Quantitation of locked nucleic acid antisense oligonucleotides in mouse tissue using a liquid-liquid extraction LC-MS/MS analytical approach

Bioanalysis. 2011 Sep;3(17):1911-21. doi: 10.4155/bio.11.100.


Background: A significant challenge of oligonucleotide bioanalysis is the selective extraction from complex tissue samples, where the molecules that distribute into the intracellular space are extensively protein bound and sit amongst a high concentration of endogenous nucleic acid material. Published analytical methodology currently purports extensive sample preparation requirements that include cell lysis steps, homogenization and dual cleanup with liquid-liquid extraction and solid-phase extraction, prior to injection.

Results: We have developed a simple liquid-liquid extraction approach to rapidly isolate antisense oligonucleotides from biological tissues with high recovery and combined these preparative steps with a robust monolithic column LC-MS/MS setup. The platform showed improved chromatographic resolution and detection sensitivity over standard reversed-phase columns and required a low sample volume.

Conclusion: The high-throughput method was sufficient to accurately quantify multiple antisense oligonucleotides in mouse tissue and plasma down to low ng/g and ng/ml levels, respectively, for pharmacokinetic determination, and exhibited a high degree of specificity.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Liquid / instrumentation
  • Chromatography, Liquid / methods*
  • Kidney / chemistry
  • Liver / chemistry
  • Mice
  • Oligonucleotides, Antisense / analysis*
  • Oligonucleotides, Antisense / pharmacokinetics
  • Reference Standards
  • Solid Phase Extraction / methods
  • Tandem Mass Spectrometry / instrumentation
  • Tandem Mass Spectrometry / methods*


  • Oligonucleotides, Antisense