An analytical procedure for the quantification of ecdysteroids (crustacean moulting hormone) in barnacles was devised so that minimum sample size could be used. A combination of solvent partitions, Sephadex chromatography, silylation and gas chromatography with electron capture detection was devised, enabling ecdysteroids to be determined down to 20 pg. This was used to determine the amount of moulting hormone in a population of barnacles over a 30 month period. Levels varied from barely detectable in winter months to a maximum value of 1.5 μg kg −1 of wet weight of barnacles in September. Polar conjugates of 20-hydroxyecdysone were detected only during the winter months. The number of barnacles moulting at any time corresponded roughly to the titre of hormone present at that time.