Amperometric detection of amino acids in high-performance liquid chromatography with a copper electrode

A copper electrode has been used as an amperometric detector for amino acids in high-performance liquid chromatography. The amino acids are separated in a reversed-phase system, using silica-based and polymer-type column materials. Neutral or alkaline buffer solutions of phosphate and carbonate can be used as mobile phases. Borate buffers are less suitable. The detection method is characterized by a high linear dynamic range, good reproducibility, the absence of electrode poisoning and a sensitivity comparable to that of UV absorption methods after derivatization of the amino acids. Detection limits with conventional-scale columns are in the range 10-100 pmoles. A reduction in the flow-rate in the flow-through cell improves the sensitivity for amino acids that give relatively low signals, such as proline. Therefore, the use of microbore columns is especially advantageous for these compounds. The absolute detection limits decrease by about one order of magnitude on changing to a miniaturized system.