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. Author manuscript; available in PMC: 2010 Jul 23.
Published in final edited form as: Glycobiol Insights. 2010 Feb 2;2010(2):1–12.

Figure 2. Liquid chromatography/mass spectrometry analysis of reduced di- and tetrasaccharides.

Figure 2

A) Heparin lyase digestion and liquid chromatography/mass spectrometry analysis. LSCH4 and OSCH2 were digested by a mixture of heparin lyase I, II, and III. OSCH2 was tagged with H-aniline and LSCH4 was tagged with D-aniline. Both tagged samples were reduced with NaBH4. An equal amount of samples was mixed and analyzed simultaneously by liquid chromatography/mass spectrometry. DA refers to a disaccharide containing an Δ4,5-unsaturated uronic acid (U) linked to N-acetylglucosamine (A). UAUH refers to a tetrasaccharide composed of uronic acid (U, GLCA or IDOA)-N-acetylglucosamine (A)-uronic acid (U)-glucosamine (H). S refers to sulfate. B) Low pH nitrous acid degradation and liquid chromatography/mass spectrometry analysis. Samples LSCH2, LSCH4, OSCH2, and S1 were degraded with nitrous acid at pH 1.5 and the resulting saccharides of LSCH2 and LSCH4 were reduced with NaBH4. OSCH2 and S1 were treated similarly and reduced with NaBD4. LSCH4 was mixed with OSCH2 whereas LSCH2 was mixed with S1. The mixtures were analyzed by liquid chromatography/mass spectrometry. UM refers to a disaccharide containing an unspecified uronic acid (U) linked to anhydromannitol (M). UAUM refers to a tetrasaccharide composed of uronic acid (U)-N-acetylglucosamine (A)-uronic acid (U)-anhydromannitol (M). S refers to sulfate.