【作者】 米贤文；

【导师】 王永生；

【作者基本信息】 南华大学 ， 卫生毒理学， 2006， 硕士

【摘要】 DNA加合物是化学毒物经生物转化后的亲电活性产物与DNA分子形成的共价结合物，它既是一种暴露标志物，又是一种效应标志物。8-羟基脱氧鸟苷是DNA损伤生物标志物中最有代表性的一种。建立8-羟基脱氧鸟苷的检测新方法具有重要的生物医学意义。 在本文第二章，用共振光散射技术建立了8-羟基脱氧鸟苷的检测新方法。研究表明：在pH9.0的R.B缓冲液中，吡啰红Y阳离子染料与8-羟基脱氧鸟苷阴离子形成离子缔合物，导致体系的共振光散射增强。在优化的实验条件下，体系的共振光散射强度随着8-羟基脱氧鸟苷的加入在339nm处急剧增强。共振光散射的增强(△I_RLS)与8-羟基脱氧鸟苷浓度成线性关系，据此建立8-羟基脱氧鸟苷的共振光散射检测新方法，方法的线性范围为50.0～226.4ng／ml，检出限为15.2ng／ml，RSD=8.8％，平均加标回收率=94.6％。 在本文第三章，用能量转移技术建立了鱼精蛋白荧光猝灭法检测8-羟基脱氧鸟苷的新方法。在25℃、pH8.2的Tris-HCl缓冲溶液中，在0.50～7.1μg／ml范围内，8-羟基脱氧鸟苷浓度与鱼精蛋白的荧光猝灭强度成线性关系，r=0.9963，方法的检出限为0.18μg／ml，RSD=6.2％，平均加标回收率为104％（n=6）。 在本文第四章，应用荧光光谱法研究了8-羟基脱氧鸟苷与鱼精蛋白的作用机制。实验表明，8-羟基脱氧鸟苷与鱼精蛋白的相互结合作用为静态荧光猝灭过程；更多还原

【Abstract】 DNA adduct is a covalent complex formed by DNA and electrophilic activity production of toxicological chemicals transformed in body, and it is not only an exposure biomarker but also an effect biomarker. 8-Hydroxy-2’-deoxyguanosine is one of the most representative biomarkers of oxidative DNA damage. To develop novel detection method for 8-hydroxy-2’-deoxyguanosine has important medical significance.In the chapter 2, a new detection method has been developed with th technology of Resonance Light Scattering. It is found that the intensity of system’s Resonance Light Scattering （RLS） is significantly enhanced duo to the formation of ion-association complex of 8-hydroxy-2’-deoxyguanosine （8-OHdG） anion with Pyronine Y cation in the pH9.0 buffer. Under the optimal experimental conditions, a new RLS determination for 8-OHdG was developed based on the enhancement(ΔI_RLS) of the system’s RLS intensity at 339nm, which is linear to the concentration of 8-OHdG in the range of 50.0²26.4ng/ml. The detection limit is 15.2ng/ml. The proposed method succeeded in the determination of spiked samples with simple and sensitive characteristic.In the chapter 3, a new detection method for 8- hydroxy-2’-deoxyguanosine has been developed with technology of fluorescence resonance energy transfer based on the fluorescence quenching effect. The fluorescent quenching intensity of protamine sulfate（Ps） at 301nm is linear to the concentration of 8-OHdG in the range of 0.50⁷.10μg/ml, at 25℃, in the pH8.2 Tris-HCl buffer solution with the detection limit of 0.18μg/ml. The relative standard deviation was 6.2% and average recovery was 104%（n=6）. The proposed method is simple and sensitive, succeeded in the更多还原