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

Synthesis of a novel fluorescence reagent of bis(1-H-benzotriazole)-ethyldione-thiosemicarbazone and its application in the 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,安徽芜湖241000

出　　处：《化学试剂》2004年第3期168-170,共3页Chemical Reagents

基　　金：安徽省教育厅基金 ( 2 0 0 3kj14 4) ;安徽省高校青年教师基金 ( 2 0 0 3jq10 4)

摘　　要：以苯并三唑、乙二醛、氨基硫脲为基本原料 ,合成了荧光试剂二 (1 H 苯并三唑 )乙二酮缩氨基硫脲席夫碱。通过对试剂进行红外光谱、元素分析确证了试剂的结构 ,并利用荧光分析法研究了它在蛋白质测定中的应用。在pH为 8 0的磷酸缓冲溶液中 ,该试剂在 394nm处 (λex=342nm)发射荧光。实验表明 ,其荧光强度随着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 ,对牛血清白蛋白、人血清白蛋白的测定 ,其线性范围分别为 0～ 1 0μg·mL-1 ,0 5 0～ 1 0 μg·mL-1 ,检测限分别为 6 3× 1 0 -3 ,1 2× 1 0 -3 μg·mL-1 。A novel fluorescence reagent bis(1 H benzotriazole) ethyldione thiosemicarbazone was synthesized with 1 H benzotriazole,biformyl,thiosemicarbazone as basic materials.The structure of the reagent was characterized by elemental analysis and infrared spectra.The application in the determination of proteins was studied by spectrophotofluorimetry.Fluorescence was emitted at 394 nm in the medium of pH 8.0 K 2HPO 4 NaOH buffer solution.A new method for the determination of trace protein was established on the basis of enhanced fluorescence by adding a certain amount of protein.Under optimal experimental conditions,the linear range for the determination of bovine sperm and human sperm was 0～10μg·mL -1 and 0.5～10μg·mL -1 ,respectively.The detection limit was 6.3×10 -3 μg·mL -1 and 1.2×10 -3 μg·mL -1 ,respectively.

关 键 词：二(1-H-苯并三唑)乙二酮缩氨基硫脲 蛋白质 荧光光度法 

分 类 号：O657[理学—分析化学]