Research On Microorganisms As Bio-SPE Materials For Extraction Of Pharmaceutical Drugs And Its Mechanism Of Extraction

Solid phase extraction(SPE)is a sample preparation method based on liquid-solid chromatography theory,which can selectively adsorb the target analytes through the interaction between the SPE materials.However,the SPE technology also has limitations,such as the simple structure of the SPE column,the limited sample analysis,and the tendency of the extraction process to be "mechanized".Therefore,it is necessary to continuously develop and improve the SPE technology.Microorganisms are the species with the largest variety,the largest number and the strongest reproduction ability in nature.The process of material exchange and information exchange with the external environment can be regarded as adsorption and elution in the process of solid phase extraction.If it can be developed into a conventional solid-phase extraction material,it will be a research direction with great development potential.The organic combination of microbial science and separation science has important guiding significance for the research of environmental pollution remediation,microbial adsorbents and sample preparation.In this paper,a new solid phase extraction method using live bacteria as the extraction materials was established.Then the method was optimized and evaluated.Finally,the possible adsorption mechanism of live bacteria was discussed by modern analytical methods,which provided a theoretical basis for broadening the selection and application of sample pretreatment.The specific research content mainly includes the following three parts:(1)Firstly,the new extraction method was developed.The experimental parameters of the new method were investigated and optimized(including the physiological state of the bacteria,the growth curve,the solvent of the analyte,the initial concentration of the analyte,the amount of bacteria,the incubation time and the extraction solvent,etc.),The adsorption effect had been significantly improved.And the method was validated,and the results showed that it is feasible to use live bacteria as the extraction materials.(2)Therefore,this thesis investigated and evaluated the enrichment and selectivity of the new method.The results showed that the method had different enrichment and selection capabilities for different analytes.Among them,E.coli had high enrichment for the analyte chlorpheniramine.At the same time,the method was applied to the analysis of rat plasma samples in vivo and in vitro,and the results showed that compared with traditional plasma sample extraction methods,this method can separate and enrich trace analytes.Therefore,it is feasible to apply this method to sample pretreatment.(3)On the basis of the above work,the possible extraction mechanism was studied.The results showed that E.coli and the six analytes all interacted with different intensities.The polysaccharides,proteins,phospholipids,etc.on the surface of E.coli cells may participate in the passive adsorption of the first stage of the adsorption process,Secondly,the nucleic acids,proteins,ribosomes,etc.inside the E.coli cells may participate in the second stage of the active adsorption of the adsorption processIn summary,the combination of microbial science and SPE to develop new bio-SPE methods provides new ideas and new directions for the development and application of microorganisms in sample pretreatment.