LC-MS Analysis of Therapeutic and Diagnostic Oligonucleotides

Library Number:
WA20735
Part Number:
WA20735
Author(s):
Martin Gilar;Ken J Fountain;John C Gebler
Source:
ISPPP 2002 Heidelberg Germany, Nov. 10-13
Content Type:
Posters
Content Subtype:
ISPPP
Compounds:
Synthetic oligonucleotides; 25mer oligoG [oligogeoxyguanidine]; TAMRA labeled 21mer diagnostic oligonucleotide
Matrix:
phosphorothioate drug digestion
Column:
XTerra MS C18 2.5 µm Steel 1 mm 50 mm
Related Products:
 
nanoACQUITY UPLC
Synthetic oligonucleotide probes are often used for highly sensitive, fluorescent-based diagnostic assays. Chemically modified oligonucleotides are also utilized for therapeutic purposes (antisense drugs). Synthesis of modified oligonucleotides is challenging and often results in a product of limited purity. Characterization of therapeutic and diagnostic compounds is a basic regulatory requirement. A method utilizing ion-pair reversed-phase chromatography with mass spectrometry detection has been developed for the analysis and characterization of chemically modified and dye-labeled oligonucleotides. We investigated synthetic by-products of oligonucleotide-based drugs and diagnostic probes such as phosphorothioate oligonucleotides, locked nucleic acid and dually dye-labeled oligonucleotides. Guidelines for chromatographic column selection and optimization of mobile phase composition are discussed.

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