甲基蓝与血红蛋白相互作用的共振瑞利散射光谱研究及小分子的影响

Investigation on the interaction between methyl blue and hemoglobin by resonance rayleigh scattering spectra and effects of small molecules

在弱酸性条件下,甲基蓝（MB）可以通过静电和疏水作用聚集到血红蛋白（Hb）上并形成离子缔合物,不仅使得吸收光谱发生变化,而且使得共振瑞利散射增强。据此发展了一种测定Hb的RRS便捷方法,其线性范围和检出限分别为0.2~20μg/mL和0.012μg/mL。与RRS法相比,分光光度法的线性范围和检出限分别为8~60μg/mL和0.688μg/mL（344 nm）,5~60μg/mL和0.398μg/mL（408 nm）。此RRS法可用于尿样中Hb的痕量检测。实验考察了体系的吸收、散射光谱性质和最佳聚合条件,并在最佳条件下研究了分析参数及作用机理。并初步讨论了有机小分子对体系的影响,结果表明分子间氢键是主要作用力不容忽视。

In weak acidic solution,methyl blue（MB） could aggregate on hemoglobin（Hb） and form ion-association complexes by electrostatic and hydrophobic interactions.It resulted in not only the changes of the absorption spectrum,but also the enhancement of resonance rayleigh scattering（RRS）.According to these phenomena,a convenient method for the determination of Hb was developed.The linear range and the detection limit for the determination of Hb were 0.2~20 μg/mL and 0.012 μg/mL,respectively.Compared with RRS method,the linear range and the detection limits for spectrophotometry（SP） were 8~60 μg/mL and 0.688 μg/mL（at 344 nm）,5~60 μg/mL and 0.398 μg/mL（at 408 nm）,respectively.In this paper,the RRS method has been successfully applied to the determination of trace Hb in urine samples.The spectral characteristics of the absorption and scattering,the optimum conditions of the aggregating were investigated.Besides,the parameters of the analytical chemistry and mechanism of interaction were investigated under the optimum conditions.Finally,the influences of small organic molecules（S.O.M.） were also discussed.The results indicated that formation of intermolecular hydrogen bond between them was the main interaction of the complex,and it was not neglectful.