CD BioGlyco provides powerful CE-based HMO analysis services for our clients. We have the ability to use multiple analysis methods alone or in combination. Based on that, we have the confidence to meet clients' requirements for HMO research.

Introduction

Capillary electrophoresis (CE) is one of the most used liquid phase separation techniques for HMO analysis. Furthermore, CE can be connected to a mass spectrometer (MS) to get structural data. It has various benefits, including minimal sample volume requirements, low buffer usage, quick, high-resolution separations, and a variety of separation modes that make it simple to change the operating parameters. Derivatization is required for CE in order to achieve adequate electromigration of the largely uncharged carbs.

Fig.1 Schematic diagram of a CE system. (Sanchez-Vega, 2008)

Analysis Methods

CE

The high sensitivity and resolving power of CE are widely recognized. Even several commercially available HMO isomeric standards, such as 2'ucosyllactose (2'FL)/3'FL or lacto-N-fucopentaose I/II (LNFP I/II), have been effectively separated using CE. Additionally, it has been noted that the analysis of HMOs using CE with laser-induced fluorescence (LIF) linked to MS (CE-LIF-MS) of milk oligosaccharides tagged with 8-amino-pyrene-1,3,6-tri sulfonic acid (APTS) is effective.

Fig.2 8-Amino-pyrene-1,3,6-trisulfonic acid. (PubChem)

CE-UV

In 2007, Bao et al. developed and validated a CE-based method for the quantification of 12 major acidic HMOs using UV detection. Later, in order to find prospective sources of these oligosaccharides as infant formula supplements, this approach was used by others to analyze the concentration of 3-sialyl-lactose (3'SL), 6-sialyl-lactose (6'SL), and DSLNT in cow, goat, and horse milk. It is crucial to remember that the UV detection limit was substantially lower than LIF, and the method's usefulness was constrained by the lengthy separation intervals.

Fig.3 Schematic diagram of sheathless CE-MS and CE-UV methods for the identification and quantification of cobra-tide drugs. (Liu, et al., 2021)

CE-LIF

The analysis of complicated biological materials using CE-LIF is appealing, but identifying the migratory analytes requires the examination of reference standards, which is sometimes costly, hard to obtain, or simply not possible. Furthermore, based purely on norms for migration timeframes, co-migration cannot be disregarded. Due to the voltage established between the cathode and anode at the MS interface, CE-LIF analysis suggests a continuing separation in the CE capillary even after the LIF-detector window, in contrast to LC-(UV/DAD)-MS.

Fig.4 Diagram of the CE system with LIF detection. (Moroz, et al., 1999)

CE-based HMO Analysis Services at CD BioGlyco

Based on the current development of CE-based analysis methods at CD BioGlyco, we establish several analysis programs such as CE, CE-UV, CE-LIF, and so on. we provide several CE-based HMO analysis services, not only using single methods but also combinate using different analysis technologies according to clients' requirements.

Advantages of Us

• We have advanced lab equipment and good research teams for analysis services.
• We have customized CE-based analysis services to meet the analysis needs of our clients, such as CE, CE-UV, CE-LIF, and so on.
• We have well-trained and experienced researchers to handle these projects.
• We have the confidence to provide powerful analysis tools, which is a helpful tool for clients to study HMOs.

CD BioGlyco aims to help customers to study the compounds, structure, and content of HMOs, we are a great choice for HMO analysis. If you are interested in our analysis services, please feel free to contact us for more detailed information.

References:

1. Moroz, L.L.; et al. Single-cell analyses of nitrergic neurons in simple nervous systems. Journal of Experimental Biology. 1999, 202(4): 333–341.
2. Sanchez-Vega, B. Capillary electrophoresis of DNA. Springer Protocols Handbooks. 2008, 65–87.
3. Liu, B.; et al. Separation, identification and quantification of associated impurities in cobratide using sheathless CE-MS and CE-UV. Analytical Methods. 2021, 13(34): 3845–385.