Brassica napus (canola) seed is a rich source of phytosterols, tocopherols and carotenoids, which all have recognized health benefits, although these are reduced or lost during crude oil refinement. Many studies are now outdated, so new research to monitor bioactive retention through current processing techniques is warranted. In this work, canola seed, in-process seed, and oil samples were collected from the major stages of five commercial canola oil processes. Analysis of phytosterols, tocopherols and carotenoids indicated seed pre-treatment enhanced bioactive concentrations in the crude oil. Although the bleaching step in each process eliminated all carotenoids, high concentrations of phytosterols and tocopherols remained in the refined oil across all processes, with losses notably lower than those found in previous reports. Moreover, crude oil samples from a two-stage cold pressing process showed greatly enriched concentrations of tocopherols (+122%), sterols (+140%) and carotenoids (+217%). The results show that modern Australian canola oil processing retains high phytosterol and tocopherol concentrations and warrants further investigation into bioactive enrichment strategies. Given the growing interest in health-enhanced foods, this study provides opportunities for nutrition and health-enhanced oil products and the potential for adding value in the edible oil industry.