The number of bees, amount of brood and haemolymph vitellogenin titre are parameters to establish the vitality of a honey bee colony. Increasing numbers of bees during summer until autumn; increasing amounts of brood in spring towards summer followed by a decrease; and low haemolymph vitellogenin titres in foraging bees and high vitellogenin titres in nurse bees and winter bees are typical courses over the bee season. In contrast to the number of bees and amount of brood, determined at the colony level, vitellogenin is determined in individual bees. To turn the factor vitellogenin from the mean of individual data into a colony parameter, a protocol was developed to determine the colony fraction vitellogenin in a pooled haemolymph sample of 30 bees, sampled from a defined location in the colony. This colony fraction vitellogenin is a parameter for honey bee colony vitality.