Distinguishing co‐existing unimolecular and bimolecular structural forms using joint analysis of Raman and UV absorption data

Standard Raman measurements of solutions require relatively high concentration of dissolved species, which supports formation of intermolecular complexes. In case of simultaneous presence of unimolecular and bimolecular structural forms, it is often difficult or even impossible to discriminate between them by using only the Raman data. We present a possible and effective way how to combine temperature dependencies of Raman scattering and UV absorption in a joint thermodynamic analysis. This approach is based on a joint fit of the results of multivariate analysis (singular value decomposition) of individual Raman and UV absorption spectral sets to a thermodynamic model. Our case study on RNA oligonucleotides, which can form imperfect intermolecular duplexes or intramolecular hairpin structures, demonstrates that this approach allows to determine thermodynamic parameters of co‐existing unimolecular and bimolecular structural forms and to isolate their Raman spectra. Copyright © 2014 John Wiley & Sons, Ltd.