This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Composition analysis by HPAECThe samples were hydrolyzed to release the monosaccharides and were analyzed by High pH-Anion-Exchange Chromatography (HPAEC). Hydrolysis of dried samples was performed with 400 L of 2 N trifluoroacetic acid (TFA) at 100 C for 4 h. Also, a 400- L blank TFA was hydrolyzed under the same conditions. After hydrolysis, the samples were lyophilized and resuspended in 150 L of nanopure water, sonicated in cold water for 7 min and transferred to an injection vial. For analysis, the autosampler was set to deliver 10 L of sample or standard solution per injection. Blanks (water) were injected between standards and the sample to prevent carry-over contamination. A mix of sugar standards with a known number of moles was hydrolyzed at the same time as the samples. Three concentrations of standards (0.5, 1.0, and 2.0 nmoles per injection) were prepared and injected to establish a calibration equation. The number of moles of each sugar was quantified by linear interpolation from the calibration equation.
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