Separation and Analysis of Nanomole Quantities of Heparin Oligosaccharides Using On-Line Capillary Isotachophoresis Coupled with NMR Detection

• Published in: Analytical chemistry (Washington) Vol. 77; no. 18; pp. 5998 - 6003
• Main Authors: Korir, Albert K, Almeida, Valentino K, Malkin, Douglas S, Larive, Cynthia K
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15-09-2005
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical chemistry; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Capillaries; Chemistry; Chromatographic methods and physical methods associated with chromatography; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Heparin - analysis; Heparin - chemistry; Heparin - isolation & purification; Magnetic Resonance Spectroscopy - methods; Molecular Structure; Nanoparticles; NMR; Nuclear magnetic resonance; Oligosaccharides - analysis; Oligosaccharides - chemistry; Oligosaccharides - isolation & purification; Online Systems; Other chromatographic methods; Spectrometric and optical methods; Detection limit; Oligosaccharide; Glycosaminoglycan; NMR spectrometry; Heparin; Isotachophoresis; Microstructure
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac050669u

Glycosaminoglycans (GAGs) are important in a number of biological processes and are structurally altered in many pathological conditions. The complete determination of GAG primary structures has been hampered by the lack of sensitive and specific analytical techniques. Nuclear magnetic resonance spectroscopy (NMR) is a powerful tool for GAG structure elucidation despite its relatively poor limits of detection. Solenoidal microcoils have greatly enhanced the mass limits of detection of NMR, enabling the on-line coupling of microseparation and concentration techniques such as capillary isotachophoresis (cITP), which can separate and concentrate analytes by 2−3 orders of magnitude. We have successfully used cITP coupled with on-line NMR detection to separate and concentrate nanomole quantities of heparin oligosaccharides. This sensitive on-line measurement approach has the potential to provide new insights into the relationships between biological function and GAG microstructures.