Low-MW Proteome Analysis On Core-shell Silica Particle-Based Nanoreactor With Size-selective Effect And A New Capillary-based SPME Method On Preconcentration And Analysis Of ?-blockers

With the development of medical science and technology,the demand for rapid and sensitive detection of biological samples is expanding.However,due to the complexity of matrix,interference,low concentration and variability in high temperature,it is necessary to purify and enrich the biological samples before detection.To find a simple,a large number of science stuffs have done much work to find a fast,low cost,high performance sample pretreatment method.The paper mainly focuses on Low-MW proteome size-selective effect analysis and human urine samples of low molecular weight protein beta blockers enrichment and purification,and two jobs were carried out in the following:1.With novel core-shell silica mesoporous(CSMS)microspheres that are synthesized using a convenient two-phase process,we report in this study the analysis of low molecular-weight(MW< 30 kDa)proteins by combining size-exclusive separation and enzyme immobilization.The particle-based enzyme reactors have been fabricated by immobilizing trypsin on the pore channels of the CSMS microspheres using either physical absorption or covalent binding via thiol or aldehyde group.Due to the unique fibrous pore structure,low molecular-weight proteins can enter the channels in the shell for interaction with immobilized trypsin,followed by analysis of the digestion products using capillary electrophoresis(CE)or MALDI-TOF MS techniques.We used various techniques to characterize the synthesized CSMS and trypsin-immobilized CSMS particles.The performance for different trypsin-immobilized CSMS particles has been systematically evaluated.The size-selective absorption and digestion of four model proteins with different MW have been investigated using the synthesized trypsin-immobilized CSMS particles.The results show that the peptide-sequence coverage of the smaller protein is enhanced by using trypsin-CSMS particles,indicating the size-dependent digestion which results from the size-exclusive interaction of the mesoporous against the high-MW proteins.The present study would pave the way for further applications of mesoporous materials in proteome analysis.2.A method for rapid and sensitive analysis of two ?-blockers(atenolol and metoprolol)in human urine was developed and validated.The method was based on combined of taperedcapillary microextraction by packed sorbent and field-amplified sample injection-capillary electrophoresis(FASI-CE)with ultraviolet detection.Various parameters of the microextraction procedure and FASI-CE analysis,which affecting extraction efficiency,dual preconcentration and CE separation,were optimized.Optimum extraction conditions were 200 ?L of sample,below 2 ?L of sorbent and the microextraction time of 6 min.Accuracies of human urine analyses were 93.7-105.5% with RSD(n=3)lower than 8.5 %.Satisfactory average enhancement of detection sensitivity about a 21-fold and 19-fold was achieved for atenolol and metoprolol,respectively.The method was environmentally friendly and allowed reusing the sorbent several times,more than 8 times,without obvious loss in its performance in the analysis of human urine samples.