Analyzing Carbohydrates by HPAE-PAD Ion Chromatography

No need for derivatization or a chromophore

Carbohydrates are carbon sources essential for cell growth and product synthesis, and they play a vital role in biological functions such as cellular communication, gene expression, immunology, and organism defense mechanisms. Characterization of glycoproteins routinely involves carbohydrate analysis, and changes in protein glycosylation are often studied in cancer research to identify potential biomarkers. There are over 30 approved glycoprotein-based biodrugs on the market, and the number is increasing rapidly.


Carbohydrate analysis

Carbohydrates are difficult to analyze using common chromatography and detection methods. They are very polar compounds, exhibit similar structural characteristics, and lack a suitable chromophore. High-Performance Anion-Exchange Chromatography (HPAE) chromatography takes advantage of the weakly acidic nature of carbohydrates for highly selective separations at high pH using strong anion exchange stationary phases. High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD) does not require modification of glycans before analysis, and specificity is ensured by the separation and the electrochemical detection that has been optimized for carbohydrate analysis.


Regulatory information

The U.S. FDA and the European Medicines Agency have increased pressure on biopharmaceutical manufacturers to demonstrate satisfactory programs for understanding, measuring, and controlling glycosylation in glycoprotein-based drugs.


Example application notes

Application Notes Sample IC Column
Rapid Screening of Sialic Acids in Glycoproteins by HPAE-PAD (AU 181) Glycoprotein Dionex CarboPac PA20 Columns
Glycoprotein Monosaccharide Analysis Using HPAE-PAD with Eluent Generation (TN 40) Glycoprotein Dionex CarboPac PA20 Columns
Evaluating Protein Glycosylation in Limited-Quantity Samples by HPAE-PAD (AN 1050) Glycoprotein Dionex CarboPac PA200 Columns
The Determination of Carbohydrates, Alcohols, and Glycols in Fermentation Broths (AN 122) Fermentation broth Dionex CarboPac MA1 Columns
HPAE-PAD Analysis of Mannose-6-Phosphate (AN 202) Glycoprotein Dionex CarboPac PA200 Columns
HPAE-PAD Determination of Carbohydrates in Honey to Evaluate Samples for Quality and Adulteration (AN1158) Honey Dionex CarboPac PA210 Fast 4µm
A Fast Method for Sugar Analysis of Instant Coffee Samples (AU202) Coffee Dionex CarboPac SA10-4µm
Determination of Carbohydrates in Kombucha Using HPAE-PAD (AN1152) Kombucha Dionex CarboPac PA20
Profiling Fructosyloligosaccharides (FOS)-containing samples by HPAE-PAD (AN 1149) Fructosyloligosaccharides Dionex CarboPac PA200
Profiling Galactosyloligosaccharide (GOS)-containing Samples by HPAE-PAD (AN1151) Galactosyloligosaccharides Dionex CarboPac PA200
HPAE-PAD Determination of Infant Formula Sialic Acid (AN253) Infant formula Dionex CarboPac PA20
Ion Chromatography: A Versatile Technique for the Analysis of Beer (AN71410) Beer Dionex CarboPac PA20
HPAE-PAD Determination of Cyclodextrins (AN73011)   Dionex CarboPac PA200
An HPAE-PAD Method for Determination of Saccharides in Atmospheric Aerosol Samples (AN73009)   Dionex CarboPac PA200
Improved Profiling of Sialylated N-linked Glycans by Ion Chromatography-Orbitrap Mass Spectrometry (AN73063)    
Carbohydrate Analysis of Agave Syrup using HPAE-PAD
(AN 73132)
Agave Syrup Dionex CarboPac PA200 Columns