Following a request for technical assistance, EFSA carried out an exposure assessment of the colouring principles bixin and norbixin of the food colour annatto extracts (E 160b), from its use as a food additive, taking into account new proposed uses and use levels. In 2016, the EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS) adopted a scientific opinion on the safety of annatto extracts (E 160b) as a food additive. In that opinion, the Panel concluded that the toxicological database was sufficient to derive an acceptable daily intake (ADI) of 6 mg bixin/kg body weight (bw) per day and an ADI of 0.3 mg norbixin/kg bw per day. Dietary exposure for annatto (E 160b), bixin- and norbixin-based annatto extracts was estimated taking into account the maximum permitted levels (MPLs) as set in the Annex II to Regulation (EC) No 1333/2008, use levels provided by food industry as well as proposed use levels from an applicant for an extension of use. Exposure estimates for bixin were below the ADI for all population groups and for all refined exposure scenarios, including the proposed extension of use. For norbixin, exceedance was observed for the extension of use at the 95th percentile for some population groups. In the current estimates, the dietary exposure for bixin does not exceed the ADI. For norbixin, dietary exposure exceeds the ADI at the high level (95th percentile) for toddlers and children.