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Cited by 52Neuroendocrine regulation and behaviorAnimal Behavior and Welfare StudiesClinical Laboratory Practices and Quality Control

Validation of an enzyme-linked immunoassay (ELISA) for plasma oxytocin in a novel mammal species reveals potential errors induced by sampling procedure

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Robinson, Kelly J., Hazon, Neil, Lonergan, Mike, Pomeroy, Patrick P.·2014·Journal of Neuroscience Methods
DOI 10.1016/j.jneumeth.2014.01.019
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Abstract

BACKGROUND: The neuropeptide oxytocin is increasingly the focus of many studies investigating human and animal social behaviours and diseases. However, interpretation and comparison of results is made difficult by a lack of consistent methodological approaches towards analysing this hormone. NEW METHOD: This study determined the sample collection and analysis protocols that cause the least amounts of protocol dependant variation in plasma oxytocin concentrations detected by ELISA. The effect of vacutainer type, sample extraction prior to analysis and capture and restraint protocol were investigated while validating an assay protocol for two novel species, grey seals (Halichoerus grypus) and harbour seals (Phoca vitulina). RESULTS: Where samples are extracted prior to analysis, vacutainer type (EDTA mean: 8.25±0.56 pg/ml, heparin mean: 8.25±0.62 pg/ml, p=0.82), time taken to obtain a sample and restraint protocol did not affect the concentration of oxytocin detected. However, concentrations of oxytocin detected in raw plasma samples were significantly higher than those in extracted samples, and varied significantly with vacutainer type (EDTA mean: 534.4±43.7 pg/ml, heparin mean: 300.9±19.6 pg/ml, p<0.001) and capture and restraint methodology. There was no relationship between oxytocin concentrations detected in raw and extracted plasma (p=0.25). COMPARISON WITH EXISTING METHOD(S): Over half the reviewed published studies analysing plasma oxytocin use raw plasma and different vacutainer types are used without consistency or justification through-out the literature. CONCLUSIONS: We caution that studies using raw plasma are likely to over estimate oxytocin concentrations, cannot be used to accurately infer true values via correlations and are susceptible to variation according vacutainer type.

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  • Three studies that compared ELISA oxytocin measurements of the same samples with and without extraction all found no correlations.

    “Three studies have compared ELISA oxytocin measurements of the same samples with and without extraction (69-71): all reported no correlations.”

    From Intranasal Oxytocin: Myths and Delusions (2016) · cites this paper as [69]

  • Three studies compared measurements of oxytocin in the same samples by the same ELISA with and without extraction; Robinson et al. found approximately 8 pg/ml in extracted seal plasma versus 300 pg/ml in unextracted heparin samples or 540 pg/ml in unextracted EDTA samples with no correlation, Christensen et al. found radioimmunoassay extracted mean levels of 1.1 pg/ml and spiked levels higher by t

    “Three studies have compared measurements of oxytocin in the same samples by the same ELISA, with and without extraction 101, 116, 117. Robinson et al. 116 measured oxytocin levels in seals: in extracted samples, they found plasma levels of approximately 8 pg/ml, compared to 300 pg/ml in unextracted samples collected in heparin vacutainers or 540 pg/ml in unextracted samples collected in ethylenedi”

    From Measuring Oxytocin and Vasopressin: Bioassays, Immunoassays and Random Numbers (2016) · cites this paper as [116]

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Research works printed but not linked: 4

  1. Baskerville TA, Douglas AJ. Dopamine and oxytocin interaction underlying behaviours: potential contributions to behavioural disorders. CNS Neurosci Ther 2010;16:e92-123.
  2. El-Masry N, Soliman A, Moety HA. Alterations of prolylendopeptidase, oxytocin and vasopressin activity in the plasma of autistic children. Curr Psychiatry 2010;17:31-7.
  3. Morin V, del Castillo JRE, Authier S, Ybarra N, Otis C, Gauvin D, et al. Evidence for nonlinear pharmacokinetics of oxytocin in anesthetized rat. J Pharm Pharmaceut Sci 2008;11:12-24.
  4. Uvnäs-Moberg K. Oxytocin may mediate the benefits of positive social interaction and emotions. Psychoneuroendocrinology 1998;23:819-35.

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Authors
Robinson, K. J., Hazon, N., Lonergan, M., & Pomeroy, P. P.
Affiliations
University of St Andrews (GB)
Corresponding author
Robinson, Kelly J.
ORCID
Venue
Journal of Neuroscience Methods
Publisher
Elsevier BV
Published
2014-04
Volume
226
Pages
73-79
ISSN
0165-0270 (ISSN-L)
DOI
10.1016/j.jneumeth.2014.01.019
OpenAlex
W2118377633
Cited by
52
References
63
Citations by year
2026: 1 · 2025: 1 · 2024: 1 · 2023: 3 · 2022: 7 · 2021: 5 · 2020: 7 · 2019: 6 · 2018: 4 · 2017: 7 · 2016: 4 · 2015: 5 · 2014: 1
FWCI
8.41
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neuroscience Methods (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Discovery Research Portal (University of Dundee) (submittedVersion, cc-by)
  • St Andrews Research Repository (St Andrews Research Repository) (submittedVersion, cc-by)
Funders
  • Natural Environment Research Council
  • SMRU Marine
Topics
  • Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
  • Health Sciences › Veterinary › Small Animals › Animal Behavior and Welfare Studies
  • Health Sciences › Medicine › Physiology › Clinical Laboratory Practices and Quality Control
Keywords
plasma oxytocin, assay validation, EDTA, heparin, sample extraction, grey seal, Phoca vitulina, methodological variability
MeSH terms
Animals, Anticoagulants, Blood Chemical Analysis, Enzyme-Linked Immunosorbent Assay, Female, Heparin, Male, Oxytocin, Plasma, Restraint, Physical, Species Specificity, Specimen Handling, Time Factors, Reproducibility of Results, Seals, Earless, Phoca

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