Only limited data on concentrations of trace elements in the blood of avian species have been published. This information can play an important role in the conservation of endangered species and their protection from environmental pollutants and can also be clinically relevant in managed individuals. Some elements are essential for the health of the animals in human care, but little is known about expected concentrations for some of these elements. Therefore, 21 elements (silver, arsenic, gold, barium, cadmium, cobalt, chromium, copper, iron, iodine, mercury, magnesium, manganese, molybdenum, nickel, platinum, antimony, selenium, thallium, vanadium, and zinc) were measured by inductively coupled plasma mass spectrometry in lithium-heparinized plasma samples from Humboldt penguins (Spheniscus humboldti) from a single zoological collection in Germany (n = 39). The plasma concentrations of cadmium, nickel, platinum, antimony, and thallium were under the limit of detection in most of the analyzed samples. The results also showed that the females had significantly (P ≤ 0.05) higher concentrations of silver, barium, cobalt, chromium, iron, mercury, magnesium, vanadium, and zinc in their plasma in comparison with males. A correlation between the element plasma concentration and the age of the individuals could not be found. The differences in plasma concentrations of individual element between males and females could be due to hormonal influences on the metabolism. Further studies in different seasons and other age groups are needed in the future to investigate environmental and host effects in more detail and to understand physiological mechanisms responsible for observed differences.