Petroleum is one of the most complex naturally occurring organic mixtures. The physical and chemical properties of petroleum in a reservoir depend on its molecular composition and the reservoir conditions (temperature, pressure). The composition of petroleum varies greatly, ranging from the simplest gas (methane), condensates, conventional crude oil to heavy oil and oil sands bitumen with complex molecules having molecular weights in excess of 1000 daltons (Da). The distribution of petroleum constituents in a reservoir largely depends on source facies (original organic material buried), age (evolution of organisms), depositional environment (dysoxic versus anoxic), maturity of the source rock (kerogen) at time of expulsion, primary/secondary migration, and in-reservoir alteration such as biodegradation, gas washing, water washing, segregation, and/or mixing from different oil charges. These geochemical aspects define the physical characteristics of a petroleum in the reservoir, including its density and viscosity. When the petroleum is released from the reservoir through an oil exploration accident like in the case of the Deepwater Horizon event, several processes are affecting the physical and chemical properties of the petroleum from the well head into the deep sea. A better understanding of these properties is crucial for the development of near-field oil spill models, oil droplet and gas bubble calculations, and partitioning behavior of oil components in the water. Section 3.1 introduces general aspects of the origin of petroleum, the impact of geochemical processes on the composition of a petroleum, and some molecular compositional and physicochemical background information of the Macondo well oil. Section 3.2 gives an overview over experimental determination of all relevant physicochemical properties of petroleum, especially of petroleum under reservoir conditions. Based on the phase equilibrium modeling using equations of state (EOS), a number of these properties can be predicted which is presented in Sect. 3.3 along with a comparison to experimental data obtained with methods described in Sect. 3.2.