Ion Pair Chromatograpy: A Critical Prespective

Abstract

Reversed phase chromatography is one of most popular modes of chromatography in use. Ion pair chromatography a subset of reverse phase chromatography even though happens due to ionic interactions, needs a reverse phase stationary phase as support. Retention of basic analytes on reverse phase columns is the most commonly encountered situation where ion pair chromatography is used. Reproducibility problems in gradient elution, slow equilibration times are some of the disadvantages of ion pair chromatography for which mixed mode columns can provide the best possible solution. In this concise review, an attempt has been made to give an overview of all aspects of ion pair chromatography.

Quantification of guanosine triphosphate and tetraphosphate in plants and algae using stable isotope-labelled internal standards

“…A limitation of this method is that the HPLC separation used prior to MS/MS detection relies on the ion-pairing reagent DMHA to retain the highly polar nucleotides on a reverse phase C18 chromatography column. Ion-pairing reagents such as DMHA can be problematic for HPLC-MS systems as they cannot be easily removed from columns and plastic tubing, and sometimes require long equilibration times [29]. Notably, there are recent examples of the successful separation of G4P and other nucleotides from both bacteria and algae using hydrophilic interaction (HILIC) chromatography [5,30].…”

Quantification of guanosine tetraphosphate and other nucleotides in plants and algae using stable isotope-labelled internal standards

“…In particular, the HPLC separation approach used here prior to MS/MS detection relies on the ion-pairing reagent DMHA to retain the highly polar nucleotides on a reverse phase C18 chromatography column. Ion-pairing reagents such as DMHA can be problematic for HPLC-MS systems as they cannot be easily removed from columns and plastic tubing, and sometimes require long equilibration times (Yerneni, 2017). Notably, there are recent examples of the successful separation of G4P and other nucleotides from both bacteria and algae using hydrophilic interaction (HILIC) chromatography (Jin et al, 2018;Zborníková et al, 2019).…”