Liu S P, Liu Q
Institute of Analytical Science, Southwest China Normal University, Chongqing, China.
Anal Sci. 2001 Feb;17(2):239-42. doi: 10.2116/analsci.17.239.
In a weak acid medium, protein reacts with a monoazo dye of chromotropic acid, such as Chromazol KS (CALKS), Acid Chrome Dark Blue (ACDB), Chrome Blue SE (CBSE), Acid Chrome Blue K (ACBK), Chlorophosphonazo I (CPAI), Arsennazo I (AAI) and Chromotrope 2R (CT2R), to form a combination product. This results in a significant enhancement of the resonance Rayleigh-scattering (RRS) intensity, and the corresponding RRS spectrum appears. The characteristics of the RRS spectra of the combination products and the optimum conditions for the reactions were investigated. The intensity of RRS is directly proportional to the concentration of protein in a certain range. The RRS methods have high sensitivity for the determination of protein, the detection limits for bovine serum albumin (BSA) are 10.0 - 30.2 ng/ml, and the sensitivity order is CALKS > AAI = CPAI > ACBK = CT2R > ACDB > CBSE. The selectivity for CALKS has been examined, and the method was applied to the determination of a microg amount of protein in a synthetic sample with satisfactory results.
在弱酸介质中，蛋白质与变色酸单偶氮染料，如铬变素KS（CALKS）、酸性铬深蓝（ACDB）、铬蓝SE（CBSE）、酸性铬蓝K（ACBK）、氯膦酸偶氮I（CPAI）、偶氮胂I（AAI）和变色酸2R（CT2R）反应，形成一种结合产物。这导致共振瑞利散射（RRS）强度显著增强，并出现相应的RRS光谱。研究了结合产物的RRS光谱特征及反应的最佳条件。在一定范围内，RRS强度与蛋白质浓度成正比。RRS方法对蛋白质的测定具有高灵敏度，牛血清白蛋白（BSA）的检测限为10.0 - 30.2 ng/ml，灵敏度顺序为CALKS > AAI = CPAI > ACBK = CT2R > ACDB > CBSE。考察了CALKS的选择性，并将该方法应用于合成样品中微量蛋白质的测定，结果令人满意。
