Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, School of Chemistry and Chemical Engineering, Southwest University, Chongqing, China.
Luminescence. 2013 May-Jun;28(3):265-9. doi: 10.1002/bio.2375. Epub 2012 Apr 30.
In Britton-Robinson (BR) buffer medium (pH 3.3), carbazochrome sodium sulfonate (CSS) can react with some aromatic amino acids such as tryptophan (Trp), tyrosine (Tyr) and phenylalanine (Phe) to form a 1:1 complex by electrostatic attraction, aromatic stacking interaction and Van der Waals' force, resulting in fluorescence quenching of these amino acids. Maximum quenching wavelengths were located at 352 nm (CSS-Trp system), 303 nm (CSS-Tyr system) and 284 nm (CSS-Phe system), respectively. The fluorescence quenching value (ΔF) was proportional to the concentration of CSS in a certain range. The fluorescence quenching method for the determination of CSS showed high sensitivity, with detection limits of 31.3 ng/mL (CSS-Trp system), 44.6 ng/mL (CSS-Tyr system) and 315.0 ng/mL (CSS-Phe system), respectively. The optimum conditions of the reaction conditions and the effect of coexisting substances were investigated and results showed that the method had good selectivity. The method was successfully applied for the rapid determination of CSS in blood and urine samples. Based on the bimolecular quenching constant Kq , the effect of temperature and Stern-Volmer plots, this study showed that quenching of fluorescence of amino acids by CSS was a static quenching process.
在 Britton-Robinson (BR) 缓冲介质（pH 3.3）中，磺基萘甲酰亚胺钠盐（CSS）可以通过静电吸引、芳香族堆积相互作用和范德华力与一些芳香族氨基酸如色氨酸（Trp）、酪氨酸（Tyr）和苯丙氨酸（Phe）反应形成 1:1 复合物，导致这些氨基酸的荧光猝灭。最大猝灭波长分别位于 352nm（CSS-Trp 体系）、303nm（CSS-Tyr 体系）和 284nm（CSS-Phe 体系）。在一定范围内，荧光猝灭值（ΔF）与 CSS 的浓度成正比。测定 CSS 的荧光猝灭法具有较高的灵敏度，检测限分别为 31.3ng/mL（CSS-Trp 体系）、44.6ng/mL（CSS-Tyr 体系）和 315.0ng/mL（CSS-Phe 体系）。考察了反应条件的最佳条件和共存物质的影响，结果表明该方法具有良好的选择性。该方法成功应用于血液和尿液样品中 CSS 的快速测定。基于双分子猝灭常数 Kq、温度和 Stern-Volmer 图的影响，本研究表明 CSS 对氨基酸荧光的猝灭是一个静态猝灭过程。
