Interactions of analytes with metal surfaces in high-performance liquid chromatography (HPLC) instruments and columns have been reported to cause deleterious effects ranging from peak tailing to a complete loss of the analyte signal. These effects are due to the adsorption of certain analytes on the metal oxide layer on the surface of the metal components.

We have developed a novel surface modification technology and applied it to the metal components in ultra HPLC (UHPLC) instruments and columns to mitigate these interactions. A hybrid organic−inorganic surface, based on an ethylene-bridged siloxane chemistry, was developed for use with reversed-phase and hydrophilic interaction chromatography. We have characterized the performance of UHPLC instruments and columns that incorporate this surface technology and compared the results with those obtained using their conventional counterparts.

We demonstrate improved performance when using the hybrid surface technology for separations of various small molecules as well as nucleotides, phosphopeptides, and oligonucleotides. The hybrid surface technology was found to result in higher and more consistent analyte peak areas and improved peak shape, particularly when using low analyte mass loads and acidic mobile phases. Reduced abundances of iron adducts in the mass spectrum of a peptide were also observed when using UHPLC systems and columns that incorporate hybrid surface technology. These results suggest that this technology will be particularly beneficial in UHPLC/mass spectrometry investigations of metal-sensitive analytes.

Join this session to:

Learn about non-specific adsorption and the challenges it can cause
Learn about current workarounds that scientists currently use to mitigate non-specific adsorption
Learn about MaxPeak HPS technology and how this prevent non-specific adsorption across many different workflows

Kim Haynes

Director of Chemistry Marketing and Product Management, Waters Corporation

Appearances