Onset of auto-ignition of premixed gas-to-liquid (GTL)/air mixture has been determined at high pressures and low temperatures over a wide range of equivalence ratios. The GTL fuel used in this study was provided by Air Force Research Laboratory (AFRL), designated by Syntroleum S-8, which is derived from natural gas via the Fischer–Tropsch (F–T) process. A blend of 32% iso-octane, 25% n-decane, and 43% n-dodecane is employed as the surrogates of GTL fuel for chemical kinetics study. A spherical chamber, which can withstand high pressures up to 400 atm and can be heated up to 500 K, was used to collect pressure rise data, due to combustion, to determine the onset of auto-ignition. A gas chromatograph (GC) system working in conjunction with specialized heated lines was used to verify the filling process. A liquid supply manifold was used to allow the fuel to enter and evaporate in a temperature-controlled portion of the manifold using two cartridge heaters. An accurate high-temperature pressure transducer was used to measure the partial pressure of the vaporized fuel. Pressure rise due to combustion process was collected using a high-speed pressure sensor and was stored in a local desktop via a data acquisition system. Measurements for the onset of auto-ignition were done in the spherical chamber for different equivalence ratios of 0.8–1.2 and different initial pressures of 8.6, 10, and 12 atm at initial temperature of 450 K. Critical pressures and temperatures of GTL/air mixture at which auto-ignition takes place have been identified by detecting aggressive oscillation of pressure data during the spontaneous combustion process throughout the unburned gas mixture. To interpret the auto-ignition conditions effectively, several available chemical kinetics mechanisms were used in modeling auto-ignition of GTL/air mixtures. For low-temperature mixtures, it was shown that auto-ignition of GTL fuel is a strong function of unburned gas temperature, and propensity of auto-ignition was increased as initial temperature and pressure increased.