Interfacial properties play important roles in processes for hydrocarbon resource production, transport, and refining. Biplex liquid crystal domains, with diameters up to 200 μm, have been identified in unreacted heavy fractions of petroleum resources, particularly asphaltenes, worldwide. These liquid crystal domains comprise thin liquid crystal shells surrounding homogeneous cores dispersed in liquid. The identification of species comprising the shells has proven elusive because they are minor constituents of species present. In a recent work, we showed that produced water is enriched in liquid crystal domains relative to a hydrocarbon resource. In this work, gram quantities of liquid-crystal-rich materials were isolated from produced water. The isolation methods are described, and their compositions are analyzed using a combination of elemental analysis, Fourier transform infrared spectroscopy, and negative-ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. The liquid-crystal-rich materials are shown to include humic substances (humic acid, fulvic acid, and humin) among the principal components. These surface-active and lyotropic liquid-crystal-forming materials are the only species known to be present that meet the constraints imposed by the analyses and self-assemble cooperatively. Validation experiments demonstrating the formation of lyotropic liquid crystal domains with solid and liquid hydrocarbon as well as aqueous cores + humic-acid-rich liquid crystalline shells dispersed in aqueous and hydrocarbon liquids were performed. The formation and behavior of liquid crystal domains in humic substance + hydrocarbon + water mixtures are analogous to those of oil−water−surfactant dispersions, where surfactants self-assemble on interfaces, forming a liquid crystal layer. Humic substances, common hydrocarbon reservoir constituents, may contribute to interfacial stability and surface deposition problems, arising in petroleum production, transport, and refining that are commonly attributed to asphaltenes because they appear to comprise a microscopic surphase on the exterior of asphaltene particles.