AbstractEcdysterone (crustecdysone; beta‐ecdysone; 20‐hydroxyecdysone) is a naturally occurring steroid hormone belonging to the ecdysteroid class. The presented study investigated the possible concentration range of ecdysterone in urine after consumption of various preparations of spinach, drinking tea (made from Rhapoiticum Carthamoides) and topical use of a cream containing Cyanotis arachnoides. It is very important to establish reference ranges reflecting concentrations compatible with dietary habits and common uses of care products. The data obtained in the research may be used in the interpretation of results of routine analyses. In addition, elimination time and observed concentrations provided by the studies conducted by the Polish Anti‐Doping Laboratory can be used by WADA. In the case of spinach, peak elimination occurred within the first few hours, followed by a rapid decline. As for the other plants, instead of clear peak concentrations, gradual elimination was observed. Individual differences were observed between volunteers depending on route of administration. Differences in ecdysterone elimination following ingestion of spinach‐based and other plant products were observed too. The highest observed ecdysterone concentration was related to the paste consumption, and it was 691 ng/ml. Finally, our findings were compared with the data collected for the samples routinely tested as part of the monitoring program. During 2.5 years, the presence of ecdysterone was confirmed in as many as 507 samples out of 11 191 total samples tested. The concentration range was very wide, from 1 ng/ml (which is the LOD for this method in the Polish Anti‐Doping Laboratory) to over 2000 ng/ml.