HIGH-PERFORMANCE CAPILLARY ELECTROPHORETIC ANALYSIS OF HYALURONAN IN EFFUSIONS FROM HUMAN-MALIGNANT MESOTHELIOMA

A procedure to quantify hyaluronan in effusions from human malignant m esothelioma using a highly sensitive and reproducible high-performance capillary electrophoresis (HPCE) method is presented. Following ethan ol precipitation, hyaluronan and galactosaminoglycans were degraded to Delta(4,5)-disaccharides with a mixture of chondroitinases ABC and AC . Heparan sulphate and proteins/glycoproteins were separated by ultraf iltration on a Centricon 3 membrane, and hyaluronan-derived disacchari des were analysed by direct injection of the filtrate into a HPCE syst em. Determination of hyaluronan in effusions from five healthy individ uals and three patients with mesothelioma gave values comparable to th ose found using the HPLC method. One of the advantages of the HPCE met hod as compared to HPLC is the low solvent consumption. The much lower detection limit (attomole level) of the HPCE method may also allow th e analysis of hyaluronan content in serum. The contribution of HPCE in diagnosis of a neoplasm, such as human malignant mesothelioma, illust rates the great potential of this technique in the field of life scien ces. (C) 1997 Elsevier Science B.V.