| Abstract [eng] |
A study of novel cationic polymer synthesis, purification was conducted for this master thesis study. Using a developed general procedure, 6 new, structurally diverse products were produced, and 3 of them were prepared for future in vitro transfection efficiency studies. Much of the attention during this study was paid to the purification of cationic polymers and identification of products that are made during the polycondensation reaction. A new reverse-phase chromatography method was proposed and developed, which helps effectively separate different products according to their size and structure. This method, compared to dialysis, which is most often used for the purification of polyamines, is faster and more selective. In order to determine polyamine nitrogen concentration, which is necessary for transfection studies, a spectrophotometric nitrogen concentration determination method was adapted. Nitrogen concentration of 3 products was successfully determined using it. During the study, 2 experimental polyamine derivatization reactions were conducted, which could possibly help solve complicated polyamine molecular weight determination and chromatographic analyses. It was determined that polyamine reaction with 2-nitrobenzyl bromide is not selective (both primary and secondary amines are reactive), therefore it is not possible to assign product peaks to any concrete structure. During the derivatization reaction with 4-fluorobenzaldehyde, a selective primary amine reaction is observed. The product of this reaction is an imine, which is observed during NMR studies. Full interpretation of the products that are produced during this reaction is complicated and demands further experimentation, however, this method has potential to be used for determination of the polyamine average molecular weight. |
