Yue Qiaoli, Song Zhenghua, Dong Faxin, Liu Yanhong, Shao Xiaodong
Department of Chemistry, Shaanxi Key Laboratory of Physico-Inorganic Chemistry, Northwest University, Xi'an 710069, China.
J Fluoresc. 2007 Sep;17(5):505-11. doi: 10.1007/s10895-007-0202-4. Epub 2007 Jun 30.
It was first found that 4-(2-pyridylazo)-resorcinol (PAR) can combine with lysozyme forming a polymolecular complex, with the molar ratio of 1:12 (lysozyme/PAR) in the medium of acetate buffer (pH 6.0). The fluorescence spectra suggested that PAR molecules may produce self-aggregation, and it was suggested that there are interactions between lysozyme and PAR possibly mainly on the cooperation of hydrophobic, electrostatic actions and chelate bond. The coordination complex of lysozyme-PAR can cause an enhanced resonance light scattering (RLS) signal with the scattering peak located at 521 nm. The increment of RLS intensity was linear with the concentration of lysozyme over the range from 1 ng mL(-1) to 30.0 mug mL(-1), with the limit of detection of 0.3 ng mL(-1) (3sigma, n = 5). Effect of experimental conditions, including temperature, pH, concentration of PAR, and the ionic strength on RLS were also tested. The proposed RLS method was successfully applied to analyze lysozyme in hen egg white, human saliva and tear samples without any special pretreatment. Compared with other methods, the proposed procedure is of high sensitivity and selectivity.
首次发现4-(2-吡啶偶氮)-间苯二酚(PAR)能与溶菌酶结合形成多分子复合物,在醋酸盐缓冲液(pH 6.0)介质中,摩尔比为1:12(溶菌酶/PAR)。荧光光谱表明PAR分子可能产生自聚集,并且表明溶菌酶与PAR之间可能存在相互作用,主要是基于疏水作用、静电作用和螯合键的协同作用。溶菌酶-PAR的配位络合物可导致共振光散射(RLS)信号增强,散射峰位于521 nm处。在1 ng mL(-1)至30.0 μg mL(-1)范围内,RLS强度的增量与溶菌酶浓度呈线性关系,检测限为0.3 ng mL(-1)(3σ,n = 5)。还测试了包括温度、pH、PAR浓度和离子强度等实验条件对RLS的影响。所提出的RLS方法成功应用于分析鸡蛋清、人唾液和泪液样品中的溶菌酶,无需任何特殊预处理。与其他方法相比,该方法具有高灵敏度和选择性。