The most important representatives of the three groups of natural steroid sex hormones that have been used as growth promoters in the intensive livestock industry are: 17ß-estradiol, progesterone and testosterone. Their effects on growth and improvement of the organoleptic properties are very positive, because that they have been applied in farm animals for a long time before their side effects were revealed. Long-term use of these substances has toxic, mutagenic, teratogenic and cancerous effects on animals. As a result of anabolic application, accumulation of residues occurs in animal tissues and the elimination of the substances is performed through the bodily fluids, including milk. Consumers could be exposed to these substances by consuming food products of animal origin (milk, meat, eggs), causing even more severe side effects and harmful repercussions in humans than in animals. Because of the mentioned reasons, anabolic use of steroid sex hormones was banned by law in 1988 in the whole EU. Their application is limited only to therapeutic purposes to treat reproductive and gestation disorders. For determining concentrations of steroid sex hormones in food of animal origin, it is essential to know physiological concentrations of the mentioned hormones, in order to know if the results are a consequence of abuse or physiological referent values. Physiological values can vary and it depends on many factors. So when we discuss the results, we have to consider anamnestic information and facts about feeding and animal accommodation. The current control systems are more detailed, the diagnostic tests are more precise, so the results are more reliable and animal food products are safer for consumers. Therefore, the constant development of new synthetic substances and possible abuse of so-called "cocktails" with low levels of active substances with anabolic effects and thus the continuous monitoring of their use is required during farm animal breeding and in slaughterhouses.