MS & NMR Center / Alfa Chemistry
GPC Center

Analysis of Oligonucleotides

Analysis of Oligonucleotides

Oligonucleotides are short single- or double-stranded fragments of DNA or RNA. Oligonucleotides can modulate gene expression through a range of processes[1]. Characterization and analysis of oligonucleotides are important components of drug development, registration, and quality control. Therefore, appropriate methods are needed for the reliable and accurate analysis and characterization.

A number of techniques exist to characterize therapeutic oligonucleotides in their native state. In fact, in traditional separation systems using organic modifiers, ion-pairing agents, and denaturing conditions, oligonucleotide duplexes (and other multiplexes) are prone to dissociation, making it difficult to analyze oligonucleotides. Gel permeation chromatography (GPC) is particularly suitable for analyzing oligonucleotides because the separation is based on the use of aqueous buffers under very mild conditions. Good separations are usually achieved due to the large molecular weight differences between single strand, duplex, and multiplexes and low molecular weight fragments.

Our Services

Given the complexity and diversity of oligonucleotide types in clinical development, including differences in length, chemical modifications, conjugation, and other characteristics, a need for a range of different analytical methods. State-of-the-art instrumentation, combined with the knowledge, experience and skills of our analysts, enables us advanced materials characterization capabilities. Use GPC to analyze the oligonucleotides, for example, when you need to:

  • To determine the identity and purity of the oligonucleotides
  • To obtain information on the polydispersity of the oligonucleotides
  • To identify the multimers in the samples
  • To detect the exact molecular mass of the main oligonucleotides

Our Technology Platforms

Our technology platforms include but are not limited to the following.

  • Gel Permeation Chromatography - Multi-Angle Static Light Scattering (GPC-MALS)
Analysis of Oligonucleotides

GPC-MALS can be used to obtain information on the polydispersity of the sample and to identify the multimers that could be present. Our GPC-MALS services of oligonucleotides include:

  • Analytical method development
  • Method validation
  • Release testing
  • In-process testing
  • Stand-alone analytical testing
  • Gel permeation chromatography - Mass Spectrometry (GPC-MS)

GPC-MS requires the use of different mobile phases due to compatibility reasons and for more detailed sample characterization. Using mass spectrometry, the presence of fragments can be detected, and the exact molecular mass of the main oligonucleotide and its fragments can be determined using ammonium acetate as the mobile phase.

Analysis of Oligonucleotides

Our Advantages

  • We have a leading technology platform and a team of experienced experts.
  • We provide customers with the most comprehensive services at the most favorable prices to help them save scientific research funds reasonably.
  • For the content entrusted by the customer, we will strictly keep the secret and sign a confidentiality agreement.
  • Even for the most complex samples, we strive to provide satisfactory results for our customers.

Alfa Chemistry is committed to providing customers with high-quality GPC services. Through our dedicated suite of identification technologies, Alfa Chemistry focuses on helping you meet industry requirements for regulatory compliance identification and quality control of oligonucleotide-based drugs. Alfa Chemistry looks forward to cooperating with you.


  1. Roberts, T.C., Langer, R. & Wood, M.J.A. Advances in oligonucleotide drug delivery. Nat Rev Drug Discov 2020, 19, 673–694.

Our services are for research use only.