Spectroscopic Studies On The Interaction Of Protein With Fluprofen And Applications Of Lomefloxacin To Fluorescence Analysis

As fluorescence analysis has the advantages of economic, simple, high sensitivity, goodselectivity and low detection limits, it is widely used on many fields, such as biochemistry,pharmacology, clinical medicine. Fluorescence spectroscopy was considered as the researchmeans in this paper, which has carried on the study of the binding mechamism between thenew animal remedy called fluprofen and bovine serum albumin, and the applications oflomefloxacin to fluorescence analysis. The research content has been divided into fivechapters:Chapter one: The progress and position of pharmaceutical analysis on the fields ofpharmacy and life science were reviewed. Accordingly, a variety of fluorescence analysismethods in pharmaceutical analysis were reviewed to provide a rich theoretical basis for theselection and development of high efficacy, wide application, toxic side effects of drugs, with59literatures quoted.Chapter two: The interaction between bovine serum albumin and fluprofen wasinvestigated at different temperatures, as well as the effect of common metal ions (Mg2+, Zn2+,Cu2+, Fe3+, Co2+, Ni2+) on this system. Results showed that fluprofen could quench theintrinsic fluorescence of bovine serum albumin significantly, and the quenching mechanismwas a static quenching process. The electrostatic force played an important role on theconjugation reaction between bovine serum albumin and fluprofen. The binding constantswere in the order of103, and the number of binding site in the binary system wasapproximately equal to1. The binding distance was about2.20nm. The values of Hill’scoefficients were slightly larger than1, which indicated that there was some positivecooperative effect. Synchronous spectra revealed that the microenvironment and theconformation of BSA were changed during the conjugation reaction.Chapter three: In the Tris-HCl buffer solution which pH was7.40, the interaction ofbovine serum albumin with fluprofen was investigated by classical fluorescence spectroscopyand improved fluorescence spectroscopy at293K,303K and310K, and the conclusions of the two methods were consistent. Results showed that fluprofen could quench the intrinsicfluorescence of bovine serum albumin, and the quenching mechanism was a static quenchingprocess. The static electricity forces attraction played an important role on the conjugationreaction between bovine serum albumin and fluprofen, the number of binding site in thebinary system was approximately to1. The values of Hill’s coefficients were more than1,which indicated very weakly positive cooperativity in this system. The binding constant thatobtained by improved fluorescence spectroscopy was much larger than the one obtained byclassical fluorescence spectroscopy, which indiciated that it was more accurate and reasonablewhen regarding the drug as the research object. At last, the scientificalness of the new methodbased on improved fluorescence spectroscopy was verified by ultraviolet spectroscopy.Chapter four: In the Acetic acid-sodium acetate buffer solution of pH6.0, terbium andlomefloxacin react and combine as steady complex under room temperature. The complexshows its sensitized fluorescence of rare earth. The maximum excitation wavelength andmaximum emission wavelength is at λex=320nm and λem=545nm, respectively. Thefluorescence intensity increase with norfloxacin added, while the locations of the excitationand emission wavelengths stay the same. According to this, a new way to determine theconcentration of norfloxacin is established.Chapter five: The energy transfer reaction between Tb3+-lomefloxacin rare earthsensitized fluorescence and alkaline dye rhodamine B in aqueous was studied by fluorescencespectroscopy. The results showed that Tb3+-lomefloxacin could quench the fluorescence ofrhodamine B significantly, and the quenching mechanism was a static quenching process. Thebinding constant between Tb3+-lomefloxacin and rhodamine B was1.63×105L/mol. Thecombined distance of the system was determined according to the Fōrster theory. In addition,the fluorescence quantum yield of the system has been investigated.