Donkeys play a crucial role as a work force in many regions worldwide. These animals may suffer from various welfare problems. The animal welfare charity The Donkey Sanctuary (DS) works towards an improvement of this situation. To evaluate the charitys effect, a standardised, animal-based welfare assessment is necessary. So far, this has been carried out subjectively by using the Hands-on Donkey Welfare assessment protocol. In the present study, parameters of existing protocols were collated and discussed with the DS team in order to develop a more elaborate protocol using a standardised approach and including qualitative behaviour assessment (QBA). The final version was tested with 107 pack donkeys in Ethiopia. The results regarding the behavioural assessment show that 45.8 % of the donkeys were apathetic. The reaction to the observer approach supports this result: 44.3 % of the animals did not move whereas 31.1 % moved away. QBA revealed three main components: the first main component described as “General mental state” showed a good overall consistency between the three different observers. The vast majority (98.5 %) of the donkeys assessed had a low body condition score. The prevalence of wounds and lesions was high with 88 % of the animals having one or more wounds including healed lesions. Additionally, inter-observer reliability was tested between three observers by assessing 32 donkeys. It showed an inconsistent rating regarding more than half of chosen parameters. Thus, more in-depth discussions on the definitions and training are highly advisable. The developed welfare assessment protocol including objective parameters and the first adaptation of QBA with working donkeys is one step to a population wide assessment of donkeys. However, further developments and additional testing in different regions and with more animals are necessary.