HILIC

The aim of this work was to find the optimal conditions to achieve sufficient limits of detection (LOD) that would permit the detection of neurotransmitters by LC–MS–MS in biological samples. An optimized HILIC–ESI–MS–MS system for the analysis of the 12 selected compounds was proposed.

The Technical Department at Crawford Scientific are regularly asked for analytical conditions or applications in the hydrophilic interaction liquid chromatography (HILIC) mode. Here?s a short primer on what you need to understand about this separation mode from a practical perspective, using some of the questions that we regularly get asked.

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.

Agilent Technologies

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.

Click here to view the complete E-Separation Solutions newsletter from November 20, 2012.

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.

Some of the newer hydrophilic interaction liquid chromatography (HILIC) columns are reviewed, and experiments performed to categorize columns for optimized performance.

A comparison of polymer- and silica-based stationary phases used in hydrophilic interaction chromatography (HILIC) for saccharide analysis.

Waters Corporation

Determination and quantification of amino acids are important in many fields. For the HPLC analysis of amino acids, cation exchange mode method has been used with post-column derivatization. Recently, use of reversed-phase mode method is also increasing. However, because of amino acids' hydrophilic nature, they are usually not well retained and have less selectivity by reversed-phase mode without pre-column derivatization steps. In contrast, HILIC mode is an ideal separation mode for hydrophilic compound analysis.

Kromasil HILIC-D is designed for the best selectivity of polar compounds. The phase is low bleed and solvents used when running in HILIC mode are fully compatible for MS detection.

The coupling of HILIC with reversed-phase HPLC and API-MS will become routine in the future, particularly in fields like metabolomics and environmental analysis that regularly involve nontargeted screening of large numbers of compounds in complex mixtures.

Some of the newer hydrophilic interaction liquid chromatography (HILIC) columns are reviewed, and experiments performed to categorize columns for optimized separations are described.

Saccharides are fundamental substances that express various bioactivities and may exist independently or form complexes with proteins or lipids.

This new column explains the essentials of key methods in separation science. This month: hydrophilic interaction chromatography.

According to law enforcement and health officials, "bath salts" are powerful synthetic stimulants, designed to be comparable in effect to cocaine or methamphetamine, carrying similar risks. Unlike cocaine or methamphetamine, these stimulants are currently legal in many parts of the United States and available at a low cost in many convenience stores and head shops.

What challenges occur when using hydrophilic interaction liquid chromatography (HILIC), and what options are available for analysis?

Click here to view the complete e-Separation Solutions newsletter from February 4, 2011.

LCGC's The Column editor, Janet Kelsey, spoke to Mark Woodruff of Fortis Technologies about the background, advantages, and applications of hydrophilic interaction chromatography and where its future lies.

Mark Woodruff of Fortis Technologies talks about the background, advantages and applications of hydrophilic interaction chromatography and where its future lies.

Site-specific analysis of protein glycosylation has become a very important issue in pharmaceutical/biotech industries, requiring specific analytical tools and methodologies.

Glycosylation is one of the most common forms of post-translational modification of eukaryotic proteins. Glycosylated proteins (glycoproteins) make up the majority of human proteins.

The separation of polar and hydrophilic compounds can be a challenge. Hydrophilic interaction liquid chromatography has proven to be a solid alternative for such applications.

The separation of mixtures of polar and hydrophilic compounds can be a significant challenge.

The thorough characterization of glycosylation is a major quality parameter in the production of biotherapeutics. Hydrophilic interaction chromatography (HILIC) is a well-recognized technique that effectively separates and quantifies isolated glycans...

Experimental data are presented to help understand HPLC column retention in a way that should make column selection and method development proceed faster when initial results with C18 columns are disappointing.

The analysis of melamine in milk and other food products became urgent when first the pet food, and later the milk scandal were discovered in the U.S. and China.

Separation of polar compounds by conventional reverse phase chromatography can be challenging due to their poor retention. Other HPLC approaches include normal phase chromatography (NPC) and its variation, hydrophilic interaction chromatography (HILIC).

An Empirical Evaluation of HILIC and Monolithic Columns for Supercritical Fluid Chromatography (SFC) Applications - Thar Apps Note