Biodegradation of a poly(ester)urea-urethane by cholesterol esterase: isolation and identification of principal...

Synthesized poly(ester)urea-urethanes with 14C-labeled toluene diisocyanate or 14C-labeled chain extender ethylene diamine were incubated with cholesterol esterase in a phosphate buffer solution at 37 degrees C. A number of biodegradation products, generated at the level of 2.8 micrograms/cm2 of polymer surface area, were isolated from this simulated physiologic system. Individual products were obtained by separation with reversed-phase high-performance liquid chromatography. The two different radiolabels were used to assist in the identification of degradation products from hard- and soft-segment domains. Approximately 20 degradation products were isolated; however, toluene diamine (TDA) was not detected from the chromatographic separation. Two principal products were identified by tandem mass spectrometry. Both products are TDA derivatives (secondary aromatic diamine) substituted with end units of the polyester segment at N and N' positions of TDA. The absence of free TDA suggests that there could be a stabilization of urethane and urea linkages within the toluene diisocyanate (TDI) segments of the polyurethanes. For TDI-synthesized polymers, this finding raises awareness to the potential biological importance of degradation products other than TDA, particularly to their interaction with surrounding cells.