This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Preparation of Partially Methylated Alditol Acetates (PMAAs) For the determination of glycosyl linkages, PMAAs were prepared from the permethylated oligosaccharides (remainder from oligosaccharide profiling). Briefly, permethylated oligosaccharides were hydrolyzed with HCl/water/acetic acid (0.5:1.5:8, by vol.) at 80oC for 18h, followed by reduction with NaBD4. The latter-freed hydroxyls after hydrolysis were acetylated with acetic anhydride/pyridine (1:1, v/v) at 100 ?C for 15 min. Gas Chromatograph-Mass Spectrometry (GC-MS) The PMAAs for neutral sugars were analyzed on an Agilent 7890A GC interfaced to a 5975C MSD equipped with a Rtx-2330 column, whereas, the PMAAs for amino sugars was analyzed on a Hewlett Packard (HP) 5890 GC interfaced to a 5970 installed with a 30-m EC 1 bonded phase fused silica capillary column (Altech). The GC conditions were the following: Agilent 7890A, Initial oven temp was 80oC, which was held for 2 min, ramped to 170oC at 30oC/min, held for 0 min, and finally ramped to 235oC at 4oC/min and held for 20 min; HP 5890 GC, oven temperature, 140?C (2.0?C/min) ? 220?C (20?C/min) ? 300?C (20?C/min);detector temperature, 280 ?C;inlet temperature, 250 ?C.
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