Methodology For Separation And Identification Of Complex Samples By HPLC And Its Hyphenated Techniques

With the development of theory, technique and instruments, high performance liquid chromatography has become the most efficient separation tool in the analysis of complex samples such as environmental samples, biological samples, medical samples and medical quality control. The complexities of real samples have showed the necessity of optimization of chromatographic separation conditions.According to traditional optimization methods, a new strategy based on uniform design, genetic algorithm and simulated annealing was established. The optimization strategy was described in details and a software package was developed. Separation conditions in different elution modes for different kinds of samples were obtained based on the proposed optimization strategy and corresponding software package. Also, the optimization strategy was applied in the resolution of main components in a typically traditional Chinese medicine of Ginkgo biloba.A comprehensive two-dimensional liquid chromatography system coupled to mass spectrometry was developed, its principle and further improvement were represented in details. Based on the connecting of different chromatography modes, some kinds of traditional Chinese medicines were separated and analyzed by the comprehensive two-dimensional system, and much more components in them were preliminary identified according to their UV and mass spectra. The experimentalresults not only demonstrated the advantages of comprehensive two-dimensional system, such as sensitivity, reliability and reproducibility, but also indicated its potentials for the resolution and identification of complex samples, such as traditional Chinese medicines and biological samples.An integration of IEC fractionation with RPLC-APCI/MS and MALDI-TOF MS system was particularly developed and successfully employed for the isolation and identification of components in the extract of traditional Chinese medicines Honeysuckle and Psoralea corylifolia. The obtained analytical results not only demonstrated the powerful resolution of integration IEC fractionation with RPLC-APCI/MS and MALDI-TOF analysis for the resolution of compounds in complex samples, but also exhibited the superiority of APCI/MS and MALDI-TOF MS for identification of low-mass compounds, such as for study of traditional Chinese medicines (TCMs) and metabolome.