Electrochemical transduction of DNA hybridization at modified electrodes by using an electroactive pyridoacridone intercalator.

A synthetic redox probe structurally related to natural pyridoacridones was designed and electrochemically characterised. These heterocycles behave as DNA intercalators due to their extended planar structure that promotes stacking in between nucleic acid base pairs. Electrochemical characterization by cyclic voltammetry revealed a quasi-reversible electrochemical behaviour occurring at a mild negative potential in aqueous solution. The study of the mechanism showed that the iminoquinone redox moiety acts similarly to quinone involving a two-electron reduction coupled with proton transfer. The easily accessible potential region with respect to aqueous electro-inactive window makes the pyridoacridone ring suitable for the indirect electrochemical detection of chemically unlabelled DNA. Its usefulness as electrochemical hybridization indicator was assessed on immobilised DNA and compared to doxorubicin. The voltamperometric response of the intercalator acts as an indicator of the presence of double-stranded DNA at the electrode surface and allows the selective transduction of immobilised oligonucleotide hybridization at both macro- and microscale electrodes.

High-precision absolute (true) density measurements on hygroscopic powders by gas pycnometry: application to determining effects of formulation and process on free volume of lyophilized products.

As density (free volume) of the amorphous solids should be related to mobility and stability, an attempt was made to develop a simple, sensitive, and reproducible method to evaluate free volume via high-precision gas pycnometry density measurements, and to apply this methodology to study the variation of free volume with formulation and thermal history (i.e., annealing). Annealed samples were prepared either by heating the product after freeze drying (postannealing) or drying at higher temperature in secondary drying than normal (in-process annealing). Density was measured using a gas pycnometer. We find that the key to high-precision density measurements is isolation of the instrument from atmospheric moisture; accordingly, all operations were carried out in a dry box. With suitable care, densities of amorphous freeze-dried products can be measured with a precision of better than 0.5% in a series of independent but nominally identical samples. Density decreased with increasing molecular weight of dextran, but density of proteins was independent of molecular weight. Small but significant increases in density upon annealing were observed for several formulations. Thus, we conclude that accurate density measurements may be made by carefully controlling residual moisture. Density may be a useful parameter to predict long-term stability.