In this study, three holders were designed, constructed and characterized to perform quality assurance on the source position and transit time in remote afterloading systems with Co-60 sources for high dose rate brachytherapy. The holders design focused on achieving accuracy, low cost, and a time efficient tool for use in clinical settings. Sensitivities greater than 0.6%/mm and maximum precisions better than 0.14 mm for the source position were obtained. The transit time was determined for the holders with a relative precision better than 19%. •Quality assurance in HDR brachytherapy strongly depends on system parameters.•Check of the dwell positions and transit time in an afterloader with a Co-60 source.•Three holders constructed for quality assurance on Co-60 HDR afterloader.•Design focused on accuracy, low cost, and a time efficiency for clinical settings.