ABSTRACT
Historically, on-line detection of biomacromolecules in postcolumn eluants has utilized measurements of the physical/compositional characteristics of the eluting molecules (e.g., measured by fluorescence, absorbance, electrochemical detection). These characteristics can even be used to detect coeluting impurities and to identify molecules that elute as pure peaks by absorbance ratio monitoring. 1-3 However, physical/compositional characteristics provide little, if any, basis for specific detection4 (i.e., the detection of a particular species of macromolecule against the background of a complex mixture). Physical/compositionalbased detection can provide specificity only when the biomolecule of interest has a unique absorbance spectrum. However, the infrequent occurrence of biomacromolecules with unique spectral characteristics limits the usefulness of this approach.
