BACKGROUND: Work-related musculoskeletal disorders are prevalent in dental hygienists. Although engineering controls and ergonomic training is available, it is unclear why this intransigent problem continues. One possible barrier is that a comprehensive, standardized protocol for evaluating dental hygiene work does not exist. OBJECTIVE: This study aimed to generate a valid and reliable observational protocol for the assessment of dental hygiene work. METHODS: An iterative process was used to establish and refine an ecologically valid video acquisition and observation protocol to assess key activities, tasks, and performance components of dental hygiene work. RESULTS: Good inter-rater reliability was achieved across all variables when the final coding scheme was completed by three independent raters. CONCLUSIONS: This work provides an exemplar of the process required to generate a comprehensive protocol for evaluating the work components of a particular job, and provides standardized nomenclature for use by scientists and practitioners interested in understanding and addressing the pervasive issue of work-related disorders in dental hygienists.