The chemical composition of cork was determined, following a sampling that covered the whole production area in Portugal (29 provenances from six regions) with samples taken at cork stripping. To analyse between population variations, a more intensive sampling was made in two locations. The overall mean chemical composition of cork was: extractives 16.2% (dichloromethane 5.8%, ethanol 5.9%, water 4.5%), suberin 42.8% (long-chain lipids 41.0%, glycerol 3.8%), and lignin 22.0% (Klason 21.1%, acid soluble 0.9%). The suberin compositional ratio of long chain lipids to glycerol, LCLip:Gly, was 11.3. The proportion of neutral sugars in the polysaccharides was: glucose 46.1%, xylose 25.1%, arabinose 18.0%, mannose 3.0%, galactose 7.3%, and rhamnose 0.5%. The range of values was large and the variation between individual trees seemed to be the major factor of the differences. Geographical location of cork production was statistically significant only in a few cases when considering site and not when considering regions. The population variation in two sites was important and the absolute difference between the site mean values was small. This research covers the natural variability of cork’s chemical composition and discusses the contribution of the structural compounds to the variation of cork properties.