PROFILING OF THE EYE AQUEOUS-HUMOR IN EXFOLIATION SYNDROME BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF HYALURONAN AND GALACTOSAMINOGLYCANS

The concentrations of hyaluronan and galactosaminoglycans i.e., chondr oitin sulfate and dermatan sulfate - were measured in the aqueous humo r of the eye from patients with exfoliation syndrome and from healthy persons. The glycosaminoglycans/proteoglycans were almost completely p recipitated (>97%) with ethanol in the presence of dextran as carrier and, following enzymic digestion, hyaluronan and galactosaminoglycans, were quantitatively converted to Delta(4.5)-disaccharides. Non-degrad ed heparan sulfate and proteins/glycoproteins were removed by ultrafil tration using a Centricon 3 membrane. Separation and determination of hyaluronan-and galactosaminoglycan-derived Delta-disaccharides were pe rformed by ion-suppression HPLC. For an accurate analysis in triplicat e, as little as 50 mu l of aqueous humor is required. Application of t his method to the analysis of samples from six patients with exfoliati on syndrome and three healthy persons showed that hyaluronan levels in patients (6.65-16.15 mu g ml(-1)) were significantly higher (3-8 rime s) than in healthy persons (2.0-2.24 mu g ml(-1)). There was no signif icant alteration in the galactosaminoglycan concentration. The obtaine d data open a new area in the deeper understanding of the exfoliation syndrome pathophysiology and in establishing a highly sensitivity and accurate HPLC method for its diagnosis and patient's follow-up. (C) 19 98 Elsevier Science B.V. All rights reserved.