眼来测定人血清中的肿瘤坏死因子。在不久的将来，这种有用的方法将应用于更广泛的实际样品。

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.