MSACL 2016 EU Abstract

Challenges in Chromatography: PART 1

Grace van der Gugten (Presenter)
St Paul's Hospital

Authorship: J. Grace van der Gugten
St. Paul’s Hospital, Department of Pathology and Laboratory Medicine, Vancouver, Canada

Short Abstract

One of the critical decisions during LC-MS/MS assay development is selection of the analytical column. This is particularly important when the analyte(s) of interest must be separated from similar compounds (particularly isobars) that cannot be uniquely identified on the basis of the mass-to-charge ratio. This presentation will review the basics of reversed phase LC and discuss column chemistries and how they affect retention and selectivity. Examples of standard solutions and extracted biological samples will be shown run on a range of vendor columns with different column chemistries.

Long Abstract

One of the critical decisions during LC-MS/MS assay development is selection of the analytical column. This is particularly important when the analyte(s) of interest must be separated from similarcompounds (particularly isobars) that cannot be uniquely identified on the basis of the mass-to-charge ratio.

Often the suggested starting point for method development is a C18 column. However, for some compounds the C18 stationary phase may inadequate for baseline separation except by use of a gradient that is impractically long. Column manufacturers are routinely developing new stationary phases and improving existing ones. Selecting the best column can be overwhelming to newcomers, especially considering the vast array of vendors and stationary phases available. Although manufacturers have slick advertising materials demonstrating the strengths of their products, it is often advisable to rely more on recommendations of colleagues and our own experience to make unbiased decisions.

This presentation will review the basics of reversed phase LC and discuss column chemistries and how they affect retention and selectivity. Examples of standard solutions and extracted biological samples will be shown run on a range of vendor columns with different column chemistries.


References & Acknowledgements:


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