Plasma adrenocorticotropic hormone concentration in horses decreases after freezing for 60 days

We investigated the stability of adrenocorticotropic hormone (ACTH) in plasma after freezing for different lengths of time. The plasma ACTH concentrations of 12 horses were measured on day 0 (baseline) and over time, after stimulation with thyrotropin-releasing hormone. Samples were stored at -80°C for 3, 7, 30, 60, and 90 d, or at -20°C for 3, 7, 30, and 60 d, or between ice packs at -20°C for 3 and 7 d prior to determination of ACTH concentration. ACTH concentrations were compared to baseline (non-frozen day 0 plasma) for each storage method using a mixed model with repeated measures in which each horse served as its own control and day was the repeated effect. Statistical significance was set at p ≤ 0.05, and 0.05 < p < 0.10 was considered a trend. Plasma ACTH frozen at -20°C or at -80°C resulted in degradation of ACTH compared to baseline samples at 60 and 90 d respectively. There was no degradation of ACTH after 7 d when stored between ice packs, or before 30 d at -20°C, or before 60 d at -80°C.