硕士研究生开题报告(师大).ppt

1、,几种ESIPT型荧光探针的合成、表征及其分析应用,姓 名：XXX 学 号：XXX 专 业：有机化学 指导老师：XXX教授,汇报提纲,铜离子荧光探针的研究意义及进展,选题背景与意义,检测铜离子的意义： 在生物体中，铜离子是继铁离子和锌离子之后的第三大必要微量元素，一定含量的铜离子在动植物的基本生理活动过程中起着至关重要的作用。摄取过量的铜离子可能引发阿尔茨海默病、肌萎缩性脊髓侧索硬化症、门克斯综合征、威尔逊疾病等 。所以，灵敏性的选择性的检测环境和活细胞中的铜离子是相当有意义的 。 设计铜离子荧光探针主要有如下几种响应机理： 1、光诱导电子转移（PET） 2、分子内电荷转移（ICT） 3、荧光

2、共振能量转移（FRET） 4、激基缔合物 本选题的创新： 本论文发展的几种铜离子荧光探针是基于激发态分子内质子转移(ESIPT)的原理设计的，通常该类探针在与被测物反应后，反应迅速，斯托克位移较大，能够很好的消除自身激发光的干扰。,研究目标、内容,选择具有ESIPT性能的苯并咪唑和苯并噻唑类荧光染料，用以合成几种新型小分子荧光探针，实现对环境或活细胞中铜离子的高灵敏度高选择性检测。,研究目标,设计流程,研究目标、内容,实验内容,一：基于ESIPT型荧光探针对铜离子和氟离子的双通道检测 二：基于ESIPT机理的增强型铜离子荧光探针的合成与应用 三：苯并噻唑类荧光探针对铜离子的高灵敏度高选择性检测

3、,Probe 1 设计原理图：,探针1本身具有强烈的荧光，与Cu2+反应螯合，导致ESIPT过程受阻从而荧光淬灭。同时，由于探针1结构中存在分子内氢键，在一定的检测条件下，氟离子能夺取探针1酰胺结构上的H，导致荧光淬灭，因此还能检测溶液中的无机氟离子。,一：基于ESIPT型荧光探针对铜离子和氟离子的双通道检测,Probe 1 合成路线：,Fig. 1. Jobs plot for determining the stoichiometry of of probe 1 and Cu2+ in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7

4、.4. The total concentration of probes and Cu2+ was 10 M.,机理研究：,Fig. 2. Partial 1H NMR spectra of probe 1 (10 mM) upon titration with various equivalents of Cu2+.,Fig. 4. pH effect on fluorescence intensity of probe 1 (10 M) in the absence and presence of Cu2+ in buffered DMSO/Tris-HCl (v/v = 1:49) s

5、olution at pH 7.4.,条件优化,Fig. 3. Kinetic study of the response of probe 1 to Cu2+ at 25 in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4. Condition: molar ratio: probe:Cu2+ = 1:2.,Fig. 5. Fluorescence spectra of probe 1 (10 M) on addition of Cu2+ (0-10 M) in buffered DMSO/Tris-HCl (v/v = 1:4

6、9) solution at pH 7.4. detection limit: 1.6 10-9 M,Fig. 6. Fluorescence intensities of probe 2 (10 M) in the presence of other related metal ions and in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4. Cu2+ was added at a concentration of 10 M, and other cations (none; Cu2+; Pb2+; Na+; Ca2+;

7、Mg2+; Fe2+; Mn2+; Ag+; Cd2+; Zn2+; Ni2+; Fe3+; Co2+; Ba2+; Hg2+) were added at a concentration of 200 M.,对Cu2+线性、选择性研究：,Fig. 7. Fluorescence spectra of probe 1 (10 M) on addition of F- (0150 M) in buffered THF/PBS (v/v = 1:49) solution at pH 7.4. detection limit: 2.1 10-7 M,Fig. 8. Selectivity of pr

8、obe 1 (10 M) upon addition of different anions (none; Cl-; Br-; I-; H2PO4-; SO42-; AcO-; F-) in buffered THF/PBS (v/v = 1:49) solution at pH = 7.4.,对F-线性、选择性研究：,Fig. 9. Fluorescence microscope images of living HeLa cells. Cells incubated with 10 M Probe 1 (A) for 30 min at 37 0C. HeLa cells pretreat

9、ed with 10 M probe 1 (B) or for 30 min at 37 0C and then further incubated with 10 M Cu2+ for another 30 min. Top: bright field image, Bottom: fluorescence image.,Probe 1 在活细胞中的应用,Probe 2 设计原理图：,探针2本身具有微弱的荧光，在水溶液中，Cu2+催化吡啶甲酸酯水解，发生激发态分子内质子转移（ESIPT）过程，导致荧光增强。,二：基于ESIPT机理增强型铜离子荧光探针的合成与应用,Probe 2 合成路线：,

10、Fig. 1. Jobs plot for determining the stoichiometry of of probe 2 and Cu2+ in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4. The total concentration of probes and Cu2+ was 10 M.,机理研究：,Fig. 2. ESI-MS spectrum of the reaction products of probe 2 with Cu2+.,Fig. 4. pH effect on fluorescence in

11、tensity of probe 2 (10 M) in the absence and presence of Cu2+ in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4.,条件考察,Fig. 3. Kinetic study of the response of probe 2 to Cu2+ at 25 in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4. Condition: molar ratio: probe:Cu2+ = 1:2.,Fig. 6. Fl

12、uorescence intensities of probe 2 (10 M) in the presence of other related metal ions and in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4. Cu2+ was added at a concentration of 10 M, and other cations (none; Cu2+; Pb2+; Na+; Ca2+; Mg2+; Fe2+; Mn2+; Ag+; Cd2+; Zn2+; Ni2+; Fe3+; Co2+; Ba2+; Hg

13、2+) were added at a concentration of 200 M.,Fig. 5. Fluorescence spectra of probe 2 (10 M) on addition of Cu2+ (0-10 M) in buffered DMSO/Tris-HCl (v/v = 1:49) solution at pH 7.4. detection limit: 1.8 10-8 M,对Cu2+线性、选择性研究：,Fig. 7. Fluorescence microscope images of living HeLa cells. Cells incubated w

14、ith 10 M Probe 2 (C) for 30 min at 37 0C. HeLa cells pretreated with 10 M Probe 2 (D) or for 30 min at 37 0C and then further incubated with 10 M Cu2+ for another 30 min. Top: bright field image, Bottom: fluorescence image.,Probe 2 在活细胞中的应用,Probe 3 设计原理：,三：苯并噻唑类荧光探针对铜离子的高灵敏性高选择性检测,Probe 3 合成路线：,prob

15、e 3 (10 M); buffered DMSO/PBS (v/v = 1:49) solution, pH 7.4. Ex/Em = 346/535 nm; Slit (5 10) nm.,现象图：,probe 3 (10 M); Cu2+ (20 M) buffered DMSO/PBS (v/v = 1:49) solution, pH 7.4. Ex/Em = 346/535 nm; Slit (5 10) nm. 结论：反应迅速，在2 min内达到平衡,反应时间,在不同pH值条件下，探针本身及与铜离子反应的荧光谱图。 Probe 3 (10 M); Cu2+ (10 M) Ex/Em = 346/