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Related Experiment Video

Updated: Jan 12, 2026

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Scatter-Free UV-Visible Spectroscopy for Accurate and Precise RNA Quantification in Complex RNA Nanoparticle

Aida López Espinar1, Eric C Le Ru2,3, Parveen Kumar1

  • 1School of Pharmacy, University College Cork, Cork T12 K8AF, Ireland.

Analytical Chemistry
|November 4, 2025
PubMed
Summary
This summary is machine-generated.

Accurately measuring ribonucleic acid (RNA) in complex nanoparticle drugs is difficult. Scatter-free absorption spectroscopy (SFAS) offers a reliable method for quantifying total RNA in intact nanoparticles, outperforming traditional fluorescence assays.

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Area of Science:

  • Biomaterials Science
  • Analytical Chemistry
  • Pharmaceutical Sciences

Background:

  • Ribonucleic acid (RNA)-based therapeutics hold promise for treating various diseases.
  • Effective clinical application necessitates complex nanoparticle (NP) formulations.
  • Accurate quantification of total RNA within these complex NPs is a significant analytical challenge.

Purpose of the Study:

  • To evaluate scatter-free absorption spectroscopy (SFAS) as a method for total RNA quantification in complex RNA-loaded nanoparticles.
  • To compare the performance of SFAS against established fluorescence-based assays (RiboGreen, SYTO 9).
  • To demonstrate SFAS's utility across diverse NP formulations resistant to disruption.

Main Methods:

  • Utilized scatter-free absorption spectroscopy (SFAS), a UV/Visible technique designed to eliminate light scattering from NP components.
  • Tested SFAS on various RNA formulations: lipid NPs, polymer/dendrimer hybrid lipid NPs, and cyclodextrin nanocomplexes.
  • Compared SFAS results with fluorescence-based assays using RiboGreen and SYTO 9 dyes.

Main Results:

  • SFAS provided accurate, precise, and reproducible total RNA quantification across all tested NP formulations.
  • SFAS outperformed fluorescence-based methods, especially for NPs exhibiting resistance to disruption.
  • RNA quantification by SFAS showed minimal influence from NP composition and measurement conditions.

Conclusions:

  • SFAS is a versatile and reliable alternative to fluorescence-based assays for quantifying total RNA in complex RNA NP formulations.
  • This method enables accurate RNA measurement in intact NPs, simplifying formulation analysis.
  • SFAS offers a robust solution for quality control and development of RNA-based nanomedicines.