Optimization of High-Performance Liquid Chromatography Parameters for Purification of Oligonucleotide-A

The modification of high-performance liquid chromatography parameters leads to a more effective oligonucleotide-A purification process. Using various experimental such as buffer, concentration, and pH, method for optimizing the an on reverse-phase C18 column was created. To purify oligonucleotide-A, High-Performance Liquid Chromatography (HPLC), Ultraviolet-Visible Spectrophotometry (UV-Vis), Chromatography-Mass Spectrometry (LC-MS), lyophilization were used. Chromatographic data collected with semi-prep HPLC system, quantified UV-Vis technique, validated LC-MS method. most optimized found obtain purity 93.0% are 40 mM triethylammonium bicarbonate (TEAB) buffer pH 7, which is approximately 6.0% higher than reported 87.0%. However, yield under these conditions reduced by about 5%. worst possible settings that resulted in lowest (84.0%) (69.0%) 10 ammonium acetate (NH4CH3CO2) 7.