Abstract
Oxytocin (OT) promotes pro-sociality, bonding, and cooperation in a variety of species. Measuring oxytocin metabolite (OTM) concentrations in urine or saliva provides intriguing opportunities to study human and animal behaviour with minimal disturbance. However, a thorough validation of analytical methods and an assessment of the physiological significance of these measures are essential. We conducted an analytical validation of a commercial Enzyme Immunoassay (EIA; Arbor OT assay kit) to measure OTM concentrations in dog, wolf, and human urine samples. To test the assay’s ability to detect changes in OTM concentrations, we administered oxytocin intranasally to 14 dogs. Assay performance with regard to parallelism was acceptable. Assay accuracy and extraction efficiency for dog and wolf samples were comparable to a previously validated assay (Enzo OT assay kit) but variation was smaller for human samples. Binding sensitivity and antibody specificity were better in the Arbor assay. Average OTM concentrations were more than twice as high as in comparable samples measured with the Enzo assay, highlighting a lack of comparability of absolute values between different assays. Changes in OTM concentrations after intranasal treatment were detected reliably. The Arbor assay met requirements of a “fit-for-purpose” validation with improvement of several parameters compared to the Enzo assay.
Highlights
Oxytocin (OT) promotes pro-sociality, bonding, and cooperation in a variety of species
All three serially diluted pools were parallel to the standard curve (dog urine: t(12) = − 0.233, P = 0.820; wolf urine: t(12) = − 0.243, P = 0.812; human urine: t(12) = − 0.351, P = 0.732) and this was confirmed by visual inspection (Fig. 2)
We aimed to evaluate the performance of a commercial Enzyme Immunoassay (EIA) kit (Arbor Assays, Ann Arbor) to measure urinary oxytocin metabolite (OTM) concentrations in dogs, wolves, and humans
Summary
Oxytocin (OT) promotes pro-sociality, bonding, and cooperation in a variety of species. We conducted an analytical validation of a commercial Enzyme Immunoassay (EIA; Arbor OT assay kit) to measure OTM concentrations in dog, wolf, and human urine samples. OT is a neuropeptide hormone that regulates physiological processes such as eating behaviour and satiety[13], heart rate and blood p ressure[14], birth, lactation, and parenting b ehaviour[15,16], and plays a crucial role in social bond formation and m aintenance[17] It is produced in the hypothalamus and released into the bloodstream by the pituitary gland, it can be measured centrally (in brain tissue by m icrodialysis[18]; in cerebrospinal fluid19) or in peripheral substrates (in plasma[20]; in milk[21]; in saliva[22]; in urine[9]). Comparison of extracted vs. unextracted samples using RIA and EIA: extraction efficiency, precision (CVs), assay accuracy/sensitivity, linearity, IR
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