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化学学报
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化学学报 ›› 2010, Vol. 68 ›› Issue (22): 2325-2330. 上一篇    下一篇

研究论文

某些芳香族氨基酸作探针荧光猝灭法测定秋水仙碱

江珊珊1,2，刘忠芳1，胡小莉1，刘绍璞*,1，许倩影1，田伦富1   

  1. (1发光与实时分析教育部重点实验室 西南大学化学化工学院 重庆 400715)
    (2国家城市供水水质检测网重庆监测站 重庆 400013)
  • 投稿日期:2010-02-04 修回日期:2010-06-26 发布日期:2010-07-25
  • 通讯作者: 刘绍璞 E-mail:liusp@swu.edu.cn
  • 基金资助:

    生物大分子间相互作用和分子识别的共振瑞利散射和共振非线性散射光谱研究

Fluorescence Quenching Method for the Determination of Colchicine with Some Aromatic Amino Acids as Probes

Jiang Shanshan1,2 Liu Zhongfang1 Hu Xiaoli1 Liu Shaopu*,1 Xu Qianying1 Tian Lunfu1   

  1. (1Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715)
    (2Chongqing Station, National Water Uality Monitoring Net of City Water Supply, Chongqing 400013)
  • Received:2010-02-04 Revised:2010-06-26 Published:2010-07-25
  • Contact: Shaopu Liu E-mail:liusp@swu.edu.cn

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1175

被引次数

25

在适当的酸性介质中, 秋水仙碱(COL)能与色氨酸(Trp)、酪氨酸(Tyr)和苯丙氨酸(Phe)等芳香族氨基酸反应并形成结合产物, 此时将引起上述氨基酸的荧光发生猝灭, 最大猝灭波长分别位于350 nm (Trp), 304 nm (Tyr), 284 nm (Phe). 其荧光猝灭值(ΔF)在一定范围内与秋水仙碱成正比. 当用Trp和Tyr作探针时, 荧光猝灭法测定秋水仙碱具有高灵敏度, 其检出限分别为15.1 ng/mL (3.78×10－8 mol/L)和19.8 ng/mL (4.96×10－8 mol/L). 文中研究了适宜的反应条件和影响因素, 考察了共存物质的影响, 表明方法有良好的选择性, 可用于秋水仙碱的测定. 文中讨论了复合物的组成、结合力和结合模式. 通过温度的影响以及Stern-Volmer作图, 判断该反应为静态猝灭反应, 它们的结合常数(K)在25 ℃时分别为9.7×106 (COL-Trp), 8.9×106 (COL-Tyr)和6.3×105 (COL-Phe). 其作用力主要是芳基堆集作用和氢键结合作用, 而芳基堆集作用是发生荧光猝灭的主要原因.

关键词: 秋水仙碱, 色氨酸, 酪氨酸, 苯丙氨酸, 荧光猝灭

In proper acid medium, colchicine could react with some aromatic amino acids such as tryptophane (Trp), tyrosine (Tyr) and phenylalanine (Phe) to form complexes, which led to fluorescence quenching of above amino acids, and the maximum quenching wavelengths were at 350 nm (Trp), 304 nm (Ty), 284 nm (Phe), respectively. Fluorescence quenching value (ΔF) was proportional to the concentration of colchicines in a certain range. Fluorescence quenching method for the determination of colchicine with Trp or Tyr as a probe had high sensitivity and the detection limits were 15.1 ng/mL (3.78×10－8 mol/L) and 19.8 ng/mL (4.96×10－8 mol/L) separately. The optimum conditions of the reaction, influencing factors and the effect of coexisting substances were investigated. The results showed that the selectivity was good, so the method could be used for the determination of colchicine. The composition of the complexes, the binding forces and binding mode were discussed in this work. Judging from the effect of temperature and the Stern-Volmer plots, the interaction was ascribed to a static quenching process. The association constants (K) were 9.7×106 (COL-Trp), 8.9×106 (COL-Tyr) and 6.3×105 (COL-Phe), respectively. The forces occurring in the reaction mainly included the aromatic stacking interaction and hydrogen bonding interaction between COL and amino acid, and the aromatic stacking interaction was the primary reason for fluorescence quenching.

Key words: colchicine, tryptophane, tyrosine, phenylalanine, fluorescence quenching