Supramolecular hydrogels formed through polymer–nanoparticle
interactions are promising biocompatible materials for translational
medicines. This class of hydrogels exhibits shear-thinning behavior
and rapid recovery of mechanical properties, providing desirable attributes
for formulating sprayable and injectable therapeutics. Characterization
of hydrogel composition and loading of encapsulated drugs is critical
to achieving the desired rheological behavior as well as tunable in vitro and in vivo payload release kinetics.
However, quantitation of hydrogel composition is challenging due to
material complexity, heterogeneity, high molecular weight, and the
lack of chromophores. Here, we present a label-free approach to simultaneously
determine hydrogel polymeric components and encapsulated payloads
by coupling a reversed phase liquid chromatographic method with a
charged aerosol detector (RPLC-CAD). The hydrogel studied consists
of modified hydroxypropylmethylcellulose, self-assembled PEG-b-PLA nanoparticles, and a therapeutic compound, bimatoprost.
The three components were resolved and quantitated using the RPLC-CAD
method with a C4 stationary phase. The method demonstrated robust
performance, applicability to alternative cargos (i.e., proteins) and was suitable for composition analysis as well as
for evaluating in vitro release of cargos from the
hydrogel. Moreover, this method can be used to monitor polymer degradation
and material stability, which can be further elucidated by coupling
the RPLC method with (1) a multi-angle light scattering detector (RPLC-MALS)
or (2) high resolution mass spectrometry (RPLC-MS) and a Fourier-transform
based deconvolution algorithm. We envision that this analytical strategy
could be generalized to characterize critical quality attributes of
other classes of supramolecular hydrogels, establish structure–property
relationships, and provide rational design guidance in hydrogel drug
product development.