HILIC

Biopharmaceuticals offer great new possibilities and potential in treating various life threatening diseases such as cancer and autoimmune diseases. However, their complexity, resulting from their heterogeneity, represents a great challenge to the analytical chemist and a suite of techniques is required for their characterization and analysis. Reversed phase chromatography, the mainstay of traditional small molecule analysis, is employed at multiple biopharmaceutical levels due to its versatility, inherent relative efficiency and ease of hyphenation to a mass spectrometer.

An excerpt from LCGC’s e-learning tutorial on biochromatography at CHROMacademy.com

A summary of what these techniques can tell us from a qualitative and quantitative perspective

Therapeutic peptides represent one of the fastest growing segments in the pharmaceutical market. To bring these products to the market in a consistent manner, high quality is a major concern and requires stringent quality control (QC) methods. This article discusses the potential of zwitterionic chiral ion-exchangers to support peptide analysis and quality control as a flexible complementary tool to monitor the stereochemical integrity and chemical modifications.

In the example shown here we seek to provide an alternative to HILIC methods by using a novel multi-mode column called the Scherzo SS-C18. This stationary phase has both reversed phase (C18) and polar components with the addition of a high density of strong anion and cation exchange ligands. This strategy for increasing polar retention is much more predictable run to run and does not require lengthy column pre-treatments.

Characterizing Cancer Tumours using HILIC–MS

Hydrophilic interaction chromatography (HILIC) is becoming more popular. Dr Tohru Ikegami from the Department of Biomolecular Engineering at the Kyoto Institute of Technology in Kyoto, Japan, reveals more about the evolution of HILIC, important new developments, and tips on how to get the most from this technique.

Hydrophilic-interaction chromatography (HILIC) is a complex system involving partition, polar, and ion‑exchange interactions. Method development can be greatly facilitated by understanding the interactions that the different stationary phases provide and applying that knowledge to the separation task at hand. Through an understanding of the main differences of the set of analytes to be separated, a column or set of columns can be judiciously chosen to screen for effective retention and selectivity.

A new chromatographic method to detect paralytic shellfish toxins (PSTs) in food has been developed by a team of scientists from the Swedish Research Agency.PSTs pose a threat to seafood and drinking water from natural contamination, but could also be used to deliberately contaminate food, according to the authors.

Due to the highly polar nature of saccharides, the analysis of sugars is typically achieved using hydrophilic interaction chromatography (HILIC).

Due to the highly polar nature of saccharides, the analysis of sugars is typically achieved using hydrophilic interaction chromatography (HILIC).

HILIC was introduced more than 50 years ago but has attracted more attention recently. Giorgia Greco from the Technische Universität Munchen in Germany discusses the different options available for combining HILIC to reversed-phase LC, and how HILIC can be hyphenated with atmospheric pressure chemical ionization MS.

Due to the highly polar nature of saccharides, the analysis of sugars is typically achieved using Hydrophilic Interaction Chromatography (HILIC). Using a high polarity packing material, separation is based on the hydrophilic interactions of the sugars on the stationary phase surface typically using an eluent with a majority of polar organic solvent.

Understanding the interactions that stationary-phase chemistries provide will help you choose the most appropriate blend of interactions to address a given separation challenge.

The retention and selectivity of stationary phases in HILIC is discussed.

Davy Guillarme, a senior lecturer at the School of Pharmaceutical Sciences at the University of Geneva, University of Lausanne, Switzerland, answers common user questions about HILIC.

Due to the highly polar nature of saccharides, the analysis of sugars is typically achieved using hydrophilic interaction chromatography (HILIC).

When you begin to now combine advances in superficially porous media to achieve high efficiency with novel and predictable new stationary phases to achieve high selectivity, then it sounds like the evolution of HPLC and UHPLC phases over the past decade or more has brought some exciting new possibilities.

Hydrophilic interaction chromatography (HILIC) provides an alternative approach for the separation of highly polar substances.

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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.

A Critical Evaluation of Hydrophilic-Interaction Chromatography (HILIC)

The modern usage and specific applications of HILIC for two other major classes of analytes - glycopeptides and glycoproteins - are discussed.

Answers to common questions about HILIC, and an overview of its application to biopharmaceutical analysis.

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.

The rising interest in HILIC can be demonstrated by the growing number of publications and applications over the last 10 years. Applications range from food and nutrition, to cosmetics, pharmaceuticals, and bioanalysis.

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