Abstract
Oligonucleotides are used in many research areas. Thus, there is a need for their successful separation methods. Ion-exchange chromatography is the most popular separation technique, but it has limitations for these compounds. For this reason, new stationary phases are developed in order to increase separation selectivity. This study aimed to apply a series of dendrimer anion exchangers with various bonded layers to separate oligonucleotides by using different mobile phases to find conditions that allow sufficient separation. The number of anion-exchange layers, type of salt, and pH significantly impacted the oligonucleotide analysis. The developed chromatographic method was characterized by adequate selectivity for oligonucleotides differing in sequence length. It is essential to underline that the number of bonded layers appeared to have a significant influence, and the three layers appeared optimal. Based on our results, it may be concluded that the dendrimer stationary phases can be successfully used as an alternative to commonly applied packing materials in ion-exchange chromatography.
Highlights
IntroductionOligonucleotides are compounds built of many nitrogen bases, sugars (ribose or deoxyribose), and phosphate groups [1]
Oligonucleotides are compounds built of many nitrogen bases, sugars, and phosphate groups [1]
Similar separation selectivity may be obtained in ion-exchange chromatography, where the retention is based on electrostatic interactions between the negatively charged oligonucleotide and positively charged stationary phase surface [7,8,9,10,11]
Summary
Oligonucleotides are compounds built of many nitrogen bases, sugars (ribose or deoxyribose), and phosphate groups [1] Their structure consists of nucleotides bounded by a phosphodiester bond in a single strand. Similar separation selectivity may be obtained in ion-exchange chromatography, where the retention is based on electrostatic interactions between the negatively charged oligonucleotide and positively charged stationary phase surface [7,8,9,10,11]. Different lengths of alkyl chains can be bounded to the quaternary nitrogen atom—usually short ones such as trimethylamine or diethylmethylamine [8,9,10,11] They were successfully applied to separate unmodified, single-stranded OGNs and modified oligonucleotides but some disadvantages were noticed—for example, a lengthy analysis time or insufficient separation [9,10,11]. Chromatographic methods of oligonucleotide analysis need improvements, such as proper, careful mobile phase composition changes or the synthesis of new and alternative stationary phases and their application
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
