Highly efficient analysis of underivatized carbohydrates using monolithic-silica-based capillary hydrophilic interaction (HILIC) HPLC

Tohru Ikegami, Kanta Horie, Nabil Saad, Ken Hosoya, Oliver Fiehn, Nobuo Tanaka

Research output: Contribution to journalArticlepeer-review

84 Scopus citations

Abstract

A polyacrylamide (PAAm)-modified monolithic silica capillary column of increased phase ratio, 200T-PAAm, for hydrophilic interaction liquid chromatography (HILIC) was prepared. The column showed high separation efficiency, with a theoretical plate height H∈=∈7-20 μm at a linear velocity, u∈=∈1-7 mm/s. From a kinetic plot analysis, it was expected that the monolithic column could provide three times faster separation than particle-packed HILIC columns under a pressure limit at 20 MPa. HILIC coupled with electrospray ionization (ESI)-mass spectrometry (HILIC-ESI-MS) using the 200T-PAAm column was employed for the analysis of underivatized carbohydrates to achieve fast and efficient separations of mixtures containing mono-, di-, and trisaccharides within 5 min. Under single MS full scan mode, 200 pg of oligosaccharides was detected by the system. The limit of detection (LOD) of the LC-ESI-MS/MS system was determined using selected reaction monitoring (SRM) to be as low as 3.2 ng/mL (attomol level) for nonreducing saccharides. The system was successfully applied to the detection of disaccharides in extracts of plant, such as corn, soybean, and Arabidopsis thaliana.

Original languageEnglish (US)
Pages (from-to)2533-2542
Number of pages10
JournalAnalytical and Bioanalytical Chemistry
Volume391
Issue number7
DOIs
StatePublished - Aug 2008

Keywords

  • HILIC
  • HILIC-ESI-MS
  • Monolithic silica
  • Polyacrylamide
  • Underivatized carbohydrates

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry

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