Study On The Determination Of Anthracycline Antibiotics By Molecular Spectroscopy

Molecular spectroscopy is a method that used to measure wavelength and intensity in the reflection,absorption or scattered radiation which is generated in the quantization energy level transition of internal material when molecular interact with the electromagnetic radiation.It is the important experimental method of determining and indentifing molecular structure.According to difference ranges of absorbing the electromagnetic wave,it can be divided into ultraviolet-visible spectrum,the near-infrared spectrum and infrared spectrum etc;according to the difference electromagnetic wave emission method,it can be divided into the molecular fluorescence,the phosphorescence and the scattering spectrum and so on.Taking the anthracycline antibiotics as examples,we have studied and developed the new methods for the determination by the molecular spectroscopy,In this thesis,we mainly investigated the interaction between anthracycline antibiotics and metal ions,dyes by the molecular spectroscopy.Besides,the spectral characteristics,the optimum conditions and the influencing factors were studied.The reaction mechanism was discussed too.Based on the above researches,the molecular spectroscopy have been established for the determination of anthracycline antibiotics.Main investigated systems are listed as follows:1 Study on the Interaction between Mitoxantrone Metal Complex and Titan Yellow by Resonance Rayleigh Scattering Spectra and Its Analytical ApplicationIn pH 2.45 Britton-Robinson(BR) buffer medium,the anthracycline antibiotics such as mitoxantrone(MXT) react with palladium(II)(Pd(II)),copper(II)(Cu(II)),cobalt(II)(Co(II))to form 1:2 cationic chelates,which further react with Titan Yellow(TY) to form 1:2 ternary ion-association complexes.As a result,not only the spectra of absorption and fluorescence axe changed,but also the resonance Rayleigh scattering(RRS) is enhanced greatly.Their spectral characteristics of RRS are similar,and the maximum RRS wavelengths are all located at 454 nm.But the increments of RRS intensity are different in the series of Pd(II)>Co(II)>Cu(II).The enhanced RRS intensities are proportional to the concentration of MXT in the range of 0.031～2.4μg/mL(Pd²⁺ system),0.032～1.2μg/mL(Co²⁺system) and 0.03～1.2μg/mL(Cu²⁺ system).The detection limits(3σr)are 9.4 ng/mL (pd²⁺system),9.7 ng/mL(Co²⁺system) and 9.8 ng/mL(Cu²⁺ system).In this paper,The optimum conditions of the reactions and the effects of foreign substances are investigated,and the composition of ion-association complexes is discussed also.Based on the ion-association reaction,a highly sensitive,simple and rapid method has been proposed to the determination of MXT.2 Study on the resonance Rayleigh scattering spectra of the interactions of Ce(IV)-mitoxantrone with Alizarin Red and their analytical applicationsIn pH 5.4 Michaelis(HAC-NaAC) buffer medium,the anthracycline antibiotics such as mitoxantrone(MXT),daunorubicin(DNR) and doxorubicin(DOX) react with Cerium(IV)(Ce(IV)) to form 1:2 cationic chelates,which further react with Alizarin Red(AR) to form 1:8 ternary ion-association complexes.As a result,the resonance Rayleigh scattering(RRS) enhanced greatly. They have similar spectral characteristics and their maximum RRS wavelengths are all located at 368 nm,and there are one smaller RRS peak in 548 nm.But the increments of RRS intensity are different in the series of MXT＞DOX＞DNR.The enhanced RRS intensities are proportional to the concentration of three anthracycline drugs in the range of 0.024～2.5μg/mL(MXT),0.067～2.0μg/mL(DOX) and 0:082～2.0μg/mL(DNR).The detection limits(3σ) are 7.2 ng/mL(MXT),12.5 ng/mL(DOX) and 15.9 ng/mL(DNR).In this paper,The optimum conditions of the reactions and the effects of foreign substances are investigated,and the composition of ion-association complexes is discussed also.Based on the ion-association reaction,a highly sensitive,simple and rapid method has been proposed to the determination of anthracycline drugs.3 Study on the interactions of Ce(IV) with mitoxantrone and their analytical applications3.1 Fluorimetric analysis of certain anthracycline antibioticsThe anthracycline antibiotics(daunorubicin,DNR;doxorubicin,DOX;mitoxantrone,MXT) can be analyzed by a simple spectrofluorimetric method based on the oxidation by cerium(IV) in the presence of proper buffer medium and monitoring the fluoresence of cerium(III) formed atλex 303 nm andλem 354 nm,while sodium tripho sphate was added the fluorescence intensity of the system could be greatly enhanced.From this,an indirect sensitive method for determining the anthracycline antibiotics was developed.The linearity ranges and the detection limits for the three anthracycline antibiotics were 0.043～3.0μg/mL and 12.8 ng/mL(DNR);0.045～3.0μg/mL and 13.4 ng/mL(DOX); 0.022～2.8μg/mL and 6.5 ng/mL(MXT),respectively.The method has been successfully applied to the determination of MXT in urine and serum samples and its recovery ranges from 94.7%to 106%.3.2 Fading spectrophotometric of certain anthracycline antibioticsIn the presence of proper buffer medium,the anthracycline antibiotics(daunorubicin,DNR; doxorubicin,DOX;mitoxantrone,MXT) react with cerium(IV) to form cerium(III),result in the great decrease of absorbance and the fading wavelengths in 316 nm,the absorption intensities(â–³A) were proportional to the concentration of the anthracycline antibiotics.From this,an indirect sensitive method for determining the anthracycline antibiotics was developed.The linearity ranges and the detection limits for the three anthracycline antibiotics were 0.88～10.0μg/mL and 0.26μg/mL(MXT);1.69～10.0μg/mL and 0.51μg/mL(DOX);1.42～10.0μg/mL and 0.43μg/mL(DNR), respectively.The method has been successfully applied to the determination of MXT in urine and serum samples and its recovery ranges from 97%to 104%.4 Simultaneous Dermination of Two anthracycline antibiotics by molecular spectroscopy4.1 Synchronous Fluorimetric Determination of Two anthracycline antibiotics by Partial Least SquaresIn the present paper,the synchronous fluorimetric spectra of doxorubicin(DOX) and daunorubicin(DNR) with△λ= 55 nm in a B-R buffer medium of pH 3.45 were measured,and the partial least squares(PLS) method was applied to the quantitative resolution of the seriously overlapped fluorimetric spectra of these compounds.Correlation coefficients of the models for DOX and DNR were 0.9897 and 0.9909,the mean recoveries were 100.97%and 101.36%,the rootmean square errors of prediction(RMSEP) were 0.1400 and 0.1395,and the standard errors of prediction (SEP) were 0.1541 and 0.1525 in the predicted range of 0.05～3.0μg/mL.The proposed method has been qpplied to the simultaneous determination of DOX and DNR in urine sample with satisfactory results.4.2 Simultaneous Dermination of Two anthracycline antibiotics by Partial Least Squares-Ultraviolet SpectrophotometryThe calibration models were established for the determination of doxorubicin(DOX) and daunorubicin(DNR) by partial least squares(PLS) and utraviolet Spectrophotometry.The absorption spectra in the range of 400～550 nm for 25 differernt mixtures of two anthracycline antibiotics in Britton-Robinson(BR) buffer medium(pH=2.46) were used as calibration set.Correlation coefficients of the models for DOX and DNR were 0.9945 and 0.9858,the mean recoveries were 100.18%and 97.63%,the rootmean square errors of prediction(RMSEP) were 1.0162 and 1.2603, and the standard errors of prediction(SEP) were 1.1347 and 1.4029 in the predicted range of 5.0～60.0μg/mL.