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高性能的超痕量传感方法，使用新的化学放大作为指示反应，具有特征的L形或S形反应曲线。特别是针对分析需求高的重要物质，两年来取得的成果如下：1.血红蛋白的测定：本法测定血红蛋白的浓度范围为12-258 μg/l，检出限（3σ）为8.13 μg/l。变异系数为2.0%（测定值），表明L型曲线传感法有助于提高分析精密度。与已知的许多分光光度法相比，该方法具有最佳的灵敏度.基于自催化反应的S形曲线的应用。(1)利用钴-吡啶偶氮染料配合物与过氧硫酸单酯的自催化反应，建立了一种简单的过氧化物视觉传感方法。可通过秒表和肉眼测定重要物质。并将该方法应用于过氧化物酶等标记物的酶免疫分析。(2)利用亚硫酸盐和过氧化氢的自催化反应，建立了一种新的传感方法。由于溴硫酚蓝（BTB）和石蕊试纸等pH指示剂可使指示反应中的pH信号可视化，因此该方法可实现秒表和肉眼测定人血清中的肿瘤坏死因子，本研究按原计划进行，获得了满意的结果。在不久的将来，这一有用的方法将被应用到广泛的实际样品。

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.