Determination of organic acid drug counter-ions by capillary electrophoresis

Summary A capillary electrophoresis method is described for the novel application of quantifying levels of the simple organic acid counter-ions of a variety of basic drugs. These counter-ions are organic acids such as succinic and maleic. The method uses indirect UV detection and an electroosmotic flow modifier. Acceptable precision and detector linearity were obtained using internal standards. Method validation was completed and acceptable data was generated. The method is now in routine use for this type of testing.     

A Series of Collaborations between Various Pharmaceutical Companies and Regulatory Authorities Concerning the Analysis of Biomolecules Using Capillary Electrophoresis: Additional Instruments/Buffer

An international project team (including members from US, Canada and UK) was formed from a number of interested biopharmaceutical companies and regulatory authorities to conduct a cross-organisation collaboration exercise. The results of the first comparison with eight different organisations that used instruments of the same equipment model, the same reagents, and the same methodology has been reported previously [1]. This report represents the addition of other instruments using a different run buffer. The relative migration times were different, as expected, prohibiting a direct comparison between companies. The within-organisation variability was low for both relative migration time (<0.34% RSD% for all companies save one) and the peak area (<5% RSD% for all companies save one) when measuring the purity of a representative IgG sample. The apparent molecular weight of bovine serum albumin was measured with good precision (less than 10% RSD% across all companies) to the theoretical value when all data is utilized (67.5 kDa compared to 66.4 kDa). For a representative IgG sample, the three main components, IgG Light Chain, IgG Non-glycosylated Heavy Chain, and IgG Heavy Chain, could not be separated, specifically the IgG Non-glycosylated Heavy Chain and IgG Heavy Chain. When the IgG Non-glycosylated Heavy Chain and IgG Heavy Chain were combined for all organisations, the fractional peak area for the IgG Light Chain and IgG Non-glycosylated Heavy Chain + IgG Heavy Chain peak also showed excellent agreement, with less than 7.5 and 3.5% RSD%, respectively. The value of this exercise is in demonstrating the reliability of CE for the determination of apparent size of biopharmaceutical proteins. This underpins the appropriate use of such CE data in support of regulatory submissions.

     

Capillary electrophoresis for the analysis of small-molecule pharmaceuticals

This paper reviews the application of CE to the analysis of small-molecule pharmaceuticals. The areas of pharmaceutical analysis covered are enantiomer separation, the analysis of small molecules such as amino acids or drug counter-ions, pharmaceutical assay, determination of related substances and physicochemical measurements such as log P and pK(a) of compounds. The different electrophoretic modes available and their advantages for pharmaceutical analysis are described. Recent applications of CE for each subject area are tabulated with electrolyte details.     

Highly efficient and selective separations of a wide range of analytes obtained by an optimised microemulsion electrokinetic chromatography method

Summary Microemulsion Electrokinetic Chromatography (MEEKC) has previously been reported to be useful for the separation of a range of hydrophobic solutes. The previous reports had tended to show relatively long separation times. In this report the separation and operation conditions were optimized and a method was successfully employed in a range of novel applications. High efficiencies, rapid separations and high repeatability were demonstrated. The applications developed included a range of both neutral and charged compounds. Both water-soluble and water-insoluble compounds were also well separated. The novel applications developed included analysis of basic drugs, various aromatic acids, a range of neutral aromatics, profiling of ink components, direct injection of urine for profiling purposes and the analysis of a range of amino acids and their derivatives. The method could also be used for hydrophobicity measurements of the resolved solutes based upon their migration times. This report clearly shows that MEEKC can be widely applied to a range of compounds and is especially useful for complex mixtures containing solutes with varying charge and hydrophobicity.