Derivatization of Oligosaccharides with 1-Pyrenemethylamine: A Novel Oligosaccharide Tag for Efficient MALDI-TOF Fragmentation
Poster Number
13C
Research or Creativity Area
Natural Sciences
Abstract
Carbohydrates are amongst the most structurally diverse and ubiquitous compounds found in nature. They play central roles in many processes such as cellular recognition, cellular adhesion, and signal transduction1. As constituents of glycoproteins, glycolipids, and proteoglycans, carbohydrates also serve as diagnostic markers for the onset and development of diseases2. Therefore, profiling of glycans and quantification is essential. However, most native carbohydrates are poorly detected by UV-VIS detectors commonly used in methods for chromatographic separation, such as high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). Therefore, chemical derivatization is a necessary step in purification of glycan pools and downstream characterization via mass spectrometry (MS). The most widely reported oligosaccharide tags are UV-active aromatic amines that are chemically introduced via reductive amination3. Our group investigated a novel oligosaccharide tag, 1-pyrenemethylamine (1-PMA) that exhibits strong UV-activity with a molar extinction coefficient of 35,000cm-1M-1 (λ=341nm). The primary amino group of 1-PMA confers three distinct properties: 1) high nucleophilicity relative to conjugated aromatic amines, 2) a λmax of 1-PMA oligosaccharide derivatives independent of pH and structure of the derivatized oligosaccharide, and 3) a pyrenemethylene moiety of derivatized oligosaccharides that is readily fragmented during analysis with matrix-assisted laser desorption-ionization (MALDI) time-of-flight (TOF) MS. Oligosaccharide standards and glycans liberated from enzymatically-digested polysaccharides have been successfully derivatized with satisfactory yield. Methods for the chromatographic separation of 1-PMA oligosaccharide derivatives on hydrophilic interaction liquid chromatography (HILIC) and C18 columns have been developed and are reported here. The lower limit of detection by photo-diode array (PDA) of 1-PMA oligosaccharide derivatives was ~5pmol.
Literature:
- Costello, C. E. & Han, L. Mass Spectrometry of Glycans. Biochemistry (Mosc), 2013, 78(7) 710-720. DOI: 10.1134/S0006297913070031
- Ohtsubo, K. & Marth, J. D. Glycosylation in cellular mechanisms of health and disease. Cell, 2006, 126(5) 855-67. DOI: 10.1016/j.cell.2006.08.019.
- Lamari, F. N., Kuhn, R., & Karamanos, N. K. Derivatization of carbohydrates for chromatographic, electrophoretic and mass spectrometric structure analysis. Journal of Chromatography B, 2003, 793(1) 15-36. DOI: 10.1016/s1570-0232(03)00362-3
- Patel, R. et al., RCS, University of the Pacific, 2024 (poster)
Location
Don and Karen DeRosa University Center (DUC) Poster Hall
Start Date
27-4-2024 10:30 AM
End Date
27-4-2024 12:30 PM
Derivatization of Oligosaccharides with 1-Pyrenemethylamine: A Novel Oligosaccharide Tag for Efficient MALDI-TOF Fragmentation
Don and Karen DeRosa University Center (DUC) Poster Hall
Carbohydrates are amongst the most structurally diverse and ubiquitous compounds found in nature. They play central roles in many processes such as cellular recognition, cellular adhesion, and signal transduction1. As constituents of glycoproteins, glycolipids, and proteoglycans, carbohydrates also serve as diagnostic markers for the onset and development of diseases2. Therefore, profiling of glycans and quantification is essential. However, most native carbohydrates are poorly detected by UV-VIS detectors commonly used in methods for chromatographic separation, such as high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). Therefore, chemical derivatization is a necessary step in purification of glycan pools and downstream characterization via mass spectrometry (MS). The most widely reported oligosaccharide tags are UV-active aromatic amines that are chemically introduced via reductive amination3. Our group investigated a novel oligosaccharide tag, 1-pyrenemethylamine (1-PMA) that exhibits strong UV-activity with a molar extinction coefficient of 35,000cm-1M-1 (λ=341nm). The primary amino group of 1-PMA confers three distinct properties: 1) high nucleophilicity relative to conjugated aromatic amines, 2) a λmax of 1-PMA oligosaccharide derivatives independent of pH and structure of the derivatized oligosaccharide, and 3) a pyrenemethylene moiety of derivatized oligosaccharides that is readily fragmented during analysis with matrix-assisted laser desorption-ionization (MALDI) time-of-flight (TOF) MS. Oligosaccharide standards and glycans liberated from enzymatically-digested polysaccharides have been successfully derivatized with satisfactory yield. Methods for the chromatographic separation of 1-PMA oligosaccharide derivatives on hydrophilic interaction liquid chromatography (HILIC) and C18 columns have been developed and are reported here. The lower limit of detection by photo-diode array (PDA) of 1-PMA oligosaccharide derivatives was ~5pmol.
Literature:
- Costello, C. E. & Han, L. Mass Spectrometry of Glycans. Biochemistry (Mosc), 2013, 78(7) 710-720. DOI: 10.1134/S0006297913070031
- Ohtsubo, K. & Marth, J. D. Glycosylation in cellular mechanisms of health and disease. Cell, 2006, 126(5) 855-67. DOI: 10.1016/j.cell.2006.08.019.
- Lamari, F. N., Kuhn, R., & Karamanos, N. K. Derivatization of carbohydrates for chromatographic, electrophoretic and mass spectrometric structure analysis. Journal of Chromatography B, 2003, 793(1) 15-36. DOI: 10.1016/s1570-0232(03)00362-3
- Patel, R. et al., RCS, University of the Pacific, 2024 (poster)
