Studies On New Methods For Medical And Pharmaceutical Analysis Using Chromatographic And Hyphenated Techniques

Pharmaceutical analysis has being boosted by the rapid development of medical and life sciences,as well as the related technologies.The research fields of pharmaceutical analysis have been expanded from the classic quality monitoring of drug products to the development of analytical techniques for the medical and pharmaceutical study.The focus has also been transformed from drug composition into body fluids.Furthermore,the accelerated intersection with information science further enlarged the fields of pharmaceutical analysis.Therefore,the innovation on the basis of integration across multiple disciplines,changing the traditional point of view on pharmaceutical analysis,has been presented,which bring tremendous challenges and opportunities for pharmaceutical analysis researchers. In this dissertation,aiming to the analysis of complex systems such as traditional Chinese medicines and bio-fluids,a series of novel methods,integrated with modern chromatographic technologies,computational intelligence and metabonomics are developed.The main innovation and academic contributions of these works are presented as follows:1.An approach to the optimization of Capillary electrophoresis(CE) separation condition based on multivariate statistics and genetic algorithms was established,and the best condition would be obtained with a lesser number of tests,which solved the issue of the CE condition optimization for the separation of complex chemical system. This method had been successfully applied to optimizing the separation of the active components in the traditional Chinese medicine 'SHUANGDAN' granule,and an analytical method for the simultaneous determination of the active components such as protocatechuic aldehyde,paeonol,danshensu,salvianolic acid B was established.2.CE has been one important technique for the separation and analysis of proteins,peptides,nucleic acids and other biological molecules,and CE mobility,as the inherent physical and chemical property for peptides,has been an important parameter in the research of peptides and proteins.In this thesis,nonlinear quantitative structure-mobility relationship models based on the molecular descriptors and machine learning algorithms were introduced for predicting the CE mobilities of peptides.The developed nonlinear models had high accuracy and stability,and had been successfully used to predict the CE mobility of a variety of peptides,resulting in the prediction accuracy higher than 90%.The developed model can not only significantly reduce the test number and practically guide the separation of peptides, but also play positive role in promoting the proteomics research.3.Pharmacokinetics of the drug products derived from traditional Chinese medicines is one of the important research areas in the current study on pharmaceutical analysis.The current attractive safety problems of injections made from traditional Chinese medicines are closely related with their pharmacokinetic characteristics.Ginseng is a common-used drug material in traditional Chinese medicines,and ginsenosides are the main active components.In this thesis,a sensitive, specific and reliable HPLC/MS method for the simultaneous determination of ginsenosides Rg1,Rf,Re,Rd and Rb1 in the rabbit plasma was developed.This method was used for the measurement of blood drug concentration-time profiles of five ginsenosides after intravenous administration of 'SHENMAI' injection,a well-sold injection made from ginseng and ophiopogonis,in rabbits.The result showed that the elimination of diol-ginsenosides in the blood was much more slowly than that of glycerol-ginsenosides,indicating that although the chemical structure of these two types of ginsensides was quite similar,a big difference was presented within their phamacokinetics.4.Metabonomics is the branch of science concerned with the quantitative understandings of the metabolite complement of integrated living systems and its dynamic responses to the changes of both endogenous factors(such as physiology and development) and exogenous factors(such as environmental factors and xenobiotics). NMR and LC/MS were two mainly used techniques in metabonomics;however,there is still lack of a real technique which can fulfill the global analysis of the whole metabonome just with only one run.In this thesis,a novel GC/MS based computational analytical method was developed for the metabonomic study.GC/MS technique and bioinformatics techniques were integrated in this method,which was capable for the metabolites profiling of various biological matrices.The developed metabonomics method was applied to the characterizing the serum metabonome profile of HBV-caused liver failure patients.This discovery indicated that the characteristic profile of serum metabonome is effective to predict the severity of liver failure and its prognosis of drug treatment,which promises to promote the personalized drug therapy of liver disease patients.Furthermore,the developed metabonomics method was also used for the evaluation of toxicity of acetaminophen and aconitine with different exposure times and doses on the mice or rats.The time related changes of metabolites profiling of the animals treated with different level of toxins have been discovered,which would be helpful to develop the more advanced safety assessment systems.