Application note: Effect of temperature on single stranded oligonucleotide analysis
Therapeutic oligonucleotides represent a recent breakthrough in the pharmaceutical industry. However, characterisation of oligos, specifically by ion-pair reversed phase liquid chromatography (IPRPLC), can be quite challenging.
Oligos are manufactured by solid phase synthesis, where nucleotides are added in a step-wise manner. As such, impurities such as n-1 and n+1 must be characterised, and this may require extensive method development to optimise. Further, it may be necessary for characterisation and quantitation of other closely related impurities,such as oxidation of phosphorothioates.