Salivary testosterone (T) and cortisol (C) concentrations were monitored across a sports competition. Data were compared using two enzyme-immunoassay (EIA) methods and two sample preparations to determine their influence on hormone concentrations.A group of male athletes (n=19) provided a saliva sample the morning before and one day after (24h post) an international rugby union match. Following an extraction procedure, the samples were analysed for T and C concentrations using a commercial kit (CM E ) and an in-house method (IH E ). Raw samples (no extraction procedure) were also tested using the commercial kit (CM R ).There were no significant changes in T and C levels from pre to post competition with each EIA method and sample preparation, but significant differences in T (IH E >CM E >CM R ) and C (CM R >IH E and CM E ) concentrations were seen when both samples were pooled. Bland–Altman analyses confirmed the presence of fixed and proportional bias. Strong and significant correlations were demonstrated between the IH E and CM E measures of salivary T (r=0.93–0.97) and C (r=0.95–0.97). The T and C values from the raw and extracted samples were also strongly correlated (r=0.93–0.96).The measurement of salivary T and C concentrations across an international sports event was influenced by different EIA methods and sample preparations, but all measures were strongly correlated with some bias. Both T and C were unresponsive to the sports event, but within the group results large individual variation was seen.