Hydrophilic Interaction Liquid Chromatography: A Technique to Separate Hydrophilic Polar Analytes Using Hydrophilic Beads

Published: April 30, 2023

Abstract

Source: Liu, D., et al. Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography. J. Vis. Exp. (2020).

This video demonstrates the principle of separation of hydrophilic polar compounds using hydrophilic interaction liquid chromatography. This technique helps in the purification of various polar analytes, including N-glycans.

Protocol

1. Hydrophilic interaction chromatography and analysis of glycans Conditioning of UPLC instruments and preparation of mobile phases Prepare mobile phases including solvent A: 100 mM ammonium formate (pH = 4.4), solvent B: 100% acetonitrile, solvent C: 90% ultra-pure water (10% methanol), and solvent D: 50% methanol (ultra-pure water). Open the software to control the mobile phases. Wash UPLC instruments at flow rate of 0.2 mL/min (50% solvent B and 50% solvent C) balanci…

Açıklamalar

The authors have nothing to disclose.

Materials

2-aminobenzamide, 2-AB Sigma, China
Acetonitrile Huihai Keyi Technology Co., Ltd, China
Ammonium formate Beijing Minruida Technology Co., Ltd.
Dextran Calibration Ladder/Glycopeptide column Watts technology Co., Ltd, China
Empower 3.0 Waters technology Co., Ltd, America
GlycoProfile 2-AB Labeling kit Sigma, China
Methanol Huihai Keyi Technology Co., Ltd, China
Milli-Q pure water meter Millipore Co., Ltd, America 202-2AB
Transfer liquid gun Smer Fell Science and Technology Co., Ltd, China Advantage A10
Ultra-low temperature refrigerator Thermo Co., Ltd, America PB-10
Ultra-performance liquid chromatography Watts technology Co., Ltd, China
Vacuum Pump Watts technology Co., Ltd, China 4672100, 0.5-10μl & 10-100μl & 20-200μl & 1000μl

Etiketler

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Bu Makaleden Alıntı Yapın
Hydrophilic Interaction Liquid Chromatography: A Technique to Separate Hydrophilic Polar Analytes Using Hydrophilic Beads. J. Vis. Exp. (Pending Publication), e21097, doi: (2023).

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