Analytical method of heparan sulfates using high-performance liquid chromatography turbo-ionspray ionization tandem...

We established a highly sensitive quantitative analytical method of heparan sulfates (HS) by LC-MS-MS. It became possible to determine the unsaturated disaccharides produced by the enzyme digestion of HS, and to perform the whole analyses on one sample within 3 min by use of a short column of CAPCELL PAK NH2 UG80 (35 mm x 2 mm I.D.). The assay method was validated and showed the satisfactory sensitivity, precision and accuracy, which enabled the quantitation up to picomol level. By employing this method, we performed the analyses of HS in mouse brain and liver, and tumor tissues of tumor-bearing mouse transplanted subcutaneously with Meth A fibrosarcoma cells. The compositions of the unsaturated disaccharide units derived from HS were found to be somewhat different among those tissues. It is assumed that the site of sulfation in HS may be controlled by certain regulatory mechanisms. The quantitative method developed in this study is believed to be a very useful method for the determination of compositional profiles of constitutive disaccharide units of tissue HS.