Effect of first dimension phase selectivity in online comprehensive two dimensional liquid chromatography (LC×LC)

Haiwei Gu, Yuan Huang, Marcelo Filgueira, Peter W. Carr

Research output: Contribution to journalArticlepeer-review

19 Scopus citations


In this study, we examined the effect of first dimension column selectivity in reversed phase (RP) online comprehensive two dimensional liquid chromatography (LC×LC). The second dimension was always a carbon clad metal oxide reversed phase material. The hydrophobic subtraction model (HSM) and the related phase selective triangles were used to guide the selection of six different RP first dimension columns. Various kinds of samples were investigated and thus two different elution conditions were needed to cause full elution from the first dimension columns. We compared LC×LC chromatograms, contours plots, and fcoverage plots by measuring peak capacities, peak numbers, relative spatial coverage, correlation values, etc. The major finding of this study is that the carbon phase due to its rather different selectivity from other reversed phases is reasonably orthogonal to a variety of common types of bonded reversed phases. Thus quite surprisingly the six different first dimension stationary phases all showed generally similar separation patterns when paired to the second dimension carbon phase. This result greatly simplifies the task of choosing the correct pair of phases for RP×RP.

Original languageEnglish (US)
Pages (from-to)6675-6687
Number of pages13
JournalJournal of Chromatography A
Issue number38
StatePublished - Sep 23 2011
Externally publishedYes


  • Carbon stationary phases
  • Hydrophobic subtraction method
  • LC×LC method development
  • Reversed phase
  • Selectivity
  • Two dimensional liquid chromatography

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry


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