Preparation Of Restricted Access Solid Phase Extraction Membrane Roll For Directly Pretreating Rat Plasma Samples

As we all know,sample preparation is a key step in the analysis of complex matrix biological samples,because this process not only consumes a lot of time,but also is of one of the important reasons resulted in the deviation of subsequent analysis result.Solid phase extraction has currently become the mainstream technology for biological sample preparation.and in most cases,the protein in the blood sample needs to be removed before the solid phase extraction treatment.Restricted solid-phase extraction media has solved this problem well.It has bifunctional groups with the external hydrophilic groups excluding the entry of biologically macromolecular matrix components(such as proteins,etc.),while allowing small molecular target analytes to get to the inner surface to interact with the internal functional groups.In this way,the retention of the target components can be achieved,and the matrix components can be effectively eliminated,which can not only simplify the sample preparation process,but also effectively improve the extraction efficiency.The development of new RA solid-phase extraction technology has become one of the current research hotspots in the analytical field.Aiming at the current research frontier of biopharmaceutical analysis technology,this thesis has carried out the research and development of restricted access solid phase extraction membrane cartridges.The prepared film roll column can be conveniently installed in a 1mL polypropylene syringe,and quick RA-SPE treatment of plasma sample be performed by manually pulling and pushing the plunger.The researchcontent of thispapermainly includesthefollowingparts:(1)LiteraturereviewIt mainly introduces the development status of biological sample pretreatment technology,focusing on the research progress of solid phase extraction,membrane extraction,and RA materials in recent years.At the same time,the application characteristics of the(ARGET-ATRP)method are also summarized.Combined with the results of literature research,the research ideas and concepts of this subject are proposed,modifying the surface of porous polypropylene film with ARGET-ATRP method to bond double-layer functional groups on the surface of the filmtoprepare the surface-RA-SPE thinfilm roll column.(2)Preparation of surface-restricted-access composite polymer film roll and its applicationinthe direct extractionof statinsfromrat plasmaFirst,using the porosity of the polypropylene film(φ15um),the pre-polymerized mixed solution(benzyl methacrylate(BZMA)and ethylene glycol dimethacrylate(EDMA))was poured into the pores of the polypropylene film to let the PP film be in the state of saturating adsorption.Then,The polypropylene film saturated with the pre-polymerized mixed solution was placed between two glass plates with the two plates pressed together.ARGET-ATRP polymerization was performed at room temperature to prepare a composite polypropylene base film(polypropulene-b-poly(benzyl methacrylate-co-ethylene glycol dimethacrylate)PP-p(BZMA-EDMA)).Following the same manner,mixed function layer(poly(benzyl methacrylate-co-2-diethylaminoethyl methacrylate),p-BZMA-DEAMEMA)and a layer of hydrophilic functional layer(linear polyhydroxyethyl methacrylate,poly-HEMA)were consecutively in turns grafted on the surface of the film.Finally,a surface-restricted-access dual-functional layer film was prepared.The SRA-film can be easily coiled into a 1mL polypropylene disposable medical syringe to form a convenient solid phase extraction device.With the help of this device,direct and rapid SPE pretreatment of rat plasma samples can be performed by manually spulling and pushing operations.In theexperiment,the film was characterized by SEM and XPS.The performance for the film column to directly extract statins from rat plasma samples was investigated.Based on the optimal SPE treatment conditions,a new SPE-HPLC/UV method was constructed.for the rapid analysis of three types of statins in rat plasma samples(rosuvastatincalcium,pravastatin,lovastatin).(3)Preparation of a hydrophobic functional film roll column based on self-assembled dopamine-polypropylene composite film and the preliminary investigationonitsabilitytoextract trimethoprim.A polypropylene-polydopamine composite film was prepared with the self-assembly polymerization of dopamine on the surface of polypropylene film.Then,the ATRP initiator dibromoisobutyryl bromide(EBIB)was grafted onto the composite film by acylation reaction.Following that,on the surface of composite film,a hydrophobic polymer layer was grafted with BZMA used as functional monomer and EDMA as cross-linking agent to construct hydrophobic function on the surface of thin film(polybenzyl methacrylate-co-ethylene glycol dimethacrylate),p-(BZMA-EDMA)).This study focused on the feasibility of for polypropylene-polydopamine composite membrane to adsorb target analytes.With trimethoprim(TMP)as the target,HPLC/UV was applied to detecting the adsorption capacity of thin filmroll towards TMP.Finally,the TMP adsorption rate ofthe composite filmroll under optimizedconditionswasmore than95.20%.