What is it? Who uses it? How much does it cost? What are thepros and cons? Kevin Altria provides the answers
Developed in the early 1990s, capillary electrophoresis (CE) is now an establishedtechnique in several areas of analysis. The use of CE is routine in many hospitals andclinics, particularly for analyzing serum proteins and disease markers. The technique hasalso dramatically increased throughput for DNA profiling in criminal investigations. CEdata have been shown to be credible evidence in law courts, and forensic testinglaboratories have published validated procedures. Pharmaceutical companies makeextensive use of CE, in particular for chiral separations, and the technique is widelyaccepted by regulatory authorities such as the US Food and Drug Administration. Outsideof these areas, however, inexperience of CE and the predominance of HPLC in manyanalytical laboratories continue to impede uptake of the technique. This is despite the factthat for many analyses CE may be easier, faster and more cost-effective.So how does it work? CE is an automated analytical technique that separates species byapplying voltage across buffer filled capillaries. It is generally used for separating ions,which move at different speeds when the voltage is applied depending on their size andcharge. The solutes are seen as peaks as they pass through the detector and the area of each peak is proportional to their concentration, which allows quantitativedeterminations. Analysis includes purity determination, assays, and trace leveldeterminations.Analysis times are in the region of 1-30 minutes depending on the complexity of theseparation. Modern instruments are relatively sophisticated and may contain fiber opticaldetection systems, high capacity auto samplers, and temperature control devices.Detection is usually by UV absorbance - often with a diode array. Other commercialdetectors include fluorescence detection and coupling to mass spectrometers. Indirect UVdetection is widely used for detecting solutes having no chromophores such as metal ionsor inorganic anions (
). Low UV wavelengths (
190-200nm) are also used to detectsimple compounds such as organic acids.
1. Inorganic Ions
CE can rapidly and efficiently separate inorganic ions such as metal ions and anions, for example chloride, sulphate and nitrate. These species have no chromophore and cannot be detected using conventional UV absorbance. To get round this problem, analysts useindirect UV detection, by adding a UV absorbing species to the electrolyte to give a large