荧光试剂二(苯并三唑-1-基)乙二酮缩氨基脲的合成及其在蛋白质测定中的应用  

Synthesis of a Fluorescence Reagent Bis(benzotriazo-1-le)-ethyldione Semicarbazone and Its Application in Determination of Proteins

作　　者：高峰[1] 朱英贵[1] 章丽[1] 谢筱娟[1] 王伦[1] 

机构地区：[1]安徽师范大学化学与材料科学学院,芜湖241000

出　　处：《应用化学》2004年第4期374-377,共4页Chinese Journal of Applied Chemistry

基　　金：安徽省教育厅基金(2003kj144);安徽省高校青年教师基金(2003jq104);安徽师范大学基金

摘　　要：由苯并三唑、乙二醛、氨基脲合成了新荧光试剂二(苯并三唑-1-基)乙二酮缩氨基脲希夫碱。通过~1H NMR、红外光谱、元素分析确证了试剂的结构。在pH=4.0的KH_2PO_4-H_3PO_4缓冲溶液中,该试剂在342 nm处(λ_(ex)=284 nm)发射荧光,其强度随蛋白质的加入明显增强,据此建立了测定痕量蛋白质的新方法。在最佳实验条件下,测定牛血清白蛋白、人血清白蛋白的线性范围分别为0～8.0×10^(-6)g/mL和1.0×10^(-5)～0.25g/mL,检测限分别为5.3×10^(-9)g/mL、1.4×10^(-9)g/mL,用于合成样品的测定,结果令人满意。A novel fluorescence reagent bis(benzotriazo-1-le) -ethyldione semicarbazone(BTES)was synthesized from 1-H-benzotriazole, biformyl and semicarbazide. The structure of the reagent was characterized by elemental analysis, IR spectrometry and ~1H NMR spectrometry. The reagent gave fluorescence emission at 342 nm in a KH_2PO_4-H_3PO_4 buffer solution(pH=4.0). A new method for the determination of trace protein has been developed based on the enhanced fluorescence emission in the presence of a certain amount of protein. Under optimal conditions the linear range concentration was at 0～8.0×10^(-6)g/mL and 1.0×10^(-5)0.25g/mL, for bovine serum albumen and human serum albumen, respectively. The detection limit was 5.3×10^(-9)g/mL and 1.4×10^(-9)g/mL, respectively. The method has been tested on the synthesized samples with satisfactory results.

关 键 词：二(苯并三唑基)乙二酮缩氨基脲 蛋白质 荧光光度法 

分 类 号：O657[理学—分析化学]