Add time:08/08/2019         Source:sciencedirect.com

The use of high-performance liquid chromatography for the quantification of glycosaminoglycan disaccharides has been hampered by the inability to isocratically resolve the chondroitinase digestion products 2-acetamido-2-deoxy-3-O-(β-d-gluco-4-enepyranosyluronic acid)-d-glucose (ΔDi-HA) and 2-acetamido-2-deoxy-3-O-(β-d-gluco-4-enepyranosyluronic acid)-d-galactose (ΔDi-OS). To overcome this limitation, we have developed a solvent system capable of resolving ΔDi-HA, ΔDi-OS, 2-acetamido-2-deoxy-3-O-(β-d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-galactose (ΔDi-6S), and 2-acetamido-2-deoxy-3-O-(β-d-gluco-4-enepyranosyluronic acid)-6-O-sulfo-d-galactose (ΔDi-4S). Integrator responses were linear from 1 μg down to 25 ng for ΔDi-HA, ΔDi-OS, and ΔDi-4S and down to 100 ng for ΔDi-6S. This method was used to examine changes in the content of urinary hyaluronic acid and chondroitin sulfates isolated from normal individuals and from patients with Lowe Syndrome, Werner Syndrome, and Hutchinson-Gilford Progeria Syndrome. We confirmed that the HPLC method gave results comparable to colorimetric methods.

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