The biological functions as well as physico-chemical properties of hyaluronan (HA), one of the most important and ubiquitous glycosaminoglycan in various organisms, are closely related to its molecular weight, molecular weight distribution and chain conformation. For this reason it is crucially important to make a reliable characterization of these parameters for HA in both chemical and clinical fields. The present work compared the application of different techniques including capillary viscometry, concentration gradient multiangle laser light scattering (CG-MALLS), size exclusion chromatography-multiangle laser light scattering (SEC-MALLS), asymmetric flow field flow fractionation-multiangle laser light scattering (AFIFFF-MALLS) and horizontal agarose gel electrophoresis (AGE), to the characterization of a range of HA samples of bacterial and animal sources. The advantage and limitation of these techniques were discussed in terms of producing accurate and reliable molecular parameters. (C) 2011 Elsevier Ltd. All rights reserved.