An innovative approach for preparing hemp fibers from hemp hurds for use in tissue and towel grades of paper is described. Hemp hurds are a low value by-product of industrial hemp processing that are generally used for animal bed litter. Tissue paper was fabricated from hemp hurd fibers by following three pulping processes: autohydrolysis (hydrothermal), sodium carbonate-based defibration, and high yield kraft pulping, and benchmarked against hardwood pulp. To meet industrial standards, hardwood and hemp pulp fibers were mixed at a dry mass ratio of 75:25, from which tissue paper sheets were prepared. Desirable tissue paper properties, such as water absorption, burst resistance, softness, and tensile strength (dry and wet), were measured and compared. Characterization of morphological and chemical properties of tissue handsheets was conducted with SEM and time-of-flight-secondary ion mass spectrometry (ToF-SIMS). The combined kraft pulped hardwood and autohydrolyzed hemp pulp fibers displayed improvements in tensile index, burst resistance, and softness of tissue handsheets compared to only kraft hardwood pulp handsheets without adversely impacting water absorption. Hardwood fibers showed a sparse distribution of surface lignin compared to hemp fibers using the same defibration. This technology can lead to a variety of eco-friendly tissue paper products that are not only highly energy efficient, but avoid harsh chemical processing.