Behavioural and cerebral lateralisation are thought to be controlled, at least in part, by prenatal testosterone (T) levels, explaining why sex differences are found in both laterality traits. The present study investigated hormonal effects on laterality using adult salivary T levels, to explore the adequacy of competing theories: the Geschwind, Behan and Galaburda, the callosal, and the sexual differentiation hypotheses. Sixty participants (15 right-handers and 15 left-handers of each sex) participated. Behavioural lateralisation was studied by means of hand preference tests (i.e., the Edinburgh Handedness Inventory and the Quantification of Hand Preference test) and a hand skill test (i.e., the Peg-Moving test) whereas cerebral lateralisation for language was studied using the consonant-vowel dichotic listening (CV-DL) test and the visual half-field lexical decision (VHLD) test. Salivary T and cortisol (C) concentrations were measured by luminescence immunoassay. Canonical correlations did not reveal significant relationships between T levels and measures of hand preference, hand skill, or language laterality. Thus, our findings add to the growing literature showing no relationship between T concentrations with behavioural or cerebral lateralisation. It is claimed that prenatal T is not a major determinant of individual variability in either behavioural or cerebral lateralisation.