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

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 w hole analyses on one sample within 3 min by use of a short column of CAPCEL L PAK NH, UG80 (35 mm x2 mm I.D.). The assay method was validated and showe d the satisfactory sensitivity, precision and accuracy, which enabled the q uantitation up to picomol level. By employing this method, we performed the analyses of HS in mouse brain and liver, and tumor tissues of tumor-bearin g mouse transplanted subcutaneously with Meth A fibrosarcoma cells. The com positions of the unsaturated disaccharide units derived from HS were found to be somewhat different among those tissues. It is assumed that the site o f sulfation in HS may be controlled by certain regulatory mechanisms. The q uantitative method developed in this study is believed to be a very useful method for the determination of compositional profiles of constitutive disa ccharide units of tissue HS. (C) 2001 Elsevier Science B.V. All rights rese rved.