Reversed-Phase Ion-Pair LC/MS Analysis of siRNA under Denaturing and Non-Denaturing Conditions

User Benefits

- The LCMS-9050 quadrupole time-of-flight mass spectrometer can be used to characterize siRNA. - The elution of siRNA in double-stranded (non-denatured) or single-stranded (denatured) state can be confirmed by changing the column temperature. - The LabSolutions Insight Biologics analysis software enables analysis of multiple oligonucleotide sequences at once.

Introduction

Among oligonucleotide therapeutics, antisense and siRNA therapeutics, which are composed of relatively short oligo nucleic acids, are attracting attention as new modalities for treating genetic and intractable diseases. Unlike single-stranded antisense, siRNA is double-stranded RNA and known to separate into single-stranded RNAs under heated or alkaline conditions. Quality control of siRNA drugs requires characterization of both single-stranded and double-stranded RNA. This application describes an example of LC/MS analysis of siRNA under denaturing and non-denaturing conditions in reversed-phase ion pair chromatography. Multiple nucleic acid sequences (sense and antisense strands) were analyzed at the same time using LabSolutions Insight Biologics, a dedicated software for oligonucleotide characterization.

July 15, 2025 GMT