| Fluorescence probes emitting at long-wavelength have strong ability of anti-interference and high penetration,which is widely used in the field of quantitative analysis and molecular imaging.Based on the principle of molecular association,the fluorescence probe of ionic associate is easy to prepare,avoiding the complexity of synthesis and the pollution produced during the synthesis process.In this thesis,the construction of metal phthalocyanine-based red-emitting fluorescent probes and their application in the quantitative analysis of heparin were carried out.This thesis has developed a new application of fluorescent phthalocyanine compounds in the analytical science.It is divided into five chapters.In the first chapter,the applications of optical probes in analytical sciences were classified,followed by a brief introduction.According to the absorption and emission in long-wavelength regions,optical probes were divided into two categories.Their advantages and application were introduced.The principle of ionic associate optical probes and its advantages compared with conventional probes are described.In the end,the biological activity of heparin,the significance and common methods for the determination of heparin were introduced.In the second chapter,according to the principle of molecular association,a phthalocyanine-based ion-associate fluorescent probe was prepared,followed by the establishment of a novel fluorimetry for the determination of heparin with high specificity at nanogram level.It was found that the fluorescence of AlS4Pc could be dramatically quenched by a structure-matched cationic copper phthalocyanie(Alcian blue 8 GX)which bears the same parent structure and oppositely charged.A fluorescent probe of AIS4Pc-Alcian blue 8 GX associate was developed in this study.Screening experiments on carbohydrates showed that the fluorescence of AIS4Pc-Alcian blue 8 GX recovered when polyanionic bio-polysaccharide existed,particularly in the presence of heparin.Based on the above findings,a highly sensitive and specific method for the determination of heparin by fluorescence enhancement was established.In the third chapter,the regularity of the fluorescence quenching of AIS4Pc by cationic surfactants was found.The discovery indicates that cationic surfactants with a conjugated cationic structure as a head,and a long carbon chain containing sixteen carbons or over as a tail have ability to quench the fluorescence of AIS4Pc with high efficiency,showing a typical "structure-activity" relationship.It was found that the poly-anionic biologic polysaccharides could restore the fluorescence of AIS4Pc-surfactant system,especially in the presence of heparin.Based on the above findings,fluorescent probe consisted of AlS4Pc-cationic surfactant was constructed,which was applied to highly specific and sensitive analysis of heparin.The calibration curves of heparin achieved from different sources were compared,it was found that these calibration curves were very close to each other,exhibiting that the response of heparins came from different sources was non-specific,which is another important discovery and advantage of this research.This work opens up a new application of phthalocyanine fluorescent compounds in the analytical sciences.In the fourth chapter,the binding constant of interaction between(Tetra(trimethylammionio)aluminum phthalocyanine,TTMAAlPc)and heparin was determined by fluorescence anisotropy.The results were in well agreement with that achieved by a plotting method constructed by our research team formerly,showing the reliability of the binding constant achieved.The fifth chapter,we introduced other studies we have engaged in during graduate studies.These studies failed to be completed for various reasons.However,some of the findings in the research process show potential value for further investigation.These works include the synthesis of cationic metal-phthalocyanine compounds and the construction of an ion-associate fluorescent probe with two phthalocyanines. |
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