Optimization Strategies for Chromatographic Analysis of Multi-Component Mixtures

Abstract

In column-chromatographic analysis of biological samples, complex chromatograms with long elution times are often encountered. Once the general separation conditions have been chosen, e.g. type and mode of use of chromatographic column and mode of detection, a number of continuously tunable variables can be used to optimize the overall analytical procedure. Necessarily a criterion has to be defined that permits an objective judgement of the resulting chromatograms. For this purpose chromatographic response functions (CRFs) used with formal strategies of experimental optimization have been developed. The strategies involve either entirely empirical search and mapping techniques or semi-empirical functions that account for specific molecular interactions. In any case the optimization procedure has to be chosen in accordance with the complexity of the separation problem, and the CRF must reflect fundamental analytical performance characteristics such as precision, accuracy and analysis time. Example in APPENDIX: thyronine I-enantiomers.