Objective The objective was to determine the sulfated glycosaminoglycans (GAGs) of the extracellular matrix (ECM) in pre- and postmenopausal women. Study design Periurethral tissue was obtained from 44 consecutive women who underwent surgery for urinary incontinence, for pelvic organ prolapse, or for other gynecologic benign conditions. Biopsy specimens were assessed by biochemical methods to characterize and quantify sulfated GAG. Measurements were made of total GAG, chondroitin sulfate, dermatan sulfate and of heparan sulfate. Data were compared using the t-test. Results Patients were divided into two groups (pre- and postmenopausal groups) and dermatan sulfate was the most predominant glycosaminoglycan. Postmenopausal women had significantly less total sulfated glycosaminoglycans ( p < 0.01), dermatan sulfate ( p < 0.01) and chondroitin sulfate ( p < 0.05) than premenopausal women. We did not observe any differences in heparan sulfate. Conclusions Postmenopausal women showed quantitative differences in the biochemical characteristics of the ECM in periurethral tissue by analysis of sulfated GAG.