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This report describes the development of a potentially clinical method to measure the cellular metabolites of zidovudine (ZDV) in patients receiving the drug. This new method combines the use of Sep-Pak cartridges to separate ZDV phosphates with radioimmunoassaying to quantitate ZDV. The detection limit is 0.02 pmol/10(6) cells, and this assay can measure a wide range of intracellular drug concentrations. The use of the cartridge-radioimmunoassay methodology should prove very useful for in vivo cellular pharmacokinetic studies of ZDV.

The use of capillary electrophoresis (CE) modified with cyclodextrin (CD) for the separation of stereoisomers of peptides is well established. To increase the solubility of β‐CD, urea is often added to the buffer which may influence the complexation of a CD with a guest molecule. The aim of the present study was to investigate the influence of urea on the complexation between dipeptides and β‐CD using Ala‐Phe and Ala‐Tyr as model compounds. For this purpose three different analytical methods were employed: capillary electrophoresis (CE), 1H‐NMR spectroscopy and molecular dynamics simulations (MD). Electropherograms of the peptide enantiomers were different in the presence and absence of urea. For example, at pH 2.5 in the absence of urea the enantiomers of Ala‐Tyr are not separated in contrast to the use of buffers containing urea. Applying “complexation‐induced chemical shift (CICS)” in NMR spectroscopy and rotating frame Overhauser enhancement spectroscopy (ROESY) revealed differences in the complexation of the peptide enantiomers by β‐CD in the absence and presence of urea suggesting the stabilization of the complex through the phenolic hydroxyl group of tyrosine. MD simulations for different complexes were carried out with consideration of both water and urea molecules in solution. Simulations were performed for 1 ns. In conclusion, NMR spectroscopy and MD methods help to understand the structure of peptide‐CD complexes and the separation and migration behavior in CE. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)

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