Abstract
A fully automated technique for the analysis of amino and imino acids allowing direct sampling of raw biological fluids is described. The procedure uses on-line membrane (ASTED™) technology to remove macromolecular interferences from samples, simultaneous pre-column derivatisation with o-phthalaldehyde and 9-fluorenylmethyl chloroformate and separation of the analytes using reversed-phase high performance liquid chromatography. Associated problems of both pre-column derivatisation reagents are also overcome. The use of membrane clean-up also permits an examination of the technique to automatically estimate non-protein bound tryptophan. Method validation includes method sensitivity, the use of enrichment to extend the analytical range and correlation with ion exchange chromatographic methodology.
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