Chios mastic gum, the resin of Pistacia lentiscus L. var. chia, is known for its acivity against gastrointestinal disorders, mainly against damage caused by the bacterium Helicobacter pylori. Triterpenes of mastic gum are considered responsible for its anti-microbial and anti-H. Pylori properties. In the present study Supercritical Fluid Extraction parameters were fine-tuned through several trials aiming at a stepwise, efficient and environmentally friendly methodology, resulting to the extraction and enrichment of the valuable groups of constituents of mastic gum: a volatile fraction, a triterpenic fraction further separated in neutral and acidic, as well as a resinous polymer. GC-MS analysis, revealed the dominance of alpha-pinene and trans-verbenol in the volatile fraction and of caryophyllene oxide, mastihadienonic and oleanonic aldehyde among others in the neutral triterpenes' fraction. HR-MS/MS analysis was implemented for the characterization of the triterpenic acid fraction, revealing the dominance of isomasticadienonic, masticadienonic, oleanonic and moronic acid.