The pyrolysis of di-n-propyl ether (DPE), a potential biofuel candidate, was studied under a temperature range of 850–1350 K in a flow reactor at low and atmospheric pressures. Synchrotron vacuum ultraviolet photoionization mass spectrometry was used for isomeric identification and mole fraction measurements of the pyrolysis products, especially the free radicals. Specific products were observed and measured for the unimolecular decomposition reactions of DPE. A detailed kinetic model of DPE pyrolysis was developed based on the n-propanol combustion model. Modeling analyses including the rate of production (ROP) and sensitivity analyses were performed to reveal the consumption pathways of DPE, as well as the formation and consumption pathways of intermediates and products. ROP and sensitivity analyses indicated that the consumption reaction of DPE was mainly controlled by the unimolecular decomposition reactions and H-abstractions at both low and atmospheric pressures. The pericyclic reaction, leading to n-propanol + propylene, was the most important unimolecular decomposition reactions, especially at low pressure. The H-abstraction reactions of DPE played crucial roles in its consumption. Further β-C–C and β-C–O scission reactions of the three primary radicals can produce some oxygenated and hydrocarbon products. Relatively high mole fractions of C1–C3 aldehydes and C1–C3 alkane, alkene, and alkyl radicals were observed, which demonstrated the structural feature of DPE. Comparison of the product distribution between DPE and n-propanol were also analyzed.