HILIC

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This article, transcribed from an LCGC web seminar presented on January 29, 2013, by Davy Guillarme of the University of Geneva and the University of Lyon, explains how HILIC works and provides keys to using the technique effectively.

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5-Fluorouracil (5-FU) is a low-molecular-weight anticancer drug in clinical use for several solid tumors in humans. Currently, the most widely used methodology for 5-FU quantitation is liquid chromatography–tandem mass spectrometry (LC–MS-MS) with either liquid–liquid extraction (LLE), protein precipitation, or a combination of both as sample cleanup procedures.

HILIC: The Pros and Cons

Davy Guillarme discussed the possibilities and limitations of HILIC in a recent web seminar. Here, he answers common questions about the method.

Davy Guillarme discussed the possibilities and limitations of HILIC in a recent web seminar. Here, he answers common questions about the method.

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Hydrophilic interaction liquid chromatography (HILIC) has recently become more important, particularly for the analysis of polar drugs, metabolites and biologically relevant compounds in glycomics, proteomics, metabolomics and clinical analysis. HILIC makes it possible to increase the retention of polar compounds, achieve orthogonal selectivity and increase mass spectrometry (MS) sensitivity, compared with reversed-phase liquid chromatography. This article discusses the advantages and limitations of HILIC in a variety of practical applications in the pharmaceutical industry.

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Reversed-phase HPLC analysis of histamine has been a challenge in the past due to the high polarity and poor retention. HILIC (hydrophilic interaction chromatography) offers a potential solution because of high polar analyte retention without the need of strong ion-pair reagents, which are not compatible with mass spectrometry (MS) detection and can cause signal suppression.

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Hot Tips on HILIC

Hydrophilic interaction chromatography (HILIC) is an alternative separation technique for highly polar analytes that are weakly retained in reversed-phase high performance liquid chromatography (HPLC). Participants in this HILIC Technology Forum are William Long of Agilent Technologies, Phil Koerner and Carl Sanchez of Phenomenex, and Nicole M. Cotta of Waters Corporation.

Hydrophilic interaction chromatography (HILIC) is an alternative separation technique for highly polar analytes that are weakly retained in reversed-phase high performance liquid chromatography (HPLC). Participants in this HILIC Technology Forum are William Long of Agilent Technologies, Phil Koerner and Carl Sanchez of Phenomenex, and Nicole M. Cotta of Waters Corp.