Combined use of phosphonium-erythrosin B-based nanoGUMBOS, UV-Vis spectroscopy, and chemometrics for discrimination and quantification of proteins

Nanoparticles derived from a group of uniform materials based on organic salts (nanoGUMBOS) are considered promising candidates for protein analysis due to their facile synthesis in aqueous media and high tunability. In this study, a phosphonium-erythrosin B-based nanoGUMBOS (i.e., [P4444]2[EB]) was prepared using an ultrasound-assisted reprecipitation method, and its ability to discriminate and quantify proteins was evaluated. Sonication time (30 s, 5 min, and 15 min) and cyclodextrin templating (alpha-, 2-HP-beta-, and gamma-CD) were investigated for their effects on discrimination performance of synthesized nanomaterial. Six proteins (albumin, hemoglobin, trypsin, catalase, lysozyme, and cytochrome c) with different abundance levels and physicochemical properties were selected as target analytes. Absorbance response patterns generated from interactions between [P4444]2[EB] nanoGUMBOS and proteins were analyzed using partial least squares discriminant analysis. Percentages of correct protein discrimination ranged from 94.6 to 99.6%, with the latter being the best result obtained using non-templated nanoGUMBOS formed after 5 min sonication. Under optimized conditions, it was possible to discriminate all protein samples with percentages of correct assignments greater than 90% for concentrations as low as 2.0 mu g mL-1. The discrimination capability of synthesized nanoGUMBOS was further evaluated using mixtures of different ratios of lysozyme, cytochrome c, and hemoglobin. Finally, partial least squares models were developed for protein quantification and the best performance was observed for albumin. Results support potential use of [P4444]2[EB] nanoGUMBOS in combination with ultraviolet-visible spectroscopy and chemo-metrics for qualitative and quantitative analyses of individual proteins and mixtures of proteins.