Monitoring activation sites on polysaccharides by GC-MS.

A method has been developed to determine the location and order of activation for potential saccharide antigens used in conjugate vaccine development. Saccharides were monitored for activation by sodium periodate oxidation and subsequent analysis by gas chromatography-mass spectrometry (GC-MS). Pneumococcal serotype polysaccharides 7F and 18C were evaluated as polysaccharides containing multiple potential sites for activation.

Addition of glycerol for improved methylation linkage analysis of polysaccharides.

A presolubilization procedure with the use of glycerol is shown to be applicable for the structural analysis of polysaccharides. Neutral, acidic, high-molecular-weight and low-molecular-weight polysaccharides were solubilized in glycerol prior to methylation and subsequent linkage analysis by GC-MS. All four types of polysaccharides showed significant increases in derivatization following presolubilization as measured by recovery of partially methylated alditol acetates.

Determination of saccharide content in pneumococcal polysaccharides and conjugate vaccines by GC-MSD.

A simple and sensitive gas chromatographic method was designed for quantitative analysis of Streptococcus pneumoniae capsular polysaccharides, activated polysaccharides, and polysaccharide conjugates. Pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F polysaccharide or conjugate were subjected to methanolysis in 3N hydrochloric acid in methanol followed by re-N-acetylation and trimethylsilylation. Derivatized samples were chromatographed and detected using gas chromatography with mass selective detector.