Simultaneous Chromatography and BIS
Its importance and potential applications
3.4.10.
Simultaneous “on column” and “post column” monitoring may seem to be a superfluous approach unless we realize that what is not eluted from the column cannot be detected by conventional chromatography.
Consequently:
- Strongly bound sample components will not be detected
- The capacity of the stationary phase will be reduced
- Subsequent assays may be compromised when stronger eluants release components of previously analyzed samples.
Yet another potential use of simultaneous “on” and “post” monitoring is the investigation of the properties of the stationary phase, either the kinetics of capture and release or the quality control of of stationary phase used in industrial biochemical separations, where Sephadex and Separaose are used in large columns for biomolecule purifications, such as insulin production.
While carefully controlled, the decision of when to regenerate the column is based on trial and error. By investigating a sample of column material by BIS or by the way discussed in the next section, the exact protocol for more economic column regeneration will be established.
