Six tree species were examined using 1H NMR spectroscopy of sap extracted by supercritical CO2. A metabolomic approach was developed to evaluate the sap extracted from sapwood of Norway spruce (Picea abies), Sitka spruce (Picea sitchensis), radiata pine (Pinus radiata), macrocarpa (Cupressus macrocarpa), and two Eucalyptus species—shining gum and mountain ash (Eucalyptus nitens and Eucalyptus regnans. The sap extraction patterns in the different species were visualised using 1H magnetic resonance imaging. In softwoods with distinct annual rings, water was first removed from the latewood bands, and then gradually from the earlywood bands. In the case of the hardwood species an almost random water redistribution, rather than water expulsion, was observed. Analysis of the principal component analysis loading plots showed that the significant differences in the sap between each species were due to the carbohydrate region. Key discriminators were identified as pinitol, sucrose, glucose, and fructose.