Sialic Acid Quantification by HPAEC-PAD

Sialic Acid Quantification by HPAEC-PAD

High-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) is an effective way to quantify sialic acids. CD BioGlyco has accumulated rich experience in the analysis of sialic acid and provides sialic acid quantification service by HPAEC-PAD for our clients worldwide.

Comparison of Sialic Acid Analysis Methods

Sialic acid is one of the most important molecules in higher animals and is present in some microorganisms. Sialic acids include a large class of N- and O-substituted neuraminic acids. The amino group of neuraminic acid is linked to an N-acetyl or N-glycolyl group to generate N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), respectively. Substituting the amino group of neuraminic acid with a hydroxyl group produces 2-keto-3-deoxy-D-glycero-D-galactonononic acid (KDN).

There have been several methods described for sialic acid analysis, such as high-performance liquid chromatography-mass spectrometry (HPLC-MS) and gas-chromatography (GC)-MS, nuclear magnetic resonance (NMR) spectrometry, and recent capillary electrophoresis (CE). However, these methods require derivatization of the hydroxyl groups used to improve sensitivity and chromatographic resolution, which become more complex, labor-intensive, and prone to contamination. HPAEC-PAD is an effective way to determine Neu5Ac and Neu5Gc without derivatization.

In HPAEC-PAD, the use of alkaline eluents causes sialic acid to be ionized, which is fit for separation by anion-exchange mechanisms. Then, the detection occurs by measuring the current generated when the carbohydrate compounds are oxidized on a gold working electrode. Compared with the methods mentioned above, the HPAEC-PAD eliminates derivatization, costly reagents, and toxic waste production and also improves safety. Moreover, it provides accurate, reproducible, and effective sialic acid analysis.

Determination of sialic acids in infant formula by HPAEC-PAD.Fig.1 Determination of sialic acids in infant formula by HPAEC-PAD. (Hurum, 2012)

Our Services

CD BioGlyco provides sialic acid quantification services for various samples using HPAEC-PAD for our clients. Our services include:

  • We use HPAEC-PAD to quantify sialic acids released from the glycoconjugate such as glycoproteins, glycolipids, trypsin glycopeptides, streptavidin, and synthetic glycoconjugates.
  • Sialic acids are determined in dairy products and infant formulas by HPAEC-PAD.
  • Identify sialic acid-containing glycosylinositol phospholipid anchor by HPAEC-PAD.
  • Determine the total sialic acid content in cells using HPAEC-PAD.
  • Purify sialic acids using HPAEC-PAD.

Advantages of Us

  • Accurate, reproducible, and effective
  • Determination of sialic acids without derivatization
  • The professional analytical team
  • Detailed data and analysis reports

CD BioGlyco is committed to providing our clients with high-quality sialic acid analysis service by HPAEC-PAD. If you are interested in our services, please feel free to contact us for more details.


  1. Hurum, D.C.; Rohrer, J.S. Determination of sialic acids in infant formula by chromatographic methods: A comparison of high-performance anion-exchange chromatography with pulsed amperometric detection and ultra-high-performance liquid chromatography methods. Journal of dairy science. 2012, 95(3): 1152-1161.
  2. Rohrer, J.S. Analyzing sialic acids using high-performance anion-exchange chromatography with pulsed amperometric detection. Analytical biochemistry. 2000, 1(283): 3-9.
This service is for Research Use Only, not intended for any clinical use.

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