A mechanistic analysis of the quantitation of α-hydroxy ketones by the bicinchoninic acid assay.

A new class of compounds amenable to quantification by the bicinchoninic acid (BCA) assay was identified, allowing an expansion of compounds quantifiable within the assay's capacity. In this article, we demonstrate that compounds containing the α-hydroxy ketone structure are easily measured under standard BCA assay conditions. A nonchromophore analyte containing the α-hydroxy ketone structure, 1,3-dihydroxypropan-2-one (commonly known as dihydroxyacetone), and various structural derivatives were explored on an equimolar basis in the BCA assay. Combined with earlier studies exploring α-hydroxy ketones within copper oxidation systems, the data support the mechanism of this class of compound's ability to enolize through an enediol intermediate to generate a strong signal in the BCA assay. This new quantification technique also highlights the potential for α-hydroxy ketones to interfere with other analytes quantified by the BCA assay.

Protein release from dihydroxyacetone-based poly(carbonate ester) matrices.

The release of therapeutics from solid polymer matrices is an important field of study in the area of controlled release. Here we report on the hydrolytic degradation of directly compressed discs comprised of statistically random polycarbonate esters based on lactic acid and dihydroxyacetone. The controlled release of two model proteins, bovine serum albumin and lysozyme, was explored using two percentage loadings (5 and 10 wt.%). A first order release pattern and a trend for faster protein release with increasing dihydroxyacetone content were observed over a time period ranging from 2.5 to 70 days. To analyze the effects of the internal polymer matrix environment on protein stability the enzymatic activity of released lysozyme was monitored. The results show a high level of enzyme activity for the polycarbonate ester ratios with more dihydroxyacetone in the backbone and at least 50% activity over the first month of release from the co-polymer ratios with more lactic acid in the backbone. Modeling of the release kinetics using the Korsmeyer-Peppas model showed a high correlation, indicating that the release of protein is a complex mechanism controlled by protein diffusion through, and erosion of, the co-polymer matrix. The outcomes show that these polycarbonate esters may be useful materials for extended controlled release of proteins.